JP2006522008A - Interferon medication for the treatment of viral diseases and liver fibrosis - Google Patents

Abstract

The present invention includes methods for treating alphavirus infection; methods for treating hepatitis C virus (HCV) infection; methods for treating West Nile virus infection; methods for treating dengue virus infection; methods for treating yellow fever virus infection; How to reduce liver fibrosis; how to increase liver function in individuals suffering from liver fibrosis; how to reduce the occurrence of complications related to HCV and cirrhosis; and in patients suffering from viral infections A method of reducing viral load, a method of reducing time for viral clearance, or a method of reducing morbidity or mortality in clinical outcome is provided. The present invention generally comprises administering a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for the treatment of viral infection or liver fibrosis.

Description

The present invention is in the field of flavivirus infections, particularly West Nile virus infection and hepatitis C virus infection, and liver fibrosis.

BACKGROUND OF THE INVENTION The United States is currently experiencing an increasing number of West Nile virus infections. West Nile virus, like hepatitis C virus, is a member of the alpha-like flavivirus. The majority of alphaviruses, including hepatitis C virus and poliovirus, are highly sensitive to type I interferon therapy. West Nile virus appears to be endemic in the United States. This is because it has a bird-carrying host and is transmitted by mosquitoes. West Nile virus can cause severe self-limiting heat, body pain, brain swelling, coma, paralysis, and death. Although it is generally accepted that West Nile virus disease results in only one death from 40,000 cases, mortality in the United States appears to be higher. In particular, five deaths have been recently reported in Louisiana. US strains may be more pathogenic or the US population may be genetically predisposed to a more severe clinical course. In Middle Eastern countries, West Nile virus has been endemic for centuries, which could allow natural selection to create populations that are resistant to the virus. There is no effective treatment for this disease.

  Hepatitis C virus (HCV) infection is an infection caused by chronic blood that is most common in the United States. Although the number of new infections has decreased, the burden of chronic infection is substantial and the Centers for Disease Control estimate is 3.9 million (1.8%) in the United States. Chronic liver disease is the 10th most common cause of death among adults in the United States, accounting for approximately 25,000 deaths annually, accounting for approximately 1% of all deaths. Studies have shown that 40% of chronic liver diseases are HCV-related, resulting in an estimated 8,000 to 10,000 deaths annually. HCV-related end-stage liver disease is the most frequent sign for liver transplantation among adults.

  Antiviral treatment of chronic hepatitis C has progressed rapidly over the last decade, with significant improvements seen in the efficiency of treatment. Nevertheless, even with combination therapy using pegylated IFN-α and ribavirin, 40% to 50% of patients fail treatment, ie unresponsive or relapsed. These patients currently have no effective alternative treatment. In particular, patients with advanced fibrosis or cirrhosis in liver biopsy are prominent developing complications of advanced liver disease, including ascites, jaundice, varicose bleeding, brain damage, and progressive liver failure There is a risk, as well as a significant increase in the risk of hepatocellular carcinoma.

  The high prevalence of chronic HCV infection has important public health speculative consequences for the future burden of chronic liver disease in the United States. Data from the United States National Health and Nutrition Examination Survey (NHANES III) is available from the late 1960s to the early 1980s, especially for generations in their 20s to 40s. It shows a large increase in the proportion of new HCV infections that occurred among humans. It is estimated that the number of individuals with long-term HCV infection over 20 years could be more than quadrupled from 1990 to 2015, from 750,000 to over 3 million. The proportional increase in individuals infected for 30 or 40 years is even greater. The risk of HCV-related chronic liver disease is related to the duration of infection, and for people who have been infected for longer than 20 years, there is a risk of progressively increasing cirrhosis, There is a significant increase in morbidity and mortality associated with cirrhosis among patients infected in between.

  Fibrosis occurs as a result of chronic toxic damage to the liver, such as chronic hepatitis C virus (HCV) infection, autoimmune injury, and chronic exposure to toxins (such as alcohol). Chronic toxic injury results in repeated cycles of hepatocyte damage and repair associated with chronic inflammation. Over a varying period, the abnormal extracellular matrix gradually accumulates as a result of the host's damage response. If left unconfirmed, this results in increased deposition of fibrous material until the liver structure is deformed and the liver's ability to regenerate is impaired. The gradual accumulation of scar tissue within the liver ultimately yields a histopathological clinical picture of cirrhosis, which is defined as the formation of a fibrous septum with the formation of micromass throughout the liver.

  There is a need in the art for improved methods for treating flavivirus infections (e.g., West Nile virus infection, hepatitis C virus infection, dengue virus infection, yellow fever virus infection) and for treating liver fibrosis. Exists in the field. The present invention addresses this need.

References

SUMMARY OF THE INVENTION This invention is a method of treating alphavirus infection; a method of treating hepatitis C virus (HCV) infection; a method of treating West Nile virus infection; a method of treating dengue virus infection; How to reduce liver fibrosis; how to increase liver function in individuals suffering from liver fibrosis; how to reduce the incidence of complications associated with HCV and cirrhosis; and whether to reduce viral load Or a method of reducing time to viral clearance or reducing morbidity or mortality in clinical outcome in patients suffering from viral infections. This method generally involves administering a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for the treatment of viral infection or liver fibrosis.

Features of the Invention The present invention generally comprises administering to an individual an IFN-γ and a type I or type III interferon receptor agonist in an amount effective to alleviate the clinical course of the disease, Features a method of treating a viral infection. The present invention also treats alphavirus infection by administering to individuals an IFN-γ and a type I or type III interferon receptor agonist in a synergistically effective amount to alleviate the clinical course of the disease. Features method.

  The present invention provides a type I or type III interferon receptor agonist, IFN-γ, in an amount effective to reduce time to viral clearance or reduce morbidity or mortality in clinical outcome. Or a method of treating West Nile virus infection, generally comprising the step of simultaneously administering to an individual an IFN-γ and a type I or type III interferon receptor agonist. The invention also provides IFN-γ and type I or type III interferon in an amount that is synergistically effective to reduce time to viral clearance or reduce morbidity or mortality in clinical outcome. Features a method of treating West Nile virus infection by administering a receptor agonist to an individual.

  The present invention provides a type I or type III interferon receptor agonist, IFN-γ, in an amount effective to reduce time to viral clearance or reduce morbidity or mortality in clinical outcome. Or a method of treating dengue virus infection, generally comprising the step of simultaneously administering to an individual an IFN-γ and a type I or type III interferon receptor agonist. The invention also provides IFN-γ and type I or type III interferon in an amount that is synergistically effective to reduce time to viral clearance or reduce morbidity or mortality in clinical outcome. Features a method of treating dengue virus infection by administering a receptor agonist to an individual.

  The present invention provides a type I or type III interferon receptor agonist, IFN-γ, in an amount effective to reduce time to viral clearance or reduce morbidity or mortality in clinical outcome. Or a method of treating yellow fever virus (YFV) infection, generally comprising the step of simultaneously administering to an individual an IFN-γ and a type I or type III interferon receptor agonist. The invention also provides IFN-γ and type I or type III interferon in an amount that is synergistically effective to reduce time to viral clearance or reduce morbidity or mortality in clinical outcome. Features a method of treating YFV infection by administering a receptor agonist to an individual.

  The present invention relates to a hepatitis C virus virus that generally comprises the step of simultaneously administering to an individual an IFN-γ and a type I or type III interferon receptor agonist in an amount effective to achieve a sustained viral response. Features a method of treating (HCV) infection. The present invention also provides a method for treating HCV infection by administering to an individual IFN-γ and a type I or type III interferon receptor agonist in a synergistically effective amount to achieve a sustained viral response. Features.

  The present invention relates to a method for reducing liver fibrosis in an individual, generally comprising the step of simultaneously administering a type I or type III interferon receptor agonist and IFN-γ in an amount effective to reduce liver fibrosis. It is characterized by. Optionally, the method of the invention comprises an amount of pirfenidone effective to enhance the antifibrotic effect or reduction of liver fibrosis achieved by treatment with a type I or type III interferon receptor agonist and / or IFN-γ. (pirfenidone) or a pirfenidone analog in combination with a combination of a type I or type III interferon receptor agonist and IFN-γ is provided. The present invention also provides a method of reducing liver fibrosis in an individual by administering a type I or type III interferon receptor agonist and IFN-γ in a synergistically effective amount to reduce liver fibrosis. Wherein the method optionally comprises an amount effective to enhance the antifibrotic effect or reduction of liver fibrosis achieved by treatment with a type I or type III interferon receptor agonist and / or IFN-γ. Of pirfenidone or an analog of pirfenidone. In some embodiments, the extent of liver fibrosis is determined by pre- and post-treatment staging of liver biopsy, wherein the stage of liver fibrosis as measured by a standardized scoring system is treatment Decrease by at least 1 unit when comparing pre- and post-treatment liver biopsies.

  The present invention relates to an individual suffering from liver fibrosis, generally comprising the step of simultaneously administering a type I or type III interferon receptor agonist and IFN-γ in an amount effective to increase liver function. Characterized by a method of increasing liver function. Optionally, the method of the invention comprises an amount of pirfenidone effective to enhance an antifibrotic effect or increased liver function achieved by treatment with a type I or type III interferon receptor agonist and / or IFN-γ, or Provided is the step of administering to the patient a combination of a type I or type III interferon receptor agonist and IFN-γ together with a pirfenidone analog. The present invention also provides for an individual suffering from liver fibrosis by administering a type I or type III interferon receptor agonist and IFN-γ in a synergistically effective amount to increase liver function. Featuring a method of increasing liver function, which method optionally enhances the antifibrotic effect or increased liver function achieved by a type I or type III interferon receptor agonist and / or IFN-γ treatment A co-administering to the patient an effective amount of pirfenidone or pirfenidone analog. Liver function includes serum transaminase level, prothrombin time, serum bilirubin level, platelet count, serum albumin level, improved portal wedge pressure, degree of ascites reduction, reduced brain damage level, and reduced internal varices It can be shown by measuring a parameter selected from the group consisting of degrees.

  The invention features a method of reducing the incidence of cirrhosis complications. This method generally involves co-administering a type I or type III interferon receptor agonist and IFN-γ in an amount effective to reduce the incidence of cirrhosis complications. Optionally, the methods of the present invention are effective to enhance the antifibrotic effect achieved by treatment with type I or type III interferon receptor agonists and / or IFN-γ or the reduction in the incidence of cirrhosis complications. A step of administering to a patient a combination of a type I or type III interferon receptor agonist and IFN-γ together with an amount of pirfenidone or a pirfenidone analog is provided. The present invention also provides for the occurrence of cirrhosis complications by administering a type I or type III interferon receptor agonist and IFN-γ in a synergistically effective amount to reduce the occurrence of cirrhosis complications. This method optionally reduces the incidence of antifibrotic effects or complications of cirrhosis achieved by treatment with a type I or type III interferon receptor agonist and / or IFN-γ. Co-administering to the patient an effective amount of pirfenidone or a pirfenidone analog to enhance. Examples of complications of cirrhosis are portal hypertension, progressive liver failure, and hepatocellular carcinoma.

  As described above, when performing a combination therapy method for alphavirus infection, hepatitis C virus infection, West Nile virus infection, dengue virus infection, YFV infection, and / or cirrhosis in an individual, type I or III An interferon receptor agonist and IFN-γ are administered to the individual. In certain embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in the same formulation. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in separate formulations. When administered in separate formulations, the Type I or Type III interferon receptor agonist and IFN-γ can be administered substantially simultaneously or within about 24 hours of each other. In many embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered subcutaneously in multiple doses. Optionally, a Type I or Type III interferon receptor agonist and / or IFN-γ is administered to the individual by a controlled drug delivery device. Optionally, a Type I or Type III interferon receptor agonist and / or IFN-γ is administered to an individual substantially continuously or continuously by a controlled drug delivery device. Optionally, the controlled drug delivery device is an implantable infusion pump that delivers a Type I or Type III interferon receptor agonist and / or IFN-γ to the individual by subcutaneous infusion.

  In certain embodiments utilizing combination therapy, IFN-γ is administered during the entire course of type I or type III interferon receptor agonist treatment. In other embodiments, IFN-γ is administered during a treatment period that overlaps with a period of type I or type III interferon receptor agonist treatment, eg, before initiation of type I or type III interferon receptor agonist treatment. IFN-γ treatment can be started and IFN-γ treatment can be completed before type I or type III interferon receptor agonist treatment is completed; after type I or type III interferon receptor agonist treatment is started IFN-γ treatment can be started and IFN-γ treatment can be completed after IFN-γ treatment is completed; IFN-γ treatment is started after type I or III interferon receptor agonist treatment is started And IFN-γ treatment can be completed before type I or type III interferon receptor agonist treatment is completed; or IFN-γ treatment can be completed before IFN-α treatment is started. Treatment can begin and IFN-γ treatment can be terminated after Type I or Type III interferon receptor agonist treatment is completed. In embodiments utilizing co-administration of pirfenidone or pirfenidone analog, the duration of treatment with pirfenidone or pirfenidone analog may be concurrent with the duration of treatment with a type I or type III interferon receptor agonist and / or IFN-γ. . In other embodiments, the course of treatment with pirfenidone or a pirfenidone analog may overlap with the course of treatment with a type I or type III interferon receptor agonist and / or IFN-γ, for example, a type I or type III interferon. Treatment with pirfenidone or a pirfenidone analog can begin before treatment with a receptor agonist and / or IFN-γ begins, and treatment with a type I or type III interferon receptor agonist and / or IFN-γ ends Treatment with pirfenidone or a pirfenidone analog may be terminated before; pirfenidone treatment may be after treatment with IFN-α and / or IFN-γ has started and a type I or type III interferon receptor agonist and / or Treatment with IFN-γ Pirfenidone treatment may be terminated before the treatment is completed; or pirfenidone treatment may be initiated before treatment with a type I or type III interferon receptor agonist and / or IFN-γ may be initiated, and type I or type III Pirfenidone treatment may be terminated after treatment with the interferon receptor agonist and / or IFN-γ is completed.

  In other embodiments, ribavirin is co-administered with a type I or type III interferon receptor agonist and IFN-γ. In yet other embodiments, ribavirin is co-administered with a type I or type III interferon receptor agonist, IFN-γ, and pirfenidone (or a specific pirfenidone analog).

  In many embodiments, any of the above methods comprise administering IFN-α and IFN-γ. In some of these embodiments, the methods of the invention comprise co-administering ribavirin, IFN-α, and IFN-γ. In other embodiments, the methods of the invention comprise co-administering IFN-α, IFN-γ, and pirfenidone or a pirfenidone analog.

  In many embodiments, any of the above methods comprise administering a PEGylated IFN-α conjugate. In certain embodiments, the PEGylated IFN-α conjugate is monoPEGylated IFN-α. In other embodiments, the monoPEGylated IFN-α conjugate is an IFN-α polypeptide covalently linked to a single PEG moiety via a lysine residue or N-terminal amino acid residue of the IFN-α polypeptide. It is. In other embodiments, the monoPEGylated IFN-α conjugate is an amide bond between either the ε-amino group or the α-amino group of the lysine residue of the IFN-α polypeptide and the activated carboxyl group of the PEG moiety. Is an IFN-α polypeptide covalently linked to a single PEG moiety. In other embodiments, the monoPEGylated IFN-α conjugate is an IFN-α polypeptide covalently linked to a single, linear PEG moiety. In other embodiments, the mono-PEGylated INF-α conjugate is an IFN-α polypeptide covalently linked to a single, linear 30 kD PEG moiety (`` mono PEG (30 kD, linear) INF -α "). In other embodiments, the monoPEGylated IFN-α conjugate is via an amide bond between the ε amino group or α amino group of the lysine residue of the IFN-α polypeptide and the activated carboxyl group of the PEG moiety. An IFN-α polypeptide covalently linked to a single, linear 30 kD PEG moiety. In other embodiments, the mono-PEGylated IFN-α conjugate is via an amide bond between the ε-amino group or α-amino group of the lysine residue of the IFN-α polypeptide and the activated propionyl group of the PEG moiety. An IFN-α polypeptide covalently linked to a single, linear 30 kD PEG moiety. In other embodiments, the monoPEGylated IFN-α conjugate is a single, IFN-α polypeptide covalently linked to monomethoxy PEG (mPEG). In other embodiments, the monoPEGylated IFN-α conjugate is a condensation reaction product between an IFN-α polypeptide and a linear succimidylpropionate activated 30 kD mPEG. In any of the foregoing methods using a PEGylated IFN-α conjugate, the INF-α polypeptide can be a consensus interferon (CIFN) polypeptide. In any of the foregoing methods using a PEGylated IFN-α conjugate, the IFN-α polypeptide can be a CIFN polypeptide that is interferon alfacon-1.

Definitions As used herein, the term “liver fibrosis” is used interchangeably herein with “liver fibrosis” and refers to scar tissue in the liver that may occur in the context of chronic hepatitis infection. Refers to proliferation.

  The terms “individual”, “host”, “subject”, and “patient” are used interchangeably herein to refer to mammals, including but not limited to primates, including apes and humans. Say.

  As used herein, the term “alphavirus” and its grammatical variants include (i) the RNA genome (ii) viral replication in the host cell cytoplasm and (iii) the DNA phase in the viral replication cycle. A group of viruses characterized by their absence.

  As used herein, the term `` liver function '' refers to the normal function of the liver, including synthetic functions (such as serum proteins (e.g. albumin, clotting factors, alkaline phosphatase, aminotransferases (e.g. , Alanine transaminase, aspartate transaminase), 5'-nucleosidase, γ-glutaminyl transpeptidase, etc.), bilirubin synthesis, cholesterol synthesis, and bile acid synthesis. ); Liver metabolic function (including but not limited to carbohydrate metabolism, amino acid and ammonia metabolism, hormone metabolism, and lipid metabolism); exogenous drug detoxification; hemodynamic function (including visceral and portal hemodynamics) But are not limited to these.

  The term “persistent viral response” (SVR; also referred to as “persistent response” or “persistent response”), as used herein, treats HCV infection in terms of serum HCV titer. The individual's response to a regimen. In general, a “persistent viral response” is a period of at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, or at least about 6 months after discontinuation of treatment Refers to undetectable HCV RNA (eg, less than about 500, less than about 200, or less than about 100 genome copies per milliliter of serum) found in the patient's serum.

  “Treatment failure patient” as used herein generally refers to an HCV infected patient (referred to as a “non-responder”) or a previous failure to respond to a previous treatment for HCV. HCV-infected patients (referred to as “relapsers”) who initially responded to treatment but did not maintain a therapeutic response in the patient. Previous treatment generally may include treatment with IFN-α monotherapy or IFN-α combination therapy, where the combination therapy may include administration of an antiviral agent such as IFN-α and ribavirin.

  As used herein, the terms “treatment”, “treating” and the like refer to obtaining a desired pharmacological and / or physiological effect. This effect can be prophylactic by completely or partially preventing the disease or its symptoms, and / or complete or partial cure for the harmful effects that can result from the disease and / or disease Can be therapeutic. `` Treatment '' as used herein encompasses any treatment of a disease in mammals, particularly humans, and (a) is diagnosed as having a risk of being susceptible to the disease. Preventing the disease from occurring in a subject who has not been treated; (b) inhibiting the disease, i.e., quiescing its occurrence; and (c) reducing the disease, i.e., causing the disease to regress. including.

  The terms “individual”, “host”, “subject”, and “patient” are used interchangeably herein and include mice, apes, humans, mammalian livestock animals, mammalian sport animals. And mammals including but not limited to mammalian pets.

  “Specific pirfenidone analogs” and all grammatical variants thereof refer to and are limited to each and every pirfenidone analog shown in Table 1.

  As used herein, the term “type I interferon receptor agonist” refers to any naturally occurring human type I interferon receptor that binds to and causes signaling through the receptor, or A non-naturally occurring ligand. Type I interferon receptor agonists include naturally occurring interferons, modified interferons, synthetic interferons, pegylated interferons, fusion proteins including interferons and heterologous proteins, interferons including shuffled interferons; antibodies specific for interferon receptors; Non-peptide chemical agonists are included.

  As used herein, the term “type III interferon receptor agonist” refers to any naturally occurring or non-naturally occurring ligand of human IL-28 receptor α (“IL-28R”). The amino acid sequence is described by Sheppard et al., Below, which binds to and causes signaling through the receptor.

  The term “medication event” as used herein refers to the administration of an antiviral agent to a patient in need thereof, and this event refers to one or more antiviral agents from a drug dispensing device. These releases can be included. Thus, the term “medication event” as used herein introduces a continuous delivery device (e.g., a pump or other controlled release injectable system); and a single subcutaneous injection followed by a continuous Including, but not limited to the introduction of a mechanical delivery system.

  `` Continuous delivery '' as used herein (e.g., in the context of continuous delivery of a substance to tissue) provides for delivery of a desired amount of substance to the tissue over a selected period of time. Is meant to refer to the transfer of drug to a delivery site (eg, tissue) in such a manner that approximately the same amount of drug is received by the patient throughout the selected time.

  “Controlled release” as used herein (eg, in the context of “controlled drug release”) is a selected, or otherwise controllable rate, interval that is substantially unaffected by the environment of use. And / or in an amount is meant to include the release of a substance (eg, a type I or type III interferon receptor agonist (eg, IFN-α)). Thus, "controlled release" includes, but is not necessarily, substantially continuous delivery and patterned delivery (e.g., intermittent delivery over a period interrupted by regular or irregular time intervals). It is not limited to.

  “Patterned” or “temporal”, when used in the context of drug delivery, for a preselected time in a fixed pattern, generally in a substantially regular pattern By delivery of the drug over a period of time (eg, other than the period associated with a bolus injection, for example). “Patterned” or “temporal” drug delivery is increased, decreased, substantially constant, or pulsed, rate or range of rates (e.g., amount of drug per unit time, Or delivery of a drug in volume of drug formulation per unit time), and further includes delivery that is continuous or substantially continuous, or long term.

  The term `` controlled drug delivery device '' means that the release (e.g., rate, timing of release) of a drug or other desired substance contained therein is controlled by or determined by the device itself, and It is meant to include any device that is substantially unaffected by the environment of use or is released at a rate that is reproducible in the environment of use.

  “Substantially continuous”, for example when used in the context of “substantially continuous infusion” or “substantially continuous delivery”, is a preselected period of drug delivery of drug delivery. Means the delivery of the drug in a substantially uninterrupted manner, where the amount of drug received by the patient during any 8 hour interval in the preselected period is never zero Will not go down. In addition, “substantially continuous” drug delivery also includes a substantially constant, preselected rate, or range of rates that is not substantially interrupted during the preselected period of drug delivery (e.g., Delivery of the drug in the amount of drug per unit time, or volume of drug formulation during unit time).

“Substantially steady state”, when used in the context of biological parameters that can vary as a function of time, indicates that the biological parameter exhibits a substantially constant value over time, so that time The area under the curve defined by the value of the biological parameter as a function of _time (AUC _{8 hours} ) for any 8 hours during the course of the biological over the 8 hours of the course of time The average area under the curve of the parameter (AUC _{8 hour average} ), not more than about 20% or less than about 20%, and preferably not more than about 15% or less than about 15%, and More preferably, it means no more than about 10% or no less than about 10%. The AUC _{8-hour average} is the quotient (q) divided by the area under the curve of biological parameters over the entire time course ( _overall AUC) divided by the number of 8-hour intervals in the time course (t _{total 1/3 day} ) That is, q = (the _whole AUC) / (the _whole t _{is 1/3 days} ). For example, in the context of drug serum concentration, the drug serum concentration is the area under the drug serum concentration curve (AUC _{8 hours} ) over any 8 hours during the time course, 8 hours during the time course. Over the mean area under the serum concentration curve of the drug (AUC _{8 hour average} ), not more than about 20% or less than about 20%, i.e. AUC _{8 hours} for the serum concentration of the drug over _time Is maintained in a substantially steady state during the time course when it does not exceed 20% or below 20% of the AUC _{8 hour average} .

  Before the present invention is further described, it is to be understood that the present invention is not limited to the specific embodiments described and, of course, can itself vary. Since the scope of the present invention is limited only by the appended claims, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. It should also be understood.

  Where a range is provided, up to one tenth of the lower limit unit, unless the context clearly dictates otherwise, each intermediate value between the upper and lower limits of the range, and the ranges mentioned Any other mentioned or intermediate values are understood to be included in the present invention. The upper and lower limits of these smaller ranges can be included independently in the smaller ranges and are also included in the present invention and are constrained to any limits specifically excluded within the stated ranges. Where the stated range includes one or both of the limits, ranges excluding either of these including the limits are also included in the invention.

  Unless defined otherwise, all technical and chemical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice and testing of the present specification, the preferred methods and materials are now described. All publications mentioned in this specification are herein incorporated by reference to disclose and describe the methods and / or materials with which the publications are cited.

  As used herein and in the appended claims, the singular forms “a”, “an”, and “the” refer to plural referents unless the context clearly dictates otherwise. Care must be taken to include. Thus, for example, reference to “a method” includes a plurality of such methods, and reference to “a dose” includes a reference to one or more doses or equivalents known to those of skill in the art. , Etc.

  The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such publication by the prior invention. Further, the dates of publication provided may be different from the actual publication dates that may need to be independently confirmed.

Detailed Description of the Invention The present invention includes methods of treating alphavirus infections (including methods of treating West Nile virus infection, methods of treating dengue virus infection, methods of treating YFV virus infection, and methods of treating HCV infection) ); And methods of treating liver fibrosis, including reducing clinical liver fibrosis, reducing the likelihood of liver fibrosis, and reducing parameters associated with liver fibrosis To do. This method generally involves administering to a subject in need thereof a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ. Of particular interest among many embodiments is human therapy.

  Liver fibrosis is a precursor to complications associated with cirrhosis (eg, portal hypertension, progressive liver failure, and hepatocellular carcinoma). Accordingly, a reduction in liver fibrosis reduces the occurrence of such future complications. Thus, the present invention further provides a method for reducing the likelihood that an individual will develop complications associated with cirrhosis.

  Some of such methods of the invention generally involve administering a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for the treatment of West Nile virus infection. including. As used herein, a “therapeutically effective amount” of a type I or type III interferon receptor agonist and IFN-γ is effective in treating West Nile virus infection, type I or The amount of type III interferon receptor agonist and IFN-γ.

  Other methods of the invention generally comprise administering a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ. As used herein, a “therapeutically effective amount” of a type I or type III interferon receptor agonist and IFN-γ is used in treating alphavirus infection or treating West Nile virus infection. Or when treating an HCV virus infection, achieving a sustained viral response in HCV infection, or reducing liver fibrosis, or the likelihood that an individual will develop liver fibrosis Amounts of type I or type III interferon receptor agonist and IFN-γ effective in reducing or reducing parameters associated with liver fibrosis or in reducing disorders associated with cirrhosis Amount.

  In certain embodiments, the methods of the invention comprise administering a synergistically effective amount of a Type I or Type III interferon receptor agonist and IFN-γ. As used herein, a “synergistically effective amount” of a type I or type III interferon receptor agonist and IFN-γ used in combination therapy for a particular disease, and any of its The grammatical equivalent is the same amount of type I or type III interferon receptor agonist used in monotherapy for the disease and the same amount of IFN-γ used in monotherapy for the disease. The amount of type I or type III interferon receptor agonist and the amount of IFN-γ that have a higher effect on the disease in combination than would be expected for a mere additive effect.

  For example, in certain embodiments, the methods of the invention comprise administering a synergistically effective amount of IFN-α and IFN-γ. As used herein, a “synergistically effective amount” of IFN-α and IFN-γ and any grammatical equivalents thereof used in combination therapy for a particular disease are: The same amount of IFN-α used in monotherapy for the disease, and the same amount of IFN-γ used in monotherapy for the disease, in the disease than expected for the mere additive effect The amount of IFN-α and the amount of IFN-γ that have a higher effect on the combination in combination.

  In certain embodiments of the invention, a selected amount of a type I or type III interferon receptor agonist and a selected amount of IFN-γ are effective when used in combination therapy for disease, but are selected. An amount of a type I or type III interferon receptor agonist and / or a selected amount of IFN-γ is not effective when used in monotherapy for a disease. Accordingly, the present invention provides: (1) When used in combination therapy for disease, a selected amount of IFN-γ is a therapeutic advantage of a selected amount of a type I or type III interferon receptor agonist. The amount of IFN-γ selected here does not provide a therapeutic benefit when used in monotherapy for disease, (2) used in combination therapy for disease Sometimes, a selected amount of a type I or type III interferon receptor agonist enhances the therapeutic benefit of a selected amount of IFN-γ, where a selected amount of a type I or type III interferon receptor An agonist is selected when used in a regimen and (3) combination therapy for disease, which does not provide a therapeutic benefit when used in monotherapy for the disease. Amount of type I or type III interferon receptor agonist and selected amount of IFN-γ enhances therapeutic benefit, wherein the selected amount of type I or type III interferon receptor agonist and IFN-γ Each includes a regimen that each provides no therapeutic benefit when used in monotherapy for the disease. As used herein, a “synergistically effective amount” of a type I or type III interferon receptor agonist and IFN-γ, and grammatical equivalents thereof are defined above as (1)-( It should be understood to include any regimen included by any of 3).

Alphavirus The present invention provides a method for treating alphavirus infection. The method generally includes administering to the individual a type I or type III interferon receptor agonist and IFN-γ in an amount effective to alleviate the clinical course of the disease.

  Whether the subject method is effective in treating alphavirus infections may include a reduction in the number or length of hospitalizations, a reduction in time to viral clearance, a reduction in morbidity or mortality in clinical outcomes, or It can be determined by other indicators of disease response.

  In general, effective amounts of type I or type III interferon receptor agonists and IFN-γ are (i) reduced time for viral clearance, (ii) morbidity or mortality in the clinical course of the disease. An amount effective for reduction, or (iii) reduction of viral load in a patient.

West Nile Virus The present invention provides a method for treating West Nile virus infection. This method generally reduces type I or type III interferon receptor agonist, or IFN-γ, or type I or type III interferon receptor agonist and IFN-γ, the time for viral clearance in an individual. And / or administering to an individual in an amount that is effective to alleviate the clinical course of the disease.

  Whether the subject method is effective in treating West Nile virus infections includes a reduction in the number or length of hospitalizations, a reduction in time to viral clearance, a reduction in morbidity or mortality in clinical outcomes, Or it can be determined by other indicators of disease response.

  In general, an effective amount of a type I or type III interferon receptor agonist, an effective amount of IFN-γ, or an effective amount of a type I or type III interferon receptor agonist and IFN-γ is due to viral clearance. An amount effective to reduce the duration of the disease, or an amount effective to reduce morbidity or mortality in the clinical course of the disease.

Dengue virus The present invention provides a method for treating dengue virus infection. This method generally reduces type I or type III interferon receptor agonist, or IFN-γ, or type I or type III interferon receptor agonist and IFN-γ, the time for viral clearance in an individual. And / or administering to an individual in an amount that is effective to alleviate the clinical course of the disease.

  Whether the subject method is effective in treating a dengue virus infection may include a reduction in the number or length of hospitalizations, a reduction in time to viral clearance, a reduction in morbidity or mortality in clinical outcome, or disease It can be determined by other indicators of response.

  In general, an effective amount of a type I or type III interferon receptor agonist, an effective amount of IFN-γ, or an effective amount of a type I or type III interferon receptor agonist and IFN-γ is up to viral clearance. An amount effective to reduce time or an amount effective to reduce morbidity or mortality in the clinical course of the disease.

Yellow Fever Virus The present invention provides a method for treating yellow fever virus (YFV) infection. This method generally reduces type I or type III interferon receptor agonists, or IFN-γ, or type I or type III interferon receptor agonists and IFN-γ, to reduce the time to viral clearance in an individual. And / or administering to an individual in an amount that is effective to alleviate the clinical course of the disease.

  Whether the subject method is effective in treating YFV infection may include a reduction in the number or length of hospitalizations, a reduction in time to viral clearance, a reduction in morbidity or mortality in clinical outcome, or disease It can be determined by other indicators of response.

  In general, an effective amount of a type I or type III interferon receptor agonist, an effective amount of IFN-γ, or an effective amount of a type I or type III interferon receptor agonist and IFN-γ is up to viral clearance. An amount effective to reduce time or an amount effective to reduce morbidity or mortality in the clinical course of the disease.

Hepatitis C virus The present invention provides a method for treating HCV. This method generally provides an individual with a type I or type III interferon receptor agonist and IFN-γ in an amount that is effective to reduce viral load in the individual and to achieve a sustained viral response. Administration step.

  Whether the subject method is effective in treating HCV infection can be determined by measuring viral load or parameters associated with HCV infection (liver fibrosis, elevated serum transaminase levels, and necrotic inflammation in the liver. Can be determined by measuring (including but not limited to sexual activity). Indicators of liver fibrosis are discussed in detail below.

  Viral load can be measured by measuring the titer or level of virus in serum. These methods include, but are not limited to, quantitative polymerase chain reaction (PCR) and branched DNA (bDNA) testing. Quantitative assays have been developed to measure the viral load (titer) of HCV RNA. Many such assays are commercially available, including quantitative reverse transcription PCR (RT-PCR) (Amplicor HCV Monitor ™, Roche Molecular Systems, New Jersey); and branched DNA (deoxyribobodies). Nucleic acid) signal amplification assay (Quantiplex ™ HCV RNA assay (bDNA), Chiron Corp., Emeryville, California). See, for example, Gretch et al. (1995) Ann. Intern. Med. 123: 321-329.

  In general, effective amounts of IFNα and IFNγ reduce viral load to undetectable levels, for example, less than about 5000, less than about 1000, less than about 500, or less than about 200 genome copies / mL serum. Is an effective amount. In certain embodiments, an effective amount of a Type I or Type III interferon receptor agonist and / or IFN-γ is an amount effective to reduce viral load to less than 100 genome copies / mL serum. In many embodiments, the methods of the invention achieve a sustained viral response, e.g., viral load is at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months after discontinuation of treatment, Decrease to a detectable level for a period of at least about 5 months, or at least about 6 months.

  As described above, whether a subject method is effective in treating HCV infection can be determined by measuring parameters associated with HCV infection, such as liver fibrosis. Methods for determining the extent of liver fibrosis are discussed in detail below. In certain embodiments, the level of liver fibrosis serum marker indicates the extent of liver fibrosis.

  As one non-limiting example, serum alanine aminotransferase (ALT) levels are measured using standard assays. In general, ALT levels below about 45 international units are considered normal. In certain embodiments, an effective amount of IFNα and IFNγ is an amount effective to reduce ALT levels to less than about 45 U / ml serum.

  A therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ is at least about 10%, at least about 20% of the level of the marker in an untreated individual, or compared to a placebo-treated individual. At least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70% At least about 75%, or at least about 80%, or more, an amount effective to reduce serum levels of markers of liver fibrosis. Methods for measuring serum markers include immunologically based methods such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, etc. using antibodies specific for a given serum marker.

Fibrosis
Whether treatment with type I or type III interferon receptor agonists and IFN-γ is effective in reducing liver fibrosis is well established for measuring liver fibrosis and liver function Determined by any of a number of techniques. Reduction in liver fibrosis is determined by analyzing liver biopsy samples. Liver biopsy analysis reflects two major components: necrotic inflammation assessed by "grade" as a measure of severity and activity of the disease being advanced, and as reflecting long-term disease progression Assessment of fibrotic lesions and parenchymal or vascular remodeling, as assessed by the “stage” of See, for example, Brunt (2000) Hepatol. 31: 241-246; and METAVIR (1994) Hepatology 20: 15-20. A score is assigned based on analysis of the liver biopsy. There are a number of standardized scoring systems that provide a quantitative assessment of the degree and severity of fibrosis. These include the scoring systems METAVIR, Knodell, Scheuer, Ludwig, and Ishak.

  The METAVIR scoring system has various features of liver biopsy (fibrosis (portal vein fibrosis, centrilobular fibrosis, and cirrhosis); necrosis (fragile and lobular necrosis, eosinophilic regression, and balloon degeneration) Inflammation (portal tract inflammation, portal lymphatic assembly, and distribution of portal venous inflammation); biliary changes; and Knodell index (periportal necrosis, lobular necrosis, portal vein inflammation, fibrosis, and overall disease activity Based on analysis). The definition of each stage in the METAVIR system is as follows: score 0: no fibrosis; score 1: astral enlargement of the portal tract but no septum formation; score: 2, star of the portal tract Enlarged and with rare septal formation; score 3, many septum without cirrhosis; and score; 4, cirrhosis.

  The Knodell scoring system, also called the hepatitis activity indicator, is divided into four categories of histological features: I. Periportal and / or bridging necrosis; II. Leaflet degeneration and focal necrosis; III. Portal vein inflammation; and IV. Classify samples based on fibrosis score. In the Knodell staging system, the scores are as follows: score: 0, no fibrosis; score: 1, mild fibrosis (fibrous portal dilation); score: 2, moderate fibrosis; score: 3, severe fibrosis (bridging fibrosis); and score 4, cirrhosis. The higher the score, the higher the severity of liver tissue damage. Knodell (1981) Hepatol. 1: 431.

  In the Scheuer scoring system, the scores are as follows: score: 0, no fibrosis; score: 1, enlarged fibrotic portal tract; score: 2, periportal or portal-portal septum There is an intact structure; score: 3, fibrosis with structural distortion but not overt cirrhosis; and score 4, possible or definitive cirrhosis. Scheuer (1991) J. Hepatol. 13: 372.

  The Ishak scoring system is described in Ishak (1995) J. Hepatol. 22: 696-699. Stage 0, no fibrosis; Stage 1, fibrosis of some portal vein areas, with or without short fibrotic septum; Stage 2, fibrosis of most portal vein areas, With or without short fibrotic septum; Stage 3, fibrous enlargement of most portal vein regions with sparsely present portal-portal (PP) bridges; Stage 4, marked bridges (PP ) As well as fibrous enlargement of the portal vein region with the portal-center (PC); stage 5, marked cross-linking (PP and / or PC) with sparsely presenting nodules (incomplete cirrhosis); stage 6 Cirrhosis, possible or definitive. The benefits of anti-fibrotic treatment also include a multi-component point system based on serum bilirubin levels, serum albumin levels, prothrombin time, presence and severity of ascites, and presence and severity of brain damage, It can be measured and evaluated by using the Pugh scoring system. Based on the presence and severity of these parameter abnormalities, patients can be placed in one of three categories of increasing severity of clinical disease: A, B, or C.

  In certain embodiments, a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ is 1 unit or more in a fibrosis stage based on a pre-treatment liver biopsy and a post-treatment liver biopsy. The amount of type I or type III interferon receptor agonist and IFN-γ that results in changes in In certain embodiments, a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ reduces liver fibrosis by at least 1 unit in the scoring systems METAVIR, Knodell, Scheuer, Ludwig, and Ishak To do.

  Second or indirectly, indicators of liver function can also be used to assess the efficacy of treatment of type I or type III interferon receptor agonists and IFN-γ. A morphometric computerized semi-automated assessment of the quantitative degree of liver fibrosis based on specific staining of collagen and / or serum markers of liver fibrosis can also be measured as an indicator of the efficacy of the subject treatment method. Secondary indicators of liver function include, but are not limited to, serum transaminase level, prothrombin time, bilirubin, platelet count, portal wedge pressure, albumin level, and Child-Pugh score. An effective amount of IFN-α and IFN-γ is an indicator of liver function in an untreated individual, or at least about 10%, at least about 20%, an indicator of liver function compared to an individual treated with placebo. At least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, An amount effective to increase by at least about 75%, or at least about 80% or more. One skilled in the art can readily measure such indicators of liver function using standard assay methods, many of which are commercially available and routinely used in clinical settings.

  Serum markers of liver fibrosis can also be measured as an indication of the efficacy of the subject treatment method. Serum markers of liver fibrosis include but are not limited to hyaluronic acid, N-terminal procollagen III peptide, type 7 collagen 7S domain, C-terminal procollagen I peptide, and laminin. Additional biochemical markers of liver fibrosis include α-2-macroglobulin, haptoglobin, gamma globulin, apolipoprotein A, and gamma glutamyl transpeptidase.

  A therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ compares the serum level of a marker of liver fibrosis to the level of the marker in an untreated individual or a placebo treated individual. At least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60 %, An amount effective to reduce by at least about 65%, at least about 70%, at least about 75%, or at least about 80% or more. One skilled in the art can readily measure such indicators of liver function using standard assay methods, many of which are commercially available and routinely used in clinical settings. Methods for measuring serum markers include immunologically based methods such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, etc. using antibodies specific for a given serum marker.

  Quantitative tests of functional liver accumulation can also be used to assess the efficacy of treatments with type I or type III interferon receptor agonists and IFN-γ. These include indocyanine green clearance (ICG), galactose removal capacity (GEC), aminopurine breath test (ABT), antipyrine clearance, monoethylglycine-xylidide (MEG-X) clearance, and caffeine clearance.

  As used herein, a "complication associated with cirrhosis" is a sequelae of decompensated liver disease, i.e. following or resulting from the occurrence of decompensated liver disease. This includes ascites activity, varicose vein bleeding, portal hypertension, jaundice, progressive liver failure, brain damage, hepatocellular carcinoma, liver failure requiring liver transplantation, and liver-related Including, but not limited to:

  A therapeutically effective amount of a Type I or Type III interferon receptor agonist and IFN-γ is at least about 10%, at least about 20%, at least about at least compared to an untreated individual or a placebo-treated individual. 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, or at least about 80% or more, an amount that is effective in reducing the occurrence of a disorder associated with cirrhosis (eg, the likelihood that an individual will develop).

  Whether a treatment with a type I or type III interferon receptor agonist and IFN-γ is effective in reducing the incidence of disorders associated with cirrhosis can be readily determined by one skilled in the art.

  Reduction in liver fibrosis increases liver function. Accordingly, the present invention provides a method for increasing liver function, which generally comprises administering a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ. including. Liver functions include, for example, serum proteins (e.g. albumin, clotting factor, alkaline phosphatase, aminotransferase (e.g. alanine transaminase, aspartate transaminase), 5'-nucleosidase, γ-glutaminyl transpeptidase, etc.) Protein synthesis, bilirubin synthesis, cholesterol synthesis, and bile acid synthesis; liver metabolic functions (including but not limited to amino acid and ammonia metabolism, hormone metabolism, and lipid metabolism); exogenous drug detoxification; hemodynamics Functions such as, but not limited to, visceral and portal hemodynamics.

  Whether liver function is increased can be readily ascertained by one skilled in the art using well-established tests of liver function. Therefore, synthesis of markers of liver function such as albumin, alkaline phosphatase, alanine transaminase, asparagine transaminase, bilirubin, measure the levels of these markers in serum using standard immunological and enzymatic assays Can be evaluated. Visceral circulation and portal hemodynamics can be measured by portal wedge pressure and / or resistance using standard methods. Metabolic function can be measured by measuring the level of ammonia in the serum.

  Whether serum proteins normally secreted by the liver are in the normal range can be determined by measuring the level of such proteins using standard immunological and enzymatic assays. One skilled in the art knows the normal range for such serum proteins. The normal level of alanine transaminase is about 45 U / ml serum. The normal range for aspartate transaminase is about 5 to about 40 units / ml serum. Bilirubin is measured using standard assays. Normal bilirubin levels are usually less than about 1.2 mg / dL. Serum albumin levels are measured using standard assays. Normal levels of serum albumin are in the range of about 35 to about 55 g / L. Prolongation of prothrombin time is measured using standard assays. Normal prothrombin time is less than about 4 seconds longer than the control.

  Therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ has a liver function of at least about 10%, at least about 20%, at least about 30%, at least about It is effective to increase 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% or more. For example, a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ provides elevated levels of serum markers of liver function of at least about 10%, at least about 20%, at least about 30%, at least An amount effective to reduce about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% or more, or to reduce the level of serum markers of liver function within the normal range . A therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ also reduces the level of serum markers of liver function by at least about 10%, at least about 20%, at least about 30%, at least about An amount effective to increase 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% or more, or to increase the level of a serum marker of liver function within the normal range.

Type I Interferon Receptor Agonist In any of the methods described above, in certain embodiments, a Type I interferon receptor agonist is administered. Type I interferon receptor agonists include IFN-α; IFN-β; IFN-τ; IFN-ω; antibody agonists specific for type I interferon receptors; and any other agonist of type I interferon receptor (non-peptidic) Including agonists).

Interferon-α
Any known IFN-α can be used in the present invention. The term “interferon-α” as used herein refers to a family of related polypeptides that inhibit viral replication and cell growth and modulate the immune response. The term “interferon-α” includes naturally occurring IFN-α; synthetic IFN-α; derivatized IFN-α (eg, PEGylated IFN-α, glycosylated IFN-α, etc.); Or synthetic IFN-α analogs; essentially any IFN-α with antiviral properties, as described for naturally occurring IFN-α, is included.

  Suitable alpha interferons include, but are not limited to: naturally occurring IFN-α (including but not limited to naturally occurring IFN-α2a, IFN-α2b); Schering Corporation, Kenilworth, Recombinant interferon α-2b such as Intron-A interferon available from NJ; Recombinant interferon α-2a such as Roferon interferon available from Hoffmann-La Roche, Nutley, NJ; Boehringer Ingelheim Pharmaceutical, Inc., Ridgefield, Recombinant interferon α-2C such as Berofor α2 interferon available from Conn .; Sumiferon available from Sumitomo, Japan or Wellferon interferon α-n1 (INS) available from Glaxo-Wellcome Ltd., London, Great Britain, etc. Interferon α-n1; a product mixture of natural α-interferon; and manufactured by Interferon Sciences and under the trade name Alferon, Purdu e Interferon α-n3, a mixture of natural α interferons available from Frederick Co., Norwalk, Conn.

The term “IFN-α” also includes consensus IFN-α. Consensus IFN-α (also referred to as `` CIFN '' and `` IFN-con '' and `` consensus interferon '') is disclosed in U.S. Pat.Nos. 4,695,623 and 4,897,471, IFN-con ₁ , IFN-con ₂ , and including but not limited to the amino acid sequence called con ₃ ; and consensus interferon is a consensus sequence determination of naturally occurring interferon alpha (e.g., Infergen®, InterMune, Inc., Brisbane, Calif.). Defined by IFN-con ₁ is a consensus interferon agent in the Infargen® alfacon 1 product. The Infargen® consensus interferon product is referred to herein by its trade name (Infargen®) or by its generic name (interferon alfacon 1). Of particular interest is the use of CIFN, where the DNA sequence encoding IFN-con can be synthesized as described in the above-mentioned patents or other standard methods.

  Also suitable for use in the present invention are fusion proteins comprising IFN-α and a heterologous polypeptide. Suitable IFN-α fusion polypeptides include Albuferon-alfa ™ (human albumin and IFN-α fusion products; Human Genome Sciences; eg, Osborn et al. (2002) J. Pharmacol. Exp. Therap. 303 : 540-548), but is not limited to this. Gene shuffled forms of IFN-α are also suitable for use in the present invention. See, for example, Masci et al. (2003) Curr. Oncol. Rep. 5: 108-113.

PEGylated interferon-α
The term “IFN-α” also includes derivatives of IFN-α that have been derivatized (eg, chemically modified) to alter certain properties, such as serum half-life. As such, the term “IFN-α” includes glycosylated IFN-α; IFN-α derivatized with polyethylene glycol (“PEGylated IFN-α”) and the like. PEGylated IFN-α and methods for making it are discussed in US Pat. Nos. 5,382,657; 5,981,709; and 5,951,974. PEGylated IFN-α includes PEG conjugates and any of the IFN-α molecules described above, which bind to interferon α-2a (Roferon, Hoffman La-Roche, Nutley, NJ) PEG, conjugated to interferon α2b (Intron, Schering-Plough, Madison, NJ), PEG conjugated to interferon α-2c (Berofor Alfa, Boehringer Ingelheim, Ingelheim, Germany); And PEG conjugated to consensus interferon as defined by determination of the consensus sequence of naturally occurring interferon alpha (Infergen®, InterMune, Inc., Brisbane, Calif.). It is not limited.

  Any of the IFN-α polypeptides described above can be modified, ie PEGylated, with one or more polyethylene glycol moieties. The PEG molecule of a PEGylated IFN-α polypeptide is conjugated to one or more amino acid side chains of the IFN-α polypeptide. In certain embodiments, PEGylated IFN-α comprises a PEG moiety on only one amino acid. In other embodiments, the PEGylated IFN-α includes a PEG moiety on two or more amino acids, for example, IFN-α is 2, 3, 4, 5, 6, 7, 8, 9 Or a PEG moiety attached to 10 different amino acid residues.

  IFN-α can be coupled directly to PEG (ie, without a linker group) through an amino group, a sulfhydryl group, a hydroxyl group, or a carboxyl group.

  In certain embodiments, the PEGylated IFN-α is PEGylated at or near the amino terminus (N-terminus) of the IFN-α polypeptide, e.g., the PEG moiety is amino acid 1 to amino acid 4 or amino acid 5 to amino acid. Up to about 10 one or more amino acid residues are conjugated to the IFN-α polypeptide.

  In other embodiments, the PEGylated IFN-α is PEGylated at about 10 to about 28 one or more amino acid residues.

  In other embodiments, the PEGylated IFN-α is at or near the carboxyl terminus (C-terminus) of the IFN-α polypeptide, eg, one or more amino acids from amino acids 156 to 166, or amino acids 150 to 155. PEGylated at the residue.

  In other embodiments, PEGylated IFN-α is PEGylated at one or more amino acid residues of amino acids 100-114.

  Polyethylene glycol derivatization of amino acid residues at or near the receptor binding and / or active site domains of the IFN-α protein can disrupt the functionality of these domains. In certain embodiments of the invention, the amino acids from which PEGylation is to be avoided include amino acid residues from amino acid 30 to amino acid 40; and amino acid residues from amino acid 113 to amino acid 149.

  In certain embodiments, PEG is attached to IFN-α via a linking group. A linking group is any biocompatible linking group, where “biocompatible” means that the compound and group are non-toxic and utilized in vitro or in vivo without causing damage, illness, disease, or death. Can be done. PEG can be attached to the linking group via a linking group, such as an ether bond, an ester bond, a thiol bond, or an amide bond. Suitable biocompatible linking groups include, but are not limited to, ester groups, amide groups, imide groups, carbamate groups, carboxyl groups, hydroxyl groups, carbohydrates, succinimide groups (e.g., succinimid groups). Rusuccinate (SS), succinimidyl propionate (SPA), succinimidyl butanoate (SBA), succinimidyl carboxymethylate (SCM), succinimidyl succinamide (SSA), Or N-hydroxysuccinimide (NHS)), epoxide groups, oxycarbonylimidazole groups (e.g. including carbonyldiimidazole (CDI)), nitrophenyl groups (e.g. nitrophenyl carbonate (NPC) or trichlorophenyl carbonate) (Including TPC)), trisylate groups, aldehyde groups, isocyanate groups, vinyl sulfone groups, tyrosines Group, a cysteine group, a histidine group or a primary amine.

  Methods for making ester activated PEGs of succimidyl propionate (SPA) and succinimidyl butanoate (SBA) are described in US Pat. No. 5,672,662 (Harris, et al.) And WO 97 / 03106.

  Methods for conjugating PEG to IFN-α polypeptides are known in the art and any known method can be used. For example, Park et al., Anticancer Res., 1: 373-376 (1981); Zaplipsky and Lee, Polyethylene Glycol Chemistry: Biotechnical and Biomedical Applications, JM Harris, Plenum Press, NY, Chapter 21 (1992); US See US Pat. No. 5,985,265; US Pat. No. 5,672,662 (Harris, et al.) And WO 97/03106.

  PEGylated IFN-α and methods for making it are discussed, for example, in US Pat. Nos. 5,382,657; 5,981,709; 5,985,265; and 5,951,974. PEGylated IFN-α includes PEG conjugates and any of the above IFN-α molecules, including but not limited to: interferon α-2a (Roferon, Hoffman LaRoche, PEG conjugated to Nutley, NJ, where PEGylated Roferon is known as Pegasys (Hoffman LaRoche); PEG conjugated to interferon α2b (Intron, Schering-Plough, Madison, NJ), Here PEGylated Intron is known as PEG-Intron (Schering-Plough); PEG conjugated to interferon alpha-2c (Berofor Alfa, Boehringer Ingelheim, Ingelheim, Germany); and naturally occurring interferon alpha PEG conjugated to consensus interferon (CIFN) as defined by determination of consensus sequence (Infergen®, InterMune, Inc., Brisbane, Calif.), Where PEGylated Infergen is PEG-Infergen Called.

  In many embodiments, the PEG is a monomethoxy PEG molecule that reacts with a primary amine group on the IFN-α polypeptide. Methods for modifying polypeptides with monomethoxy PEG via reductive alkylation are known in the art. See, for example, Chamow et al. (1994) Bioconj. Chem. 5: 133-140.

  In one non-limiting example, PEG is attached to IFN-α via a SPA linking group. The SPA ester of PEG and methods for making it are described in US Pat. No. 5,672,662. SPA binding provides binding to a free amine group on the IFN-α polypeptide.

  For example, the PEG molecule is covalently linked via a bond comprising an amide bond between the propionyl group of the PEG moiety and the ε-amino group of the lysine residue exposed on the surface in the IFN-α polypeptide. Such a bond can be formed, for example, by condensation of α-methoxy, ωpropanoic acid activated ester of PEG (mPEGspa).

As one non-limiting example, one preferred monoPEGylated CIFN conjugate for use herein is an approximately 30 kD linear PEG moiety attached via covalent linkage to a CIFN polypeptide. Where the covalent bond is an amide bond between the propionyl group of the PEG moiety and the ε-amino group of the surface exposed lysine residue in CIFN, where the surface exposed lysine residue Is selected from lys ³¹ , lys ⁵⁰ , lys ⁷¹ , lys ⁸⁴ , lys ¹²¹ , lys ¹²² , lys ¹³⁴ , lys ¹³⁵ , and lys ¹⁶⁵ , and the amide bond is activated by α-methoxy, ω propionic acid activated ester of PEG It is formed.

Polyethylene glycol
Polyethylene glycols suitable for conjugation to an IFN-α polypeptide are soluble in water at room temperature and have the general formula R (O—CH ₂ —CH ₂ ) _n OR, where R is Hydrogen or a protecting group (eg an alkyl or alkanol group), where n is an integer from 1 to 1000; When R is a protecting group, it generally has 1 to 8 carbon atoms.

  In many embodiments, PEG has at least one hydroxyl group (e.g., a terminal hydroxyl group), which is an amino group (e.g., the ε-amino group of a lysine residue, the free amino group at the N-terminus of a polypeptide). Or any other amino group such as the amino group of asparagine, glutamine, arginine, or histidine).

In other embodiments, PEG is derivatized to be reactive with a free carboxyl group in an IFN-α polypeptide (eg, a free carboxyl group at the carboxyl terminus of an IFN-α polypeptide). Suitable derivatives of PEG that are reactive with the free carboxyl group at the carboxyl terminus of IFN-α include, but are not limited to, PEG-amine, and hydrazine derivatives of PEG (eg, PEG-NH-NH ₂ ). Not.

  In other embodiments, the PEG is derivatized to include a terminal thiocarboxylic acid group, —COSH, that selectively reacts with an amino group to produce an amide derivative. Due to the reactive nature of thioacids, certain amino group selectivity over other groups is achieved. For example, -SH exhibits sufficient free radical properties in the reaction with the N-terminal amino group at appropriate pH conditions so that the ε amino group in the lysine residue is protonated and remains non-nucleophilic. is there. On the other hand, reactions under appropriate pH conditions can create several accessible lysine residues to react with selectivity.

In other embodiments, the PEG comprises a reactive ester such as N-hydroxysuccinimidate at the end of the PEG chain. Such N-hydroxysuccinimidate-containing PEG molecules react with selected amino groups at specific pH conditions (eg, at neutral 6.5-7.5). For example, the N-terminal amino group can be selectively modified under neutral pH conditions. However, if the reagent reactivity is extreme, the accessible NH ₂ group of lysine can also react.

  PEG can be conjugated to the IFN-α polypeptide directly or through a linker. In certain embodiments, a linker is added to the IFN-α polypeptide to form a linker-modified IFN-α polypeptide. Such linkers provide various functionalities (eg, reactive groups (such as sulfhydryl groups, amino groups, or carboxyl groups) for coupling PEG reagents to linker-modified IFN-α polypeptides).

  In certain embodiments, the PEG conjugated to the IFN-α polypeptide is linear. In other embodiments, the PEG conjugated to the IFN-α polypeptide is branched. Branched PEG derivatives are multi-arms such as those described in US Pat. No. 5,643,575, “star-PEG”, and those described in Shearwater Polymers, Inc. catalog “Polyethylene Glycol Derivatives 1997-1998”. PEG. star PEG is described in the art, for example, in US Pat. No. 6,046,305.

  PEG having a molecular weight in the range of about 2 kDa to about 100 kDa is commonly used, where the term “about” refers to a molecule in a polyethylene glycol preparation in the context of PEG. Indicates that the molecular weight is somewhat higher or lower than the molecular weight. For example, suitable PEGs for conjugation to IFN-α are about 2 kDa to about 5 kDa, about 5 kDa to about 10 kDa, about 10 kDa to about 15 kDa, about 15 kDa to about 20 kDa, about 20 kDa to about 25 kDa, about 25 kDa. To about 30 kDa, about 30 kDa to about 40 kDa, about 40 kDa to about 50 kDa, about 50 kDa to about 60 kDa, about 60 kDa to about 70 kDa, about 70 kDa to about 80 kDa, about 80 kDa to about 90 kDa, or about 90 kDa to about 100 kDa. .

Preparation of PEG-IFN-α conjugates As discussed above, the PEG moiety can be directly or via a linker at or near the N-terminus of the IFN-α polypeptide, within, or at or near the C-terminus. Can be attached to an amino acid residue. Conjugation can be performed in a liquid or in a solid phase.

N-terminal bond
Methods for attaching a PEG moiety to an amino acid residue at or near the N-terminus of an IFN-α polypeptide are known in the art. See, for example, US Pat. No. 5,985,265.

In some embodiments, known methods for selectively obtaining N-terminally chemically modified IFN-α are used. For example, methods of protein modification by reductive alkylation are used that take advantage of the differential reactivity of different types of primary amino groups available for derivatization in specific proteins (e.g., lysine vs. N-terminus). obtain. Under appropriate reaction conditions, substantially selective derivatization of the protein at the N-terminus with a carbonyl group-containing polymer is achieved. The reaction is performed at pH which allows one to take advantage of the difference in the pK _a between the ε-amino group of a lysine residue, the α-amino group of the N-terminal residue of the protein. Such selective derivatization controls the attachment of the PEG moiety to IFN-α: Conjugation with the polymer occurs predominantly at the N-terminus of IFN-α and other reactive groups (e.g., No significant modification of the lysine side chain amino group occurs.

C-terminal coupling N-terminal specific coupling procedures such as those described in US Pat. No. 5,985,265 predominately provide mono-PEGylated products. However, the purification procedure is aimed at removing excess reagents, and a few multiplexed PEGylated products will remove N-terminally blocked polypeptides. From a therapeutic point of view, such a process results in a significant increase in manufacturing costs. For example, the well-characterized Infergen® Alfacon-1 CIFN polypeptide amino acid sequence is approximately 5% shortened at the carboxyl terminus, so there is only one major C-terminal sequence. Thus, in one embodiment, N-terminal PEGylated IFN-α is not used and instead the IFN-α polypeptide is C-terminal PEGylated.

  Therefore, an effective synthetic and therapeutic approach to obtain monoPEGylated Infergen products is envisioned as follows.

  PEG reagents that are selective for the C-terminus can be prepared with or without a spacer. For example, polyethylene glycol modified with methyl ether or the like at one end and having an amino functional group at one end can be used as a starting material.

  Preparing or obtaining a water-soluble carbodiimide as a condensing agent can be performed. Coupling IFN-α (e.g., Infergen® Alfacon-1 CIFN or consensus interferon) with a water soluble carbodiide as a condensing agent is generally performed in an aqueous medium using a suitable buffer system. Performed at an optimum pH to provide an amide bond. High molecular weight PEG can be covalently added to proteins to increase molecular weight.

  The reagent selected depends on the process optimization study. A non-limiting example of a suitable reagent is EDAC or 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide. The water solubility of EDAC allows for direct addition to the reaction without the need for prior organic solvent dissolution. Both excess reagent and isourea formed as a by-product of the cross-linking reaction are water soluble and can be easily removed by dialysis or gel filtration. A concentrated solution of EDAC in water is prepared to facilitate the addition of low molar amounts to the reaction. Stock solutions are prepared and used immediately in view of the labile nature of the reagents. Many of the synthetic protocols in the literature suggest that the optimal reaction medium is in the pH range between 4.7 and 6.0. However, the condensation reaction proceeds without significant loss of yield up to pH 7.5. Water can be used as a solvent. In view of Infargen's intended use, preferably the medium is a 2- (N-morpholino) ethane sulfonate buffer pre-titrated to a pH between 4.7 and 6.0. However, 0.1M phosphoric acid at pH 7-7.5 can also be used in view of the fact that the product is in the same buffer. The ratio of PEG amine to IFN-α molecules is optimized so that the C-terminal carboxyl residue is selectively PEGylated to give a monoPEGylated derivative.

Even though the use of PEG amine is referred to above by name or structure, such derivatives are meant to be exemplary only, and hydrazine derivatives as in PEG-NH-NH ₂ that also condense with the carboxyl group of the IFN-α protein Other groups such as can also be used. In addition to the aqueous phase, the reaction can also be carried out on a solid phase. The polyethylene glycol can be selected from a list of compounds with molecular weights ranging from 300 to 40,000. The choice of the various polyethylene glycols is also determined by the coupling efficiency and in vitro and in vivo biological performance of the purified derivative, ie, circulation time, viral activity, and the like.

In addition, an appropriate spacer can be added to the C-terminus of the protein. This spacer may have a reactive group such as SH, NH ₂ , or COOH to couple with a suitable PEG reagent to provide a high molecular weight IFN-α derivative. A combined solid / aqueous phase methodology can be devised for the preparation of C-terminal PEGylated interferon. For example, the C-terminus of IFN-α is extended on a solid phase using a Gly-Gly-Cys-NH ₂ spacer and then mono-PEG in solution using the appropriate molecular weight dithiopyridyl-PEG reagent. It becomes. As coupling at the C-terminus is independent of blocking at the N-terminus, the envisaged processes and products are cost-effective (less than one third of the protein is consumed as in the N-terminal PEGylation method) and chronic C Contributes to the economics of treatment for treating hepatitis B infection, cirrhosis, etc.

  There may be a more reactive carboxyl group of the amino acid residue elsewhere in the molecule to react with the PEG reagent, resulting in monoPEGylation at that site or -COOH at the C-terminus of IFN-α In addition to the group, it results in multiple PEGylations. These reactions are assumed to be the best and minimal due to steric freedom at the C-terminus of the molecule and steric hindrance imposed by carbodiimide and PEG reagents (e.g. in branched chain molecules). . This is therefore the preferred mode of PEG modification for Infergen and similar such proteins, either native or expressed in host systems, which improves efficiency and increases in vivo biology. The N-terminus may have been blocked to varying degrees to maintain specific activity.

  Another way to achieve C-terminal PEGylation is as follows. Selectivity for C-terminal PEGylation is achieved using sterically hindered reagents that exclude reactions at carboxy residues either embedded in the helix or internally in IFN-α. For example, one such reagent can be a branched PEG with a molecular weight of ˜40 kd and the agent can be synthesized as follows.

OH ₃ C- (CH ₂ CH ₂ O) _n -CH ₂ CH ₂ NH ₂ + glutamic acid, i.e. HOCO-CH ₂ CH ₂ CH (NH ₂ ) -COOH, with a suitable agent such as dicyclohexylcarbodiimide or water-soluble EDC Condensed and branched PEG drug OH ₃ C- (CH ₂ CH ₂ O) _n -CH ₂ CH ₂ NHCOCH (NH ₂ ) CH ₂ OCH _3- (CH ₂ CH ₂ O) _n -CH ₂ CH ₂ NHCOCH Provide _two .

  This reaction can be used in excess to couple the amino group with the free and flexible carboxyl group of IFN-α to form a peptide bond.

  If desired, the PEGylated IFN-α is not PEGylated using any known method including, but not limited to, ion exchange chromatography, size exclusion chromatography, and combinations of the above. Separated from. For example, if the PEG-IFN-α conjugate is monoPEGylated IFN-α, the products are first separated by ion exchange chromatography to produce a material having the charge characteristics of a monoPEGylated material (with the same apparent charge). Other multi-PEGylated materials may also be present) and then the monoPEGylated material is separated using size exclusion chromatography.

IFN-β
The term interferon-β (`` IFN-β '') refers to a naturally occurring IFN-β polypeptide; a non-naturally occurring IFN-β polypeptide; and a parent naturally occurring IFN-β or a non-naturally occurring IFN Includes naturally occurring IFN-β or non-naturally occurring analogs of IFN-β that retain antiviral activity of -β.

  Any of a variety of beta interferons can be delivered by the continuous delivery method of the present invention. Suitable β-interferons include naturally occurring IFN-β; IFN-β1a, such as Avonex® (Biogen, Inc.) and Rebif® (Serono, SA); IFN-β1b (Betaseron ( Registered trademark); Berlex) and the like.

  The IFN-β formulation may contain N-block species, where the N-terminal amino acid is acylated with an acyl group such as a formyl group, acetyl group, malonyl group, and the like. Consensus IFN-β is also suitable for use.

  IFN-β polypeptides can be produced by any known method. The DNA sequence encoding IFN-β can be synthesized using standard methods. In many embodiments, the IFN-β polypeptide is transformed or transfected into a bacterial host (e.g., E. coli) or a eukaryotic host cell (e.g., yeast; mammalian cells such as CHO cells), It is an expression product of the produced DNA sequence. In these embodiments, IFN-β is “recombinant IFN-β”. If the host cell is a bacterial host cell, IFN-β is modified to include an N-terminal methionine.

  It should be understood that IFN-β as described herein may include one or more modified amino acid residues, eg, glycosylation, chemical modification, and the like.

IFN-τ
The term IFN-τ refers to the antiviral activity of a naturally occurring IFN-τ polypeptide; a non-naturally occurring IFN-τ polypeptide; and the parent naturally occurring IFN-τ or non-naturally occurring IFN-τ. Contains a naturally occurring or non-naturally occurring analog of IFN-τ that is retained.

  Suitable τ interferons include, but are not limited to, naturally occurring IFN-τ; Tauferon® (Pepgen Corp.) and the like.

  IFN-τ as shown in any one of GenBank accession numbers P15696; P56828; P56832; P56829; P56831; Q29429; Q28595; Q28594; S08072; Q08071; Q08070; Q08053; P56830; P28169; P28172; and P28171 Amino acid sequences. The sequence of any known IFN-τ polypeptide can be altered in various ways known in the art to produce targeted changes in the sequence. Variant polypeptides are typically substantially similar to the sequences provided herein, ie, differ by at least 1 amino acid and can differ by at least 2 amino acids, but not more than about 10 amino acids. The sequence change can be a substitution, insertion, or deletion. Conservative amino acid substitutions typically include substitutions in the following groups: (glycine, alanine); (valine, isoleucine, leucine); (aspartic acid, glutamic acid); (asparagine, glutamine); Serine, threonine); (lysine, arginine); or (phenylalanine, tyrosine).

  Modifications of interest that may or may not change the primary amino acid sequence include chemical derivatization of polypeptides (e.g., acetylation or carboxylation; amino acids that introduce or remove glycosylation sites). Sequence changes; amino acid sequence changes that render the protein sensitive to PEGylation, etc.). Modifications of glycosylation, such as those made by modifying the glycosylation pattern of a polypeptide during its synthesis and processing, or in further processing steps; such as mammalian glucosylation or deglycosylation enzymes Also included are those made by exposing a polypeptide to an enzyme that affects glycosylation. Also included are sequences having phosphorylated amino acid residues, eg, phosphotyrosine, phosphoserine, or phosphothreonine.

  The IFN-τ formulation can contain N-block species, where the N-terminal amino acid is acylated with an acyl group such as a formyl group, acetyl group, malonyl group, and the like. A consensus IFN-τ is also suitable for use.

  IFN-τ polypeptides can be produced by any known method. The DNA sequence encoding IFN-τ can be synthesized using standard methods. In many embodiments, the IFN-τ polypeptide is transformed or transfected into a bacterial host (e.g., E. coli) or a eukaryotic host cell (e.g., yeast; mammalian cells such as CHO cells), It is an expression product of the produced DNA sequence. In these embodiments, IFN-τ is “recombinant IFN-τ”. If the host cell is a bacterial host cell, IFN-τ is modified to include an N-terminal methionine.

  It should be understood that IFN-τ as described herein may include one or more modified amino acid residues, eg, glycosylation, chemical modification, and the like.

IFN-ω
The term interferon-omega (`` IFN-omega '') refers to a naturally occurring IFN-omega polypeptide; a non-naturally occurring IFN-omega polypeptide; and a parent naturally occurring IFN-omega or non-naturally occurring IFN Includes naturally occurring IFN-ω or non-naturally occurring IFN-ω analogs that retain the antiviral activity of -ω.

  Any known omega interferon can be delivered by the continuous delivery method of the present invention. Suitable omega interferons include, but are not limited to, naturally occurring IFN-omega; recombinant IFN-omega, such as Biomed 510® (BioMedicines.).

  IFN-ω may comprise an amino acid sequence as shown in GenBank Accession Number NP — 002168; or AAA70091. The sequence of any known IFN-ω polypeptide can be altered in various ways known in the art to produce targeted changes in the sequence. Variant polypeptides are typically substantially similar to the sequences provided herein, ie, differ by at least 1 amino acid and can differ by at least 2 amino acids, but not more than about 10 amino acids. The sequence change can be a substitution, insertion, or deletion. Conservative amino acid substitutions typically include substitutions in the following groups: (glycine, alanine); (valine, isoleucine, leucine); (aspartic acid, glutamic acid); (asparagine, glutamine); Serine, threonine); (lysine, arginine); or (phenylalanine, tyrosine).

  Modifications of interest that may or may not change the primary amino acid sequence include chemical derivatization of polypeptides (e.g., acetylation or carboxylation; amino acids that introduce or remove glycosylation sites). Sequence changes; amino acid sequence changes that render the protein sensitive to PEGylation, etc.). Modifications of glycosylation, such as those made by modifying the glycosylation pattern of a polypeptide during its synthesis and processing, or in further processing steps; such as mammalian glucosylation or deglycosylation enzymes Also included are those made by exposing a polypeptide to an enzyme that affects glycosylation. Also included are sequences having phosphorylated amino acid residues, eg, phosphotyrosine, phosphoserine, or phosphothreonine.

  The IFN-ω formulation may contain N-block species, where the N-terminal amino acid is acylated with an acyl group such as a formyl group, acetyl group, malonyl group, and the like. A consensus IFN-ω is also suitable for use.

  IFN-ω polypeptides can be produced by any known method. The DNA sequence encoding IFN-ω can be synthesized using standard methods. In many embodiments, the IFN-ω polypeptide is transformed or transfected into a bacterial host (e.g., E. coli) or a eukaryotic host cell (e.g., yeast; a mammalian cell such as a CHO cell), It is an expression product of the produced DNA sequence. In these embodiments, IFN-ω is “recombinant IFN-ω”. If the host cell is a bacterial host cell, IFN-ω is modified to include an N-terminal methionine.

  It should be understood that IFN-ω as described herein may include one or more modified amino acid residues, eg, glycosylation, chemical modification, and the like.

Type III Interferon Receptor Agonist In any of the methods and apparatus described above, the interferon receptor agonist is in some embodiments an agonist of a type III interferon receptor (eg, a “type III interferon agonist”). Type III interferon agonists include IL-28b polypeptide; IL-28a polypeptide; and IL-29 polypeptide; an antibody specific for type III interferon receptor; and any other agonist of type III interferon receptor (non-peptide Sex agonists).

  IL-28A, IL-28B, and IL-29 (collectively referred to herein as “type III interferon” or “type III IFN”) are Sheppard et al. (2003) Nature 4: 63-68. Described in. Each polypeptide binds a heterodimeric receptor consisting of IL-10 receptor β chain and IL-28 receptor α. Sheppard et al. (2003), supra. The amino acid sequences of IL-28A, IL-28B, and IL-29 are found under GenBank accession numbers NP_742150, NP_742151, and NP_742152, respectively.

  The amino acid sequence of a type III IFN polypeptide can be altered in various ways known in the art to produce targeted changes in the sequence. Variant polypeptides are typically substantially similar to the sequences provided herein, ie, differ by at least 1 amino acid and can differ by at least 2 amino acids, but not more than about 10 amino acids. The sequence change can be a substitution, insertion, or deletion. Scanning mutations that systematically introduce alanine or other residues can be used to determine key amino acids. Specific amino acid substitutions of interest include conservative changes and non-conservative changes. Conservative amino acid substitutions typically include substitutions in the following groups: (glycine, alanine); (valine, isoleucine, leucine); (aspartic acid, glutamic acid); (asparagine, glutamine); Serine, threonine); (lysine, arginine); or (phenylalanine, tyrosine).

  Modifications of interest that may or may not change the primary amino acid sequence include chemical derivatization of polypeptides (e.g., acetylation or carboxylation; amino acids that introduce or remove glycosylation sites). Sequence changes; amino acid sequence changes that render the protein sensitive to PEGylation, etc.). Modifications of glycosylation, such as those made by modifying the glycosylation pattern of a polypeptide during its synthesis and processing, or in further processing steps; such as mammalian glucosylation or deglycosylation enzymes Also included are those made by exposing a polypeptide to an enzyme that affects glycosylation. Also included are sequences having phosphorylated amino acid residues, eg, phosphotyrosine, phosphoserine, or phosphothreonine.

Polypeptides modified using conventional chemical techniques to improve the resistance of the polypeptide to proteolysis, to optimize solubility properties, or to make the polypeptide more suitable as a therapeutic agent It is included in the present invention. For example, the peptide backbone can be cyclized to enhance stability (eg, Friedler et al. (2000) J. Biol. Chem. 275: 23783-23789). Analogs containing residues other than naturally occurring L-amino acids, eg, D-amino acids or non-naturally occurring synthetic amino acids may be used. The protein can be PEGylated to enhance stability. The polypeptide can be fused with albumin.

  Polypeptides can be prepared by in vitro synthesis, by recombinant methods, using conventional methods as known in the art, or isolated from cells that are derived or naturally produce proteins. Can be done. The particular sequence or mode of preparation is determined by convenience, economy, purity required, etc. If desired, various groups can be introduced into the polypeptide during synthesis or during expression, which allows linkage to other molecules or surfaces. Thus, cysteine is used to make thioesters, histidine is used to link to metal ion complexes, carboxyl groups form amides or esters, amino groups form amides, etc. obtain.

Interferon gamma
Nucleic acid sequences encoding IFN-γ polypeptides can be accessed from public databases (eg, Genbank, academic paper publications, etc.). Although various mammalian IFN-γ polypeptides are of interest, human proteins are generally used for the treatment of human diseases. The human IFN-γ coding sequence can be found in Genbank accession numbers X13274; V00543; and NM_000619. The corresponding genomic sequence can be found in Genbank accession numbers J00219; M37265; and V00536. See, for example, Gray et al. (1982) Nature 295: 501 (Genbank X13274); and Rinderknecht et al. (1984) J. Biol. Chem. 259: 6790.

  IFN-γ1b (Actimmune®; human interferon) is a 140 amino acid single chain polypeptide. It is made recombinantly in E. coli and is not glycosylated. Rinderknecht et al. (1984) J. B. C. 259: 6790-6797.

  The IFN-γ used in the method of the present invention is any of natural IFN-γ, recombinant IFN-γ, and derivatives thereof as long as they have IFN-γ activity, particularly human IFN-γ activity. possible. Human IFN-γ exhibits the antiviral and antiproliferative properties of interferon, as well as a number of other immunomodulatory activities as are known in the art. Although IFN-γ is based on sequences as provided above, protein production and proteolytic processing can result in processing the variant. The unprocessed sequence provided by Gray et al., Supra consists of 166 amino acids (aa). Recombinant IFN-γ produced in E. coli was originally thought to be 146 amino acids, but native human IFN-γ was cleaved after residue 23 (starting at amino acid 20) and 143aa It was next found to produce 144aa in the presence of terminal methionine as required for protein or bacterial expression. During purification, the mature protein can be further cleaved at the C-terminus after residue 162 (see Gray et al. Sequence), 139 amino acids, or first, for example, if required for bacterial expression In the presence of methionine, 140 amino acids are produced. The N-terminal methionine is an artifact encoded by the mRNA translation “start” signal AUG, which is not processed away in the specific case of E. coli expression. In other microbial or eukaryotic expression systems, methionine cannot be removed.

  For use in the subject methods, any of the native IFN-γ peptides, modifications, and variants and combinations of one or more peptides can be used. The IFN-γ peptides of interest include fragments and can be variously truncated at the carboxyl terminus relative to the entire sequence. Such fragments continue to exhibit the characteristic properties of human interferon as long as amino acids 24 to about 149 (numbering from unprocessed polypeptide residues) are present. An extra sequence can be substituted for the sequence after amino acid 155 without loss of activity. See, for example, US Pat. No. 5,690,925. The native IFN-γ moiety includes moieties extending from amino acid residues 24-150; 24-151; 24-152; 24-153; 24-155; and 24-157. Any of these variants, and other variants known in the art and having IFN-γ activity, can be used in the methods of the invention.

  The sequence of an IFN-γ polypeptide can be altered in various ways known in the art to produce targeted sequence changes. Variant polypeptides are typically substantially similar to the sequences provided herein, ie, differ by at least 1 amino acid and can differ by at least 2 amino acids, but not more than about 10 amino acids. The sequence change can be a substitution, insertion, or deletion. Scanning mutations that systematically introduce alanine or other residues can be used to determine key amino acids. Specific amino acid substitutions of interest include conservative changes and non-conservative changes. Conservative amino acid substitutions typically include substitutions in the following groups: (glycine, alanine); (valine, isoleucine, leucine); (aspartic acid, glutamic acid); (asparagine, glutamine); (Serine, threonine); (lysine, arginine); or (phenylalanine, tyrosine).

  Modifications of interest that may or may not change the primary amino acid sequence include chemical derivatization of polypeptides (e.g., acetylation or carboxylation; amino acids that introduce or remove glycosylation sites). Sequence changes; amino acid sequence changes that render the protein sensitive to PEGylation, etc.). In one embodiment, the present invention provides glycosyl or PEG derivatized polypeptides, such as IFN-γ polypeptide variants described in International Patent Publication No. WO 01/36001, with reduced serum clearance. Contemplated is the use of IFN-γ having one or more non-naturally occurring glycosylation and / or PEGylation sites that have been engineered as follows. Modifications of glycosylation, such as those made by modifying the glycosylation pattern of a polypeptide during its synthesis and processing, or in further processing steps; such as mammalian glucosylation or deglycosylation enzymes Also included are those made by exposing a polypeptide to an enzyme that affects glycosylation. Also included are sequences having phosphorylated amino acid residues, eg, phosphotyrosine, phosphoserine, or phosphothreonine.

  Polypeptides modified using conventional chemical techniques to improve the resistance of the polypeptide to proteolysis, to optimize solubility properties, or to make the polypeptide more suitable as a therapeutic agent It is included in the present invention. For example, the peptide backbone can be cyclized to enhance stability (eg, Friedler et al. (2000) J. Biol. Chem. 275: 23783-23789). Analogs containing residues other than naturally occurring L-amino acids, eg, D-amino acids or non-naturally occurring synthetic amino acids may be used. The protein can be PEGylated to enhance stability.

  Polypeptides can be prepared by in vitro synthesis, by recombinant methods, using conventional methods as known in the art, or isolated from cells that are derived or naturally produce proteins. Can be done. The particular sequence or mode of preparation is determined by convenience, economy, purity required, etc. If desired, various groups can be introduced into the polypeptide during synthesis or during expression, which allows linkage to other molecules or surfaces. Thus, cysteine is used to make thioesters, histidine is used to link to metal ion complexes, carboxyl groups form amides or esters, amino groups form amides, etc. obtain.

  The polypeptides of the invention can also be isolated and synthesized according to recombinant synthesis of conventional methods. Expression host lysates can be prepared, and the lysates are purified using HPLC, exclusion chromatography, gel electrophoresis, affinity chromatography, or other purification techniques. For the most part, the composition used is at least 20% by weight of the desired product, more usually at least about 75% by weight relative to the contaminants with respect to the method of preparation of the product and its purification. Preferably it contains at least about 95% weight and, for therapeutic purposes, usually at least about 99.5% weight. Usually the percentage is based on total protein.

ピルフェニドンアナログ

Pirfenidone and its analogs Pirfenidone (5-methyl-1-phenyl-2- (1H) -pyridone) and pirfenidone analogs have been disclosed for the treatment of fibrotic conditions. A “fibrotic condition” is one that is susceptible to treatment by administration of a compound having antifibrotic activity.
Pirfenidone

Pirfenidone analog

Description of substituents R ₁ , R ₂ , X
R ₁ : carbocyclic (saturated and unsaturated), heterocyclic (saturated and unsaturated), alkyl (saturated and unsaturated). Examples include phenyl, benzyl, pyrimidyl, naphthyl, indolyl, pyrrolyl, furyl, thienyl, imidazolyl, cyclohexyl, piperidyl, pyrrolidyl, morpholinyl, cyclohexenyl, butadienyl and the like.

R ₁ is further substituted with substituents such as halogen, nitro, amino, hydroxyl, alkoxy, carboxy, cyano, thio, alkyl, aryl, heteroalkyl, heteroaryl, and combinations thereof, such as 4-nitrophenyl, 3-chlorophenyl , Carbocyclic or heterocyclic moieties having 2,5-dinitrophenyl, 4-methoxyphenyl, 5-methyl-pyrrolyl, 2,5-dichlorocyclohexyl, guanidinyl-cyclohexenyl, and the like.

R ₂ : alkyl, carbocyclic, aryl, heterocyclic. Examples include methyl, ethyl, propyl, isopropyl, phenyl, 4-nitrophenyl, thienyl and the like.

  X: can be any number (1 to 3) of substituents on the carbocycle or heterocycle. The substituents can be the same or different. Substituents can include hydrogen, alkyl, heteroalkyl, aryl, heteroaryl, halo, nitro, carboxyl, hydroxyl, cyano, amino, thio, alkylamino, haloaryl, and the like.

  The substituents may optionally be substituted with 1-3 substituents from the group consisting of alkyl groups, nitro groups, alkoxy groups, hydroxyl groups, and halo groups. Examples include methyl, 2,3-dimethyl, phenyl, p-tolyl, 4-chlorophenyl, 4-nitrophenyl, 2,5-dichlorophenyl, furyl, thienyl and the like.

  Specific examples include:

(Table 1)

  U.S. Pat. Methods for synthesis and formulation are described.

Dosage, formulation, and route of administration
A Type I or Type III interferon receptor agonist and IFN-γ are administered to an individual in a formulation having a pharmaceutically acceptable excipient. A wide variety of pharmaceutically acceptable excipients are known in the art and need not be discussed in detail herein. Pharmaceutically acceptable excipients are described in various publications, including, for example, A. Gennaro (2000) “Remington: The Science and Practicce of Pharmacy” 20th edition, Lippincottt, Williams. , &Wilkins; Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) HC Ansel et al., 7th edition, Lippincottt, Williams, &Wilkins; and Handbook of Pharmaceutical Excipients (2000) AH Kibbe et al., 3rd edition , Amer. Pharmaceutical. Assoc.

  Pharmaceutically acceptable excipients such as vehicles, adjuvants, carriers, or diluents are readily available publicly. In addition, pharmaceutically acceptable auxiliary substances such as pH adjusting and buffering agents, isotonic preparations, wetting agents and the like are readily available publicly.

  In the subject methods, the active agent can be administered to the host using any convenient means capable of producing the desired therapeutic effect. Thus, this agent can be incorporated into various formulations for therapeutic administration. More particularly, the agents of the invention can be formulated into pharmaceutical compositions by combination with suitable pharmaceutically acceptable carriers or diluents, tablets, capsules, powders, granules, ointments, solutions, suppositories. Formulations may be formulated in the form of solids, semi-solids, liquids or gases such as agents, injections, inhalants, and aerosols.

  As such, drug administration can be accomplished in a variety of ways, including oral, buccal, rectal, parenteral, intraperitoneal, intradermal, subcutaneous, intramuscular, transdermal, intratracheal, etc. Administration. In certain embodiments, two different routes of administration are used. For example, in certain embodiments, IFN-α is administered subcutaneously and pirfenidone or a pirfenidone analog is administered orally.

  Subcutaneous administration of a Type I or Type III interferon receptor agonist is accomplished using standard methods and devices (eg, needles and syringes, subcutaneous injection port delivery systems, etc.). See, for example, U.S. Pat. Nos. 3,547,119; 4,755,173; 4,531,937; 4,311,137; and 6,017,328. The combination of subcutaneous injection port and device for administration of IFN-α to the patient through the port is referred to herein as a “subcutaneous injection port delivery system”. In certain embodiments, subcutaneous administration is achieved by a combination of devices (eg, needle and syringe) followed by delivery using a continuous delivery system.

  In certain embodiments, the Type I or Type III interferon receptor agonist is delivered by a continuous delivery system. The term “continuous delivery system” is used interchangeably herein with “control delivery device” and is a continuous (eg, controlled) delivery device (eg, pump) in combination with a catheter, injection device, etc. A wide variety of these are known in the art.

  Mechanical or electromechanical infusion pumps may also be suitable for use with the present invention. Examples of such devices include those described, for example, in U.S. Patent Nos. 4,692,147; 4,360,019; 4,487,603; 4,360,019; 4,725,852; 5,820,589; 5,643,207; In general, the method of drug delivery can be accomplished using any of a variety of refillable pump systems. The pump provides consistent controlled release over time. Typically, the drug (eg, a type I or type III interferon receptor agonist, eg, IFN-α) is in a liquid formulation in a drug-impermeable reservoir and is delivered to the individual in a continuous manner. .

  In one embodiment, the drug delivery system is an at least partially implantable device. The implantable device can be implanted at any suitable implantation site using methods and devices well known in the art. The implantation site is the site in the subject's body where the drug delivery device is introduced and placed. Implantation sites include, but are not necessarily limited to, subdermal, subcutaneous, intramuscular, or other suitable sites in the subject's body. Subcutaneous implantation sites are generally preferred due to the convenience of implantation and removal of the drug delivery device.

  Drug delivery devices suitable for use in the present invention can be based on any of a variety of modes of operation. For example, drug release devices can be based on diffusion systems, convection systems, or erosion systems (eg, erosion-based systems). For example, the drug delivery device can be an electrochemical pump, osmotic pump, electroosmotic pump, vapor pressure pump, or osmotic burst matrix, where the drug is incorporated into the polymer and the polymer is a polymer filled with the drug It provides for the release of a drug formulation associated with a substance (eg, a biodegradable, drug-filled polymeric substance). In other embodiments, the drug release device is based on an electrical diffusion system, an electrolytic pump, an effervescent pump, a piezoelectric pump, a hydrolysis system, and the like.

などに記載されるものが含まれるがこれらに必ずしも限定されない。 Drug release devices based on mechanical or electromechanical infusion pumps may also be suitable for use with the present invention. Examples of such devices include, for example, those described in US Pat. Nos. 4,692,147; 4,360,019; 4,487,603; 4,360,019; 4,725,852, and the like. In general, the inventive method of drug delivery can be accomplished using any of a variety of fillable non-exchangeable pump systems. Pumps and other convective systems are generally preferred because of their generally controlled release that is consistent over time. Osmotic pumps are particularly preferred due to their more consistent controlled release and their combined advantages of relatively small size (see, e.g., PCT Application Publication Nos. WO 97/27840 and US Patent Nos. 5,985,305 and 5,728,396). Wanna). Suitable exemplary osmotic drive devices for use in the present invention include:

However, it is not necessarily limited to these.

  In certain embodiments, the drug delivery device is an implantable device. The drug delivery device can be implanted at any suitable implantation site using methods and devices known in the art. As described below, the implantation site is the site within the subject's body where the drug delivery device is introduced and positioned. Implantation sites include, but are not necessarily limited to, subdermal, subcutaneous, intramuscular, or other suitable site within the subject's body.

  In certain embodiments, a Type I or Type III interferon receptor agonist is delivered using an implantable drug delivery system (eg, a system programmable to provide administration of a Type I or Type III interferon receptor agonist). . An exemplary programmable implantable system includes an implantable infusion pump. Exemplary implantable infusion pumps or devices useful in connection with such pumps are, for example, U.S. Pat.Nos. 4,350,155; 5,443,450; 5,814,019; 5,976,109; 6,017,328; 6,171,276; 6,241,704; 6,464,687 No .; 6,475,180; and 6,512,954. A further exemplary device that can be adapted to the present invention is a Synchromed infusion pump (Medtronic).

  In pharmaceutical dosage forms, the agents can be administered in the form of their pharmaceutically acceptable salts, or the agents can also be alone or in appropriate association as well as in combination with other pharmaceutically active compounds. Can be used in The following methods and excipients are merely exemplary and are in no way limiting.

  For oral preparations, the drug can be used alone or in a suitable adjunct for making tablets, powders, granules, or capsules (e.g., conventional adducts such as lactose, mannitol, root starch, or potato starch). Binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch, or gelatin; disintegrants such as corn starch, potato starch, or sodium carboxymethyl cellulose; lubricants such as talc or magnesium stearate; and dilution if desired Agents, buffers, wetting agents, preservatives, and perfumes).

  Agents are desired by dissolving, suspending, or emulsifying them in an aqueous or non-aqueous solvent such as vegetable oil or other similar oils, synthetic fatty acid glycerides, esters of higher fatty acids, or propylene glycol; and desired In some cases, solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizing agents, and preservatives may be used to formulate a preparation for injection.

  In addition, the drug can be made into suppositories by mixing with a variety of bases such as emulsifying bases or water-soluble bases. The compounds of the present invention can be administered rectally via a suppository. Suppositories can include vehicles such as cocoa butter, carbowaxes, and polyethylene glycols, which melt at body temperature but solidify at room temperature.

  Unit dosage forms for oral or rectal administration such as syrups, elixirs, and suspensions may be provided, where each dosage unit (e.g., 1 teaspoon, 1 tablespoon, tablet, or suppository) ) Comprises a predetermined amount of a composition comprising one or more inhibitors. Similarly, unit dosage forms for injection or intravenous administration may contain the inhibitor in a composition as a solution in sterile water, normal saline, or another pharmaceutically acceptable carrier.

  The term “unit dosage form” as used herein refers to physically separate units suitable as unit dosages for human and animal subjects, each unit being pharmaceutically acceptable. A predetermined amount of a compound of the present invention, calculated in an amount sufficient to produce the desired effect, together with a diluent, carrier, or vehicle to be included. The details of the new unit dosage form of the present invention depend on the particular compound utilized and the effect achieved and the pharmacodynamics associated with each compound in the host.

  In certain embodiments, at least one dose of IFN-γ is administered concurrently with at least one dose of a Type I or Type III interferon receptor agonist. As used herein, the term “simultaneously” means that IFN-α and a Type I or Type III interferon receptor agonist are administered separately and are within about 5 seconds to about 15 seconds of each other, from about 15 seconds to Within about 30 seconds, within 30 seconds to 60 seconds, within 1 minute to 5 minutes, within 5 minutes to 15 minutes, within 15 minutes to 30 minutes, within 30 minutes to 60 minutes, Administered within about 1 hour to about 2 hours, about 2 hours to about 6 hours, about 6 hours to about 12 hours, about 12 hours to about 24 hours, or about 24 hours to about 48 hours.

  In certain embodiments, IFN-γ is administered during the entire course of type I or type III interferon receptor agonist treatment. In other embodiments, the IFN-γ is administered for a period of time that overlaps that of a type I or type III interferon receptor agonist treatment, eg, the IFN-γ treatment is, eg, a type I or type III interferon receptor agonist. IFN-γ treatment can begin before treatment begins, and IFN-γ treatment can end before type I or type III interferon receptor agonist treatment is completed; type I or type III interferon receptor agonist treatment IFN-γ treatment can begin after IFN has started, and IFN-γ treatment can end after IFN-γ treatment has ended; IFN- after the start of type I or type III interferon receptor agonist treatment IFN-γ treatment can be completed and IFN-γ treatment can be terminated before type I or type III interferon receptor agonist treatment is completed; or type I or III The IFN-γ treatment can begin before the type of interferon receptor agonist treatment begins, and the IFN-γ treatment can end after the type I or type III interferon receptor agonist treatment ends.

  In connection with each of the methods described herein, the present invention provides embodiments in which a Type I or Type III interferon receptor agonist and / or IFN-γ is administered to a patient by a controlled delivery device. In certain embodiments, a Type I or Type III interferon receptor agonist and / or IFN-γ is provided to a patient substantially continuously or continuously by a controlled delivery device. Optionally, an implantable infusion pump is used to deliver a type I or type III interferon receptor agonist and / or IFN-γ to the patient substantially continuously or continuously by subcutaneous infusion. .

  In other embodiments, the Type I or Type III interferon receptor agonist and / or IFN-γ is in a substantially steady state during the period of treatment of the Type I or Type III interferon receptor agonist and / or IFN-γ. A patient is administered to achieve and maintain a desired daily mean serum concentration of a type I or type III interferon receptor agonist and / or IFN-γ. Optionally, the implantable infusion pump may provide a type I or type III interferon receptor agonist and / or a type I or type III interferon receptor agonist that is substantially steady during the period of IFN-γ treatment and / or Or, to achieve and maintain the desired daily average serum concentration of IFN-γ, it is used to deliver a type I or type III interferon receptor agonist and / or IFN-γ to the patient by subcutaneous injection.

Effective dosages of IFN-γ can range from about 0.5 μg / m ² to about 500 μg / m ² , usually from about 1.5 μg / m ² to about 200 μg / m ² , depending on the size of the patient. This activity is based on 10 ⁶ international units (U) per 50 μg protein. IFN-γ can be administered daily, every other day, three times a week, or substantially continuously or continuously.

  In particular embodiments of interest, IFN-γ is administered to an individual in a unit dosage form of about 25 μg to about 500 μg, about 50 μg to about 400 μg, or about 100 μg to about 300 μg. In a particularly interesting embodiment, the dose is about 200 μg IFN-γ. In many embodiments of interest, IFN-γ1b is administered.

  When the dosage is 200 μg IFN-γ per dose, the amount of IFN-γ per body weight (assuming the range of body weight is about 45 kg to about 135 kg) is 4.4 μg IFN-γ / kg body weight to 1.48 μg IFN- γ / kg body weight.

The body surface area of the subject individual generally ranges from about 1.33 m ² to about 2.50 m ² . Thus, in many embodiments, the IFN-γ dosage ranges from about 150 μg / m ² to about 20 μg / m ² . For example, the IFN-γ dosage is about 20 μg / m ² to about 30 μg / m ² , about 30 μg / m ² to about 40 μg / m ² , about 40 μg / m ² to about 50 μg / m ² , about 50 μg / m ² About 60 μg / m ² , about 60 μg / m ² to about 70 μg / m ² , about 70 μg / m ² to about 80 μg / m ² , about 80 μg / m ² to about 90 μg / m ² , about 90 μg / m ² to about 100 μg / m ² , about 100 μg / m ² to about 110 μg / m ² , about 110 μg / m ² to about 120 μg / m ² , about 120 μg / m ² to about 130 μg / m ² , about 130 μg / m ² to about 140 μg / m ² , or in the range of about 140 μg / m ² to about 150 μg / m ² . In certain embodiments, dosage groups range from about 25 μg / m ² to about 100 μg / m ² . In other embodiments, the dosage group ranges from about 25 μg / m ² to about 50 μg / m ² .

  Type I or III interferon receptor agonists are daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, the type I interferon receptor agonist is IFN-α. Effective dosages of IFN-α range from about 1 μg to about 30 μg, about 3 μg to about 27 μg, about 1 MU to about 20 MU, about 3 MU to about 10 MU, about 90 μg to about 180 μg, or about 18 μg to about 90 μg. obtain.

  Effective dosages of Infergen® consensus IFN-α include about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg of drug per dose. Effective dosages of IFN-α2a and IFN-α2b can range from 3 million units (MU) to 10 MU per dose. An effective dosage of PEGylated IFN-α2a may comprise about 90 μg to about 180 μg, or about 135 μg of drug per dose. An effective dosage of PEGylated IFN-α2b may comprise an amount of drug of about 0.5 μg to 1.5 μg per kg body weight per dose. Effective dosages of PEGylated consensus interferon (PEG-CIFN) may comprise CIFN amino acids in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per kg body weight per dose of PEG-CIFN . IFN-α can be administered daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously.

  In certain embodiments, a Type I or Type III interferon receptor agonist is administered in a first dosing regimen followed by a second dosing regimen. A first dosing regimen of a Type I or Type III interferon receptor agonist (also referred to as a “transduction regimen”) generally involves administration of a higher dosage of a Type I or Type III interferon receptor agonist. For example, in the case of Infergen® consensus IFN-α (CIFN), the first dosing regimen comprises administering CIFN at about 9 μg, about 15 μg, about 18 μg, or about 27 μg. The first dosing regimen can include a single dosing event, or at least two or more dosing events. The first dosing regimen of a type I or type III interferon receptor agonist is daily, every other day, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  A first dosing regimen of a Type I or Type III interferon receptor agonist is administered for a first period, which can be at least about 4 weeks, at least about 8 weeks, or at least about 12 weeks.

  A second dosing regimen of a type I or type III interferon receptor agonist (also referred to as a “maintenance regimen”) generally involves the administration of a lower dosage of a type I or type III interferon receptor agonist. For example, in the case of CIFN, the second dosage regimen comprises administering CIFN at least about 9 μg, at least about 15 μg, or at least about 18 μg. The second dosing regimen can include a single dosing event, or at least two or more dosing events.

  A second dosing regimen of a type I or type III interferon receptor agonist is daily, every other day, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, an “introduction” / “maintenance” dosing regimen of a Type I or Type III interferon receptor agonist is administered and a “priming” dose of IFN-γ is included. In these embodiments, the IFN-γ is about 1 day to about 14 days, about 2 days to about 10 days, or about 3 days to about 3 days prior to initiation of treatment with a Type I or Type III interferon receptor agonist. Can be administered for a period of 7 days. This period is called the “priming” phase. In some of these embodiments, IFN-γ treatment continues throughout the entire period of treatment with a Type I or Type III interferon receptor agonist. In other embodiments, IFN-γ treatment is discontinued prior to the end of treatment with a Type I or Type III interferon receptor agonist. In these embodiments, the total time of treatment with IFN-γ (including the “priming” phase) is about 2 days to about 30 days, about 4 days to about 25 days, about 8 days to about 20 days, about 10 days to about 18 days, or about 12 days to about 16 days. In yet other embodiments, IFN-γ therapy is discontinued once type I or type III interferon receptor agonist treatment is initiated.

  In other embodiments, the Type I or Type III interferon receptor agonist is administered in a single dosing regimen. For example, in the case of CIFN, the dose of CIFN is generally in the range of about 3 μg to about 15 μg, or about 9 μg to about 15 μg. Type I or III interferon receptor agonists are generally administered daily, every other day, three times a week, every other week, three times a month, once a month, or substantially continuously. The type I or type III interferon receptor agonist is administered for a period of time, which can be, for example, at least about 24 weeks to at least about 48 weeks, or longer.

  In certain embodiments, when a single dosage regimen of a Type I or Type III interferon receptor agonist is administered, a “priming” dose of IFN-γ is included. In these embodiments, the IFN-γ is about 1 day to about 14 days, about 2 days to about 10 days, or about 3 days to about 7 days prior to initiation of treatment with a Type I or Type III interferon receptor agonist. Administered for a period of days. This period is called the “priming” phase. In some of these embodiments, IFN-γ treatment continues throughout the entire period of treatment with a Type I or Type III interferon receptor agonist. In other embodiments, IFN-γ treatment is discontinued prior to the end of treatment with a Type I or Type III interferon receptor agonist. In these embodiments, the total time of treatment with IFN-γ (including the “priming” phase) is about 2 days to about 30 days, about 4 days to about 25 days, about 8 days to about 20 days, about 10 days to about 18 days, or about 12 days to about 16 days. In yet other embodiments, IFN-γ therapy is discontinued once type I or type III interferon receptor agonist treatment is initiated.

  Those skilled in the art will readily recognize that dose levels can vary as a function of the particular compound, the severity of symptoms, and the subject's sensitivity to side effects. The preferred dosage for a given compound can be readily determined by one skilled in the art by various means.

  Those skilled in the art will readily recognize that dose levels can vary as a function of the particular compound, the severity of symptoms, and the subject's sensitivity to side effects. The preferred dosage for a given compound can be readily determined by one skilled in the art by various means. A preferred means is to measure the physiological potency of a given compound.

  In certain embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in the same formulation and are administered simultaneously. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered separately, eg, in separate formulations. In some of these embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered separately and administered simultaneously. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered separately and within about 5 seconds to about 15 seconds, within about 15 seconds to about 30 seconds, from about 30 seconds to about Within about 60 seconds, within about 1 minute to about 5 minutes, within about 5 minutes to about 15 minutes, within about 15 minutes to about 30 minutes, within about 30 minutes to about 60 minutes, about 1 hour to about Within 2 hours, within about 2 hours to about 6 hours, within about 6 hours to about 12 hours, within about 12 hours to about 24 hours, or within about 24 hours to about 48 hours.

  Multiple doses of a Type I or Type III interferon receptor agonist and IFN-γ are administered, for example, a Type I or Type III interferon receptor agonist and IFN-γ are about 1 day to about 1 week, about 2 weeks to About 4 weeks, about 1 month to about 2 months, about 2 months to about 4 months, about 4 months to about 6 months, about 6 months to about 8 months, about 8 months to about 1 year, about 1 year to about 2 Once a month, twice a month, three times a month, once a week, twice a week, over a period of time, or about 2 years to about 4 years or more, It can be administered three times a week, four times a week, five times a week, six times a week, or daily. In certain embodiments of interest, the Type I or Type III interferon receptor agonist and IFN-γ are administered three times a week over a period of about 48 weeks.

  In certain embodiments where IFN-α and IFN-γ are administered in combination therapy, the IFN-α and IFN-γ are a single liquid contained in a single reservoir for use in a drug delivery device. It is co-formulated in the formulation. Accordingly, the present invention provides a pharmaceutical formulation comprising a liquid formulation comprising a single dose of IFN-α and a single dose of IFN-γ. Accordingly, the present invention provides a drug reservoir or other container comprising IFN-α and IFN-γ that is co-formulated in a liquid, wherein both IFN-α and IFN-γ are each once Is present in the formulation in an amount appropriate for the dose of. Dosages are described herein. The reservoir may be provided in any of a variety of forms, including but not limited to cartridges, syringes, continuous delivery device reservoirs, and the like. The invention further provides a drug delivery device comprising a reservoir (eg, preloaded using the reservoir) comprising a liquid formulation comprising a single dose of IFN-α and a single dose of IFN-γ. Exemplary, non-limiting drug delivery devices include injection devices such as pen injectors, needle / syringe devices, continuous delivery devices, and the like. Any of the dosages disclosed herein including a synergistically effective amount can be used in a pharmaceutical formulation, in a reservoir or in a drug delivery device.

  In other embodiments where IFN-α and IFN-γ are administered in combination therapy, IFN-α and IFN-γ are in a pharmaceutical formulation contained in separate reservoirs in the same drug delivery device. The present invention further provides a drug delivery device that is preloaded using separate reservoirs, one reservoir comprising a liquid formulation comprising a single dose of IFN-α, and a second reservoir comprising a single dose of IFN. Includes liquid formulations containing -γ. Any dosage described herein, including a synergistically effective amount, can be used in a pharmaceutical formulation, reservoir, or drug delivery device.

  In certain embodiments, in the therapeutic methods described herein, the interferon receptor agonist is IFN-α and the subject method provides an effective amount of IFN-γ to the patient during the period of IFN-α treatment. Co-administering. In one embodiment, IFN-α and IFN-γ are administered to a patient by a drug delivery device. Optionally, this device is used to deliver IFN-α to the patient by substantially continuous or continuous administration, and IFN-γ to the patient by bolus administration tiw, biw, qod, or qd Used to deliver. Optionally, the device is used to deliver IFN-α and IFN-γ to the patient in the same mode of administration and pattern (eg, in substantially continuous or continuous administration). Optionally, IFN-α and IFN-γ are contained in separate reservoirs in the drug delivery device. Optionally, IFN-α and IFN-γ are co-formulated in a single liquid formulation contained in a single reservoir in the drug delivery device.

  Where the agent is a polypeptide, polynucleotide (e.g., a type I or type III interferon receptor agonist or IFN-γ), this may be by any number of routes including viral infection, microinjection, or vesicle fusion. It can be introduced into a tissue or host cell. Jet injection as described by Furth et al. (1992) Anal. Biochem. 205: 365-368 can also be used for intramuscular administration. DNA is described in the literature (see, e.g., Tang et al. (1992) Nature 356: 152-154) (where gold microparticle bullets are coated with therapeutic DNA and then skin cells Can be coated onto gold microparticles and delivered intradermally through a particle bombardment device, or “gene gun”. In these embodiments, of particular interest is the use of liver-specific promoters to preferentially drive transcription of operably linked IFN-α and IFN-γ coding sequences in hepatocytes.

Additional therapeutic agents In certain embodiments, the methods of the invention further comprise administration of pirfenidone and a pirfenidone analog. Pirfenidone and pirfenidone analogs are about 1 day to about 1 week, about 2 weeks to about 4 weeks, about 1 month to about 2 months, about 2 months to about 4 months, about 4 months to about 6 months, about 6 months to Once a month, twice a month, over a period ranging from about 8 months, about 8 months to about 1 year, about 1 year to about 2 years, or about 2 years to about 4 years, or more, 3 times a month, once a week, 2 times a week, 3 times a week, 4 times a week, 5 times a week, 6 times a week, daily, or once daily It can be administered in divided daily doses ranging up to 5 times.

  Effective dosages of pirfenidone and specific pirfenidone analogs are dosages based on body weight ranging from about 5 mg / kg / day to about 125 mg / kg / day, or from about 400 mg to about 3600 mg per day, or from about 800 mg per day Contains a fixed dosage of about 2400 mg, or about 1000 mg to about 1800 mg per day, or about 1200 mg to about 1600 mg per day, and is administered orally. Other doses and formulations of pirfenidone and specific pirfenidone analogs suitable for use in the treatment of fibrotic diseases are described in US Pat. Nos. 5,310,562; 5,518,729; 5,716,632; and 6,090,822.

  In certain embodiments, pirfenidone and pirfenidone analogs are administered throughout the entire course of a Type I or Type III interferon receptor agonist and / or IFN-γ treatment. In other embodiments, the pirfenidone and pirfenidone analog may be a type I or type III interferon receptor agonist and / or in the course of less than full treatment of IFN-γ, e.g., a type I or type III interferon receptor agonist and / or Only during the first phase of IFN-γ treatment, type I or III interferon receptor agonist and / or only during the second phase of treatment of IFN-γ, or type I or III interferon receptor agonist And / or administered in other parts of the IFN-γ treatment regimen.

  In certain embodiments, the methods of the invention further comprise administration of ribavirin. Ribavirin, 1-β-D-ribofuranosyl-1H-1,2,4-triazole-3-carboxamide, available from ICN Pharmaceuticals, Inc., Costa Mesa, Calif., Is Merck Index, 11th Edition, Compound No. It is described in 8199. Its manufacture and formulation is described in US Pat. No. 4,211,771. The present invention also contemplates the use of derivatives of ribavirin (see, eg, US Pat. No. 6,277,830). Ribavirin can be administered orally in capsule or tablet form, or in the same or different dosage form, and the same or different route as a type I or type III interferon receptor agonist. Of course, other types of administration of both medications by nasal spray, transdermal, intravenous, suppository, sustained release dosage form, etc. are contemplated, making both medications available. Any route of administration is used so long as the appropriate dosage is delivered without destroying the active ingredient.

  Ribavirin is about 30 mg to about 60 mg, about 60 mg to about 125 mg, about 125 mg to about 200 mg, about 200 mg to about 300 mg, about 300 mg to about 400 mg, about 400 mg to about 1200 mg, about 600 mg to about 1000 mg, or about 700 mg per day. It is generally administered in an amount ranging from about 900 mg or about 10 mg / kg body weight per day.

  In certain embodiments, ribavirin is administered throughout the course of treatment of a type I or type III interferon receptor agonist and / or IFN-γ. In other embodiments, ribavirin is administered in a less than complete course of treatment of a type I or type III interferon receptor agonist and / or IFN-γ, such as a type I or type III interferon receptor agonist and / or IFN-γ. Only during the first phase of treatment of type I or type III interferon receptor agonist and / or during the second phase of treatment of IFN-γ, or type I or type III interferon receptor agonist and / or It is administered in other parts of the IFN-γ treatment regimen.

Method of treatment
1. Treatment of Alphavirus Infection The present invention relates to a method of treating alphavirus infection by administering a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ to an individual in need thereof. I will provide a. Individuals treated according to the methods of the present invention include individuals who have been clinically diagnosed as having an alphavirus infection and have one or more signs and symptoms of clinical infection but have not yet been diagnosed with an alphavirus infection. No individuals are included.

  In performing a combination therapy method for alphavirus infection in an individual as described above, a Type I or Type III interferon receptor agonist and IFN-γ are administered to the individual. In certain embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in the same formulation. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in separate formulations. When administered in separate formulations, the Type I or Type III interferon receptor agonist and IFN-γ can be administered substantially simultaneously or within about 24 hours of each other. In many embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered subcutaneously in multiple doses.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Type I or III interferon receptor agonists are daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, the type I interferon receptor agonist is IFN-α. Effective dosages of IFN-α range from about 1 μg to about 30 μg, about 3 μg to about 27 μg, about 1 MU to about 20 MU, about 3 MU to about 10 MU, about 90 μg to about 180 μg, or about 18 μg to about 90 μg. obtain.

  Effective dosages of Infergen® consensus IFN-α can include an amount of drug of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) may comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN.

  In many embodiments, the Type I or Type III interferon receptor agonist and / or IFN-γ is about 1 day to about 7 days, or about 1 week to about 2 weeks, or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks, or about 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 It can be administered for a period of months, or at least one year, and can be administered over a longer period of time. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly.

  In certain embodiments, the present invention provides methods of using a synergistically effective amount of a type I or type III interferon receptor agonist and IFN-γ in the treatment of alphavirus infection in a patient. In certain embodiments, the present invention provides methods of using synergistically effective amounts of IFN-α and IFN-γ in the treatment of alphavirus infections in patients. In one embodiment, the present invention provides a method of using synergistically effective amounts of consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient.

  Generally, a synergistically effective amount of consensus interferon (CIFN) and IFN-γ, suitable for use in the methods of the present invention, is provided at a dosage ratio of 1 μg CIFN: 10 μg IFN-γ, Here, CIFN and IFN-γ are both non-PEGylated and non-glycosylated species.

  In one embodiment, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: During the desired treatment period, a dose of INFERGEN® containing about 1 μg to about 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, Dosage of IFN-γ containing drug in an amount of about 10 μg to about 300 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day (Subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Including subcutaneous administration.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: During the desired treatment period, a dose of INFERGEN® containing about 1 μg to about 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, Dosage of IFN-γ containing drug in an amount of about 10 μg to about 100 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day (Subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Including subcutaneous administration.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: During the desired treatment period, a dose of INFERGEN® containing approximately 1 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 month. IFN-γ dosage (subcutaneous) containing an amount of drug of about 10 μg to about 50 μg per dose of IFN-γ, three times, once a month, or substantially continuously or continuously per day In combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Including stages.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: During the desired treatment period, a dose of INFERGEN® containing approximately 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 month. A dosage of IFN-γ containing about 90 μg to about 100 μg of drug per dose of IFN-γ (subcutaneously, three times, once a month, or substantially continuously or daily) In combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Including stages.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: During the desired treatment period, a dose of INFERGEN® containing approximately 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 month. IFN-γ dosage (subcutaneously) containing about 200 μg to about 300 μg of drug per dose of IFN-γ, three times, once a month, or substantially continuously or continuously per day In combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Including stages.

  In another aspect, the present invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient, wherein the method is desired During the treatment period, a dose of PEG-CIFN containing CIFN-amino acids in an amount of about 4 μg to about 60 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN) is about 30 μg to about 1,000 μg / week. Qw in combination with the entire weekly dosage of IFN-γ (qd, qod, tiw, biw, or substantially continuously or continuously in divided doses) containing the amount of drug Qow, 3 times a month, or monthly, including subcutaneous administration to the patient.

  In another aspect, the present invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of alphavirus infection in a patient, wherein the method is desired During the treatment period, a dose of PEG-CIFN containing CIFN-amino acids in an amount of about 18 μg to about 24 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN) is about 100 μg to about 300 μg / week. Qw, in combination with a total weekly dosage of IFN-γ containing an amount of drug (qd, qod, tiw, biw, or administered substantially continuously or continuously in divided doses) qow, including subcutaneous administration to the patient three times a month or monthly.

  In general, a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ suitable for use in the methods of the invention is one million units (MU) IFN-α 2a or 2b. Or 2c: provided at a dosage ratio of 30 μg IFN-γ, where IFN-α 2a or 2b or 2c and both IFN-γ are non-PEGylated and non-glycosylated species.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: During the treatment period to be administered, a dosage of IFN-α2a containing an amount of drug of about 1 MU to about 20 MU per dose of IFN-α 2a or 2b or 2c is about 30 μg to about 600 μg per dose of IFN-γ. Qd, qod, tiw, biw in combination with a dosage of IFN-γ containing the amount of drug (subcutaneous, qd, qod, tiw, biw, or substantially continuously or continuously per day) Or subcutaneously administered to a patient substantially continuously or continuously per day.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: IFN-α2a containing a dose of about 3 MU per dose of IFN-α 2a or 2b or 2c, and IFN containing an amount of about 100 μg of drug per dose of IFN-γ -in combination with γ dosage (subcutaneous, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or daily Or subcutaneously to the patient continuously or continuously.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: IFN-α2a containing a dose of about 10 MU per dose of IFN-α 2a or 2b or 2c, and IFN containing a dose of about 300 μg of drug per dose of IFN-γ -in combination with γ dosage (subcutaneous, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or daily Or subcutaneously to the patient continuously or continuously.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing about 90 μg to about 360 μg of drug per dose of PEGASYS® is administered at an amount of about 30 μg to about 1,000 μg / week. Total weekly dosage of IFN-γ containing drug (subcutaneously administered in qd, qod, tiw, or biw in divided doses, or substantially continuously or continuously ) In combination with qw, qow, 3 times a month or monthly.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of alphavirus infection in a patient, the method comprising: A dose of PEGASYS® containing about 180 μg of drug per dose of PEGASYS® during the desired treatment period, IFN containing about 100 μg to about 300 μg / week of drug -in combination with a weekly total dosage of γ (subcutaneously administered in divided doses qd, qod, tiw, or biw, or substantially continuously or continuously) Qw, qow, 3 times a month or every month.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of alphavirus infection in a patient, wherein The method comprises administering a dosage of PEG-INTRON® comprising a drug in an amount of about 0.75 μg to about 3.0 μg / kg body weight per dose of PEG-INTRON® during a desired treatment period. Total weekly IFN-γ dosage containing about 30 μg to about 1,000 μg / week of drug (subcutaneously administered in divided doses qd, qod, tiw, or biw, or substantially Qw, qow, 3 times a month, or monthly, in combination with a patient.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of alphavirus infection in a patient, wherein The method comprises administering a dosage of PEG-INTRON® comprising about 100 μg to about 1.5 μg / kg body weight of drug per dose of PEG-INTRON® for a desired treatment period. Total weekly IFN-γ dosage containing 300 μg / week of drug (subcutaneously administered in qd, qod, tiw, or biw in divided doses, or substantially continuously or Qw, qow, 3 times a month, or monthly, in combination with a patient.

  The present invention also provides a method for the treatment of alphavirus infection, wherein ribavirin treatment is added to any of the combination treatments of type I or type III interferon receptor agonists and IFN-γ described above. In certain embodiments, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 800 mg to 1200 mg ribavirin oral qd for a particular treatment period. In other embodiments, a combination therapy of a type I or type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 1000 mg ribavirin oral qd for a particular treatment period. In a further embodiment, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include an oral qd ribavirin regimen of about 10 mg ribavirin / kg body weight for a particular period of treatment. The daily ribavirin dosage can be administered in one dose per day, or in divided doses (including once, twice, three, or four doses per day).

  In one embodiment, the invention provides for treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment, and administering a ribavirin regimen orally administered in qd to an individual having an alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering to a subject with alphavirus infection a 50 μg Actimmune® human IFN-γ1b administered subcutaneously at 1 mg; and a ribavirin regimen administered orally at qd. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering a 100 μg Actimmune® human IFN-γ1b administered subcutaneously at a dose; and a ribavirin regimen administered orally at qd to an individual having an alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and Administering a 50 μg Actimmune® human IFN-γ1b regimen administered subcutaneously at tiw to an individual with alphavirus infection.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and Administering a 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously at tiw to an individual with HCV infection.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering a 25 μg Actimmune® human IFN-γ1b administered subcutaneously at pH 5; and a ribavirin regimen administered orally at qd to an individual having an alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administration of 200 μg Actimmune® human IFN-γ1b administered subcutaneously at a dose; and a ribavirin regimen administered orally at qd to an individual with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and administering a 25 μg Actimmune® human IFN-γ1b regimen administered subcutaneously at tiw to an individual with alphavirus infection.

  In one embodiment, the invention provides for the treatment of alphavirus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and Administering a 200 μg Actimmune® human IFN-γ1b regimen administered subcutaneously at tiw to an individual with alphavirus infection.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 100 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Administering a consensus IFN-α and a ribavirin regimen administered orally in qd or tiw to an individual having an alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 100 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 100 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; 100 μg Actimmune® human IFN-γ1b administered subcutaneously with tiw; and ribavirin regimen administered orally with qd include administering to an individual with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 100 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously with consensus IFN-α; and tiw to an individual with alphavirus infection.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 100 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; and 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously at tiw, comprising administering to an individual with alphavirus infection.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 150 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α and ribavirin regimen administered orally in qd include administering to an individual with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 150 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 150 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; 100 μg Actimmune® human IFN-γ1b administered subcutaneously with tiw; and ribavirin regimen administered orally with qd include administering to an individual with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 150 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α and a 50 μg Actimmune® human IFN-γ1b regimen administered subcutaneously with tiw are included in individuals with alphavirus infection.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 150 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; and 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously at tiw, comprising administering to an individual with alphavirus infection.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 200 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α and ribavirin regimen administered orally in qd include administering to an individual with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 200 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with alphavirus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 200 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; 100 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd include administering to an individual with viral infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 200 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously with consensus IFN-α; and tiw to an individual with HCV infection.

  In one embodiment, the present invention provides for the treatment of alphavirus infection, which method comprises 200 μg monoPEG (30 kD, linearized) administered subcutaneously every 10 days or qw for the desired duration of treatment. Consensus IFN-α; and 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously at tiw, comprising administering to an individual with alphavirus infection.

2. Treatment of West Nile Virus Infection The present invention relates to West Nile virus infection by administering a therapeutically effective amount of a type I or type III interferon receptor agonist and / or IFN-γ to an individual in need thereof. Provide a method of treating. Individuals treated according to the methods of the present invention are individuals who have been clinically diagnosed as having West Nile virus infection and have one or more signs and symptoms of clinical infection but are still diagnosed with West Nile virus infection Individuals that have not been included are included.

  In carrying out the monotherapy method for West Nile virus infection as described above, a Type I or Type III interferon receptor agonist or IFN-γ is administered to an individual in need of such treatment.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Type I or III interferon receptor agonists are daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, the type I interferon receptor agonist is IFN-α. Effective dosages of IFN-α range from about 1 μg to about 30 μg, about 3 μg to about 27 μg, about 1 MU to about 20 MU, about 3 MU to about 10 MU, about 90 μg to about 180 μg, or about 18 μg to about 90 μg. obtain.

  Effective dosages of Infergen® consensus IFN-α include drugs in an amount of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) may comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN. In many embodiments, the IFN-α or IFN-γ is from about 1 day to about 7 days, or from about 1 week to about 2 weeks, or from about 2 weeks to about 3 weeks, or from about 3 weeks to about 4 weeks, or A period of about 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months, or at least 1 year And can be administered over a longer period of time. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly. In certain embodiments, the dosage is administered subcutaneously.

  In performing the combination therapy method for West Nile virus infection in an individual as described above, a Type I or Type III interferon receptor agonist and IFN-γ are administered to the individual. In certain embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in the same formulation. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in separate formulations. When administered in separate formulations, the Type I or Type III interferon receptor agonist and IFN-γ can be administered substantially simultaneously or within about 24 hours of each other. In many embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered subcutaneously in multiple doses.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Effective dosages of Infergen® consensus IFN-α can include an amount of drug of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) may comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN. In many embodiments, the IFN-α and / or IFN-γ is about 1 day to about 7 days, or about 1 week to about 2 weeks, or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks. About 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months, or at least 1 year It can be administered for a period of time and can be administered over a longer period. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly.

  In certain embodiments, the present invention provides methods of using a synergistically effective amount of a type I or type III interferon receptor agonist and IFN-γ in the treatment of West Nile virus infection in a patient. In certain embodiments, the present invention provides methods of using synergistically effective amounts of IFN-α and IFN-γ in the treatment of West Nile virus infection in a patient. In one embodiment, the present invention provides a method of using synergistically effective amounts of consensus IFN-α and IFN-γ in the treatment of West Nile virus infection in a patient.

  Generally, a synergistically effective amount of consensus interferon (CIFN) and IFN-γ, suitable for use in the methods of the present invention, is provided at a dosage ratio of 1 μg CIFN: 10 μg IFN-γ, Here, CIFN and IFN-γ are both non-PEGylated and non-glycosylated species.

  In one embodiment, the present invention provides a method of using synergistically effective amounts of INFERGEN® consensus IFN-α and IFN-γ in the treatment of West Nile virus (WNV) infection in a patient, This method involves administering a dosage of INFERGEN® containing an amount of drug of about 1 μg to about 30 μg per dose of INFERGEN® during the desired treatment period, qd, qod, tiw, biw, qw, qow, IFN- containing drug in an amount of about 10 μg to about 300 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day Combined with γ dosage (subcutaneous, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) And subcutaneously administering to the patient.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of WNV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg to about 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 A dosage of IFN-γ comprising an amount of drug of about 10 μg to about 100 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day ( Subcutaneously in combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Administration step.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of WNV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. IFN-γ dosage containing about 10 μg to about 50 μg of drug per dose of IFN-γ (subcutaneously, once or once a month or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of WNV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 IFN-γ dosage containing about 90 μg to about 100 μg of drug per dose of IFN-γ (subcutaneously, once or once a month, or substantially continuously or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of WNV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. Dosages of IFN-γ containing about 200 μg to about 300 μg of drug per dose of IFN-γ, subcutaneously, once a month, or substantially continuously or daily. , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of WNV infection in a patient, wherein the method comprises a desired treatment During the period, a dose of PEG-CIFN containing CIFN-amino acids in an amount of about 4 μg to about 60 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN) is about 30 μg to about 1,000 μg / week. Qw, in combination with a total weekly dosage of IFN-γ containing an amount of drug (qd, qod, tiw, biw, or administered substantially continuously or continuously in divided doses) qow, including subcutaneous administration to the patient three times a month or monthly.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of WNV infection in a patient, wherein the method comprises a desired treatment During the period, a dose of PEG-CIFN comprising CIFN-amino acids in an amount of about 18 μg to about 24 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN), an amount of about 100 μg to about 300 μg / week. Qw, qow in combination with a total weekly dosage of IFN-γ containing qd, qod, tiw, biw, or substantially continuously or continuously in divided doses Including subcutaneous administration to the patient three times a month or monthly.

  In general, a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ suitable for use in the methods of the invention is one million units (MU) IFN-α 2a or 2b. Or 2c: provided at a dosage ratio of 30 μg IFN-γ, where IFN-α 2a or 2b or 2c and both IFN-γ are non-PEGylated and non-glycosylated species.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of WNV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing an amount of drug of about 1 MU to about 20 MU per dose of IFN-α 2a or 2b or 2c, an amount of about 30 μg to about 600 μg per dose of IFN-γ Qd, qod, tiw, biw, in combination with IFN-γ dosages (subcutaneously, qd, qod, tiw, biw, or substantially continuously or continuously per day) Alternatively, the method includes subcutaneous administration to the patient substantially continuously or continuously per day.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of WNV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 3 MU of drug per dose of IFN-α 2a, 2b or 2c, and IFN-α containing about 100 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of WNV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 10 MU of drug per dose of IFN-α 2a, 2b or 2c, and IFN-α containing about 300 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of WNV infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of drug of about 90 μg to about 360 μg per dose of PEGASYS® is administered to an amount of drug of about 30 μg to about 1,000 μg / week. 1 week total dosage of IFN-γ (administered subcutaneously in qd, qod, tiw, or biw in divided doses, or substantially continuously or continuously) In combination with qw, qow, 3 times a month or monthly administered to the patient.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of WNV infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of about 180 μg of drug per dose of PEGASYS® will result in an IFN − containing about 100 μg to about 300 μg / week of drug. in combination with a weekly total dosage of γ (subcutaneously administered in divided doses qd, qod, tiw, or biw, or administered substantially continuously or continuously), qw, qow, including subcutaneous administration to the patient three times a month or monthly.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of WNV infection in a patient, the method Is about PEG-INTRON® dosage containing about 0.75 μg to about 3.0 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period. Total weekly IFN-γ dosage containing 30 μg to about 1,000 μg / week of drug (subcutaneously administered in divided doses, qd, qod, tiw, or biw, or substantially Qw, qow, 3 times a month, or monthly, in combination with a patient.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of WNV infection in a patient, the method PEG-INTRON® containing about 1.5 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period, from about 100 μg to about 300 μg 1 week total dosage of IFN-γ with weekly dose of drug (subcutaneously administered in divided doses qd, qod, tiw, or biw, or substantially continuously or continuously Qw, qow, 3 times a month, or monthly, in combination with a patient.

  The present invention also provides a method for the treatment of West Nile virus infection, wherein ribavirin treatment is added to any of the combination treatments of type I or type III interferon receptor agonists and IFN-γ described above. In certain embodiments, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 800 mg to 1200 mg ribavirin oral qd for a particular treatment period. In other embodiments, a combination therapy of a type I or type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 1000 mg ribavirin oral qd for a particular treatment period. In a further embodiment, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include an oral qd ribavirin regimen of about 10 mg ribavirin / kg body weight for a particular period of treatment. The daily ribavirin dosage can be administered in one dose per day, or in divided doses (including once, twice, three, or four doses per day).

  In one embodiment, the present invention provides for the treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw or tiw for the duration of the desired treatment. And a ribavirin regimen administered orally at qd is administered to an individual with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering a subcutaneously administered 50 μg Actimmune® human IFN-γ1b; and a ribavirin regimen administered orally in qd to an individual having WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 100 μg Actimmune® human IFN-γ1b administered subcutaneously; and a ribavirin regimen administered orally in qd to an individual having WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 50 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with WNV infection.

  In one embodiment, the present invention provides for treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with WNV infection.

  In one embodiment, the present invention provides for the treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 25 μg Actimmune® human IFN-γ1b administered subcutaneously; and a ribavirin regimen administered orally in qd to an individual with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 200 μg Actimmune® human IFN-γ1b administered subcutaneously; and a ribavirin regimen administered orally in qd to an individual with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 25 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with WNV infection.

  In one embodiment, the present invention provides for treatment of WNV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering 200 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with WNV infection.

  In one embodiment, the present invention provides for the treatment of WNV infection, the method comprising 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a ribavirin regimen administered qd orally is administered to an individual with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, the method comprising 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, the method comprising 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 100 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, the method comprising 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with WNV infection.

  In one embodiment, the present invention provides for the treatment of WNV infection, the method comprising 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with WNV infection.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a ribavirin regimen administered qd orally is administered to an individual with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 100 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with WNV infection.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with WNV infection.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a ribavirin regimen administered qd orally is administered to an individual with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 100 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with WNV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with WNV infection.

  In one embodiment, the present invention provides for the treatment of WNV infection, which comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with WNV infection.

3. Treatment of Dengue Virus Infection The present invention relates to a method of treating dengue virus infection by administering a therapeutically effective amount of a type I or type III interferon receptor agonist and / or IFN-γ to an individual in need thereof. I will provide a. Individuals treated according to the methods of the present invention include individuals who have been clinically diagnosed as having a dengue virus infection, and individuals who have one or more signs and symptoms of clinical infection but have not yet been diagnosed with a dengue virus infection Is included.

  In carrying out the monotherapy method for dengue virus infection as described above, a type I or type III interferon receptor agonist or IFN-γ is administered to an individual in need of such treatment.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Type I or III interferon receptor agonists are daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, the type I interferon receptor agonist is IFN-α. Effective dosages of IFN-α range from about 1 μg to about 30 μg, about 3 μg to about 27 μg, about 1 MU to about 20 MU, about 3 MU to about 10 MU, about 90 μg to about 180 μg, or about 18 μg to about 90 μg. obtain.

  Effective dosages of Infergen® consensus IFN-α include drugs in an amount of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) may comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN. In many embodiments, the IFN-α or IFN-γ is from about 1 day to about 7 days, or from about 1 week to about 2 weeks, or from about 2 weeks to about 3 weeks, or from about 3 weeks to about 4 weeks, or A period of about 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months, or at least 1 year And can be administered over a longer period of time. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly. In certain embodiments, the dosage is administered subcutaneously.

  In performing the combination therapy method for dengue virus infection in an individual as described above, a Type I or Type III interferon receptor agonist and IFN-γ are administered to the individual. In certain embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in the same formulation. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in separate formulations. When administered in separate formulations, the Type I or Type III interferon receptor agonist and IFN-γ can be administered substantially simultaneously or within about 24 hours of each other. In many embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered subcutaneously in multiple doses.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Effective dosages of Infergen® consensus IFN-α can include an amount of drug of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) may comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN. In many embodiments, the IFN-α and / or IFN-γ is about 1 day to about 7 days, or about 1 week to about 2 weeks, or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks. About 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months, or at least 1 year It can be administered for a period of time and can be administered over a longer period. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly.

  In certain embodiments, the present invention provides methods of using a synergistically effective amount of a type I or type III interferon receptor agonist and IFN-γ in the treatment of dengue virus infection in a patient. In certain embodiments, the present invention provides methods of using synergistically effective amounts of IFN-α and IFN-γ in the treatment of dengue virus infection in a patient. In one embodiment, the present invention provides a method of using synergistically effective amounts of consensus IFN-α and IFN-γ in the treatment of dengue virus infection in a patient.

  Generally, a synergistically effective amount of consensus interferon (CIFN) and IFN-γ, suitable for use in the methods of the present invention, is provided at a dosage ratio of 1 μg CIFN: 10 μg IFN-γ, Here, CIFN and IFN-γ are both non-PEGylated and non-glycosylated species.

  In one embodiment, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of dengue infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg to about 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 A dosage of IFN-γ comprising an amount of drug of about 10 μg to about 300 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day ( Subcutaneously in combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Administration step.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of dengue infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg to about 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 A dosage of IFN-γ comprising an amount of drug of about 10 μg to about 100 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day ( Subcutaneously in combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Administration step.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of dengue infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. IFN-γ dosage containing about 10 μg to about 50 μg of drug per dose of IFN-γ (subcutaneously, once or once a month or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of dengue infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 IFN-γ dosage containing about 90 μg to about 100 μg of drug per dose of IFN-γ (subcutaneously, once or once a month, or substantially continuously or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of dengue infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. Dosages of IFN-γ containing about 200 μg to about 300 μg of drug per dose of IFN-γ, subcutaneously, once a month, or substantially continuously or daily. , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of dengue infection in a patient, the method comprising a desired treatment During the period, a dose of PEG-CIFN containing CIFN-amino acids in an amount of about 4 μg to about 60 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN) is about 30 μg to about 1,000 μg / week. Qw, in combination with a total weekly dosage of IFN-γ containing an amount of drug (qd, qod, tiw, biw, or administered substantially continuously or continuously in divided doses) qow, including subcutaneous administration to the patient three times a month or monthly.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of dengue infection in a patient, the method comprising a desired treatment During the period, a dose of PEG-CIFN comprising CIFN-amino acids in an amount of about 18 μg to about 24 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN), an amount of about 100 μg to about 300 μg / week. Qw, qow in combination with a total weekly dosage of IFN-γ containing qd, qod, tiw, biw, or substantially continuously or continuously in divided doses Including subcutaneous administration to the patient three times a month or monthly.

  In general, a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ suitable for use in the methods of the invention is one million units (MU) IFN-α 2a or 2b. Or 2c: provided at a dosage ratio of 30 μg IFN-γ, where IFN-α 2a or 2b or 2c and both IFN-γ are non-PEGylated and non-glycosylated species.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of dengue infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing an amount of drug of about 1 MU to about 20 MU per dose of IFN-α 2a or 2b or 2c, an amount of about 30 μg to about 600 μg per dose of IFN-γ Qd, qod, tiw, biw, in combination with IFN-γ dosages (subcutaneously, qd, qod, tiw, biw, or substantially continuously or continuously per day) Alternatively, the method includes subcutaneous administration to the patient substantially continuously or continuously per day.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of dengue infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 3 MU of drug per dose of IFN-α 2a, 2b or 2c, and IFN-α containing about 100 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of dengue infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 10 MU of drug per dose of IFN-α 2a, 2b or 2c, and IFN-α containing about 300 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of dengue infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of drug of about 90 μg to about 360 μg per dose of PEGASYS® is administered to an amount of drug of about 30 μg to about 1,000 μg / week. 1 week total dosage of IFN-γ (administered subcutaneously in qd, qod, tiw, or biw in divided doses, or substantially continuously or continuously) In combination with qw, qow, 3 times a month or monthly administered to the patient.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of dengue infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of about 180 μg of drug per dose of PEGASYS® will result in an IFN − containing about 100 μg to about 300 μg / week of drug. in combination with a weekly total dosage of γ (subcutaneously administered in divided doses qd, qod, tiw, or biw, or administered substantially continuously or continuously), qw, qow, including subcutaneous administration to the patient three times a month or monthly.

  In another aspect, the present invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of dengue infection in a patient, the method Is about PEG-INTRON® dosage containing about 0.75 μg to about 3.0 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period. Total weekly IFN-γ dosage containing 30 μg to about 1,000 μg / week of drug (subcutaneously administered in divided doses, qd, qod, tiw, or biw, or substantially Qw, qow, 3 times a month, or monthly, in combination with a patient.

  In another aspect, the present invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of dengue infection in a patient, the method PEG-INTRON® containing about 1.5 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period, from about 100 μg to about 300 μg 1 week total dosage of IFN-γ with weekly dose of drug (subcutaneously administered in divided doses qd, qod, tiw, or biw, or substantially continuously or continuously Qw, qow, 3 times a month, or monthly, in combination with a patient.

  The present invention also provides a method for the treatment of dengue virus infection, wherein ribavirin therapy is added to any of the combination therapy of type I or type III interferon receptor agonist and IFN-γ described above. In certain embodiments, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 800 mg to 1200 mg ribavirin oral qd for a particular treatment period. In other embodiments, a combination therapy of a type I or type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 1000 mg ribavirin oral qd for a particular treatment period. In a further embodiment, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include an oral qd ribavirin regimen of about 10 mg ribavirin / kg body weight for a particular period of treatment. The daily ribavirin dosage can be administered in one dose per day, or in divided doses (including once, twice, three, or four doses per day).

  In one embodiment, the present invention provides for the treatment of dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment, and qd Administering a ribavirin regimen that is orally administered to an individual having dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for treatment of dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering a subcutaneously administered 50 μg Actimmune® human IFN-γ1b; and ribavirin regimen administered orally in qd to an individual with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for treatment of dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 100 μg Actimmune® human IFN-γ1b administered subcutaneously; and a ribavirin regimen administered orally in qd to an individual having dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment for dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 50 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with dengue virus infection.

  In one embodiment, the present invention provides a treatment for dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously to an individual having dengue virus infection.

  In one embodiment, the present invention provides for treatment of dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 25 μg Actimmune® human IFN-γ1b administered subcutaneously; and a ribavirin regimen administered orally in qd to an individual with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for treatment of dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 200 μg Actimmune® human IFN-γ1b administered subcutaneously; and ribavirin regimen administered orally in qd to an individual with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment for dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 25 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with dengue virus infection.

  In one embodiment, the present invention provides a treatment for dengue virus infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering 200 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with dengue virus infection.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. Injecting an IFN-α and ribavirin regimen administered orally in qd to an individual with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; 50 μg Actimmune® human IFN-γ1b administered subcutaneously with tiw; and ribavirin regimen administered orally with qd are included in individuals with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. Administering 100 μg Actimmune® human IFN-γ1b administered subcutaneously with IFN-α; tiw; and ribavirin regimen administered orally with qd to individuals with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with dengue virus infection.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with dengue virus infection.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. Injecting an IFN-α and ribavirin regimen administered orally in qd to an individual with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; 50 μg Actimmune® human IFN-γ1b administered subcutaneously with tiw; and ribavirin regimen administered orally with qd are included in individuals with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. Administering 100 μg Actimmune® human IFN-γ1b administered subcutaneously with IFN-α; tiw; and ribavirin regimen administered orally with qd to individuals with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a 50 μg Actimmune® human IFN-γ1b regimen administered subcutaneously with tiw is administered to an individual with dengue virus infection.

  In one embodiment, the present invention provides for the treatment of dengue virus infection, which comprises a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously with tiw is administered to an individual with dengue virus infection.

  In one embodiment, the present invention provides a treatment for dengue virus infection, the method comprising a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. Injecting an IFN-α and ribavirin regimen administered orally in qd to an individual with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment for dengue virus infection, the method comprising a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; 50 μg Actimmune® human IFN-γ1b administered subcutaneously with tiw; and ribavirin regimen administered orally with qd are included in individuals with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment for dengue virus infection, the method comprising a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. Administering 100 μg Actimmune® human IFN-γ1b administered subcutaneously with IFN-α; tiw; and ribavirin regimen administered orally with qd to individuals with dengue virus infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment for dengue virus infection, the method comprising a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with dengue virus infection.

  In one embodiment, the present invention provides a treatment for dengue virus infection, the method comprising a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with dengue virus infection.

4. Treatment of Yellow Fever Virus Infection The present invention relates to yellow fever virus (I) by administering a therapeutically effective amount of a type I or type III interferon receptor agonist and / or IFN-γ to an individual in need thereof. YFV) provides a method of treating infection. Individuals treated according to the methods of the present invention include individuals who have been clinically diagnosed as having YFV infection and individuals who have one or more signs and symptoms of clinical infection but have not yet been diagnosed as YFV infection Is included.

  In carrying out the monotherapy method for YFV infection as described above, a type I or type III interferon receptor agonist or IFN-γ is administered to an individual in need of such treatment.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Type I or III interferon receptor agonists are daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, the type I interferon receptor agonist is IFN-α. Effective dosages of IFN-α range from about 1 μg to about 30 μg, about 3 μg to about 27 μg, about 1 MU to about 20 MU, about 3 MU to about 10 MU, about 90 μg to about 180 μg, or about 18 μg to about 90 μg. obtain.

  Effective dosages of Infergen® consensus IFN-α include drugs in an amount of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) may comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN. In many embodiments, the IFN-α or IFN-γ is from about 1 day to about 7 days, or from about 1 week to about 2 weeks, or from about 2 weeks to about 3 weeks, or from about 3 weeks to about 4 weeks, or A period of about 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months, or at least 1 year And can be administered over a longer period of time. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly. In certain embodiments, the dosage is administered subcutaneously.

  In performing the combination therapy method for YFV infection in an individual as described above, a Type I or Type III interferon receptor agonist and IFN-γ are administered to the individual. In certain embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in the same formulation. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in separate formulations. When administered in separate formulations, the Type I or Type III interferon receptor agonist and IFN-γ can be administered substantially simultaneously or within about 24 hours of each other. In many embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered subcutaneously in multiple doses.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Effective dosages of Infergen® consensus IFN-α can include an amount of drug of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) may comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN. In many embodiments, the IFN-α and / or IFN-γ is about 1 day to about 7 days, or about 1 week to about 2 weeks, or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks. Or about 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months, or at least 1 year For a period of time, and may be administered over a longer period of time. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly.

  In certain embodiments, the present invention provides methods of using a synergistically effective amount of a type I or type III interferon receptor agonist and IFN-γ in the treatment of YFV infection in a patient. In certain embodiments, the present invention provides methods of using synergistically effective amounts of IFN-α and IFN-γ in the treatment of YFV infection in a patient. In one embodiment, the present invention provides a method of using synergistically effective amounts of consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient.

  Generally, a synergistically effective amount of consensus interferon (CIFN) and IFN-γ, suitable for use in the methods of the present invention, is provided at a dosage ratio of 1 μg CIFN: 10 μg IFN-γ, Here, CIFN and IFN-γ are both non-PEGylated and non-glycosylated species.

  In one embodiment, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg to about 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 A dosage of IFN-γ comprising an amount of drug of about 10 μg to about 300 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day ( Subcutaneously in combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Administration step.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg to about 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 A dosage of IFN-γ comprising an amount of drug of about 10 μg to about 100 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day ( Subcutaneously in combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Administration step.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. IFN-γ dosage containing about 10 μg to about 50 μg of drug per dose of IFN-γ (subcutaneously, once or once a month or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 IFN-γ dosage containing about 90 μg to about 100 μg of drug per dose of IFN-γ (subcutaneously, once or once a month, or substantially continuously or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. Dosages of IFN-γ containing about 200 μg to about 300 μg of drug per dose of IFN-γ, subcutaneously, once a month, or substantially continuously or daily. , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient, the method comprising a desired treatment During the period, a dose of PEG-CIFN containing CIFN-amino acids in an amount of about 4 μg to about 60 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN) is about 30 μg to about 1,000 μg / week. Qw, in combination with a total weekly dosage of IFN-γ containing an amount of drug (qd, qod, tiw, biw, or administered substantially continuously or continuously in divided doses) qow, including subcutaneous administration to the patient three times a month or monthly.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of YFV infection in a patient, the method comprising a desired treatment During the period, a dose of PEG-CIFN comprising CIFN-amino acids in an amount of about 18 μg to about 24 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN), an amount of about 100 μg to about 300 μg / week. Qw, qow in combination with a total weekly dosage of IFN-γ containing qd, qod, tiw, biw, or substantially continuously or continuously in divided doses Including subcutaneous administration to the patient three times a month or monthly.

  In general, a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ suitable for use in the methods of the invention is one million units (MU) IFN-α 2a or 2b. Or 2c: provided at a dosage ratio of 30 μg IFN-γ, where IFN-α 2a or 2b or 2c and both IFN-γ are non-PEGylated and non-glycosylated species.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of YFV infection in a patient, which method is desirable. During the treatment period, a dosage of IFN-α2a containing an amount of drug of about 1 MU to about 20 MU per dose of IFN-α 2a, 2b or 2c, and an amount of about 30 μg to about 600 μg per dose of IFN-γ Qd, qod, tiw, biw, in combination with IFN-γ dosages (subcutaneously, qd, qod, tiw, biw, or substantially continuously or continuously per day) Alternatively, the method includes subcutaneous administration to the patient substantially continuously or continuously per day.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of YFV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 3 MU of drug per dose of IFN-α 2a, 2b or 2c, and IFN-α containing about 100 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of YFV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 10 MU of drug per dose of IFN-α 2a, 2b or 2c, and IFN-α containing about 300 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of YFV infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of drug of about 90 μg to about 360 μg per dose of PEGASYS® is administered to an amount of drug of about 30 μg to about 1,000 μg / week. 1 week total dosage of IFN-γ (administered subcutaneously in qd, qod, tiw, or biw in divided doses, or substantially continuously or continuously) In combination with qw, qow, 3 times a month or monthly administered to the patient.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of YFV infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of about 180 μg of drug per dose of PEGASYS® will result in an IFN − containing about 100 μg to about 300 μg / week of drug. in combination with a weekly total dosage of γ (subcutaneously administered in divided doses qd, qod, tiw, or biw, or administered substantially continuously or continuously), qw, qow, including subcutaneous administration to the patient three times a month or monthly.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of YFV infection in a patient, the method Is about PEG-INTRON® dosage containing about 0.75 μg to about 3.0 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period. Total weekly IFN-γ dosage containing 30 μg to about 1,000 μg / week of drug (subcutaneously administered in divided doses, qd, qod, tiw, or biw, or substantially Qw, qow, 3 times a month, or monthly, in combination with a patient.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of YFV infection in a patient, the method PEG-INTRON® containing about 1.5 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period, from about 100 μg to about 300 μg 1 week total dosage of IFN-γ with weekly dose of drug (subcutaneously administered in divided doses qd, qod, tiw, or biw, or substantially continuously or continuously Qw, qow, 3 times a month, or monthly, in combination with a patient.

  The present invention also provides a method for the treatment of YFV infection, wherein ribavirin treatment is added to any of the combination treatments of type I or type III interferon receptor agonists and IFN-γ described above. In certain embodiments, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 800 mg to 1200 mg ribavirin oral qd for a particular treatment period. In other embodiments, a combination therapy of a type I or type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 1000 mg ribavirin oral qd for a particular treatment period. In a further embodiment, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include an oral qd ribavirin regimen of about 10 mg ribavirin / kg body weight for a particular period of treatment. The daily ribavirin dosage can be administered in one dose per day, or in divided doses (including once, twice, three, or four doses per day).

  In one embodiment, the present invention provides a treatment for YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment, and qd Administering a ribavirin regimen administered orally in an individual having YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering a subcutaneously administered 50 μg Actimmune® human IFN-γ1b; and a ribavirin regimen administered orally in qd to an individual with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 100 μg Actimmune® human IFN-γ1b administered subcutaneously; and ribavirin regimen administered orally in qd to individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 50 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with YFV infection.

  In one embodiment, the present invention provides a treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with YFV infection.

  In one embodiment, the present invention provides a treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering a subcutaneously administered 25 μg Actimmune® human IFN-γ1b; and a ribavirin regimen administered orally in qd to an individual with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α; tiw administered subcutaneously with qd or tiw for the desired duration of treatment. Administering 200 μg Actimmune® human IFN-γ1b administered subcutaneously; and a ribavirin regimen administered orally in qd to an individual with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides a treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering a 25 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with YFV infection.

  In one embodiment, the present invention provides a treatment of YFV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw for the desired duration of treatment; and tiw Administering 200 μg Actimmune® human IFN-γ1b regimen administered subcutaneously to an individual with YFV infection.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a ribavirin regimen administered orally in qd is included in individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 100 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with YFV infection.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with YFV infection.

  In one embodiment, the present invention provides for the treatment of YFV infection, the method comprising a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a ribavirin regimen administered orally in qd are included in individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, the method comprising a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, the method comprising a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 100 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, the method comprising a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a 50 μg Actimmune® human IFN-γ1b regimen administered subcutaneously with tiw is included in individuals with YFV infection.

  In one embodiment, the present invention provides for the treatment of YFV infection, the method comprising a 150 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a 100 μg Actimmune® human IFN-γ1b regimen administered subcutaneously with tiw is included in individuals with YFV infection.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α and a ribavirin regimen administered orally in qd are included in individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 50 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. IFN-α; including 100 μg Actimmune® human IFN-γ1b administered subcutaneously in tiw; and ribavirin regimen administered orally in qd to individuals with YFV infection. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 50 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with YFV infection.

  In one embodiment, the present invention provides for the treatment of YFV infection, which method comprises a 200 μg monoPEG (30 kD, linear) consensus administered subcutaneously every 10 days or qw for the desired duration of treatment. And administering a regimen of 100 μg Actimmune® human IFN-γ1b administered subcutaneously at IFN-α; and tiw to an individual with YFV infection.

5. Treatment of Hepatitis C Virus Infection The present invention treats HCV infection by administering a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ to an individual in need thereof. Provide a method. Individuals treated according to the methods of the present invention include individuals who have been clinically diagnosed as having HCV infection. Individuals infected with HCV are identified as having HCV RNA in the individual and / or having anti-HCV antibodies in the individual.

  Individuals clinically diagnosed as infected with HCV include untreated individuals (e.g., individuals not previously treated for HCV) and individuals who have failed previous treatment for HCV ( ]). “Treatment failure patients” include non-responders (eg, individuals whose HCV titers have not been significantly or sufficiently reduced by previous treatment for HCV) and relapsed individuals (eg, those who have been previously treated for and have been treated for HCV). Individuals whose HCV titers are reduced and subsequently increased).

In embodiments of particular interest, the individual has an HCV titer of HCV genomic copies of at least about 10 ⁵ , at least about 5 × 10 ⁵ , or at least about 10 ⁶ , or at least about 2 × 10 ⁶ per milliliter of serum. . Patients may be infected with any HCV genotype (including genotype 1 (including 1a and 1b, 2, 3, 4, 6, etc.) and subtypes (e.g. 2a, 2b, 3a, etc.)) and genotype ( For example, it is particularly difficult to treat HCV genotype 1 and certain HCV subtypes and similar species).

  Severe fibrosis or early cirrhosis (decompensated, Child's-Pugh class A or milder) or more advanced cirrhosis (compensated, Child's-Pugh class B or C) due to chronic HCV infection HCV positive individuals (as described above) and individuals who are viremia or cannot become resistant to IFN-α based therapy despite prior antiviral treatment with IFN-α based therapy Or individuals with contraindications to such treatment are also of particular interest. In an embodiment of particular interest, HCV positive individuals with stage 3 or 4 liver fibrosis following the METAVIR scoring system are suitable for treatment using the methods of the invention. In other embodiments, individuals suitable for treatment using the methods of the invention are patients with decompensated cirrhosis with clinical signs, including patients with much more advanced cirrhosis, and liver transplantation Includes patients waiting. In yet other embodiments, individuals suitable for treatment using the methods of the present invention include patients with a milder degree of fibrosis, including patients with early fibrosis (scoring systems METAVIR, Ludwig, And steps 1 and 2 in Scheuer; or steps 1, 2, or 3) in the Ishak scoring system.

  In performing the combination therapy method for hepatitis C virus infection in an individual as described above, a type I or type III interferon receptor agonist and IFN-γ are administered to the individual. In certain embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in the same formulation. In other embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered in separate formulations. When administered in separate formulations, the Type I or Type III interferon receptor agonist and IFN-γ can be administered substantially simultaneously or within about 24 hours of each other. In many embodiments, the Type I or Type III interferon receptor agonist and IFN-γ are administered subcutaneously in multiple doses.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Type I or III interferon receptor agonists are daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, the type I interferon receptor agonist is IFN-α. Effective dosages of IFN-α range from about 1 μg to about 30 μg, about 3 μg to about 27 μg, about 1 MU to about 20 MU, about 3 MU to about 10 MU, about 90 μg to about 180 μg, or about 18 μg to about 90 μg. obtain.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Effective dosages of Infergen® consensus IFN-α can include an amount of drug of about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg per dose. Effective dosages of IFN-α2a and IFN-α2b can include an amount of drug from about 3 million units (MU) to about 10 MU per dose. An effective dosage of PEGASYS® PEGylated IFN-α2a may comprise an amount of drug from about 90 μg to about 180 μg, or about 135 μg per dose. An effective dosage of PEG-INTRON® PEGylated IFN-α2b may comprise an amount of drug from about 0.5 μg to about 1.5 μg per kg body weight per dose. An effective dosage of PEGylated consensus interferon (PEG-CIFN) can comprise a CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of PEG-CIFN. In many embodiments, the IFN-α and / or IFN-γ is about 1 day to about 7 days, or about 1 week to about 2 weeks, or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks. Or about 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months, or at least 1 year For a period of time, and may be administered over a longer period of time. Dosage regimens may include tid, bid, qd, qod, biw, tiw, qw, qow, three times a month, or monthly.

  In certain embodiments, the specific regimen of drug therapy used in the treatment of HCV patients is the specific disease parameters (initial viral load, genotype of HCV infection in the patient, liver histology, and / or Or the stage of liver fibrosis in the patient). In one embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) Advanced or severe as measured by a Knodell score of 3 or 4 Identifying a patient with stage liver fibrosis, then (2) about 24 weeks to about 60 weeks, or about 30 weeks to about 1 year, or about 36 weeks to about 50 weeks, or about 40 weeks to about 48 A therapeutically effective amount of IFN for a period of weeks, or at least about 24 weeks, or at least about 30 weeks, or at least about 36 weeks, or at least about 40 weeks, or at least about 48 weeks, or at least about 60 weeks -administering α and IFN-γ to the patient.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) Advanced or severe stage liver fibrosis as measured by a Knodell score of 3 or 4 Identifying a patient with the disease, and then (2) a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN- for a period of about 40 weeks, about 50 weeks, or about 48 weeks administering γ to the patient.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV genotype 1 infection and more than 2 million viral genome copies per ml patient serum Identifying a patient with an initial viral load, and then (2) about 24 weeks to about 60 weeks, or about 30 weeks to about 1 year, or about 36 weeks to about 50 weeks, or about 40 weeks to about 48 A therapeutically effective amount of I for a period of weeks, or at least about 24 weeks, or at least about 30 weeks, or at least about 36 weeks, or at least about 40 weeks, or at least about 48 weeks, or at least about 60 weeks Administering a type I or type III interferon receptor agonist and IFN-γ to the patient.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV genotype 1 infection and more than 2 million viral genome copies per ml patient serum Identifying a patient with an initial viral load, and then (2) a therapeutically effective amount of a type I or type III interferon receptor agonist for a period of about 40 weeks, about 50 weeks, or about 48 weeks And administering IFN-γ to the patient.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV genotype 1 infection, and more than 2 million viral genome copies per ml patient serum Identifying a patient with an initial viral load of, and absence of liver fibrosis as measured by a Knodell score of 0, 1, or 2 or early stage liver fibrosis, and then (2) from about 24 weeks About 60 weeks, or about 30 weeks to about 1 year, or about 36 weeks to about 50 weeks, or about 40 weeks to about 48 weeks, or at least about 24 weeks, or at least about 30 weeks, or at least about 36 weeks, or Administering to the patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of at least about 40 weeks, or at least about 48 weeks, or at least about 60 weeks.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV genotype 1 infection, and more than 2 million viral genome copies per ml patient serum Identifying a patient with an initial viral load of, and absence of liver fibrosis or an early stage liver fibrosis as measured by a Knodell score of 0, 1, or 2, and then (2) about 40 weeks, Administering to the patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of about 50 weeks, or about 48 weeks.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV genotype 1 infection, and initial less than 2 million viral genome copies per ml patient serum And (2) about 20 weeks to about 50 weeks, or about 24 weeks to about 48 weeks, or about 30 weeks to about 40 weeks, or up to about 20 weeks, or about Administering to a patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of up to 24 weeks, or up to 30 weeks, or up to 36 weeks, or up to about 48 weeks .

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV genotype 1 infection, and initial less than 2 million viral genome copies per ml patient serum Identifying patients with a viral load of, and then (2) patients with a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of about 20 weeks to about 24 weeks To the stage of administration.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV genotype 1 infection, and initial less than 2 million viral genome copies per ml patient serum Identifying patients with a viral load of, and then (2) patients having a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of about 24 weeks to about 48 weeks To the stage of administration.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) identifying a patient with an infection of HCV genotype 2 or 3, and then (2 ) About 24 weeks to about 60 weeks, or about 30 weeks to about 1 year, or about 36 weeks to about 50 weeks, or about 40 weeks to about 48 weeks, or at least about 24 weeks, or at least about 30 weeks, or at least Administering a patient with a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of about 36 weeks, or at least about 40 weeks, or at least about 48 weeks, or at least about 60 weeks Stage to do.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) identifying a patient with an infection of HCV genotype 2 or 3, and then (2 ) About 20 weeks to about 50 weeks, or about 24 weeks to about 48 weeks, or about 30 weeks to about 40 weeks, or up to about 20 weeks, or up to about 24 weeks, or up to 30 weeks, or up to 36 weeks, or Administering to the patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of up to about 48 weeks.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) identifying a patient with an infection of HCV genotype 2 or 3, and then (2 ) Administering to the patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of about 20 weeks to about 24 weeks.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) identifying a patient with an infection of HCV genotype 2 or 3, and then (2 ) Administering to the patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of at least about 24 weeks.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) identifying a patient having an infection with HCV genotype 4, and then (2) about 24 weeks to about 60 weeks, or about 30 weeks to about 1 year, or about 36 weeks to about 50 weeks, or about 40 weeks to about 48 weeks, or at least about 24 weeks, or at least about 30 weeks, or at least about 36 Administering to a patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of weeks, or at least about 40 weeks, or at least about 48 weeks, or at least about 60 weeks .

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV characterized by any of HCV genotypes 5, 6, 7, 8 or 9 Identifying a patient having an infection, and then (2) administering to the patient a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of about 20 weeks to about 50 weeks Stage to do.

  In another embodiment, the present invention provides a method for the treatment of HCV infection comprising the following steps: (1) HCV characterized by any of HCV genotypes 5, 6, 7, 8 or 9 Identifying a patient having an infection, and then (2) a patient with a therapeutically effective amount of a type I or type III interferon receptor agonist and IFN-γ for a period of at least about 24 weeks to about 48 weeks To the stage of administration.

  In practicing the methods of the invention specific for a patient's HCV genotype and / or viral load as described above, the clinician is sufficient to achieve a sustained viral response over the course of treatment. At a dosage, a type I or type III interferon receptor agonist and IFN-γ are administered to the patient. In one embodiment, such methods include qd, qod, tiw, biw, qw, qow, about 3 μg, about 9 μg, about 15 μg administered subcutaneously three times a month or monthly for a particular treatment period. About 18 μg, or about 27 μg of Infergen® consensus IFN-α IFN-α regimen, and qd, qod, tiw, biw, qw, qow, 3 times a month or 25 μg administered monthly It is provided to treat a patient with an IFN-γ regimen of 300 μg IFN-γ.

  In another embodiment, such methods include qd, qod, tiw, biw, qw, qow, about 3 μg, about 9 μg, about 15 μg administered subcutaneously three times a month or monthly for a particular treatment period. About 18 μg, or about 27 μg of Infergen® consensus IFN-α IFN-α regimen, and qd, qod, tiw, biw, qw, qow, 3 times a month or 100 μg administered monthly It is provided to treat a patient with an IFN-γ regimen of 200 μg IFN-γ.

  In another embodiment, such a method comprises from about 3 million units (MU) administered qd, qod, tiw, biw, qw, qow, 3 times a month, or monthly subcutaneously for a specified treatment period. About 10 MU IFN-α2a or IFN-α2b IFN-α regimen and qd, qod, tiw, biw, qw, qow, 25 μg to 300 μg IFN-γ IFN-γ administered subcutaneously three times a month or monthly It is provided to treat a patient using a gamma regimen.

  In another embodiment, such a method comprises from about 3 million units (MU) administered qd, qod, tiw, biw, qw, qow, 3 times a month, or monthly subcutaneously for a specified treatment period. About 10 MU IFN-α2a or IFN-α2b IFN-α regimen and qd, qod, tiw, biw, qw, qow, 100 μg to 200 μg IFN-γ IFN-γ administered subcutaneously three times a month or monthly It is provided to treat a patient using a gamma regimen.

  In another embodiment, such a method comprises about 90 μg per dose of PEGASYS® PEGylated IFN-α2a administered qw, qow three times a month, or monthly subcutaneously for a specified treatment period. PEGASYS® PEGylated IFN-α2a IFN-α regimen that delivers drug in an amount of ~ 180 μg, or about 135 μg, and qd, qod, tiw, biw, qw, qow, 3 times a month, or It is provided to treat patients with an IFN-γ regimen of 25 μg to 300 μg of IFN-γ administered subcutaneously every month.

  In another embodiment, such a method comprises about 90 μg per dose of PEGASYS® PEGylated IFN-α2a administered qw, qow three times a month, or monthly subcutaneously for a specified treatment period. PEGASYS® PEGylated IFN-α2a IFN-α regimen that delivers drug in an amount of ~ 180 μg, or about 135 μg, and qd, qod, tiw, biw, qw, qow, 3 times a month, or It is provided to treat a patient with an IFN-γ regimen of 100 μg to 200 μg of IFN-γ administered subcutaneously every month.

  In another embodiment, such a method comprises about 0.5 μg PEG-INTRON® PEG per kg body weight administered qw, qow three times a month, or subcutaneously for a specified treatment period. IFN-α2b to about 1.5 μg of PEG-INTRON® IFN-α regimen, and qd, qod, tiw, biw, qw, qow, 3 times a month, or 25 μg administered monthly It is provided to treat a patient with an IFN-γ regimen of 300 μg IFN-γ.

  In another embodiment, such a method comprises about 0.5 μg PEG-INTRON® PEG per kg body weight administered qw, qow three times a month, or subcutaneously for a specified treatment period. IFN-α2b to about 1.5 μg of PEG-INTRON® IFN-α regimen, and qd, qod, tiw, biw, qw, qow, 3 times a month or 100 μg administered monthly It is provided to treat a patient with an IFN-γ regimen of 200 μg IFN-γ.

  In another embodiment, such a method comprises from about 18 μg per dose of PEGylated consensus interferon (PEG-CIFN) administered qw, qow, 3 times a month, or monthly subcutaneously for a particular treatment period. PEG-CIFN IFN-α regimen that provides CIFN amino acid weight in an amount of about 90 μg, or about 27 μg to about 60 μg, or 45 μg, and qd, qod, tiw, biw, qw, qow, 3 times a month, or It is provided to treat patients with an IFN-γ regimen of 25 μg to 300 μg of IFN-γ administered subcutaneously every month.

  In another embodiment, such a method comprises from about 18 μg per dose of PEGylated consensus interferon (PEG-CIFN) administered qw, qow, 3 times a month, or monthly subcutaneously for a particular treatment period. PEG-CIFN IFN-α regimen that provides CIFN amino acid weight in an amount of about 90 μg, or about 27 μg to about 60 μg, or 45 μg, and qd, qod, tiw, biw, qw, qow, 3 times a month, or It is provided to treat a patient with an IFN-γ regimen of 100 μg to 200 μg of IFN-γ administered subcutaneously every month.

  In certain embodiments, the present invention provides methods of using a synergistically effective amount of a type I or type III interferon receptor agonist and IFN-γ in the treatment of hepatitis C virus infection in a patient. In certain embodiments, the present invention provides methods of using synergistically effective amounts of IFN-α and IFN-γ in the treatment of hepatitis C virus infection in a patient. In one embodiment, the present invention provides a method of using synergistically effective amounts of consensus IFN-α and IFN-γ in the treatment of hepatitis C virus infection in a patient.

  Generally, a synergistically effective amount of consensus interferon (CIFN) and IFN-γ, suitable for use in the methods of the present invention, is provided at a dosage ratio of 1 μg CIFN: 10 μg IFN-γ, Here, CIFN and IFN-γ are both non-PEGylated and non-glycosylated species.

  In one embodiment, the invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of hepatitis C virus (HCV) infection in a patient. The method comprises administering a dosage of INFERGEN® comprising an amount of drug of about 1 μg to about 30 μg per dose of INFERGEN® during the desired treatment period, qd, qod, tiw, biw Qw, qow, IFN containing drug in an amount of about 10 μg to about 300 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day -γ dosage (subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) In combination, and subcutaneously administering to the patient.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of HCV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg to about 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 1 A dosage of IFN-γ comprising an amount of drug of about 10 μg to about 100 μg per dose of IFN-γ, three times a month, once a month, or substantially continuously or continuously per day ( Subcutaneously in combination with qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day) Administration step.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of HCV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing about 1 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. IFN-γ dosage containing about 10 μg to about 50 μg of drug per dose of IFN-γ (subcutaneously, once or once a month or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of HCV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 9 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 IFN-γ dosage containing about 90 μg to about 100 μg of drug per dose of IFN-γ (subcutaneously, once or once a month, or substantially continuously or daily) , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the present invention provides a method of using a synergistically effective amount of INFERGEN® consensus IFN-α and IFN-γ in the treatment of HCV infection in a patient, the method comprising: During the treatment period to be administered, the dosage of INFERGEN® containing approximately 30 μg of drug per dose of INFERGEN® is qd, qod, tiw, biw, qw, qow, 3 per month. Dosages of IFN-γ containing about 200 μg to about 300 μg of drug per dose of IFN-γ, subcutaneously, once a month, or substantially continuously or daily. , Qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or in combination with daily, substantially continuously or continuously) including.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of HCV infection in a patient, the method comprising a desired treatment During the period, a dose of PEG-CIFN containing CIFN-amino acids in an amount of about 4 μg to about 60 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN) is about 30 μg to about 1,000 μg / week. Qw, in combination with a total weekly dosage of IFN-γ containing an amount of drug (qd, qod, tiw, biw, or administered substantially continuously or continuously in divided doses) qow, including subcutaneous administration to the patient three times a month or monthly.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGylated consensus IFN-α and IFN-γ in the treatment of HCV infection in a patient, the method comprising a desired treatment During the period, a dose of PEG-CIFN comprising CIFN-amino acids in an amount of about 18 μg to about 24 μg weight per dose of PEGylated consensus IFN-α (PEG-CIFN), an amount of about 100 μg to about 300 μg / week. Qw, qow in combination with a total weekly dosage of IFN-γ containing qd, qod, tiw, biw, or substantially continuously or continuously in divided doses Including subcutaneous administration to the patient three times a month or monthly.

  In general, a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ suitable for use in the methods of the invention is one million units (MU) IFN-α 2a or 2b. Or 2c: provided at a dosage ratio of 30 μg IFN-γ, where IFN-α 2a or 2b or 2c and both IFN-γ are non-PEGylated and non-glycosylated species.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of HCV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing an amount of drug of about 1 MU to about 20 MU per dose of IFN-α 2a or 2b or 2c, an amount of about 30 μg to about 600 μg per dose of IFN-γ Qd, qod, tiw, biw, in combination with IFN-γ dosages (subcutaneously, qd, qod, tiw, biw, or substantially continuously or continuously per day) Alternatively, the method includes subcutaneous administration to the patient substantially continuously or continuously per day.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of HCV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 3 MU of drug per dose of IFN-α 2a or 2b or 2c, and IFN-α containing about 100 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another embodiment, the present invention provides a method of using a synergistically effective amount of IFN-α 2a or 2b or 2c and IFN-γ in the treatment of HCV infection in a patient, which method is desirable. During the treatment period, a dose of IFN-α2a containing about 10 MU of drug per dose of IFN-α 2a or 2b or 2c and IFN-α containing about 300 μg of drug per dose of IFN-γ combined with a dosage of γ (subcutaneously, qd, qod, tiw, biw, or substantially daily or continuously), qd, qod, tiw, biw, or substantially daily The method includes the step of administering to a patient subcutaneously continuously or continuously.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of HCV infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of drug of about 90 μg to about 360 μg per dose of PEGASYS® is administered to an amount of drug of about 30 μg to about 1,000 μg / week. 1 week total dosage of IFN-γ (administered subcutaneously in qd, qod, tiw, or biw in divided doses, or substantially continuously or continuously) In combination with qw, qow, 3 times a month or monthly administered to the patient.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEGASYS® PEGylated IFN-α2a and IFN-γ in the treatment of HCV infection in a patient, the method comprising: During the desired treatment period, a dosage of PEGASYS® containing an amount of about 180 μg of drug per dose of PEGASYS® will result in an IFN − containing about 100 μg to about 300 μg / week of drug. in combination with a weekly total dosage of γ (subcutaneously administered in divided doses qd, qod, tiw, or biw, or administered substantially continuously or continuously), qw, qow, including subcutaneous administration to the patient three times a month or monthly.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of HCV infection in a patient, the method Is about PEG-INTRON® dosage containing about 0.75 μg to about 3.0 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period. Total weekly IFN-γ dosage containing 30 μg to about 1,000 μg / week of drug (subcutaneously administered in divided doses, qd, qod, tiw, or biw, or substantially Qw, qow, 3 times a month, or monthly, in combination with a patient.

  In another aspect, the invention provides a method of using a synergistically effective amount of PEG-INTRON® PEGylated IFN-α2b and IFN-γ in the treatment of HCV infection in a patient, the method PEG-INTRON® containing about 1.5 μg / kg body weight of drug per dose of PEG-INTRON® during the desired treatment period, from about 100 μg to about 300 μg 1 week total dosage of IFN-γ with weekly dose of drug (subcutaneously administered in divided doses qd, qod, tiw, or biw, or substantially continuously or continuously Qw, qow, 3 times a month, or monthly, in combination with a patient.

  In one embodiment, the present invention provides for the treatment of HCV infection, comprising: 9 μg INFERGEN® consensus IFN-α administered subcutaneously qd or tiw, and ribavirin regimen administered orally qd. Is administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; 50 μg Actimmune administered subcutaneously with tiw. Trademarked human IFN-γ1b; and a ribavirin regimen administered qd orally to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; 100 μg Actimmune (registered) administered subcutaneously with tiw. Trademarked human IFN-γ1b; and a ribavirin regimen administered qd orally to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; and 50 μg Actimmune administered subcutaneously with tiw ( Comprising administering a regimen of human IFN-γ1b to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; and 100 μg Actimmune administered subcutaneously with tiw ( Comprising administering a regimen of human IFN-γ1b to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; 25 μg Actimmune (registered) administered subcutaneously with tiw. Trademarked human IFN-γ1b; and a ribavirin regimen administered qd orally to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; 200 μg Actimmune administered subcutaneously with tiw. Trademarked human IFN-γ1b; and a ribavirin regimen administered qd orally to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method comprises 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; and 25 μg Actimmune administered subcutaneously with tiw ( Comprising administering a regimen of human IFN-γ1b to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the present invention provides for the treatment of HCV infection, comprising: 9 μg INFERGEN® consensus IFN-α administered subcutaneously with qd or tiw; and 200 μg Actimmune administered subcutaneously with tiw ( Comprising administering a regimen of human IFN-γ1b to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is orally administered with 100 μg monoPEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw, and qd. Administering the ribavirin regimen to be administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is administered subcutaneously at 100 μg monoPEG (30 kD, linear) consensus IFN-α; tiw administered subcutaneously every 10 days or qw. And a ribavirin regimen administered orally in qd is administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. The 50 μg Actimmune® human IFN-γ1b; In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is administered subcutaneously at 100 μg monoPEG (30 kD, linear) consensus IFN-α; tiw administered subcutaneously every 10 days or qw. 100 μg Actimmune® human IFN-γ1b; and a ribavirin regimen administered orally in qd is administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw; and tiw subcutaneously. Administering an administered 50 μg Actimmune® human IFN-γ1b regimen to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the present invention provides for the treatment of HCV infection, which method comprises 100 μg monoPEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw; and tiw subcutaneously. Administering an administered 100 μg Actimmune® human IFN-γ1b regimen to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method is orally administered with 150 μg monoPEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw, and qd. Administering the ribavirin regimen to be administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is administered subcutaneously at 150 μg monoPEG (30 kD, linear) consensus IFN-α; tiw administered subcutaneously every 10 days or qw. And a ribavirin regimen administered orally in qd is administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. The 50 μg Actimmune® human IFN-γ1b; In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is administered subcutaneously at 150 μg monoPEG (30 kD, linear) consensus IFN-α; tiw administered subcutaneously every 10 days or qw. 100 μg Actimmune® human IFN-γ1b; and a ribavirin regimen administered orally in qd is administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is subcutaneously administered 150 μg monoPEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw, and tiw. Administering an administered 50 μg Actimmune® human IFN-γ1b regimen to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is subcutaneously administered 150 μg monoPEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw, and tiw. Administering an administered 100 μg Actimmune® human IFN-γ1b regimen to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the present invention provides for the treatment of HCV infection, wherein the method is administered orally with 200 μg monoPEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw, and qd. Administering the ribavirin regimen to be administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method is administered subcutaneously at 200 μg monoPEG (30 kD, linear) consensus IFN-α; tiw administered subcutaneously every 10 days or qw. And a ribavirin regimen administered orally in qd is administered to an individual with HCV infection, wherein the duration of treatment is 48 weeks. The 50 μg Actimmune® human IFN-γ1b; In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the invention provides for the treatment of HCV infection, wherein the method is administered subcutaneously at 200 μg monoPEG (30 kD, linear) consensus IFN-α; tiw administered subcutaneously every 10 days or qw. 100 μg Actimmune® human IFN-γ1b; and a ribavirin regimen orally administered qd is administered to an individual with viral infection, wherein the duration of treatment is 48 weeks. In this embodiment, ribavirin is administered in an amount of 1000 mg for individuals weighing less than 75 kg and 1200 mg for individuals weighing 75 kg or more.

  In one embodiment, the present invention provides for the treatment of HCV infection, which method comprises 200 μg mono-PEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw; and tiw subcutaneously. Administering an administered 50 μg Actimmune® human IFN-γ1b regimen to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  In one embodiment, the present invention provides for the treatment of HCV infection, which method comprises 200 μg mono-PEG (30 kD, linear) consensus IFN-α administered subcutaneously every 10 days or qw; and tiw subcutaneously. Administering an administered 100 μg Actimmune® human IFN-γ1b regimen to an individual with HCV infection, wherein the duration of treatment is 48 weeks.

  Any of the above treatment regimens can be administered to individuals who have failed previous treatment for HCV ("treatment failure patients" including non-responders and relapsers). Accordingly, in certain embodiments, the present invention provides a method of treating HCV infection in a treatment failure patient, wherein the method administers an effective amount of IFN-α and an effective amount of IFN-γ for 48 hours. Including stages. In other embodiments, the present invention provides a method for treating HCV infection in patients with treatment failure, wherein the method comprises an effective amount of IFN-α and an effective amount of IFN-γ for 48 hours, And administering an effective amount of ribavirin. In other embodiments, the present invention provides a method for treating HCV infection in non-responder patients, wherein the method provides an effective amount of IFN-α and an effective amount of IFN-γ for 48 hours. Administration step. In other embodiments, the invention provides a method for treating HCV infection in non-responder patients, wherein the method comprises an effective amount of IFN-α and an effective amount of IFN-γ for 48 hours. And administering an effective amount of ribavirin. In other embodiments, the invention provides a method for treating HCV infection in relapsed patients, wherein the method administers an effective amount of IFN-α and an effective amount of IFN-γ for 48 hours. Including the steps of: In other embodiments, the invention provides a method for treating HCV infection in relapsed patients, wherein the method comprises an effective amount of IFN-α and an effective amount of IFN-γ for 48 hours, And administering an effective amount of ribavirin.

  Any of the above therapeutic regimens can be administered to an untreated patient infected with HCV genotype 1. Accordingly, in certain embodiments, the present invention provides a method of treating HCV infection in an untreated patient having an infection with HCV genotype 1, wherein the method comprises an effective amount of IFN-α and Administering an effective amount of IFN-γ. In other embodiments, the present invention provides a method for treating HCV infection in an untreated patient having an infection with HCV genotype 1, wherein the method comprises an effective amount of IFN-α for 48 hours. And administering an effective amount of IFN-γ and an effective amount of ribavirin.

  Any of the above therapeutic regimens can be administered to an untreated patient infected with HCV genotype 4. Accordingly, in certain embodiments, the invention provides a method of treating HCV infection in an untreated patient having an infection with HCV genotype 4, wherein the method comprises an effective amount of IFN-α and Administering an effective amount of IFN-γ. In other embodiments, the present invention provides a method for treating HCV infection in an untreated patient having an infection with HCV genotype 4, wherein the method comprises an effective amount of IFN-α for 48 hours. And administering an effective amount of IFN-γ and an effective amount of ribavirin.

Any of the above therapeutic regimens can be administered to an untreated patient infected with HCV genotype 1, which patient has a high viral load (HVL), where “HVL” is 2 × per mL of serum. 10 ⁶ Refers to a higher HCV viral load than HCV genome copies. Accordingly, in certain embodiments, the present invention provides a method of treating HCV infection in an untreated patient having an infection with HCV genotype 1 and having a high viral load, wherein the method is effective for 48 hours. Administering an effective amount of IFN-α and an effective amount of IFN-γ. In other embodiments, the present invention provides a method for treating HCV infection in an untreated patient having an infection with HCV genotype 1 and having a high viral load, the method comprising a period of 48 hours. Administering an effective amount of IFN-α, and an effective amount of IFN-γ, and an effective amount of ribavirin.

  The present invention also provides a method for the treatment of HCV infection, wherein ribavirin treatment is added to any of the combinations of type I or type III interferon receptor agonists and IFN-γ described above. In certain embodiments, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 800 mg to 1200 mg ribavirin oral qd for a particular treatment period. In other embodiments, a combination therapy of a type I or type III interferon receptor agonist and IFN-γ is modified to include a ribavirin regimen of 1000 mg ribavirin oral qd for a particular treatment period. In a further embodiment, a combination therapy of a Type I or Type III interferon receptor agonist and IFN-γ is modified to include an oral qd ribavirin regimen of about 10 mg ribavirin / kg body weight for a particular period of treatment. The daily ribavirin dosage can be administered in one dose per day, or in divided doses (including once, twice, three, or four doses per day).

7. Treatment of liver fibrosis Individuals with liver fibrosis that are suitable for treatment according to the invention of the present invention include individuals who have been clinically diagnosed as having liver fibrosis, as well as clinical liver fibrosis. Includes individuals who have not yet developed but are considered at risk for developing liver fibrosis. Such individuals include, but are not limited to: individuals infected with HCV; individuals infected with HBV; individuals infected with Schistosoma mansoni; known to cause liver fibrosis Individuals exposed to known chemical agents; individuals diagnosed with Wilson disease; individuals diagnosed with hemochromatosis; individuals with alcoholic liver disease; individuals with nonalcoholic steatohepatitis; autoimmune hepatitis An individual having primary sclerosing cholangitis, primary biliary cirrhosis, or α-1-antitrypsin deficiency.

  In one aspect, the present invention treats liver fibrosis in a patient comprising administering a type I or type III interferon receptor agonist and IFN-γ in an amount effective to reduce liver fibrosis. Features method. Optionally, the method comprises an effective amount of pirfenidone or a specific pirfenidone analog to enhance the antifibrotic effect or reduction of liver fibrosis achieved by a type I or type III interferon receptor agonist and IFN-γ treatment And providing a patient with a combination of a type I or type III interferon receptor agonist and IFN-γ.

  In another aspect, the present invention provides a method of increasing liver function in a patient suffering from liver fibrosis, wherein the method comprises an amount of type I or type III effective to increase liver function. Administering to the patient an interferon receptor agonist and IFN-γ. Optionally, the method of the invention comprises an amount of pirfenidone effective to reduce the antifibrotic effect achieved by treatment of a type I or type III interferon receptor agonist and IFN-γ or to increase liver function. A step of administering to a patient a combination of a type I or type III interferon receptor agonist and IFN-γ along with a specific pirfenidone analog is provided.

  In another aspect, the present invention provides a method for reducing the incidence of cirrhosis complications in a patient suffering from cirrhosis, wherein the method comprises an amount effective to reduce the occurrence of cirrhosis complications. Administering a type I or type III interferon receptor agonist and IFN-γ to the patient. Optionally, the method of the present invention is effective in enhancing the antifibrotic effect achieved by treatment with a type I or type III interferon receptor agonist and IFN-γ or reducing the incidence of complications of cirrhosis. A step of administering to a patient a combination of a type I or type III interferon receptor agonist and IFN-γ together with pirfenidone or a specific pirfenidone analog is provided.

  Effective dosages of IFN-γ can range from about 25 μg / dose to about 300 μg / dose, from about 10 μg / dose to about 100 μg / dose, or from about 100 μg / dose to about 1000 μg / dose.

  Type I or III interferon receptor agonists are daily, every other day, once a week, three times a week, every other week, three times a month, once a month, substantially continuously, or continuously Can be administered.

  In certain embodiments, the type I interferon receptor agonist is IFN-α. Effective dosages of IFN-α range from about 1 μg to about 30 μg, about 3 μg to about 27 μg, about 1 MU to about 20 MU, about 3 MU to about 10 MU, about 90 μg to about 180 μg, or about 18 μg to about 90 μg. obtain.

  In one embodiment, the method of the invention for the treatment of liver fibrosis described above comprises administering IFN-γ comprising an amount of drug from about 25 μg to about 300 μg, or from about 100 μg to about 200 μg per dose of IFN-γ. Amount (subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day), and INFERGEN (registration) A dosage of INFERGEN® containing about 3 μg, about 9 μg, about 15 μg, about 18 μg, or about 27 μg of drug per dose (subcutaneously qd, qod, tiw, biw, qw, qow, three times a month, once a month, or substantially continuously or continuously per day). The antifibrotic effects or other therapeutic benefits of such regimens range from about 5 mg / kg body weight to about 125 mg / kg body weight, or from about 400 mg to about 3600 mg, or from about 800 mg to about 2400 mg, or from about 1000 mg to about A dose based on the weight of 1800 mg, or a fixed dosage of pirfenidone or a specific pirfenidone analog of about 1200 mg to about 1600 m, co-administered to the patient via oral qd during the desired period of treatment of IFN-α and IFN-γ Can be enhanced.

  In another embodiment, the method of the invention for the treatment of liver fibrosis as described above comprises administering an IFN-γ comprising an amount of a drug from about 25 μg to about 300 μg, or from about 100 μg to about 200 μg per dose of IFN-γ. Amount (subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day), and IFN-α2a Or a dosage of IFN-α2a or IFN-α2b containing 3 million units (MU) to about 10 MU of drug per dose of IFN-α2b (subcutaneously, qd, qod, tiw, biw, qw, qow, 1 Can be performed by administering to the patient three times a month, once a month, or substantially continuously or continuously per day). The antifibrotic effects or other therapeutic benefits of such regimens range from about 5 mg / kg body weight to about 125 mg / kg body weight, or from about 400 mg to about 3600 mg, or from about 800 mg to about 2400 mg, or from about 1000 mg to about A dose based on the weight of 1800 mg, or a fixed dosage of pirfenidone or a specific pirfenidone analog of about 1200 mg to about 1600 m, co-administered to the patient via oral qd during the desired period of treatment of IFN-α and IFN-γ Can be enhanced.

  In another embodiment, the method of the invention for the treatment of liver fibrosis as described above comprises administering an IFN-γ comprising an amount of a drug from about 25 μg to about 300 μg, or from about 100 μg to about 200 μg per dose of IFN-γ. Amount (subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day), and IFN-α2a Or a dosage of PEGASYS® PEGylated IFN-α2a containing about 90 μg to about 180 μg, or about 135 μg of drug per dose of PEGASYS® (subcutaneously, qw, qow, 3 per month Once or monthly) to the patient. The antifibrotic effects or other therapeutic benefits of such regimens range from about 5 mg / kg body weight to about 125 mg / kg body weight, or from about 400 mg to about 3600 mg, or from about 800 mg to about 2400 mg, or from about 1000 mg to about A dose based on the weight of 1800 mg, or a fixed dosage of pirfenidone or a specific pirfenidone analog of about 1200 mg to about 1600 m, co-administered to the patient via oral qd during the desired period of treatment of IFN-α and IFN-γ Can be enhanced.

  In another embodiment, the method of the invention for the treatment of liver fibrosis as described above comprises administering an IFN-γ comprising an amount of a drug from about 25 μg to about 300 μg, or from about 100 μg to about 200 μg per dose of IFN-γ. Amount (subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day), and PEG-INTRON Dosage of PEG-INTRON® PEGylated IFN-α2b containing drug in an amount of about 0.5 μg to about 1.5 μg per kg body weight per dose of subcutaneous (qw, qow, 1 month 3 times or monthly) to the patient. The antifibrotic effects or other therapeutic benefits of such regimens range from about 5 mg / kg body weight to about 125 mg / kg body weight, or from about 400 mg to about 3600 mg, or from about 800 mg to about 2400 mg, or from about 1000 mg to about A dose based on the weight of 1800 mg, or a fixed dosage of pirfenidone or a specific pirfenidone analog of about 1200 mg to about 1600 m, co-administered to the patient via oral qd during the desired period of treatment of IFN-α and IFN-γ Can be enhanced.

  In another embodiment, the method of the invention for the treatment of liver fibrosis as described above comprises administering an IFN-γ comprising an amount of a drug from about 25 μg to about 300 μg, or from about 100 μg to about 200 μg per dose of IFN-γ. Amount (subcutaneously, qd, qod, tiw, biw, qw, qow, 3 times a month, once a month, or substantially continuously or continuously per day), and PEGylation consensus Dosage of PEG-CIFN (subcutaneously qw, qow, 1 month) containing CIFN amino acid weight in an amount of about 18 μg to about 90 μg, or about 27 μg to about 60 μg, or 45 μg per dose of interferon (PEG-CIFN) 3 times or monthly) to the patient.

  The antifibrotic effects or other therapeutic benefits of such regimens range from about 5 mg / kg body weight to about 125 mg / kg body weight, or from about 400 mg to about 3600 mg, or from about 800 mg to about 2400 mg, or from about 1000 mg to about A dosage based on the weight of 1800 mg, or a fixed dosage of pirfenidone or a specific pirfenidone analog of about 1200 mg to about 1600 m is administered orally during the desired period of treatment of a type I or type III interferon receptor agonist and IFN-γ. Can be enhanced by co-administering to patients with qd.

  In many embodiments, the Type I or Type III interferon receptor agonist and / or IFN-γ is about 1 day to about 7 days, or about 1 week to about 2 weeks, or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks, about 1 month to about 2 months, or about 3 months to about 4 months, or about 4 months to about 6 months, or about 6 months to about 8 months, or about 8 months to about 12 months Or for a period of at least one year and may be administered over a longer period of time. In embodiments utilizing co-administration of pirfenidone or specific pirfenidone analogs, the duration of treatment with pirfenidone or specific pirfenidone analogs is concurrent with the duration of treatment with a type I or type III interferon receptor agonist and / or IFN-γ. It can be.

EXAMPLES The following examples are presented to provide one of ordinary skill in the art with a complete disclosure and description of how to make and use the invention and which we regard as our invention. It is not intended to limit the scope of what is present, and is not intended to represent that the experiment below is all performed or that the experiment is alone. Efforts have been made to ensure accuracy with respect to numbers used (eg amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.

Example 1: CIFN and IFN-γ antiviral activity properties materials and methods
Hela cells were treated with 10% heat-inactivated serum (bovine or fetal as needed) (Hyclone Laboratories, Inc., Logan, UT), L-glutamine (2 mM), streptomycin (100 μg / ml), and penicillin (100 u / ml) and supplemented in DMEM or RPMI-1640 medium. Cells were grown at 37 ° C. in a 5% CO ₂ humidified incubator. HeLa cells (2 × 10 ⁴ cells per well in a 96 well microtiter plate) were treated with IFN 24 hours prior to virus addition. 0.1 MOI of virus was added and the plates were incubated for an additional 48 hours, at which point the cell monolayer was stained with 0.5% crystal violet in 20% (volume / volume) methanol. All cytopathic effect (CPE) inhibition assays were performed in duplicate. One unit of IFN was defined as the amount of IFN that inhibited CPE by 50%. All IFN activities were measured against NIH standard human IFN-α (Namalwa / Sendai) (Ga23-901-532).

Results The data generated in the assay is shown in Table 2 below.

^*ここで使用した値は別々の毒性研究からである。なぜなら、毒性効果を実証するために十分に高い希釈はこのプレート上では実行されなかったからである。
EC50=有効な濃度。抗ウイルス効果の測定。
IC50=阻害濃度。薬物のみ、ウイルスなし。薬物の毒性の測定。
SI=選択性指数。IC50/EC50。高SIはさらに試験する価値のある薬物の標準的指数である。 (Table 2) Infergen, Intron A, and Actimmune activity against West Nile virus

^* Values used here are from separate toxicity studies. This is because dilutions high enough to demonstrate toxic effects were not performed on this plate.
EC50 = effective concentration. Measurement of antiviral effect.
IC50 = inhibitory concentration. Drug only, no virus. Measurement of drug toxicity.
SI = selectivity index. IC50 / EC50. High SI is a standard index of drugs worth further testing.

  The results of the HeLa / VSV assay indicate that Infergen and IFN-γ show highly synergistic antiviral growth inhibitory activity against West Nile virus. Given the role of interferon in the naturally occurring immune system, the antiviral properties of interferon may not be specific to or limited to a particular virus. It is believed that Infergen (and other interferon-α) and IFN-γ exhibit synergistic antiviral activity against hepatitis C virus and other alphaviruses.

Example 2: Effect of IFN-α and interferon inducer against West Nile virus in cell culture and mouse and hamster models Materials and methods Viruses and animals
WNV New York Crow brain extract (NY, CDC 996625, V1 D3 11/10/1999) was used. Dengue virus, yellow fever virus, and West Nile virus were grown in African green monkey kidney (Vero 76, ATCC CCL1587) cells using medium 199 with 5% fetal bovine serum (FBS), 0.1% NaHCO ₃ . Cells were maintained during antiviral experiments using MEM with 1% FBS, 0.1% NaHCO ₃ , and 50 μg / mL gentamicin (Sigma, St. Lois, MO). Virus was titered in sub-confluent Vero cells in 96 well microtiter plates. Female 4-5 week old BALB / c mice (Simonsen Laboratories, Gilroy, CA or Harlan Sprague Dawley, Indianapolis, IN) and male 4-5 or> 7 week old Syrian golden hamsters were used. Animals were challenged subcutaneously with 10 ⁴ 50% cell culture infectious dose (CCID ₅₀ ) diluted in MEM. Animals were randomly assigned to cages.

Compounds Human hybrid interferon-α B / D (IFN-αB / D) was obtained from either David Gangemi (Clemson University) or Johnson Lau (Ribapharm, Inc Costa Mesa, California). IFN-αB / D is known to have activity in mice. Wintergerst et al. (1999) Antiviral Res 44 (1): 75-8; Gangemi et al. (1989) 9 (3): 275-83; Brooks et al. (1999) Antiviral Res 41 (1): 57-64; and Gangemi et al. (1989) 9 (2): 227-37. InfergenTM (lot number P002586, interferon alfacon-1, Intermune, Inc., Brisbane, CA) is the first engineered to contain the most frequently occurring amino acids among non-allelic interferon alpha subtypes. It is a two generation cytokine. In a cell culture model, Infergen demonstrates increased strength when compared to naturally occurring type I interferon, and in a comparative clinical trial using naturally occurring type I interferon, It is a potent inhibitor. Infergen is active in hamsters but not in mice. Fish et al. (1986) Antimicrob Agents Chemother 30 (1): 52-6. Interferon-α2b (IFN-α2b) and interferon-γ (IFN-γ) were also obtained from Intermune, Inc. Infergen and ribavirin with appropriate doses were mixed together for combination studies in the same injection bottle and stored at 4 ° C. during the course of the experiment. The drug was co-injected subcutaneously. Aqueous Ampligen ™ (30 μg / mL) was obtained from William M. Mitchell (School of Medical Pathology, Vanderbilt University). Ampligen ™ is an RNA-like molecule composed of double-stranded poly I: poly C12U, where uridylic acid substitution in the poly C chain creates a non-hydrogen bonding region. Ampligen ™ induces and activates the production of 2-5A synthetase, which in turn activates RNAseL, which destroys viral RNA. Since Ampligen ™ is an RNA-like molecule, care was taken to prevent RNase contamination by using RNase-free material and DEPC-treated water. Ampligen was aliquoted and frozen until use. Aldara ™ (imiquimod) was obtained from a pharmacy in a topical cream at a concentration of 12.5 mg imiquimod per 250 mg cream package. One package of cream was spread over the shaved back of 9 mice or 10 hamsters over a 2.2 cm ² surface area. This resulted in dosages of 1.4 and 1.3 mg / 2.2 cm ² for mice and hamsters, respectively.

Cell culture assay
A cytopathic effect (CPE) assay for WNV has been previously described. Morrey et al. (2002) 55: 107-116; Smee et al. (1988) 10: 253-262. Briefly, serial dilutions of test compounds are added to subconfluent Vero 76 cells in a 96-well microplate, followed by _5-50 % cell culture infectious dose (CCID ₅₀ ) of WNV and 50 CCID ₅₀ of yellow fever virus. And dengue virus were added. Controls on each 96 well plate were uninfected cells, infected cells without drug, and uninfected drug treated cells. Duplicate toxicity controls at each drug concentration and triplicate test samples were run. Cells were visually scored for CPE. The 50% effective concentration (EC ₅₀ ) and 50% inhibitory cytotoxic concentration (IC ₅₀ ) were determined by regression analysis using the mean CPE titer at each concentration of compound. The selectivity index (SI) was calculated as IC ₅₀ ÷ EC ₅₀ . Neutral red (NR) virus staining was used to confirm the visual CPE assay and to provide more quantitative results. Morrey et al. (2002) supra; and Smee et al. (1988) supra. After visual reading of CPE, cells were incubated with neutral red dye at 37 ° C. for 2-3 hours. Free dye was washed from the wells and dye uptake was quantified at 540 and 405 nm using a macroplate reader (Bio-Tek EL 1309, BioTek, Burlington, VT). Absorption values were expressed as a percentage of control and EC ₅₀ and IC ₅₀ values were calculated by regression analysis.

  Viral titers in tissue or plasma are assayed using a virus yield cell culture assay (Morrey et al. (2000), supra), where either a specific volume of tissue homogenate or plasma is run in a series. Added to the first tube of the dilution tube. Serial dilutions were made and added to Vero cells. After 6 days, the end point of infection was identified using CPE. Sidwell et al. (1971) 22: 797-801. Four replicate experiments were used to calculate the infection dose per mL of serum or gram of tissue.

Climbing assay
The hamster was placed at the lower end of a ramp with a slope of 42 degrees (4 inch internal width, 6 inch internal height). The distance traveled by the hamster over a specified 15 second period was recorded. Measurements of uninfected hamsters were taken at night and during the day after shaking the cage to excite the hamsters to determine that both measurements were similar. All hamsters, including sick hamsters, seemed to have a run up the ramp. This assay reflects signs of disease such as muscle weakness, rotational behavior, balancing behavior, and hind limb paralysis. Xiao et al. (2001) Emerg Infect Dis, 7 (4): 714-21; and Chowers et al. (2001) Emerg Infect Dis. 7 (4): 675-8.

Treatment of mice -1 day or -4 to -6 hours after virus injection Female BALB / c mice (4-5 weeks old) were intraperitoneally (ip) Ampligen (13 mg / kg) every other day (eod) to 9 dpi Qd × 7 days topical imiquimod (1.4 mg / 2.2 cm ² ) or qd × 7 days ip IFN-αB / D (6.5 × 10 ⁷ U / kg) (Johnson Lau, Ribapharm, Inc.) -1 treatment after virus injection (dpi). Saline was used as vehicle treatment for Ampligen and IFN-α B / D. Ten animals were used in each WNV infected and non-infected-non-treated group, and 6 animals were used in the toxic control group. The percent survival was calculated based on the number of animals still alive at 11 dpi.

Female BALB / c mice (4-5 weeks old) are also treated with ip Ampligen (15 or 4.8 mg / kg) every other day (eod) up to 6 dpi, or q Id × 5 days ip IFN-α B / D (1.5 × 10 ⁷ , or 1.5 × 10 ⁶ U / kg) (David Gangemi, Clemson University), treated at -4 to -6 hours post-virus injection (hpi). Saline was used as the vehicle. The percent survival was calculated based on the number of animals still alive at 13 dpi.

Treatment of hamsters -4 to -6 hours after virus injection Female hamsters (4-5 weeks of age) are administered every other day (eod) ip Ampligen (10 or 3.2 mg / kg) up to 6 dpi, or qd x 7 days sc Infergen (10 or 1 μg / kg) was treated to -4 to -6 hpi. Saline was used for the vehicle control group. Percent survival and weight change were calculated based on the number of animals still alive at 13 dpi and the difference between -2 dip and 7 dpi, respectively. Nine hamsters were used in the highest dosage group of therapeutic agents tested and in the saline-infected group. Seven hamsters were used in the lower dosage group and the toxicity control group.

Infergen and ribavirin combination study Subcutaneous treatment of hamsters was started 2 days after virus challenge (2 dpi). Average percent weight gain was calculated based on the difference between 3 and 7 dpi and survival was determined up to 12 dpi. All combinations of Infergen (5, 0.5, 0.05, and 0 μg / kg) and ribavirin (75, 7.5, and 0 mg / kg) are injected subcutaneously simultaneously using 10 7-8 week old hamsters per group did. The placebo-infected control group in Study 1 used 17 animals. Non-infected controls using 3 hamsters per group included all combinations of 5, 0.5, and 0 μg / kg Infergen and ribavirin (75 and 0 mg / kg). A replication test protocol (Study 2) was performed using 5 μg / kg Infergen and 75 mg / kg ribavirin.

Results interferons (Table ^** (Table 3), using both visual CPE assay and NR assay, were all active against WNV in culture when administered one day before virus challenge 3).

^aWNV-西ナイルウイルス、DV-デングウイルス、YFV-黄熱病ウイルスワクチン。
^b50%有効濃度。
^c50%細胞毒性濃度。
^ｄ選択性指数(IC₅₀÷EC₅₀)。 TABLE 3 Interferon activity against flavivirus, West Nile virus, Dengue virus, and yellow fever virus using Vero 76 cells.

^a WNV-West Nile virus, DV-dengue virus, YFV-yellow fever virus vaccine.
^b 50% effective concentration.
^c 50% cytotoxic concentration.
^d Selectivity index (IC ₅₀ ÷ EC ₅₀ ).

Infergen was most active with an average EC ₅₀ = 0.026 ng / mL. IFN-α B / D and IFN-α 2b were 3-5 times less active than Infergen. IFN-γ was the least active among interferons with EC ₅₀ = 2.8 ng / μL. The nucleoside analog, 6-azauridine (Table 3) had almost the same strength activity as IFN-γ. Ribavirin was the least active. The same pattern of activity is similar for other flaviviruses, dengue viruses, and yellow fever viruses in Vero cells (Table 3), that is, Infergen is more active than IFN-α B / D against these viruses Met. Since these interferons were found to be active in cell culture, animal studies were performed using Infergen and IFN-α B / D. Dose and treatment schedules in animals are for Infergen in hamsters (Fish et al. (1986) Antimicrob Agents Chemother. 30 (1): 52-6); for IFN-α B / D in mice (Gangemi et al. (1989) J Interferon Res. 9 (3): 275-83; and Gangemi et al. (1989) J Interferon Res. 9 (2): 227-37); Ampligen in mice (Leyssen et al. (2003) Antimicrob Agents Chemother. 47 (2): 777-82; and Sidwell et al. (1994) Antiviral Res. 25 (2): 105-22); and imiquimod package inserts based on previously published studies .

IFN-α B / D (6.5 × 10 ⁷ U / kg) and Ampligen (13 mg / kg) administered ip starting 1 day before the subcutaneous virus challenge, prevented 100% of mouse deaths (P ≦ 0.01) ), The mortality of mice treated with saline-infected controls was 20% and 10%, respectively. The biological response modifier imiquimod, which has been shown to induce IFN-α, IFN-β, and other cytokines when administered topically, was effective in preventing death in 70% of infected mice (Table 4).

^aウイルス注射後11日まで。
^b治療は-1dpi(ウイルス注射後日数)、qd(毎日1回)、eod(隔日)で開始した。
^c生理食塩水ip、-1dpi、eodで治療した感染生理食塩水対照動物と統計学的に比較。
^**それぞれの感染対照と比較してP≦0.01。 Table 4 Effect of interferon-α B / D and interferon inducer administered 1 day prior to subcutaneous injection of West Nile virus on survival of 4-5 week old female BALB / c mice.

up to 11 days after ^a virus injection.
^b Treatment started at -1 dpi (days after virus injection), qd (once daily), and eod (every other day).
^c saline ip, -1dpi, statistically compared to infected saline control animals treated with eod.
^** P ≦ 0.01 compared to each infection control.

  Plasma virus titers from mice treated with either IFN-α B / D or Ampligen and collected 3 days after virus injection correlate with drug efficacy by reducing the virus titer to below the detection limit Whereas (P ≦ 0.001), the saline placebo sample had a 5 log of WNV titer.

In another experiment also using BALB / c mice, IFN-α B / D or Ampligen administered -4 to -6 hours post virus infection (hpi) ip did not improve survival statistically ( Table 5), mice treated with both Ampligen doses (15 and 4.8 mg / kg) and the lower of the two doses of IFN-α B / D (1.5 × 10 ⁶ U / kg) had a slightly increased survival Had a rate. Treatment delay from -1dpi to -4 to -6hpi greatly reduced the efficacy of IFN-α B / D and Ampligen.

^aウイルス注射後13日まで。
^b治療はhpi(ウイルス注射後時間)、qd(毎日1回)、eod(隔日)で開始した。
感染対照動物と感染治療動物の統計学的な違いはなかった。 Table 5: Effects of interferon-α B / D and Ampligen administered 4 to 6 hours prior to subcutaneous injection of West Nile virus on survival of 4-5 week old female BALB / c mice.

until 13 days after ^a virus injection.
^b Treatment started with hpi (time after virus injection), qd (once daily), and eod (every other day).
There was no statistical difference between infected control animals and infected treated animals.

  Infergen, Ampligen, and imiquimod administered at -4 to -6 hpi appeared to improve survival in 4-5 week-old hamsters compared to saline placebo controls, but 66% of imiquimod Only the survival rate was statistically different from the 11% survival rate of the saline control (P ≦ 0.05) (Table 6).

^a治療はhpi(ウイルス注射後時間)、qd(毎日1回)、sc(皮下)、ip(腹腔内)、eod(隔日)で開始した。
^bウイルス注射後13日まで。
^c統計学的分析は、-2と7dpiの個々のハムスターの重量を比較することによって行った。各個体の同一性は追跡せず、従って、各個体の平均重量変化は決定できなかった。
^*カイ二乗分析を使用して、生理食塩水感染対照と比較してP≦0.05。 Table 6: Effects of Infergen, Ampligen and Imiquimod administered 4-6 hours prior to subcutaneous injection of West Nile virus on survival and body weight changes in 4-5 week old female hamsters.

^a treatment hpi (after virus injection time), qd (1 once daily), sc (subcutaneous), ip (intraperitoneally), were initiated in eod (every other day).
^b Up to 13 days after virus injection.
^c Statistical analysis was performed by comparing the weight of each hamster -2 and 7 dpi. The identity of each individual was not followed and therefore the average weight change for each individual could not be determined.
^* P ≦ 0.05 compared to saline infected control using chi-square analysis.

  Hamsters had a significant weight loss when infected with the virus, which was a useful parameter for monitoring efficacy. All treatments improved body weight changes compared to the saline control group (Table 6). The change in body weight of animals treated with Ampligen and imiquimod was statistically different when compared to controls. Hamster weight change was another parameter that demonstrated the anti-WNV effects of these therapeutic agents in addition to survival.

  To determine whether IFN-α administered after virus challenge changed morbidity or mortality, 7-8 week old hamsters were administered Infergen 2 days after WMN injection. These older hamsters were used because older hamsters have more pronounced disease signs than younger mice or hamsters. Xiao et al. (2001) Emerg Infect Dis. 7 (4): 714-21. Ribavirin combinations were evaluated in this study. Infergen and ribavirin combination therapy started after two viral challenge and co-infected subcutaneously did not act synergistically to improve survival, ability to run up the ramp, or weight change (Table 7).

^aInfergenおよびリバビリンは、ウイルス注射+2日後に開始する単回sc注射、qd×7日のために一緒に混合した。
^b12dpiまで。
^cウイルス注射の7日後と3日後の間の重量(g)の変化。
^d15秒間で42度傾斜を上がって移動したインチ。
不活性プラセボ対照値と比較して^*p≦0.05、^**p≦0.01、^***p≦0.001。 Table 7: Combined study of Infergen and ribavirin administered subcutaneously +2 days after subcutaneous injection of West Nile virus for survival and disease symptoms of> 7 week old female Syrian golden hamsters ^a

^a Infergen and ribavirin were mixed together for virus injection + single sc injection starting 2 days, qd x 7 days.
^b Up to 12dpi.
^c Change in weight (g) between 7 and 3 days after virus injection.
^d An inch that moved 42 degrees uphill in 15 seconds.
^* P ≦ 0.05, ^** p ≦ 0.01, ^*** p ≦ 0.001 compared to inactive placebo control values.

  However, Infergen treatment, which increased the dose to 5 μg / kg without ribavirin, had a dose-responsive tendency to improve all measured parameters. Hamsters treated with the highest dose (5 μg / kg) Infergen had 90% survival, whereas hamsters treated with placebo showed only 59% survival, the difference being statistical Was not significant. A second study was performed to confirm the apparent activity of Infergen when treatment was initiated at 2 dpi and to confirm the apparent adverse effects of ribavirin in infected animals. In Study 2, approximately the same survival rate was observed compared to Study 1, which was 90% in Infergen treated animals and 50% in placebo animals, confirming the efficacy of Infergen treatment.

  In addition to the low dosage of ribavirin (7.5 mg / kg), animals treated with 5 μg / kg Infergen showed less weight loss than control animals (P ≦ 0.001) (Table 7). Animals treated with 5 or 0.5 μg / kg Infergen and without ribavirin ran up the 42 ° tilt table significantly better than control animals (P ≦ 0.01). Overall, Infergen treatment improved morbidity and mortality. However, all groups treated with the highest dosage (75 mg / kg) of ribavirin had significantly higher mortality than all other groups. Survival was much lower when WNV-infected animals were treated with the highest dosage of ribavirin. This result was confirmed in trial 2 (Table 7). This increased mortality was not due to ribavirin toxicity in uninfected animals. This is because uninfected treated control animals had the same survival, weight gain, and ability to run up as uninfected untreated controls.

In Study 2, WNV titers in plasma, brain, kidney, and spleen were assayed to determine if these parameters correlated with Infergen potency. Plasma WNV titers (<3.9 log ₁₀ cell culture infectious units / mL) obtained at 3 dpi from Infergen-treated hamsters were lower (<6.2 log ₁₀ cell cultures) than animals treated with saline. Infectious units / mL) (Table 8).

^aInfergenおよびリバビリンは、ウイルス注射+2日後に開始する単回注射、qd×7日のために一緒に混合した。
^b血漿は3dpiおよび最初の治療の1日後に収集した。
^c組織は7dpiに収集した。
^d斜字体は検出限界未満の力価を示す。細胞培養力価は、血漿または細胞単層に加えられたホモジナイズされた組織の毒性から制限された。 Table 8. Effects of Infergen and ribavirin administered subcutaneously +2 days after subcutaneous injection of West Nile virus on viral titers of> 7-week-old female Syrian golden hamsters

^a Infergen and ribavirin were mixed together for a single injection, qd × 7 days, starting 2 days after virus injection.
^b Plasma was collected at 3 dpi and 1 day after the first treatment.
^c Tissue was collected at 7 dpi.
^d Italics indicate a titer below the detection limit. Cell culture titers were limited by the toxicity of homogenized tissue added to plasma or cell monolayers.

  A slight trend of lower titers and a lower number of WNV positive tissues obtained at 7 dpi were observed in Infergen treated animals, but the differences were not statistically different. Plasma and tissue WNV titers appeared to correlate with Infergen treatment.

Example 3: Active materials and methods of the combination of IFN alfacon 1 (IFN-alfacon 1) and IFN γ 1b (IFN-γ1b) Interferon:
Interferon alfacon-1 or IFN alfacon 1, IFN alfacon 1 (Intermune, Brisbane, CA) as a sterile, clean, colorless, preservative-free liquid at 30 μg / mL (0.5 mL total fill) Obtained at drug concentration. Interferon γ-1b or IFN-γ1b (Intermune, Brisbane, CA) was obtained as a sterile, clean, colorless solution at a drug concentration of 200 μg / mL (0.5 mL total loading).

Cytotoxicity and stability studies:
To establish a dose dilution series, we utilized the Cmax concentrations of IFN-γ1b and IFN-alfacon 1 based on doses of 100 μg and 15 μg, respectively. These Cmax concentrations were determined in a human phase 1 clinical study. Using these data in experiments designed to test combinations of two IFNs for cytotoxicity studies at doses of 100-fold Cmax concentration is useful for both IFN alfacon 1 and IFN-γ1b Used. The Celltiter 96 Aqueous 1 solution cell proliferation assay was used to determine the number of viable cells in the cytotoxicity assay (Promega, Madison, Wisconsin). MTS was added to A549 cells in 96 plates after various concentrations of IFN alfacon 1 and IFN-γ1b alone and in accordance with the manufacturer's protocol. Cells were treated for 24 hours.

Cell culture methodology:
Approximately 4 × 10 ⁴ A549 cells are seeded in 96-well plates and 5% in culture medium supplemented with 10% fetal bovine serum (Gibco BRL), penicillin G (100 U / ml), and streptomycin (100 μg / ml). Maintained at 37 ° C. with CO ₂ .

Virus stock preparation:
The vesicular stomatitis Indiana virus used in this study was a gift from Dr. Milton Taylor, a professor of microbiology at Indiana University. The virus stock was obtained by inoculating a monolayer of Vero cells in a 75 cm ² tissue culture flask with virus diluted 1: 500 in 1 ml DMEM / 10% FBS. The cells and supernatant were then harvested by gentle pipetting after producing a cytopathic effect (CPE). Cell debris was removed by centrifugation and the virus supernatant was stored as aliquots and stored at -80 ° C.

Virus titration:
Virus production was titrated by plaque assay using Vero cells as the host. Vero cells were seeded in 6-well (6 × 10 ⁵ cells / well) plates in DMEM with 10% FBS at 37 ° C. for 24 hours. The medium was removed and serial dilutions of virus supernatant were added to DMEM with 5% FBS and incubated for 1 hour at 37 ° C. When the adsorption step was complete, DMEM with 5% FBS and 1% low melting point agarose (3 ml / well for 6 well plates) were added and the plates were incubated at 37 ° C. for 30-48 hours. When the virus formed a visible plaque, removal of the agarose plug was completed and the plate was briefly stained with 1% crystal violet solution in 20% ethanol. Virus concentration was determined as PFU per milliliter.

Viral cell infection and inhibition assays:
One day after seeding, the cells were washed once with PBS, and for single dose studies, medium was diluted with various dose concentrations of IFN alfacon 1 (1: 2 from 40 pg / ml to 0.3125 pg / ml serially diluted) and Replaced with medium with IFN-γ1b (1: 2 serial dilution from 4800 pg / ml to 18.75 pg / ml). For combination studies, IFN alfacon 1 and IFN-γ1b were added at a ratio of 1: 120 and 1:60. After 24 hours incubation, the cells were washed 3 times with PBS and VSV was added to each plate at MOI = 0.1. Each treatment regimen was performed in quadruplicate.

(MGBNFQ:分子溝結合非蛍光クエンチャー) Oligonucleotide primer design and probe design for RT-PCR:
Primer Express oligo design software (Applied Biosystems, Foster City, California) was used to identify primer / probe pairs corresponding to the VSV sequences. Using these primers and probes, the VSV Indiana serotype (J02428) amplicon was positioned between nucleotides 1764 and 1810, and the following primer / probe set was used:

(MGBNFQ: Molecular groove binding non-fluorescent quencher)

RNA extraction and standard viral RNA:
VSV viral RNA was collected from different concentrations of infected cells and total RNA was extracted using the QIAamp Viral RNA Kit (Qiagen Corp, Valencia, California) according to the manufacturer's protocol. Viral titer was expressed as plaque forming units (PFU) per ml, equal to the number of copies per ml. Serial dilutions of VSV RNA samples from infected HeLa cells were used as standards. The viral titer of these samples was estimated at 6-6 × 10 ⁶ pfu / ml (10) in advance by plaque assay.

RT-PCR for real-time quantification:
A 5 μL aliquot of VSV viral RNA was used for quantification using real-time PCR. Single tube reactions were performed using the TaqMan Gold RT-PCR kit (Applied Biosystems, Foster City, California). Triplicate reactions on RNA standards and samples were performed and a 50 μl volume with 5 μl viral RNA was set up for each reaction. The RT step was carried out at 48 ° C. for 30 minutes followed by 95 ° C. for 10 minutes, followed by 40 cycles of 95 ° C. for 15 seconds and 60 ° C. for 1 minute.

Synergy analysis:
Data were analyzed assuming that the drugs are not mutually exclusive, ie, the two drugs have different or independent mechanisms of action. The percent inhibition of virus replication for each combination and monotherapy combination was calculated by measuring virus copy number compared to their respective controls. The experiment was repeated at least 3 times to confirm the results. Analysis of synergistic, additive, or antagonistic effects of antiviral drugs was performed using CalcuSyn (Biosoft, Cambridge, United Kingdom) software and the Chau-Talalay (11) methodology (CI value <0.1 Represents a very strong synergism, CI values of 0.1-0.3 represent strong synergism and CI values of 0.3-0.7 represent synergistic activity) to determine the combination index (CI). Drug combinations that return CI = 1 have an additive effect, whereas CI values> 1 represent an antagonistic effect. The size of the CI value is directly proportional to the amount of interaction in the combination. To provide a more complete analysis of the overall effect surface, the data was also analyzed using a conservative isobologram methodology to quantify the degree of synergy or antagonism. Specifically, a specific effect level is selected, which includes the maximum 50%, 75%, and 90%, and the dose of IFN alfacon 1 and IFN-γ1b (each alone) that gives this effect is the Cartesian plot Plotted as axis points. The upwardly convex curve connecting IFN alfacon 1 and IFN-γ1b (conservative) is simply the point where this effect occurs in an additive combination. If the combination data points are on the curve, an additive effect is shown; if the point is on the lower left side, synergy is shown; and if the point is on the upper right side, antagonism is shown .

result
VSV research
The dose response for monotherapy with IFN-alfacon 1 and IFN-γ1b in the VSV / A549 system is shown in FIG.

As seen in Figure 2, both IFN-alfacon 1 and IFN-γ produced a dose response against VSV-infected A549 cells, with a 50% effective concentration (EC-50) for IFN alfacon 1 of approximately 5 pg / mL. Yes, and the EC-50 for IFN-γ1b was about 300 pg / mL. The same concentration used in the antiviral assay to demonstrate that the apparent antiviral effects of IFN-alfacon 1 and IFN-γ1b are not due to the direct antiproliferative effects of these cytokines A cell proliferation assay was performed. Approximately 90% maximum inhibition (1 log ₁₀ reduction) was achieved at the maximum dose for each IFN monotherapy, and> 90% reduction was achieved at any dose of monotherapy at 24 hours It is important to note that it was not possible.

  FIG. 3 demonstrates that there is no antiproliferative effect at each IFN dose used in the antiviral assay, indicating that the antiviral response to VSV is a direct antiviral by each IFN. Indicates that this is due to the introduction of a state.

  To determine whether the combination of IFN-alfacon 1 and IFN-γ1b produces a synergistic, additive or antagonistic antiviral effect, these two IFNs were used at 1: 120 and Combined at a 1:60 ratio and evaluated in the VSV / A549 antiviral assay system (FIGS. 4a and 4b).

As seen in FIGS. 4a and 4b, combination therapy of IFN-alfacon 1 and IFN-γ against VSV in A549 cells produced a dramatic dose response effect in vitro. More specifically, a reduction of> 1 log ₁₀ could be achieved, unlike monotherapy treatment. For example, a reduction of 2, 3, and 5 log ₁₀ could be obtained at a ratio of 1: 120 (see FIG. 4a).

  Analysis of synergy for the combination of IFN-alfacon 1 and IFN-γ1b in the VSV / A549 system using Chau-Talalay methodology demonstrated very strong synergy at a wide range of doses (CI < 0.1). In this methodology, Chau-Talalay analysis calculates a combination index value (CI). CI values <1 indicate synergistic combinations, while CI values> 1 indicate antagonistic interactions. Furthermore, CI values <0.01 indicate a very strong synergy, whereas CI values <0.1 indicate a strong synergy. The CI values calculated in the IFN-alfacon 1 and IFN-γ1b dose ranges used in the VSV / A549 assay (FIGS. 4a and 4b) are shown in Table 9 below.

(Table 9)

  As seen in Table 9, IFN-alfacon 1 and IFN- in the ratio of 1: 120 (IFN-alfacon 1: IFN-γ1b) and 1:60 (IFN-alfacon 1: IFN-γ1b) in the VSV / A549 system The γ1b combination yielded CI values that were all <1. Over these dose ranges, all values were returned as either very strong synergism or strong synergy (Table 9).

  In addition to Chau-Talalay analysis, isobologram plots are also used in synergy evaluation to determine whether a combination of therapeutic agents is additive, synergistic, or antagonistic. FIGS. 5a and 5b are isobologram plots of IFN-γ at concentrations of EC-50, EC-75, and EC 90 for the combination of IFN-alfacon 1 and IFN-γ1b in the VSV / A549 model of antiviral activity. It is. EC values below the dose concentration line (on the left) for each individual IFN are considered synergistic concentrations, those on the line are additive and those above the line are antagonistic It is. As seen in FIGS. 5a and 5b, all EC values at a ratio of 1: 120 (IFN-alfacon 1: IFN-γ1b) or 1:60 (IFN-alfacon 1: IFN-γ1b) are monotherapy dose response Below the curve, this shows a strong synergy of these two drugs by isobologram analysis.

  Although the invention has been described with reference to specific embodiments thereof, those skilled in the art will recognize that various changes can be made and equivalents can be substituted without departing from the true spirit and scope of the invention. Should be understood by. In addition, many modifications may be made to adapt a particular situation, material, composition, process, process step, objective, spirit, and scope of the invention. All such modifications are intended to be within the scope of the specification and the appended claims.

FIG. 2 is a graph showing the effect of interferon-α B / D and Ampligen ™ administered 1 day prior to subcutaneous injection of West Nile virus on plasma WNV titers in 4-5 week old female BALB / c mice. Plasma was collected at 3pi. It is a graph which shows the effect of the dosage of IFN-alfacon 1 and IFN-γ1b on VSV in A549 cells. It is a graph which shows the result of the cytotoxicity assay about the interferon in A549 cell. Figures 4a and 4b show that the IFN-alfacon 1 / IFN-γ1b combination (Figure 4a: 1: 120 ratio; Figure 4b: 1:60 ratio) directs antiviral effects on VSV in A549 cells. It is a graph to show. Figures 5a and 5b show the EC-50 for the combination of IFN-alfacon 1 and IFN-γ1b (Figure 5a: 1: 120 ratio; Figure 5b: 1:60 ratio) in the VSV / A549 model of antiviral activity. It is a graph which shows the isobologram analysis about the density | concentration of EC-75 and EC90.

Claims (43)

  A method of reducing liver fibrosis in an individual comprising administering IFN-α and IFN-γ in an effective amount to reduce liver fibrosis.   A method of treating hepatitis C virus infection in an individual comprising administering an effective amount of IFN-α and IFN-γ to achieve a sustained viral response.   2. The method of claim 1, wherein the degree of liver fibrosis is determined by staging, wherein the stage of liver fibrosis is reduced by at least one unit as measured by a standardized scoring system.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are co-administered. The method according to claim 1 or claim 2.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are within about 24 hours of each other 3. The method of claim 1 or claim 2, wherein the method is administered.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are administered in multiple doses 3. The method according to claim 1 or claim 2, wherein:   A method of increasing liver function in an individual suffering from liver fibrosis comprising administering IFN-α and IFM-γ in an amount effective to increase liver function.   8. The method of claim 7, wherein liver function is indicated by measuring a parameter selected from the group consisting of serum transaminase level, prothrombin time, serum bilirubin level, platelet count, viral load, and serum albumin level.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are administered in consecutive doses 8. The method of claim 7, wherein   Reduces the incidence of cirrhosis complications, including administering IFN-α and IFN-γ combinations to individuals suffering from liver fibrosis in an amount effective to reduce the incidence of cirrhosis complications How to make.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are administered in multiple doses 11. The method of claim 10, wherein:   A method of treating hepatitis C virus (HCV) in an individual comprising administering a therapeutically effective amount of IFN-α and IFN-γ.   13. The method of claim 12, wherein the level of alanine aminotransferase is reduced to less than about 45 international units per milliliter of serum.   13. The method of claim 12, wherein the individual's viral load is reduced to less than about 500 HCV genome copies per milliliter of serum.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are administered simultaneously. The method according to claim 12.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are administered separately. 13. The method according to claim 12.   IFN-γ is administered subcutaneously in an amount of about 25 μg to about 300 μg per dose, IFN-α is administered subcutaneously in an amount of about 3 μg to about 27 μg, and IFN-α and IFN-γ are administered in consecutive doses 13. The method of claim 12, wherein:   A method of reducing viral load in an individual suffering from hepatitis C virus, comprising administering an amount of IFN-α and IFN-γ effective to reduce viral load.   (a) identifying an individual suffering from genotype 1 hepatitis C virus (HCV) infection and having an initial viral titer of HCV genomic copies greater than 2 million copies per milliliter of serum; and (b) about A method of treating hepatitis C virus in an individual comprising administering to the individual an effective amount of IFN-α and IFN-γ for a period of 48 weeks.   Identification of the individual in stage (a) further requires that the individual does not have liver fibrosis as measured by a Knodell score of 0, 1, or 2 or has early stage liver fibrosis 20. The method of claim 19.   (a) identifying individuals suffering from genotype 1 hepatitis C virus (HCV) infection and having an initial viral load of no more than 2 million copies of HCV genomic copies per milliliter of serum; and (b) about 24 A method of treating hepatitis C virus in an individual comprising administering to the individual an effective amount of IFN-α and IFN-γ during a period of 48 weeks.   (a) identifying an individual suffering from genotype 2 or 3 hepatitis C virus (HCV) infection, and (b) an effective amount of IFN-α during a period of about 24 to 48 weeks and A method of treating hepatitis C virus in an individual comprising administering IFN-γ to the individual.   The method according to any one of claims 12 to 22, wherein IFN-α is a consensus interferon.   24. The method of any one of claims 12-23, further comprising co-administering to the individual an effective amount of ribavirin during the period of IFN-α and IFN-γ treatment.   25. The method of any one of claims 12-24, wherein the individual receives a synergistic amount of IFN-α and IFN-γ.   The method according to any one of claims 12 to 25, wherein IFN-α is PEGylated IFN-α.   27. A method according to any one of claims 12 to 26, wherein IFN-α is administered subcutaneously by pump-controlled continuous infusion.   A method of treating an alphavirus infection in an individual comprising administering to the individual an effective amount of IFN-α and IFN-γ.   29. The method of claim 28, wherein the individual receives a synergistic amount of IFN-α and IFN-γ.   30. The method of claim 28 or 29, further comprising co-administering to the individual an effective amount of ribavirin during the period of IFN-α and IFN-γ treatment.   31. The method according to any one of claims 28 to 30, wherein IFN-α is consensus interferon.   The method according to any one of claims 28 to 31, wherein IFN-α is PEGylated IFN-α.   A method of treating West Nile virus infection in an individual comprising administering to the individual an effective amount of consensus interferon (CIFN).   A method of treating West Nile virus infection in an individual comprising administering to the individual an effective amount of IFN-γ.   A method of treating West Nile virus infection in an individual comprising administering to the individual an effective amount of IFN-α and IFN-γ.   36. The method of claim 35, wherein the individual receives a synergistic amount of IFN-α and IFN-γ.   37. The method of claim 35 or 36, wherein IFN-α is a consensus interferon.   The method according to any one of claims 35 to 37, wherein IFN-α is PEGylated IFN-α.   39. The method of any one of claims 33-38, further comprising co-administering an effective amount of ribavirin to the individual during the period of treatment of IFN-α and IFN-γ.   12. The method of any one of claims 1-11, wherein the individual receives a synergistic amount of IFN-α and IFN-γ.   41. The method of claim 40, wherein IFN-α is consensus interferon.   42. The method according to claim 40 or 41, wherein IFN-α is PEGylated IFN-α.   43. Any of claims 1 and 40-42, further comprising co-administering to the individual an effective amount of pirfenidone or a specific pirfenidone analog during the period of treatment of IFN-α and IFN-γ The method according to one item.

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