WO2024169694A1

WO2024169694A1 - Drug against skin fibrosis

Info

Publication number: WO2024169694A1
Application number: PCT/CN2024/075808
Authority: WO
Inventors: 张文杰; 杨廷佳
Original assignee: 上海萨丽斐生物科技有限公司
Priority date: 2023-02-13
Filing date: 2024-02-04
Publication date: 2024-08-22
Also published as: CN118001374A

Abstract

A drug against skin fibrosis. A use of an annexin in preparation of a composition or preparation against skin fibrosis. A use of an annexin, wherein the annexin can be used for ameliorating skin fibrosis, reducing the number of neutrophils, avoiding tissue structure damage caused by continuous accumulation of ECM components caused by tissue repair reaction disorder, and preparing a pharmaceutical composition and preparation against skin fibrosis.

Description

A drug for treating skin fibrosis

Technical Field

The invention belongs to the field of biomedicine, and in particular, relates to a drug for resisting skin fibrosis.

Background Art

Fibrosis is not a disease, but the result of a disordered tissue repair response in various types of tissue damage, especially in chronic inflammatory diseases. It can affect any organ. When tissue is damaged, local fibroblast activation, secretion of inflammatory mediators, and synthesis of extracellular matrix (ECM) (such as collagen and fibronectin) occur. These changes together initiate the healing response. However, when the injury is severe or recurrent, ECM components will continue to accumulate, leading to tissue structural destruction, organ dysfunction and even failure.

Fibrotic skin diseases involve the accumulation of ECM components in the dermis, including scleroderma, hypertrophic scars, keloids, etc.; in mild cases, sweat gland secretion is reduced, thereby affecting body temperature regulation; in severe cases, joint contracture leads to dysfunction. To date, the pathogenesis of fibrotic skin diseases has not been thoroughly elucidated, and there is a lack of a complete cure. Clinically, drugs used to treat skin fibrosis have not achieved good therapeutic effects. In summary, there is an unmet medical need for specific treatments for fibrotic diseases with extremely high social impacts, and this field urgently needs to develop new drugs for anti-skin fibrosis.

Summary of the invention

The purpose of the present invention is to provide a medicine for resisting skin fibrosis.

The first aspect of the present invention provides a use of annexin in preparing a composition or preparation for resisting skin fibrosis.

In another preferred embodiment, the annexin is annexin A5.

In another preferred embodiment, the skin fibrosis is selected from the group consisting of keloid, hypertrophic scar or scleroderma.

In another preferred embodiment, the skin fibrosis occurs during the process of scleroderma and skin scar formation, and is mainly characterized by thickening of the dermis, obstruction of skin appendages, hair follicles, sweat glands, infiltration of inflammatory cells around small blood vessels in the dermis, significant thickening of blood vessel walls, and reduction or occlusion of organ cavities.

In another preferred embodiment, the fibrosis is the necrosis of organ parenchymal cells, A pathological process in which there is an abnormal increase and excessive deposition of matrix.

In another preferred embodiment, the skin fibrosis is caused by disordered skin repair response.

In another preferred embodiment, the skin fibrosis is caused by the continuous accumulation of extracellular matrix (ECM) components due to disordered skin repair response.

In another preferred embodiment, the annexin also reduces the number of neutrophils.

In another preferred embodiment, the annexin improves bleomycin-induced skin fibrosis.

The second aspect of the present invention provides a composition or preparation, wherein the composition or preparation comprises:

(a) an annexin as the first active ingredient;

(b) an additional drug that inhibits skin fibrosis as a second active ingredient; and

(c) a pharmaceutically acceptable carrier.

In another preferred embodiment, the composition or preparation is used for preventing and/or treating skin fibrosis.

In another preferred embodiment, the composition or preparation further comprises a pharmaceutically acceptable salt.

In another preferred embodiment, the dosage form of the composition or preparation is selected from the following group: liquid dosage form, solid dosage form.

In another preferred embodiment, the dosage form of the composition or preparation is selected from the following group: tablets, capsules, powders, granules, lozenges, patches, suppositories, pills, ointments, injections; preferably, the dosage form of the composition or preparation is selected from the following group: intravenous injections, intramuscular injections or subcutaneous preparations.

In another preferred embodiment, the additional drug for inhibiting skin fibrosis includes: glucocorticoids.

The third aspect of the present invention provides a kit, the kit comprising:

(i) the composition or formulation according to the second aspect of the present invention; and

(ii) Detection reagents for detecting extracellular matrix (ECM) components in the skin.

In another preferred embodiment, the kit further comprises:

(iii) instructions, which state that when the detection reagent detects that the extracellular matrix (ECM) components in the skin continue to accumulate, resulting in a content significantly higher than that of the normal control, the skin is defined as a skin type in which the tissue structure is destroyed due to the continuous accumulation of ECM components caused by the disordered tissue repair response, and the application of membrane annexin for treatment is suggested.

In another preferred embodiment, the detection reagent includes: ELISA enzyme label plate (detachable), freeze-dried standard, standard/sample diluent, concentrated biotinylated antibody, biotinylated antibody diluent, concentrated HRP enzyme conjugate, enzyme conjugate diluent, concentrated washing solution, substrate solution (TMB), reaction stop solution or sealing film.

A fourth aspect of the present invention provides a method for preventing or treating skin fibrosis, comprising the steps of: An effective amount of annexin is administered to the subject.

In another preferred embodiment, the method comprises the step of: applying the composition or preparation described in the second aspect of the present invention to the skin fibrosis area of the subject.

In another preferred embodiment, the subject or object is a human or non-human mammal.

In another preferred embodiment, the non-human mammals include primates and rodents (such as mice and rats).

It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here.

DETAILED DESCRIPTION

After extensive and in-depth research and a large number of screenings, the inventors have developed a novel drug for anti-skin fibrosis for the first time. The inventors unexpectedly found that annexin can improve skin fibrosis phenomena such as scleroderma, and can avoid the continuous accumulation of ECM components caused by tissue repair response disorders and thus cause tissue structure damage, so it is very suitable for preparing drugs for anti-skin fibrosis. On this basis, the present invention was completed.

the term

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, when used in reference to a specific recited numerical value, the term "about" means that the value may vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

As used herein, the term "comprising" or "including (comprising)" may be open, semi-closed and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

As used herein, the term "room temperature" or "normal temperature" refers to a temperature of 4-40°C, preferably, 25±5°C.

Active ingredients

Annexins are a class of widely distributed calcium-dependent phospholipid-binding proteins. Based on molecular phylogenetics and comparative genomics analysis, this protein family was uniformly named annexin (abbreviated as Anx). The capital letters represent different species: A, B, C, D, and E represent vertebrates, invertebrates, fungi and myxomycetes, plants, and protists, respectively. So far, there are 12 members in the Anx A family: A1-A11 and A13 (A12 is unspecified).

Typically, the annexin in the present invention is Anx A. More preferably, the annexin includes annexin A5. Annexin A5 (AnxA5) is widely distributed in various tissue cells of the body. In vitro studies have found that AnxA5 has multiple functions such as anti-phospholipase, anti-coagulation, and anti-kinase, but its specific in vivo function is not very clear. It is currently mainly used as a reagent for detecting cell apoptosis.

The annexin that can be used in the present invention is not particularly limited, and can be an annexin derived from any organism, preferably from a mammal (such as a primate), more preferably from a human, a monkey or a mouse. In addition, it should be understood that "annexin" includes wild-type or mutant (including truncated) annexin, as long as the mutant annexin retains or maintains the detoxification activity of the wild-type annexin.

Pharmaceutical compositions and uses

The pharmaceutical composition of the present invention comprises the above active ingredients and a pharmaceutically acceptable carrier.

Experiments have shown that the active ingredients of the present invention can effectively improve skin fibrosis, reduce the number of neutrophils, and avoid tissue structure damage caused by continuous accumulation of ECM components caused by tissue repair response disorders, and are therefore very suitable for preparing drugs against skin fibrosis.

In the present invention, the "fibrosis" is a pathological change, which is manifested as the activation and proliferation of fibroblasts, the increase of fibrous connective tissue in tissues and organs, and the decrease of parenchymal cells. Continuous progression can lead to the destruction of tissue and organ structure and loss of function. Skin fibrosis forms scar tissue. Scar tissue is a fibrous connective tissue in the aging stage formed by the remodeling and maturation of granulation tissue. Under trauma and other conditions, fibroblasts divide, proliferate, migrate to the damaged part, produce extracellular matrix, form scar tissue, and repair the trauma. The formation of scars is the process of gradual fibrosis of granulation tissue. At this time, there are more and more reticular fibers and collagen fibers, the reticular fibers are collagenized, and the collagen fibers become thicker. At the same time, there are fewer and fewer fibroblasts, and the few remaining ones are transformed into fibroblasts; the fluid in the interstitium is gradually absorbed, and neutrophils, macrophages, lymphocytes and plasma cells disappear one after another; capillaries close, degenerate, and disappear, leaving very few arterioles and venules. In this way, the granulation tissue is transformed into scar tissue with few blood vessels, which is mainly composed of collagen fibers, white to the naked eye, and tough in texture. Systemic sclerosis (SSc), also known as scleroderma, is a systemic autoimmune disease characterized by localized or diffuse skin thickening and fibrosis. The lesions are characterized by skin fibrosis and onion-like changes in blood vessels, which eventually lead to skin hardening and vascular ischemia. The disease is clinically characterized by localized or diffuse skin thickening and fibrosis. In addition to skin involvement, it can also affect internal organs (heart, lungs, digestive tract, etc.). official).

Preferably, the skin fibrosis is selected from the group consisting of keloid, hypertrophic scar or scleroderma.

"Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" refer to substances that facilitate formulation and/or administration of an active agent and/or absorption by a subject, and may be included in the compositions of the present disclosure without causing significant adverse toxicological effects to the subject.

In certain embodiments, the pharmaceutical compositions of the present invention may be in solid or liquid form.

The active ingredient can be administered to a subject by any suitable route, including orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted cartridge. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. Preferably, the composition is administered orally, subcutaneously or intravenously.

Pharmaceutical compositions of the invention suitable for oral administration will typically be discrete units in solid form, such as tablets, capsules, cachets, granules, lozenges, pills, or in liquid form, such as liquids, injectable or infusible solutions or suspensions.

Preferably, the pharmaceutical composition of the present invention is suitable for targeted local administration to the skin fibrosis site. The pharmaceutical composition suitable for local administration is typically in the form of powder, patch, injection, ointment, etc.

The exact amount of the compound that provides a therapeutically effective amount to an individual will depend on the mode of administration, the type and severity of the disease and/or condition, and the characteristics of the individual, such as general health, age, sex, weight, and tolerance to drugs. One of ordinary skill in the art will be able to determine an appropriate dose based on these and other factors. When administered in combination with other therapeutic agents, the "therapeutically effective amount" of any other therapeutic agent will depend on the type of drug used. Suitable doses are known for approved therapeutic agents and can be adjusted by one of ordinary skill in the art based on the individual's condition, the type of condition being treated, and the amount of the compound of the invention used by, for example, those reported in the literature and recommended in Physician's Desk Reference (57th edition, 2003). Preferably, the compositions should be formulated so that an inhibitor dose of 0.01-100 mg/kg body weight/day can be administered to patients receiving these compositions. In certain embodiments, the compositions of the present invention provide a dose of 0.01 mg to 50 mg. In other embodiments, a dose of 0.1 mg-25 mg or 5 mg-40 mg is provided.

Examples of subjects to whom the pharmaceutical composition or therapeutic agent of the present invention can be administered include mammals (eg, humans, mice, rats, hamsters, rabbits, cats, dogs, cows, sheep, monkeys, etc.).

The present invention also provides a method of treatment, which comprises the steps of administering the active ingredient of the present invention, or administering the pharmaceutical composition or preparation of the present invention to a subject in need of treatment, for preventing and/or Treat skin fibrosis.

The main advantages of the present invention include:

The present invention provides a use of annexin, which can be used to improve skin fibrosis, reduce the number of neutrophils, and avoid tissue structure damage caused by continuous accumulation of ECM components caused by tissue repair response disorders. It is very suitable for preparing pharmaceutical compositions and preparations for resisting skin fibrosis.

The present invention finds that annexin can effectively improve bleomycin-induced skin fibrosis.

The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

Unless otherwise specified, the experimental materials and reagents used in the following examples can be obtained from commercial channels.

Example 1. Anti-skin fibrosis animal experiment

This experiment will conduct an animal experiment on the anti-skin fibrosis of annexin A5. The specific method is:

(1) Animal grouping

SPF C57bl6 mice aged 6-8 weeks were randomly divided into 4 groups according to body weight.

(2) Model preparation

The mice in the normal control group were not treated in any way; each mouse in the model control, positive control, and drug-treated groups was subcutaneously injected with 100 μL of bleomycin (0.5 mg/mL) on the back skin once every other day.

(3) Administration

Each group of mice was intraperitoneally injected with bleomycin every day starting from the second day after the model was established, once every other day.

(4) Specimen collection

At the 3rd and 6th week after modeling, 2-3 animals were randomly selected from each group to observe the changes in the back skin of the mice with the naked eye. After the observation, the mice were killed and the back skin tissues were obtained.

(5) Detection method

HE staining was used to observe the thickness of the dermis, Masson staining, myofibroblast count (α-SMA positive), CD31 staining, immunohistochemistry (expression levels of TGF-β and type I collagen), and inflammatory staining.

In summary, annexin A5 can play a good anti-inflammatory role, reduce the number of neutrophils, avoid the continuous accumulation of ECM components caused by the disorder of tissue repair response, and then cause tissue structure destruction, organ dysfunction and even failure, so it can be used as a good anti-skin fibrosis drug.

All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

A use of annexin in preparing a composition or preparation for resisting skin fibrosis.
The use according to claim 1, characterized in that the annexin is annexin A5.
The use according to claim 1, characterized in that the skin fibrosis is selected from the group consisting of keloid, hypertrophic scar or scleroderma.
The use as claimed in claim 1 is characterized in that the skin fibrosis occurs in the process of scleroderma and skin scar formation, and is mainly characterized by thickening of the dermis, obstruction of hair follicles and sweat glands of skin appendages, infiltration of inflammatory cells around small blood vessels in the dermis, significant thickening of blood vessel walls, and reduction or occlusion of organ cavities.
The use according to claim 1 is characterized in that the fibrosis is a pathological process in which organ parenchymal cells undergo necrosis and extracellular matrix in the tissue increases abnormally and deposits excessively.
The use according to claim 1, characterized in that the annexin also reduces the number of neutrophils.
The use according to claim 1, characterized in that the annexin improves bleomycin-induced skin fibrosis.
A composition or preparation, characterized in that the composition or preparation comprises:

(a) an annexin as the first active ingredient;

(b) an additional drug that inhibits skin fibrosis as a second active ingredient; and

(c) a pharmaceutically acceptable carrier.
The composition or preparation according to claim 8, characterized in that the additional drug for inhibiting skin fibrosis comprises: glucocorticoid.
A kit, characterized in that the kit contains:

(i) the composition or formulation of claim 8; and

(ii) Detection reagents for detecting extracellular matrix (ECM) components in the skin.
A method for preventing or treating skin fibrosis, characterized in that it comprises the step of administering an effective amount of annexin to a subject.