KR20120068035A - Compositions and methods of delivery of pharmacological agents - Google Patents

Abstract

The present invention provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises a protein, such as human serum albumin and / or deferoxamine. It is about. Human serum albumin is present in an amount effective to reduce one or more side effects associated with administration of the pharmaceutical composition. The invention also provides a method of reducing one or more side effects associated with the administration of a pharmaceutical composition, a method of inhibiting the growth and oxidation of microorganisms in a pharmaceutical composition, and a method of enhancing the transport and binding of pharmaceutical substances to cells. To provide.

Description

Compositions and methods of delivery of pharmacological agents

Cross reference of related patent application

This application is directed to U.S. Provisional Application No. 60 / 432,317, filed Dec. 9, 2002, U.S. Provisional Application (attorney docket no. 225519), filed Dec. 3, 2003, and U.S. Provisional Application No. : 225549), and US Provisional Application (attorney docket number: 225585), filed December 5, 2003.

Technical field

The present invention relates to a pharmaceutical composition comprising a pharmaceutically active substance for parenteral or oral use which has the effect of reducing undesirable side effects upon administration compared to available formulations of similar drugs.

Many drugs used parenterally, especially intravenously, can cause undesirable side effects such as vascular irritation, phlebitis, burning and pain in the injection site, venous thrombosis, extravasation, and other administration related side effects. Many of these drugs are insoluble in water and therefore formulated with solubilizers, surfactants, solvents, and / or emulsifiers that are irritating, allergenic, or toxic upon administration to a patient (eg, Briggs et al. , Anesthesis 37, 1099 (1982), and Waugh et al., Am. J. Hosp.Pharmacists, 48, 1520 (1991). Often, the free drug present in the formulation causes pain or irritation upon administration. For example, phlebitis has been observed in 50% of patients receiving peripheral iv, ifosfamide and vinorelbine, the primary chemotherapy agents for severe non-small cell lung cancer (eg, Vallejo et al. , Am. J. Clin .. Oncol., 19 (6), 584-8 (1996). Vancomycin has also been observed to cause side effects such as phlebitis (see, eg, Lopes Rocha et al., Braz. J. Infect. Diss., 6 (4), 196-200 (2002)). Patients with solid cancers developed severe venous thrombosis and phlebitis when using cisplatin, gemcitabine, and SU5416 (eg Kuenen et al., J. Clin. Oncol., 20 (6), 1657-67 (2002) ) Reference). In addition, propofol, an anesthetic, may cause pain upon injection, especially burning and venous irritation when administered as a lecithin-stabilized fat emulsion (eg Tan et al., Anathesia, 53, 468-76, (1998)). Other drugs that show side effects associated with administration include, for example, Taxol (paclitaxel) (see, for example, Taxol IV's instructions), Codarone (amiodarone hydrochloride) (see, eg, Codarone IV's instructions), thyroid hormone T3 Or liothyronine (trade name: Triostat), thioteta, bleomycin, and diagnostic radiographic contrast agents, but are not limited thereto.

Another problem associated with the preparation of injectables, especially injectables of water-insoluble drugs, is the securing of sterility. The sterile production of the emulsion / suspension of the drug can be accomplished by absolute aseptic techniques at all stages of manufacture, after all the components prior to manufacture have been completely sterilized. However, such a method is time consuming and costly. In addition, oxidation of the drug formulation by exposure to air during manufacture or storage of the drug results in, for example, pH reduction, drug degradation, and discolouration, thereby destabilizing the drug formulation and / or shortening the shelf life.

To avoid such problems with side effects associated with the administration of drug formulations, alternative formulations have been tried. For propofol, for example, a method of reducing propofol-induced pain may include the fat content of the solvent (eg, long chain triglycerides (LCT)), non-steroidal drugs, local anesthetics, pretreatment of opioid drugs, pretreatment, lidocaine Addition, addition of cyclodextrin, and microfiltration (eg, Mayer et al., Anaesthesist, 45 (11), 1082-4 (1996), Davies, et al. Anaesthesia, 57, 557 -61 (2002), Doenicke, et al., Anaesth.Analg., 82, 472-4 (1996), Larsen et al., Anaesthesitis 50, 842-5 (2001), Lilley et al., Anaesthesia, 51, 815-8 (1996), Bielen et al., Anesth.Analg., 82 (5), 920-4 (1996), and Knibbe et al., Br. J. Clin.Pharmacol., 47 (6), 653 -60 (1999)). However, these agents cause other side effects (eg cardiovascular complications) or cause destabilization of the propofol emulsion.

To overcome the problem of bacterial infections, propofol preparations have been developed with antibiotics such as EDTA equivalents (e.g. edetate), pentate, or sulfite-containing drugs, or formulated to have lower pH (e.g., US 5,714,520, 5,731,355, 5,731,356, 6,028,108, 6,100,302, 6,147,122, 6,177,477, 6,399,087, 6,469,069 and International Patent Application WO 99/39696. However, since dates and pentates are metal ion chelators, there is also a possibility that they may be dangerous by removing essential metal ions from the body system. Moreover, adding sulfite to drug formulations has the potential to have deleterious effects on the pediatric population and the general population allergic to sulfur.

Thus, there remains a need for methods to reduce or eliminate the side effects associated with parenteral or in vivo administration of drugs. There is also a need for sterile pharmaceutical compositions, methods of making such compositions. In addition, there is a need for a pharmaceutical composition and a method of preparing the same that reduce or eliminate the oxidation of the pharmaceutical composition to prevent destabilization of the drug.

The present invention provides such compositions and methods. These and other advantages and additional inventive features of the present invention may be apparent from the description of the invention described herein.

Brief summary of the invention

The present invention provides various embodiments of the pharmaceutical composition. One, some, or all features of the various embodiments can be found in different embodiments of the invention and fall within the scope of the appended claims.

The present invention provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is an amount of protein, such as albumin, effective to reduce one or more side effects when administering the pharmaceutical composition to a human. , More preferably human serum albumin, and the pharmaceutically acceptable carrier provides a pharmaceutical composition comprising an amount of deferoxamine effective to inhibit the growth of microorganisms in the pharmaceutical composition. The present invention also provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is an amount such as albumin in an amount effective to reduce one or more side effects when administering the pharmaceutical composition to a human. A protein, wherein the pharmaceutically acceptable carrier provides a pharmaceutical composition comprising an amount of deferoxamine effective to inhibit oxidation in the pharmaceutical composition.

The invention also provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises administering to a human a pharmaceutical composition comprising albumin and deferoxamine A method of reducing one or more side effects associated with administering a pharmaceutical composition to a human is provided. The present invention also provides a method of inhibiting the growth of microorganisms, or a method of inhibiting oxidation, or a method of inhibiting the growth of microorganism growth and oxidation in a pharmaceutical composition. These methods are pharmaceutical compositions comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is effective in inhibiting oxidation in a pharmaceutical composition or in an amount effective to inhibit the growth of microorganisms in the pharmaceutical composition. Preparing a pharmaceutical composition comprising an effective amount of deferoxamine.

The invention also provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises albumin, wherein the ratio of albumin to the pharmaceutical substance in the pharmaceutical composition is about 18 Provided are methods for enhancing delivery of a pharmaceutical substance to a disease site, comprising administering a pharmaceutical composition of less than or equal to: 1 to a human. The invention also provides a pharmaceutical composition comprising a pharmaceutical agent and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises albumin and the ratio of albumin to pharmaceutical agent in the pharmaceutical composition is about 18: 1. Provided are methods for enhancing binding of pharmaceutical drugs to in vitro or in vivo cells, comprising administering the following pharmaceutical compositions to in vitro or in vivo cells.

The present invention is also a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises albumin in an amount effective to enhance delivery of the drug to a disease site in humans, The pharmaceutical composition provides a pharmaceutical composition having a ratio of about 18: 1 or less.

The invention also provides a method of enhancing the delivery of a pharmaceutical drug to cells in vitro or in vivo by binding a pharmaceutical agent to a protein, wherein the protein binds to a specific cell-surface receptor on a cell and the protein-pharmaceutical Binding of the substance combination with the receptor causes delivery of the pharmaceutical substance, and enhances the delivery of the pharmaceutical substance to an in vitro or in vivo cell, wherein the ratio of the protein to the pharmaceutical substance is about 18: 1 or less. Provide a method.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises an effective amount of albumin and an effective amount to reduce one or more side effects when administering the pharmaceutical composition to a human. A pharmaceutical composition comprising the same protein, preferably human serum albumin, wherein the pharmaceutically acceptable carrier comprises an amount of deferoxamine effective to inhibit the growth of microorganisms in the pharmaceutical composition. The present invention also provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is an effective amount of albumin in reducing one or more side effects when administering the pharmaceutical composition to a human. A pharmaceutical composition comprising a protein such as, wherein the pharmaceutically acceptable carrier comprises an amount of deferoxamine effective to inhibit oxidation in the pharmaceutical composition.

Any suitable pharmaceutical substance can be used in the pharmaceutical composition of the present invention. Suitable pharmaceutical agents include anticancer or anti-neoplastic agents, antimicrotuble agents, immunosuppressants, anesthetics, hormones, cardiovascular agents, antiarrhythmics, antibiotics, antifungals, antihypertensives, antihistamines, analgesics, anti-inflammatory agents , Anti-arthritis agents, and vascular active agents. The present invention can also be usefully used in many other drug classes. More specifically, pharmaceutical substances include taxanes (e.g. Taxol ^R (paclitaxel), and Taxotere ^™ (docetaxel)), epithyron, camptothecin, colchicine, amiodarone, thyroid hormones, vascular active peptides (e.g., blood vessels). Active small intestinal peptide), amphotericin, corticosteroids, propofol, melatonin, cyclosporine, rapamycin (cyrrolimus), tacrolimus, mycophenolic acid, ifosfamide, vinorelbine, vancomycin, gemcitabine, SU5416, thiopeta , Bleomycin, diagnostic radiation contrast agents, and derivatives thereof, but are not limited thereto. Other drugs useful in the compositions of the present invention are described, for example, in US Pat. No. 5,916,596 and co-pending US patent application 09 / 446,783. Preferably, the pharmaceutical substance is propofol, paclitaxel, or docetaxel. More preferably, the pharmaceutical agent is propofol or paclitaxel. Most preferably, the pharmaceutical substance is propofol.

Taxol ^R (Paclitaxel) (Bristol-Myers Squibb) is active against ovarian, breast, lung, esophagus, and head and neck carcinomas. However, Taxol has been shown to cause toxicity associated with administration as well as significant and cumulative toxicity such as myelosuppression, neutrophenic fever, anaphylactic reactions, and peripheral neuropathy. Paclitaxel is very insoluble in water, and cremophores are typically used as solvents, requiring large injection volumes and special tubing and filters. Cremophores are associated with potentially serious side effects, including anaphylaxis and other hypersensitivity reactions that may require pretreatment with corticosteroids, antihistamines, and H2 blockers (eg, Gelderblom et al. , Eur. J. of Cancer, 37, 1590-1598, (2001)). Taxotere ^™ (docetaxel) is used for the treatment of anthracycline-resistant breast cancers, but has also been shown to induce side effects of hypersensitivity and fluid retention that may already be severe. Epotyrone (and its derivatives) has also typically been shown to cause severe neutropenia, hypersensitivity, and neuropathy.

Propofol (2,6-diisopropylphenol) is a hydrophobic, water-insoluble oil, which is widely used as an intravenous anesthetic to induce and maintain general anesthesia and sedation in humans. Propofol is typically administered directly into the bloodstream and crosses the blood-brain barrier. Pharmaceutical compositions comprising propofol must have sufficient fat solubility to penetrate these barriers and inhibit the relevant mechanisms of the brain. Propofol has a maximum water solubility of 1.0 +/- 0.02 μM at 22.5 ° C. (see, eg, Tonner et al., Anesthesiology, 77, 926-931 (1992)). Propofol by itself is formulated as an emulsion containing a solubilizer, a surfactant, a solvent, or as an oil-in-water emulsion (see, eg, US 6,150,423, 6,326,406, and 6,362,234). In addition to the active pharmaceutical substance, the compositions of the present invention comprise a pharmaceutical carrier or excipient. The choice of carrier is not necessarily important and any carrier known in the art can be used in the composition. The choice of carrier is preferably determined in part by the particular site to which the pharmaceutical composition is to be administered and the particular method used to administer the pharmaceutical composition. Preferably, the pharmaceutically acceptable carrier comprises a protein. Any suitable protein can be used. Examples of suitable proteins include, but are not limited to, albumin, immunoglobulins including IgA, lipoproteins, apolipoprotein B, beta-2-macroglobulin, thyroglobulin, and the like. Most preferably, the pharmaceutically acceptable carrier comprises albumin, most preferably human serum albumin. Proteins containing albumin suitable for the present invention are originally natural or can be prepared synthetically.

Human serum albumin (HSA) is a globular protein with high solubility of Mr 65K and consists of 585 amino acids. HSA is the most abundant protein in plasma and contributes 70-80% of colloidal osmotic pressure in human plasma. The amino acid sequence of HSA contains a total of 17 disulfide bonds, one free thiol (Cys 34), and a single tryptophan (Trp 214). Intravenous use of HSA solution is useful for the prevention and treatment of hypovolumetric shock (Tullis, JAMA, 237, 355-360, 460-463m (1997) and Hauser et al., Surgery, Gynecology and Obsterics, 150, 811). -816 (1980)) and exchange infusion (eg, Finlayson, Seminars in Thrombosis and Hemostasis, 6,85-120, (1980)) in the treatment of hyperbilirubinemia in newborns.

Human serum albumin (HSA) has several hydrophobic binding sites (total of eight endogenous ligands of HSA to fatty acids) and binds to various sets of drugs, especially neutral and negatively charged hydrophobic compounds (Goodman et al., The Pharmacological Basis of Therapeutics, 9 ^th ed, McGraw-Hill New York (1996)). Two binding sites with high affinity have been proposed in subdomains IIA and IIIA of HSA, such subdomains being long hydrophobic with charged lysine and arginine residues near the surface serving as an attachment point for polar ligand characterization. Pocket (Fehske et al., Biochem. Pharmacol., 30, 687-92 (1981), Vorum, Dan. Med. Bull., 46, 379-99 (1999), Kragh-Hansen, Dan. Med. Bull. , 1441, 131-40 (1990), Curry et al., Nat. Struct. Biol., 5, 827-35 (1998), Sugio et al., Protein.Eng, 12, 439-46 (1999), He et al., Nature, 358, 209-15 (1992), and Carter et al., Adv. Protein.Chem., 45, 153-203 (1994)). In addition, docetaxel has been shown to bind to human plasma proteins (eg, Urien et al., Invest. New Drugs, 14 (2), 147-51 (1996)). Thus, although not wishing to be bound by any particular theory, the inclusion of a protein, such as albumin, in the pharmaceutical composition of the present invention, wherein at least a portion of the human serum albumin binds to any free drug present in the composition of the present invention. It is believed to cause a reduction in the side effects associated with administration of the pharmaceutical composition resulting from the.

The amount of albumin included in the pharmaceutical composition of the present invention will vary depending on the pharmaceutically active substance, other excipients, and intended route and site of administration. Preferably, the amount of albumin included in the composition is an amount effective to reduce one or more side effects of the pharmacologically active substance resulting from administering the pharmaceutical composition of the present invention to a human. Typically, the pharmaceutical composition of the present invention is prepared in the form of a liquid, and then albumin is added to the liquid. Preferably, the pharmaceutical composition in liquid form comprises from about 0.1% to about 25% by weight (eg, about 0.5%, about 5%, about 10%, about 15%, about 20%) Contains albumin. Most preferably, the pharmaceutical composition in liquid form comprises about 0.5% to about 5% by weight albumin. The pharmaceutical composition can be dehydrated, for example, by lyophilization, spray drying, fluidized bed drying, wet granulation, and other suitable methods known in the art. When the composition is prepared in solid form, such as by wet granulation, fluidized bed drying, and other suitable methods known in the art, albumin is preferably applied as a solution to the active pharmaceutical substance and other excipients, if present. The HSA solution is preferably in the range of about 0.1 wt% to about 25 wt% (about 0.5 wt%, about 5 wt%, about 10 wt%, about 15 wt%, or about 20 wt%).

In addition to albumin, the composition of the present invention preferably comprises deferoxamine. Deferoxamine is a natural substance liberated from Streptomyces pilosus and can form iron complexes. For example, USP injectable deferoxamine mesylate can be used for intramuscular, subcutaneous, and intravenous administration. Diferoxamine mesylate USP is a white to off-white powder. It is easily soluble in water and has a molecular weight of 656.79. The chemical name of diferoxamine mesylate is N- [5- [3-[(5-aminopentyl) -hydroxycarbamoyl] -propion-amido] pentyl] -3 [[5-((N-hydroxyacet Amido) pentyl] -carbamoyl] propionohydroxysamic acid monomethanesulfonate (salt), the structural formula of which is C ₂₅ H ₄₈ N ₆ O ₈ .CH ₃ SO ₃ H. As described in the Examples, Diferoxamine or analogs, derivatives, or salts thereof (such as mesylate salts) inhibit microbial growth and oxidation in pharmaceutical compositions, which are believed to bind the free drug of the composition. (See, eg, Juven et al., J. Appl. Bacteriol., 76 (6), 626-31 (1994)) Paclitaxel, docetaxel, propofol and the like are phenol-like compounds or phenol or phenyl Therefore, deferoxamine may be combined with the free drug or free drug in the composition of the present invention. Reducing the amount of and, and also to reduce the irritation or pain upon injection or mitigate thereby.

The amount of deferoxamine or its preferred salt, ie, deferoxamine mesylate salt, included in the compositions of the present invention will depend on the active pharmaceutical substance and other excipients. Preferably, the amount of deferoxamine, salts thereof, and analogs thereof in the composition is an amount effective to inhibit the growth of and / or inhibit the proliferation of microorganisms. As noted above, pharmaceutical compositions are typically prepared in the form of a liquid, and then deferoxamine, its salts, and analogs thereof are added to the solution. Preferably, the pharmaceutical composition in liquid form comprises about 0.1% by weight of deferoxamine mesylate. When the composition is prepared in solid form, as described above, by wet granulation, fluidized bed drying, and other methods known to those skilled in the art, deferoxamine mesylate is preferably an active agent. It is applied as a solution to pharmaceutical substances and other excipients, if present. The deferoxamine mesylate solution preferably contains from about 0.0001% to about 0.5% (eg, about 0.005%, about 0.1%, about 0.25% by weight) of deferoxamine.

In the present invention, the pharmaceutical composition may include other substances, excipients, or stabilizers to enhance the characteristics of the composition. For example, certain negatively charged constituents may be added to increase stability by increasing the negative zeta potential of the nanoparticles or nanodroplets. Such negatively charged additional components include glycolic acid, cholic acid, chenodeoxycholic acid, taurocholic acid, glycodenodeoxycholic acid, taurocenodeoxycholic acid, lithocholic acid, urosodeoxycholic acid, dehydrocholic acid, and Bile salts of bile acids consisting of others; Phosphatidylcholine, Palmitoyl OleoylPhosphatidylcholine, PalmitoylolinoylPhosphatidylCholine, StearoyloleoylPhosphatidylCholine, StearoylOleoylPhosphatidylCholine, StearoylArachidylPhosphatidylCholine, and DipalmitoylPhosphoryl-containing Pylinthylcholine Phospholipids including the yolk-based phospholipids, but are not limited thereto. Other phospholipids include L-α-dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), distearioolphosphatidylcholine (DSPC), hydrogenated soy phosphatidylcholine (HSPC), D-α-phosphatidylcholine, β-acetyl -γ-O-hexadecyl, L-α-phosphatidylcholine, β-acetyl-γ-O-hexadecyl, DL-α-phosphatidylcholine, β-acetyl-γ-O-hexadecyl, L-α-phosphatidylcholine, β- Acetyl-γ-O-octadecyl, L-α-phosphatidylcholine, β-arachidonoyl-γ-O-hexadecyl, L-α-phosphatidylcholine, β-acetyl-γ-O- (octadec-9-cis- Enyl), D-α-phosphatidylcholine, β-arachidonoyl-γ-O-palmitoyl, 3-sn-phosphatidylcholine, 2-arachidinoyl-1-stearoyl, L-α-phosphatidylcholine, β-arachido Noyl-γ-stearoyl, L-α-phosphatidylcholine, diarachidonoyl, L-α-phosphatidylcholine, dibehenyl, L-α-phosphatidylcholine, β- (cis-8,11,14-eicosatriee Noyl) -γ-O-hexadecyl, L-α- Phosphatidylcholine, β-oleoyl-γ-myristoyl, L-α-phosphatidylcholine, β- (phylen-1-yl) decanoyl-γ-palmitoyl, 3-sn-phosphatidyl-N, N-dimethylethanolamine, 1,2-dipalmitoyl, L-α-phosphatidylethanolamine, diheptadecanoyl, 3-sn-phosphatidylethanolamine, 1,2-dilauroyl, 3-sn-phosphatidylethanolamine, 1,2- Dimyristoyl, 3-sn-phosphatidylethanolamine, 1,2-dioleoyl, 3-sn-phosphatidylethanolamine, 1,2-dipalmitoyl, L-α-phosphatidylethanolamine, dipalmitoyl, L -α-phosphatidylethanolamine, dipalmitoyl, L-α-phosphatidylethanolamine, dipalmitoyl, N-silyl, L-α-phosphatidylethanolamine, dipalmitoyl, N, N-dimethyl, L-α-di Myristoylphosphatidylglycerol (sodium salt) (DMPG), dipalmitoylphosphatidylglycerol (sodium salt) (DMPG), dipalmitoylphosphatidylglycerol (sodium salt) (DPPG), distearoylphosphatidylglycerol (sodium salt) ( DSPG), N- ( Carbonyl-methoxypolyethylene glycol 2000) -1,2-dstearoyl-sn-glycero-3-phosphoethanolamine sodium (MPEG-DSPE), L-α-phosphatidic acid, didecanoyl sodium salt , L-α-phosphatidic acid, diheptadecanoyl sodium salt, 3-sn-phosphatidic acid, 1,2-dimyristoyl sodium salt, L-α-phosphatidic acid, dioctanoyl sodium salt, L α-phosphatidic acid, dioleoyl sodium salt, L-α-phosphatidic acid, dipalmitoyl sodium salt, L-α-phosphatidyl-DL-glycerol, dimyristoyl sodium salt, L-α-phosphatidyl- DL-glycerol, dioleoyl sodium salt, L-α-phosphatidyl-DL-glycerol, dipalmitoyl ammonium salt, L-α-phosphatidyl-DL-glycerol, dimyristoyl sodium salt, L-α-phosphatidyl-DL- Glycerol, dipalmitoyl ammonium salt, L-α-phosphatidyl-DL-glycerol, distearoyl ammonium salt, L-α-phosphatidyl-DL-glycerol, β-oleyl-γ-palmitoyl ammonium salt, L-α-phosphatidyl inositol Ammonium salt, L-α-phosphatidylinositol sodium salt, L-α-phosphatidyl-L-serine, dioleoyl sodium salt, L-α-phosphatidyl-L-serine, dipalmitoyl sodium salt and the like. For example, emulsifiers which are negatively charged surfactants such as sodium cholesteryl sulfate and the like are also suitable as additives.

Pharmaceutical substances (eg propofol) can be used alone or dissolved in an aqueous solvent. A wide range of water insoluble solvents can be used, such as soybean, safflower, cottonseed, corn, sunflower, peanut, chestnut, or olive oil. Preferred oils are vegetable oils, with soybean oil being most preferred. Soybean oil may be used in the range of 1% to 10% by weight in the composition. Preferably soybean oil is present in the pharmaceutical composition in an amount of about 3% by weight.

The pharmaceutical composition of the present invention may be stabilized with a pharmaceutically acceptable surfactant. The term "surfactant" as used herein refers to the surfactant group (s) of the amphoteric molecule. Any surfactant can be anionic, cationic, nonionic, and zwitterionic. Any suitable surfactant can be included in the pharmaceutical composition of the present invention. Suitable surfactants can be used, for example, phosphatides, polyoxyethylene sorbitan esters, and tocopheryl polyethylene glycol succinate. Suitable surfactants are egg lecithin, tween 80, and vitamin E-t d-α-tocopheryl polyethylene glycol-1000 succinate (TPGS). In soybean oil-containing preparations, egg lecithin is preferred and contains at least 1.2% by weight, preferably at least 1.1% by weight, in formulations containing 3% soybean oil. In formulations without soybean oil, 0.1-1.5 wt.% Tween 80 or 0.5-4 wt.% Vitamin E-TPGS is suitable. Preferably, 1.5 wt% Tween 80 or 1 wt% Vitamin E-TPGS is used. Examples of other suitable surfactants are described, for example, in Becher, Emulsions: Theory and Practice, Robert E. Kriger Publishing, Malabar, Fla. (1965).

There are a wide variety of suitable formulations of the compositions of the present invention (see, eg US Pat. No. 5,916,596). The following formulations and methods are illustrative only and in no way limiting. Formulations suitable for oral administration include (a) a liquid solution such as an effective amount of a compound dissolved in a diluent such as water, saline, or orange juice, (b) a capsule or sachet containing a predetermined amount of the active ingredient as a solid or granules. Or tablets, (c) suspending agents in suitable liquids, and (d) suitable emulsions. Tablet forms include one or more lactose, mannitol, corn starch, potato starch, microcrystalline cellulose, acacia, gelatin, colloidal silicon dioxide, crocamellose sodium, talc, magnesium stearate, stearic acid, and other excipients, colorants, diluents, buffers Agents, wetting agents, preservatives, fragrances, and pharmaceutically suitable excipients. Lozenge forms may include active ingredients in fragrances, usually sucrose and acacia, or tragacanth, and pastilles include gelatin and glycerin containing the active ingredient as well as excipients known in the art, Or sucrose and an inert base such as acacia, emulsions, gels and the like.

Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic sterile injectable solutions, and suspending agents, solubilizers, which may contain antioxidants, buffers, bacteriostatics, and solutes that cause the formulation to isotonic with the blood of the intended recipient. , Aqueous and non-aqueous sterile suspending agents, which may include thickeners, stabilizers, and preservatives. The formulations may be present in single-dose or multiple-dose sealed containers such as ampoules and vials, stored under lyophililzed conditions, and sterile liquid excipients (e.g. water for injection) immediately prior to use. Only needs to be added. Instant injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described. Injectable preparations are preferred.

Formulations suitable for aerosol administration may include antioxidants, buffers, bacteriostatics, as well as aqueous and non-aqueous sterile suspensions, which may be included alone or in combination with other suitable ingredients, as suspending, solubilizing, thickening, stabilizing and preservatives. Aqueous and non-aqueous, isotonic, sterile liquids which may contain solutes are included alone or in combination with other suitable components that may be prepared in an aerosol formulation for administration by inhalation. Such aerosol formulations may be added to a suitable pressurized propellant such as dichlorofluoromethane, propane, nitrogen and the like. They may also be formulated as drugs of unpressurized formulations such as nebulizers or nebulizers.

Other suitable formulations are also possible, for example suppositories can be prepared by the use of various bases, such as emulsifying bases or water-soluble bases. Formulations suitable for vaginal administration may be present as pessaries, tampons, creams, gels, pastas, foams, or sprays containing, in addition to the active ingredient, carriers known to be suitable in the art.

In a preferred embodiment of the invention, the pharmaceutical composition is formulated to have a pH of 4.5 to 9.0, more preferably pH 5.0 to 8.0. Pharmaceutical compositions may also be prepared for isotonicity with blood by the addition of suitable tonicity modifiers. Moreover, the pharmaceutically acceptable carrier may also comprise water or USP injectable water, preferably free of pyrogenic substances. Preferably, the compositions of the present invention are prepared as sterile aqueous formulations, nanoparticles, oil-in-water (o / w) emulsions, or water-in-oil (w / o) emulsions. Most preferably, the pharmaceutical composition is oil-in-water emulsion.

In pharmaceutical compositions comprising propofol, according to the present invention, oil-in-water emulsions are prepared by dissolving propofol in an aqueous solvent alone, followed by preparing an aqueous phase containing albumin, deferoxamine, surfactants, and other water-soluble components. Step, and then mixing the oil and the water phase. The crude oil is formed by homogenizing at a high pressure of 10,000 to 25,000 psi and recycling 5 to 20 cycles. Preferred pressures are 15,000 psi to 20,000 psi, more preferably 10,000 psi. The stabilizer may be recycled for 7 to 15 cycles, preferably for 15 cycles. Alternatively, passage of material through the homogenizer may be used.

Preferably, the compositions of the present invention may have a particle or droplet size of less than about 200 nm. For example, for paclitaxel, docetaxel, rapamycin, cyclosporin, propofol, and other drugs, the average size of such dispersants is less than 200 nm.

The invention also provides a method of reducing one or more side effects associated with administering a pharmaceutical composition to a human. The method comprises a pharmaceutical composition comprising a pharmaceutical composition and a pharmaceutically acceptable carrier to a human, wherein the pharmaceutically acceptable carrier comprises administering to the human a pharmaceutical composition comprising albumin and deferoxamine. The descriptions of the pharmaceutical compositions, pharmaceutical substances, and pharmaceutically acceptable carriers, and their components, indicated above in connection with the pharmaceutical compositions of the present invention, are also applicable to the same aspects of the methods of the present invention.

The dosage of the pharmaceutical composition of the present invention to be administered to a human in the present invention will vary depending on the particular pharmaceutical composition, the method of administration, and the particular site to be treated. The dosage should be sufficient to elicit a desired response, such as a therapeutic or prophylactic response to a particular disease, or if the pharmaceutical agent is an anesthetic such as propofol, to elicit an anesthetic response within the desired time frame.

Although any suitable means of administering the pharmaceutical composition to a human can be used within the context of the present invention, preferably the composition of the present invention is administered intravenously, intraarterally, intrapulmonally, orally, inhaledly to a human. , Intraalveolar administration, intramuscular administration, intratracheal administration, subcutaneous administration, intraocular administration, intrathecal administration, or transdermal administration. For example, the pharmaceutical compositions of the present invention can be administered by inhalation to treat the condition of the respiratory system. Because albumin is a natural component in the inner wall of the respiratory tract and sprays, there are minimal side effects associated with inhalation of the compositions of the present invention. The composition of the present invention can be used to treat respiratory diseases such as pulmonary fibrosis, obstructive bronchiolitis pneumonia, lung cancer, bronchoalveolar cancer and the like.

The methods of the invention cause a reduction in one or more side effects associated with administering the pharmaceutical composition to a human. Such side effects include, for example, myelosuppression, neurotoxicity, hypersensitivity, inflammation, vascular irritation, phlebitis, pain, skin irritation, and combinations thereof. However, these side effects are very exemplary and other side effects or combinations of side effects associated with various pharmaceutical substances may be reduced or eliminated by the use of the new compositions and methods of the present invention.

The invention also provides a method of inhibiting the growth of microorganisms in a pharmaceutical composition. "Inhibition of microbial proliferation" means complete removal of microorganisms from a pharmaceutical composition or reduction of the amount or rate of growth of microorganisms in a pharmaceutical composition. The method of the present invention comprises a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is an amount of diferoxamine, salts thereof, effective to inhibit the growth of microorganisms in the pharmaceutical composition; Preparing a pharmaceutical composition comprising analogs thereof, and combinations thereof. The present invention also provides a method of inhibiting oxidation of a pharmaceutical composition. This method comprises a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier is in an amount effective to inhibit oxidation of the pharmaceutical composition, diferoxamine, salts thereof, analogs thereof, and Preparing a pharmaceutical composition comprising a combination thereof. The descriptions of the pharmaceutical compositions, pharmaceutical substances, and pharmaceutically acceptable carriers, and their components, indicated above in connection with the pharmaceutical compositions of the present invention, are also applicable to the same aspects of the methods of the present invention.

The amount of deferoxamine, or a preferred salt thereof, deferoxamine mesylate salt included in the composition of the present invention will vary depending on the active pharmaceutical substance and other excipients. Preferably, the amount of deferoxamine, salts thereof, and analogs thereof in the composition is an amount effective to inhibit the growth and / or oxidation of the microorganisms. As noted above, pharmaceutical compositions are typically prepared in the form of a liquid, and then deferoxamine, its salts, and analogs thereof are added in solution. Preferably, the pharmaceutical composition in liquid form comprises from about 0.0001% to about 0.5% by weight (eg, about 0.005%, about 0.1%, or about 0.25% by weight) of deferoxamine, salts thereof, or the like It includes. More preferably, the composition in liquid form comprises a similar amount of diferoxamine mesylate, which is a preferred diferoxamine salt. Most preferably, the pharmaceutical composition in liquid form comprises about 0.5% by weight of dipropoxamine mesylate. When the pharmaceutical composition of the present invention is prepared in solid form, as described above, by wet granulation, fluidized bed drying, and other methods known in the art, deferoxamine mesylate is preferably used as the active pharmaceutical substance and If present, apply as a solution to other excipients. The deferoxamine mesylate solution is preferably from about 0.0001 wt% to about 0.5 wt% (eg, about 0.005 wt%, about 0.1 wt%, or about 0.25 wt%) which is diferoxamine.

The invention also provides a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises albumin, wherein the ratio of albumin to the pharmaceutical substance in the pharmaceutical composition is about 18 A method of enhancing delivery of a pharmaceutical agent to a disease site, comprising administering to a human a pharmaceutical composition of less than or equal to: 1. The present invention relates to a pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises albumin and the ratio of albumin to the pharmaceutical substance in the pharmaceutical composition is 18: 1 or less. Provided are methods for enhancing binding of a pharmaceutical drug to in vitro or in vivo cells, comprising administering the pharmaceutical composition to cells in vitro or in vivo. The descriptions of the pharmaceutical compositions, pharmaceutical substances, and pharmaceutically acceptable carriers, and their components, shown above in connection with the pharmaceutical compositions and methods of the present invention, also apply to the same aspects of the methods of transport and binding of the present invention. It is possible.

In a method of enhancing the transport of a pharmaceutical substance to a disease site or in a method of enhancing the binding of a pharmaceutical substance to cells, the pharmaceutically acceptable carrier preferably comprises albumin, most preferably human serum albumin. . Without wishing to be bound by any particular theory, the ratio of proteins, such as, for example, human serum albumin to pharmaceutical substances, in pharmaceutical compositions is dependent on the ability of the pharmaceutical substance to bind cells and the transport of pharmaceutical substances to cells. Affect ability In this regard, higher ratios of protein to pharmaceutical substances are associated with cell binding and transport of the poorer pharmaceutical substances, which is probably the result of competition for receptors on the cell surface. The ratio of protein (eg albumin) to pharmacologically active substance should be such that the pharmaceutical substance is sufficient to bind or be delivered by the cell. An exemplary ratio of protein-drug formulation is from 0.01 to about 100: 1 protein: drug. More preferably, the ratio is from 0.02: 1 to about 40: 1. The ratio of protein to pharmaceutical substance should be optimized for the combination of different proteins and pharmaceutical substances, but in general, the ratio of protein (eg albumin) to pharmaceutical substance is about 18: 1 or less (eg about 15 : 1, about 10: 1, about 5: 1, or about 3: 1). More preferably, the ratio is about 0.2: 1 to about 12: 1. Most preferably, the ratio is about 1: 1 to about 9: 1. Preferably, the preparations do not necessarily contain cremophors, more preferably do not contain Cremophor EL ^R (BASF). Cremophor EL ^R is a polyether of chestnut oil and a nonionic emulsifier that is ethylene oxide. As noted above, cremophores are typically used as solvents of paclitaxel and are associated with potentially serious side effects (see, eg, Gelderblom et al., Supra).

The pharmaceutical substance may be any suitable substance shown herein, such as propofol, paclitaxel, or docetaxel. In addition, the pharmaceutical agent may be a nucleic acid sequence, preferably a DNA sequence. In this regard, the pharmaceutical compositions of the invention can be used to transport genes to cells by receptor mediated / caveolar / alveolar transport. Preparation of pharmaceutical compositions comprising albumin with genetic material to transport DNA sequences, such as, but not limited to, plasmids or c-DNAs, such as genes or other genetic material, into cells (eg, endothelial cells or tumor cells) can do. Since tumor cells and other cells at the site of inflammation have high protein uptake, the genetic material is preferably absorbed into this cell type and can be integrated into the genetic material of the cell for useful therapeutic effects. The use of proteins such as human serum albumin acts as a nonviral vector for the delivery of genetic material without the risk of virus-related diseases or side effects. For example, the pharmaceutical composition may be prepared with a pharmaceutical composition comprising a nucleic acid sequence encoding β-galagotosidase or green fluorescent protein (GFP), which is derived from human umbilical vein or human lung vesicles. Contact with endothelial cells may facilitate integration of nucleic acid sequences into endothelial cells. Integration of nucleic acid sequences can be detected using methods known in the art, such as fluorescence or staining.

In the method of the present invention for enhancing the transport of a pharmaceutical substance to a disease site, the disease may be a suitable disease or condition. Preferably, the disease is cancer, cardiovascular disease, or arthritis.

In the methods of the present invention for delivering pharmaceutical substances to cells in vitro or in vivo, the pharmaceutical composition is administered to cells in vitro or in vivo. Preferably, the cell is an animal cell. More preferably, the cell is a mammalian cell, most preferably the cell is a human cell. The pharmaceutical composition is preferably administered to cells in vivo. The cell may be any suitable cell that is a preferred target for administration of the pharmaceutical composition. For example, the cells may be located or derived from tissues of the digestive organs, including, for example, the esophagus, stomach, small intestine, large intestine, rectum, anus, liver, gallbladder, and pancreas. The cells may be located in or derived from tissues of the respiratory system, including, for example, the larynx, lungs, and bronchus. The cells are located in the bladder, kidney, renal pelvis, and ureter, including, for example, the cervix, uterine corpus, ovaries, vagina, prostate, testes, and penis, and the urinary constituents of the female and male genitalia. Or derived therefrom. The cells may be located in or derived from tissues of the cardiovascular system, including, for example, endothelial cells and cardiomyocytes. The cells may also be located in or derived from the lymphatic system (eg lymphocytes), nervous system (eg neurons or glial cells), and endocrine system (eg thyroid cells). Preferably, the cells may be located in or derived from tissue of the cardiovascular system. Most preferably, the cells are endothelial cells. In the context of the present invention for enhancing the transport of a pharmaceutical substance to cells and for enhancing the binding of the pharmaceutical substance to cells, the pharmaceutical composition is preferably in contact with one or more cells.

In another aspect of the invention, the methods of the invention for enhancing the transport and binding of pharmaceutical agents can be used to treat tumor cells. Tumor cells show an increase in protein uptake, including albumin and transferrin, for example, compared to normal cells. Because tumor cells divide at a rapid rate, such tumor cells require additional nutrition compared to normal cells. Tumor studies of the pharmaceutical compositions of the invention containing paclitaxel and human serum albumin showed high uptake of albumin-paclitaxel into the tumor. This was found to be due to albumin-drug transport by the glycoprotein 60 ("gp60") receptor specific for albumin, which was not previously recognized.

Thus, according to another aspect of the invention, albumin-specific gp60 receptors and other protein transport receptors present in tumor cells can be used as targets for inhibiting tumor proliferation. proteins to these cells by blocking the gp60 receptor and blocking other protein transport receptors on tumor cells or tumor endothelial cells with antibodies to the gp60 receptor or other large or small molecules that bind to, block or inactivate gp60 It is possible to block their transport, thereby reducing their rate of proliferation and inducing cell death. Blocking this mechanism thus leads to the treatment of cancer or another disease in an individual (eg a human). Confirmation of blocking / binding of specific protein receptors is made by screening the number of compounds against free gp60 or other receptors such as gp16 or gp30 or by whole cell preparation. In addition, suitable animal models such as, for example, mice that contain "knock-out" mutations in genes encoding gp60 or caveolin-1 or contain other proteins specific for transport are also suitable for this purpose. Can be used. Thus, methods of identifying compounds that block or bind gp60, gp16, gp30, or other protein receptors are within the scope of the present invention.

In addition, compounds that block or bind the gp60 receptor or other protein receptors can be used to treat a variety of diseases, including cancer. With respect to cancer treatment, blocking or binding compounds can be used in combination with a single drug or other standard chemotherapy or chemotherapy. For example, it is useful to treat cancer with conventional chemotherapy or with albumin-drug pharmaceutical compositions of the invention (compositions that exhibit high accumulation in tumors) and then with compounds that block the transport of proteins to tumor cells. Do. The blocking compound is administered before or in combination with the other chemotherapeutic agent or other anticancer agent. Thus, any drug that blocks or binds to gp60 or other protein receptors is within the scope of the present invention.

Albumin-drug compositions of the invention (eg, albumin-paclitaxel, albumin-doxitaxel, albumin-epotyrone, albumin-camptothecin, or albumin-rapamycin) are useful for the treatment of diseases. Such drug compositions are effective because of the increased receptor mediated transport of the protein-drug composition to the desired site, eg, a tumor. While not wishing to be bound by any theory, the transport of protein-drug compositions by receptor mediated transport that induces a therapeutic effect is believed to be a transport mechanism, eg, of albumin-paclitaxel compositions, to tumors, as well as albumin-paclitaxel and It is thought to be a mechanism of transport through the lungs of albumin-rapamycin. Transport is affected by the presence of gp60, gp16, or gp30 in such tissues. Thus, drugs and protein-drug compositions in which delivery to a disease site such as, for example, inflammation (eg arthritis) or a tumor are associated with the gp60, gp16, or gp30 receptor and induce a therapeutic effect are considered as compositions of the present invention. .

According to another aspect of the invention, endothelial cells can be co-cultured with cells having a particular function. Incubation of endothelial cells with islet cells, hepatocytes, neuroendocrine cells, and other cells allows for the required delivery of components such as proteins and other beneficial components to these cells. Endothelial cells are characterized by the in vivo conditions, that is, these cell types are usually in close proximity to the endothelial cells, so that these components can be cultivated to mimic conditions that depend on endothelial cells for the delivery of nutrients, growth factors, and hormonal signals. Provide delivery. In the absence of endothelial cells, it was not possible to properly culture these different cell types and obtain physiological performance. The presence of endothelial cells in culture with the desired cell type allows for the differentiation and proper functioning of islets, hepatocytes, or neuroendocrine tissues in vitro or ex vivo. Thus, coculture of endothelial cells with islets leads to islet cells with improved physiological properties, eg insulin secretion, compared to cultured without endothelial cells. These tissues can then be used ex vivo or transplanted in vivo, resulting in diseases caused by lack of proper cell function (e.g. diabetes in islets, hepatic dysfunction in hepatocytes, neuroendocrine in neuroendocrine cells). Disease or pain relief). Cells derived from other tissues and organs (above) may be cocultured with endothelial cells to provide the same benefits. Coculture can also be used to integrate genetic material into target cell types. The presence of albumin in this culture has been found to be very beneficial.

The following examples are intended to further illustrate the invention and, of course, are not intended to limit the scope of the invention in any way.

Example  One

This example illustrates the preparation of a pharmaceutical composition comprising paclitaxel and albumin. The preparation of the paclitaxel-albumin composition is described in US 5,439,686 and 5,916,596, which are hereby incorporated by reference in their entirety. Specifically, 30 mg of paclitaxel was dissolved in 3.0 mL of methylene chloride. The solution was added to 27.0 ml of human serum albumin solution (2% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest I.Q.) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the methylene chloride was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting suspension is translucent and the typical average diameter of the resulting paclitaxel particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization.

It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of paclitaxel dissolved in the cremophore formulation, the pharmaceutical compositions of the invention containing albumin exhibited substantially lower toxicity.

Example  2

This example illustrates the preparation of a pharmaceutical composition comprising amiodarone and albumin. 30 mg of amiodarone were dissolved in 3.0 mL of methylene chloride. The solution was added to 27.0 ml of human serum albumin solution (1% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest I.Q.) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the methylene chloride was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting amiodarone particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization.

It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of amiodarone dissolved in the Tween formulation, the pharmaceutical compositions of the present invention containing albumin exhibited substantially lower toxicity.

Example  3

This example illustrates the preparation of a pharmaceutical composition comprising liotyrone and albumin. Riotyronin (or a salt thereof) was dissolved in an alcohol or alkaline aqueous solution at a concentration of 0.5-50 mg / ml. The alcohol (or alkali) solution was added to the albumin solution (1-25% w / v) and stirred. Agitation was performed with a high shear force using a sonicator or homogenizer with a low shear force using a stirrer. When the concentration of liothyronine was low, a clear solution of (5-1000 µg / ml) was obtained. As the concentration increased, a milky stable suspension was obtained. This solution or suspension was filtered through sterile filter. The organic solvent was removed by evaporation or other suitable method.

Example  4

This example illustrates the preparation of a pharmaceutical composition comprising rapamycin and albumin. 30 mg of rapamycin was dissolved in 2 mL of chloroform / ethanol. The solution was then added to 27.0 ml of human serum albumin solution (3% w / v). The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the methylene chloride was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way.

Example  5

This example illustrates the preparation of a pharmaceutical composition comprising epotyrone B and albumin. 30 mg of epotyrone B was dissolved in 2 mL of chloroform / ethanol. The solution was then added to 27.0 ml of human serum albumin solution (3% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to Rotavap and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of epotyrone B dissolved in the cremophore formulation, the pharmaceutical composition of the present invention comprising albumin exhibited substantially lower toxicity.

Example  6

This example illustrates the preparation of a pharmaceutical composition comprising colchicine dimer and albumin. 30 mg of colchicine dimer was dissolved in 2 mL of chloroform / ethanol. The solution was then added to 27.0 ml of human serum albumin solution (3% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to Rotavap and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of colchicine dimers dissolved in Tween preparations, the pharmaceutical compositions of the invention comprising albumin exhibited substantially lower toxicity.

Example  7

This example illustrates the preparation of a pharmaceutical composition comprising docetaxel and albumin. 30 mg of docetaxel was dissolved in 2 mL of chloroform / ethanol. The solution was then added to 27.0 ml of human serum albumin solution (3% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to Rotavap and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of docetaxel dissolved in tween / ethanol, the pharmaceutical composition of the invention comprising albumin exhibited substantially lower toxicity.

Example  8

This example illustrates the preparation of a pharmaceutical composition comprising docetaxel and albumin. 150 mg of docetaxel was dissolved in 1 mL of ethyl acetate / butyl acetate and 0.5 mL of an oil such as soybean oil or vitamin E oil. Different proportions of solvents and oils were used and such compositions are also part of the present invention. Small amounts of negatively charged constituents such as benzoic acid (0.001% -0.5%) were also optionally added. The solution was then added to 27.0 ml of human serum albumin solution (5% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to Rotavap and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of docetaxel dissolved in tween / ethanol, the pharmaceutical composition of the invention comprising albumin exhibited substantially lower toxicity.

Example  9

This example illustrates the preparation of a pharmaceutical composition comprising taxane IDN5390 and albumin. 150 mg of taxane IDN5390 was dissolved in 1 mL of ethyl acetate / butyl acetate and 0.5 mL of oil, such as soybean oil or vitamin E oil. Different proportions of solvents and oils were used and such compositions are also part of the present invention. Small amounts of negatively charged constituents such as benzoic acid (0.001% -0.5%) were also optionally added. The solution was then added to 27.0 ml of human serum albumin solution (5% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to Rotavap and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of IDN5390 dissolved in Tween, the pharmaceutical compositions of the present invention comprising albumin exhibited substantially lower toxicity.

Example  10

This example illustrates the preparation of a pharmaceutical composition comprising taxane IDN5109 and albumin. 150 mg of taxane IDN5109 was dissolved in 2 mL of chloroform / ethanol. Different proportions of solvents and oils were used and such compositions are also part of the present invention. Small amounts of negatively charged constituents such as benzoic acid (0.001% -0.5%) were also optionally added. The solution was then added to 27.0 ml of human serum albumin solution (5% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to Rotavap and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of IDN5109 dissolved in Tween, the pharmaceutical compositions of the present invention comprising albumin exhibited substantially lower toxicity.

Example  11

This example illustrates the preparation of a pharmaceutical composition comprising 10-hydroxy camptothecin (10HC) and albumin. 30 mg of 10-HC was dissolved in 2.0 mL of DMF / methylene chloride / soybean oil. The solution was then added to 27.0 ml of human serum albumin solution (3% w / v). The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to Rotavap and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way.

Example  12

This example illustrates the preparation of a pharmaceutical composition comprising cyclosporin and albumin. 30 mg of cyclosporine was dissolved in 3.0 mL of methylene chloride. The solution was then added to 27.0 ml of human serum albumin solution (1% w / v). The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the methylene chloride was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization.

Example  13

This example illustrates the preparation of a pharmaceutical composition comprising an oil and cyclosporin and albumin. 30 mg of cyclosporine was dissolved in 3.0 mL of an appropriate oil (sesame oil containing 10% orange oil). The solution was then added to 27.0 ml of human serum albumin solution (1% w / v). The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. Typical average diameters of the resulting dispersions ranged from 50-220 nm (Z-means, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours by optionally adding an appropriate cryoprotectant. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way.

Example  14

This example illustrates the preparation of a pharmaceutical composition comprising amphotericin and albumin. 30 mg of amphotericin was dissolved in 3.0 mL of methyl pyrrolidinone / methylene chloride. The solution was added to 27.0 ml of human serum albumin solution (1% w / v). The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest IQ) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the solvent was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting amphotericin particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. The addition of other components, such as lipids, bile salts and the like, also resulted in appropriate formulations.

Example  15

This example shows preclinical pharmacokinetics and pharmacokinetics of pharmaceutical compositions comprising paclitaxel and albumin.

Several preclinical pharmacokinetic studies in mice and rats were performed to evaluate the possible benefits of albumin-paclitaxel pharmaceutical compositions as compared to the Cremophor-Paclitaxel pharmaceutical compositions. These studies indicate that (1) the pharmacokinetics of albumin-paclitaxel in rats is linear with dose, while the pharmacokinetics of Taxol is nonlinear, and (2) pharmaceutical compositions comprising albumin and paclitaxel are lower. Plasma AUC and C _max , indicating that the distribution of the albumin-paclitaxel composition to tissue is faster than taxol, (3) pharmaceutical compositions comprising albumin and paclitaxel exhibit lower C _max , Demonstrates a decrease in toxicity associated with peak blood concentrations compared to Taxol, (4) the half-life of the pharmaceutical composition comprising albumin and paclitaxel is about 2 times higher in rats and about 4 times higher in mice with tumors, (5) The metabolism of paclitaxel in pharmaceutical compositions comprising albumin and paclitaxel was lower than in taxol pharmaceutical compositions . Twenty four hours after injection into rats, 44% of total radioactivity is still bound to paclitaxel in pharmaceutical compositions comprising albumin and paclitaxel, whereas only 22% is bound for taxol. A final effect of the pharmacokinetics of the pharmaceutical composition comprising albumin and paclitaxel, i.e. increased intracellular uptake, prolonged half-life, and decreased metabolic rate, increased 1.7 times the tumor AUC compared to Taxol in mice with tumors. , A 1.2-fold increase in tumor C _max , and a 1.7-fold increase in tumor half-life.

Example  16

This example shows a reduction in side effects and a reduction in toxicity associated with pharmaceutical compositions comprising paclitaxel and albumin.

Due to the unique properties of pharmaceutical compositions comprising paclitaxel and albumin in the absence of cremophores, the toxicity of pharmaceutical compositions comprising paclitaxel and albumin is substantially lower than taxol. In preclinical studies in mice and rats, acute toxicity studies of single doses in mice showed that the LD ₅₀ dose was about 59 times higher in pharmaceutical compositions comprising paclitaxel and albumin compared to Taxol. In multi-dose toxicity studies in mice, the LD ₅₀ dose was found to be about 10 times higher in pharmaceutical compositions comprising paclitaxel and albumin compared to Taxol. In a further study, the degree of myelosuppression in rats treated with Taxol and a pharmaceutical composition comprising paclitaxel and albumin was evaluated. As a result, at equivalent doses, pharmaceutical compositions comprising paclitaxel and albumin showed significantly lower myelosuppression in rats than Taxol. In acute toxicity studies in rats, cerebral cortical necrosis or acute neurotoxicity was observed in animals receiving Taxol at 9 mg / kg, but compositions containing paclitaxel and albumin at doses of up to 120 mg / kg It did not appear in animals. Thus, the presence of albumin in the composition comprising paclitaxel causes substantial side effects and reduced toxicity compared to conventional pharmaceutical compositions comprising paclitaxel.

Example  17

This example demonstrates the clinical effects in humans of pharmaceutical compositions comprising paclitaxel and albumin.

Clinical studies in over 500 human patients provide evidence supporting that pharmaceutical compositions comprising paclitaxel and albumin have reduced toxicity and side effects compared to cremophor-paclitaxel compositions (taxols). In a Phase I study of 19 patients, the maximum internal dose of albumin-paclitaxel administered every three weeks was 300 mg / m ² . This is substantially higher than the generally administered dose of cremophor-paclitaxel administered at 175 mg / m ² once every three weeks. Hematologic toxicity in these patients did not show hypersensitivity, mild neuropathy, and no administration-related side effects such as vascular irritation.

In another Phase I study of 27 patients, the maximum content of albumin-paclitaxel administered weekly was 125-150 mg / m ² . This is substantially higher than the generally administered dose of cremophor-paclitaxel administered once every week at 80 mg / m ² . Hematologic toxicity in these patients did not show hypersensitivity, mild neuropathy, and no administration-related side effects such as vascular irritation.

In the Phase II study where albumin-paclitaxel was administered to 43 and 63 patients every three weeks at 175 or 300 mg / m ² , the hematologic toxicity was ANC <at 175 mg / m ² and 300 mg / m ^{2 respectively} . It was as low as only 7% and 24% of patients with 500 / mm ³ . Severe neuropathy occurred in 0% and 14% of patients receiving 175 mg / m ² and 300 mg / m ² , respectively. No serious hypersensitivity reactions occurred and no adverse events related to administration such as blood irritation or injection site pain occurred. These side effects were substantially lower than those seen in Taxol.

In Phase III experiments in which the albumin-paclitaxel composition ABI-007 was compared to Taxol (containing Cremophor-Paclitaxel), the dose of ABI-007 was substantially higher (260 mg / m ² vs. Taxol administration). Amount 175 mg / m ² ), indicating that ABI-007 has a higher content. Albumin-paclitaxel compositions also exhibited significantly lower neutropenia compared to cremophor-paclitaxel.

Example  18

This example shows an increase in preclinical efficacy when using a pharmaceutical composition comprising paclitaxel and albumin.

In vitro cytotoxicity studies comparing albumin-paclitaxel and Taxol's effects on cervical squamous cell carcinoma A431 showed an IC ₅₀ of 0.0038 0.012 μg / ml for albumin-paclitaxel and 0.012 μg / ml for Taxol. An increase in cytotoxic activity of paclitaxel was shown.

MTD of ABI-007 in five different human xenograft tumor models (MX-1 breast, NCI-H522 lung, SK-OV-3 ovary, PC-3 prostate, and HT-29 colon) in athymic mice Or equivalent toxic doses were 1.5-3.4 times higher than Taxol, and statistically significant improvement in tumor growth retardation (p <0.05) was found in all tumors except lung tumors (p = 0.15).

In the MX 1 breast model, 100% of albumin-paclitaxel treated animals survived for 103 days, compared to only 20-40% in the group treated with equivalent doses of Taxol.

Example  19

This example shows an increase in clinical efficacy with pharmaceutical compositions comprising albumin and paclitaxel when administered intraarterally.

As indicated herein, in a Phase I / II study of intraarterial administration of a pharmaceutical composition comprising albumin and paclitaxel, the patient was treated for head & neck cancer (N = 31) and anal cancer (N = 12). Registered. The dose was increased to 120-300 mg / m ² by transdermal superselective intraarterial infusion over 30 minutes. Head and neck cancer patients had a 76% response rate (N = 29), while anal cancer patients had a 64% response rate (N = 11).

Example  20

This example shows the preparation of a pharmaceutical composition containing 3% oil and comprising propofol and albumin.

Oil-in-water emulsions containing 1% by weight of propofol were prepared as follows. The aqueous phase was prepared by adding glycerol (22% by weight) and human serum albumin (0.5% by weight) to water for injection and stirring until dissolved. The aqueous phase was passed through a filter (0.2 um filter). An oily phase was prepared by dissolving egg lecithin (0.4% by weight) and propofol (1% by weight) in soybean oil (3% by weight) and stirring until dissolved. The oil phase was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen. The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.5-3%; Soybean oil 0.5-3.0%; Egg lecithin 0.12-1.2%; Glycerol 2.25%; Add water for injection to 100; pH 5-8. Appropriate chelators such as deferoxamine (0.001-0.1%) were optionally added.

Example  21

This example shows the preparation of a pharmaceutical composition containing 5% oil and comprising propofol and albumin.

Oil-in-water emulsions containing 1% by weight of propofol were prepared as follows. The aqueous phase was prepared by adding glycerol (22% by weight) and human serum albumin (0.5% by weight) to water for injection and stirring until dissolved. The aqueous phase was passed through a filter (0.2 um filter). An oily phase was prepared by dissolving egg lecithin (0.8% by weight) and propofol (1% by weight) in soybean oil (5% by weight) and stirring until dissolved. The oil phase was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen. The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.5-3%; Soybean oil 0.5-10.0%; Egg lecithin 0.12-1.2%; Glycerol 2.25%; Add water for injection to 100; pH 5-8. Appropriate chelators such as deferoxamine (0.001-0.1%) were optionally added.

Example  22

This example illustrates the preparation of a pharmaceutical composition comprising propofol and albumin.

Using a method similar to that shown in Example 18, a propofol composition containing albumin and tween 80 was prepared. The aqueous phase was prepared by adding glycerol (2.25 wt%), human serum albumin (0.5 wt%), tween 80 (1.5 wt%), and deferoxamine mesylate (0.1 wt%) to water for injection and stirring until dissolved. . The aqueous phase was passed through a filter (0.2 μm filter). Propofol (1% by weight) was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen. The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.5-3%; Tween 80 0.1-1.5%; Deferoxamine mesylate 0.0001-0.1%; Glycerol 2.25%; Add water for injection to 100; pH 5-8.

Example  23

This example illustrates the preparation of a pharmaceutical composition comprising propofol, albumin, and oil-free vitamin E-TPGS.

Using a method similar to that shown in Example 19, a propofol composition containing albumin and vitamin E-TPGS was prepared. The aqueous phase was added to water for injection by adding glycerol (2.25 wt%), human serum albumin (0.5 wt%), vitamin E-TPGS (1 wt%), and deferoxamine mesylate (0.1 wt%) and stirring until dissolved. Prepared. The aqueous phase was passed through a filter (0.2 μm filter). Propofol (1% by weight) was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen. The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.5-3%; Vitamin E-TPGS 0.5-4.0%; Optionally deferoxamine mesylate 0.0001-0.1%; Glycerol 2.25%; Add water for injection to 100; pH 5-8.

Example  24

This example shows the preparation of a pharmaceutical composition comprising propofol, albumin, vitamin E-TPGS, and 1% oil.

An emulsion containing 1% by weight of propofol was prepared by the following method. The aqueous phase was prepared by adding glycerol (2.25 wt%) and human serum albumin (0.5 wt%) to water for injection and stirring until dissolved. The aqueous phase was passed through a filter (0.2 μm filter). Surfactant such as vitamin E-TPGS (0.5%) was added to the aqueous phase. The oil phase consisted of propofol (1% by weight) and 1% soybean oil. The oil phase was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen.

The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; Soybean oil or other oils 0.1% -5%; Glycerol 2.25%; Add water for injection to 100; pH 5-8. Diferoxamine mesylate was optionally added (0.001-0.1 wt.%).

Example  25

This example shows the preparation of a pharmaceutical composition comprising propofol, albumin, vitamin E-TPGS, 1% oil, and a negatively charged component.

An emulsion containing 1% by weight of propofol was prepared by the following method. The aqueous phase was prepared by adding glycerol (2.25 wt%) and human serum albumin (0.5 wt%) to water for injection and stirring until dissolved. The aqueous phase was passed through a filter (0.2 μm filter). Surfactant such as vitamin E-TPGS (0.5%) was added to the aqueous phase. The oil phase consisted of propofol (1% by weight) and 1% soybean oil. A small amount of negatively charged constituents (0.001% -1%), for example phospholipids or bile salts, was added. The oil phase was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen.

The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; Soybean oil or other oils 0.1% -5%; Glycerol 2.25%; Add water for injection to 100; pH 5-8. Diferoxamine was optionally added (0.001-0.1 wt.%).

Example  26

This example shows the preparation of a pharmaceutical composition comprising propofol, albumin, vitamin E-TPGS, 1% oil, and a negatively charged component (sodium deoxycholate).

An emulsion containing 1% by weight of propofol was prepared by the following method. The aqueous phase was prepared by adding glycerol (2.25 wt%) and human serum albumin (0.5 wt%) to water for injection and stirring until dissolved. The aqueous phase was passed through a filter (0.2 μm filter). Surfactant such as vitamin E-TPGS (0.5%) was added to the aqueous phase. The oil phase consisted of propofol (1% by weight) and 1% soybean oil. A small amount of negatively charged constituent (0.001% -1%), for example sodium deoxycholate, was added. The oil phase was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen.

The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; Soybean oil or other oils 0.1% -5%; Glycerol 2.25%; Add water for injection to 100; pH 5-8. Diferoxamine was optionally added (0.001-0.1 wt.%).

Example  27

This example shows the preparation of a pharmaceutical composition comprising propofol, albumin, vitamin E-TPGS, 1% oil, and negatively charged components (phospholipids, bile salts, polyamino acids, etc.).

An emulsion containing 1% by weight of propol was prepared as follows. The aqueous phase was prepared by adding glycerol (2.25 wt%) and human serum albumin (0.5 wt%) to water for injection and stirring until dissolved. The aqueous phase was passed through a filter (0.2 μm filter). Surfactant such as vitamin E-TPGS (0.5%) was added to the aqueous phase. The oil phase consisted of propofol (1% by weight) and 1% soybean oil. A small amount of negatively charged component (0.001% -1%), for example phosphatidyl choline, was added. The oil phase was added to the aqueous phase and homogenized at 10,000 RPM for 5 minutes. The crude was homogenized to a high pressure of 20,000 psi and recycled at 5 ° C. for 15 cycles. Alternatively, discrete passes through the homogenizer were used. The final emulsion was filtered (0.2 μm filter) and stored under nitrogen.

The resulting pharmaceutical composition contained the following general range of ingredients (% by weight): propofol 0.5-5%; Human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; Soybean oil or other oils 0.1% -5%; Glycerol 2.25%; Add water for injection to 100; pH 5-8. Diferoxamine was optionally added (0.001-0.1 wt.%).

Example  28

This example shows the binding of propofol to albumin.

The binding of propofol to albumin was determined as follows. Solubility of propofol was tested in water and in albumin containing solutions. 250 μl propofol was added to 10 mL of water or albumin solution and stirred for 2 hours in a scintillation vial. The solution was then transferred to a 15 mL polyethylene centrifuge tube and stored at 40 ° C. for 16 hours. Samples of water and albumin solution were assayed for propofol. The water solubility of propofol was determined to be 0.12 mg / mL. The solubility of propofol in albumin solution was dependent on albumin concentration and increased to 0.44 mg / ml when albumin concentration was 2% (20 mg / mL). The solution was ultrafiltered through a 30 kD MWCO filter and the filtrate was assayed against propofol. In the propofol / water solution, 61% of propofol was recovered from the filtrate, whereas only 14% of the propofol / albumin solution was recovered from the filtrate, indicating substantial binding of propofol to albumin. Based on these results, the addition of albumin to a pharmaceutical composition comprising propofol causes a reduction in the amount of free propofol by albumin binding of propofol.

Example  29

This example demonstrates the reduction of free propofol in the pharmaceutical composition by filtration / membrane contact.

As observed in the experiments shown in Example 28, filtration or ultrafiltration of the pharmaceutical composition comprising propofol results in a decrease in the amount of free propofol. The pharmaceutical compositions prepared according to the invention comprising Diprivan and albumin, each containing 1% propofol (10 mg / mL), were ultrafiltered using a 30 kD membrane. The amount of free propofol in the filtrate was measured using HPLC. The concentration of free propofol in the filtrate was about 17 μg / mL for Diprivan, while 7 μg / mL for the pharmaceutical composition of the present invention. This result corresponds to an effective reduction of free propofol by more than two times for pharmaceutical compositions comprising propofol and albumin.

Example  30

This example illustrates the administration of a pharmaceutical composition comprising propofol and albumin to a human.

To compare the harmful skin detection of pharmaceutical compositions comprising propofol and albumin with those of the commercially available propofol formulation Diprivan, a randomized double-blind trial was conducted. The trial was conducted according to Good Clinical Practice, and consent was obtained from the subjects. Adults are eligible for this test as long as the back of the hand is clearly normal skin without any fracture of any gender.

The formulation, which was originally stored in the refrigerator, was allowed to come to room temperature, and then 10 μl of the formulation was slowly added simultaneously to the back of both hands of the subject. The overall response and feeling in the hand to the formulation was recorded. The results of this study are shown in Table 1.

For objects
Trial order % Of object for ABI-propofol detection % Of objects for Diprivan detection Mild fever or pain or pain stupor Mild fever or pain or pain stupor 1st application 0.0 100.0 75 25

Example  31

This example illustrates the use of diperoxamine as an antioxidant in a pharmaceutical composition comprising propofol.

Pharmaceutical compositions comprising propofol and albumin and containing tween or TPGS were stored at 4 ° C., 25 ° C., or 40 ° C. to test the effect of deferoxamine mesylate in inhibiting the oxidation of propofol. The concentration of propofol in these formulations was measured over time to determine the antioxidant activity of deferoxamine. The data is shown in Tables 2 and 3 as% potency relative to time zero.

Albumin / Twin Formulation 1 month storage Temperature 4 ℃ 25 ℃ 40 ℃ Control group 100% 88% 48% 0.01% Def 100% 89% 61% 0.1% Def 100% 89% 64%

Albumin / TPGS Formulation 1 month storage Temperature 4 ℃ 25 ℃ 40 ℃ Control group 99% 73% 42% 0.01% Def 99% 87% 55% 0.1% Def 99% 85% 58%

Under these conditions, deferoxamine is effective to reduce the oxidation level of propofol. The effect was enhanced at higher temperatures. There was no significant oxidation at 4 ° C. This study was performed using a non-inert or Teflon coated stopper.

Example  32

This example shows intrapulmonary delivery of a pharmaceutical composition (ABI-007) comprising paclitaxel and albumin.

The purpose of this study was to perform intratracheal instillation in Sprague Dawley rats and then determine the time course of [ ³ H] ABI-007 in blood and tissues.

The target volume of the endotracheal dose formulation administered to the animal was calculated based on a dose volume of 1.5 mL per kg of body weight. The dosing device consisted of a Penn-Century microspray attached to a 1-mL gas-tight luer-lock syringe (Model 1A-1B; Penn-Century, Inc., Philadelphia, PA; Purchased from Delong Distributors, Long Branch, NJ). The appropriate volume of the dosage formulation was drawn into the dosing device and the filled device was weighed to record the weight. The catheter is placed in the trachea of the anesthetized animal, the microspray portion of the administration device is placed in the trachea via the catheter and then dosed once. After administration of a single dose, the empty dose device was reweighed to calculate the dose administered as the weight difference before and after dose administration. The average dose of all animals was 4.7738 ± 0.0060 (CV 1.5059) mg paclitaxel per kg body weight.

About 250 μl of blood samples were collected from the intrinsic jugular vein cannula of JVC rats at the following previously determined dose-time points: 1, 5, 10, 15, 30, and 45 minutes (min), and 1, 4, 8 , And 24 hours (h). Blood samples obtained from animals sacrificed at 10 minutes, 45 minutes, and 2 hours, as well as 24-h blood samples were collected by cardiac puncture from anesthetized rats at sacrifice. All blood samples analyzed for total radioactivity were dispensed into pre-weighed sample tubes, and the sample tubes were weighed again and the weight of each sample was calculated by subtraction. Blood samples obtained from the jugular vein as well as 250 μl aliquots of blood collected from each animal at sacrifice were assayed for total tritium content.

In all rats, the maximum concentration of tritium in blood was observed 5 minutes (0.0833 hr) after administration. The elimination half-life of tritium, determined at time intervals of 4 to 24 hours, ranges from 19.73 hours to 43.02 hours. It should be noted that this interval contains only three data points, which can explain the variety of these parameters. The clear clearance of tritium from the blood is 0.04 L / h. The results of these experiments are shown in Table 4 below.

^[3 H] After my infusion engine of ABI-007 blood tritium concentration in rats vs. Profiles of non-part-time analysis of time parameter Average +/- SD C _max (mg-eqL) 1.615 +/- 0.279 T _max (hr) 0.0833 +/- 0.0 t1 / 2beta (hr) 33.02 +/- 1.99 AUC _Final (mg-eq x hr / L) 7.051 +/- 1.535 Cl / F (L / hr) 0.0442 +/- 0.0070 Fa (bioavailability) 1.229 +/- 0.268

To assess the bioavailability of tritium derived from intratracheal administration of [ ³ H] ABI-007, the mean blood concentration of [ ³ H] ABI-007-derived radioactivity was analyzed as a function of time. This analysis showed an AUC (AUC _final ) of 6.1354 mg-eq hr / L. Based on these data, radioactivity derived from intratracheal administration of [ ³ H] ABI-007 is highly bioavailable. This assay is based on total radioactivity.

Tritium derived from [ ³ H] ABI-007 is rapidly absorbed after intratracheal instillation. The mean absorption and elimination half-life (k ₀₁ half-life and k ₁₀ half-life) (mean +/- SD) for tritium in blood after intratracheal administration of [ ³ H] ABI-007 (0.055 +/- 0.0058 and 4.738 +, respectively) /-0.360 L / hr. The apparent clearance of tritium from the mean blood was 0.1235 +/- 0.0180 L / hr (see Table 4 above).

Tritium derived from [ ³ H] ABI-007 was absorbed and distributed after intratracheal administration. The time course of tritium in blood is well illustrated by a two-compartment model with mean absorption and elimination half-lives of 0.0155 and 4.738 hr, respectively. About 28% of the administered dose was recovered in the lung 10 minutes after intratracheal administration. At all time points, less than 1% of the dose was recovered from tissues other than the gastrointestinal tract.

Based on results from an intravenous dosing study already performed with [ ³ H] Capsol ^™ , the bioavailability of tritium derived from intratracheal administration was 1.229 ± 0.268 (mean ± ±) in three animals in this dosing group. SD). However, it should be understood that this assessment of bioavailability is based on total radioactivity. Surprisingly, paclitaxel administered to the pulmonary route with albumin in combination with the present invention allows for rapid bioavailability, indicating excellent transport through the pulmonary endothelial. No toxicity was seen in animals, which is surprising because intrapulmonary administration of cytotoxic substances is known to cause pulmonary toxicity.

Appropriate amount of radioactivity was present in the gastrointestinal tract (including the contents) 24 hours after administration (27% of intratracheal dose). The amount of tritium in the gastrointestinal tract may be due to bile excretion or clearance of tritium from the respiratory tract with mucus ciliary clearance and subsequent swallowing.

Example  33

This example shows the investigation of Arerotech II and Pari nebulizer for intrapulmonary administration of pharmaceutical compositions comprising paclitaxel and albumin.

This study was performed using paclitaxel-albumin containing pharmaceutical composition ABI-007 under the following conditions: room temperature (20-23 ° C.), relative humidity (48-54%), ambient pressure (629 mmHg), nebulizer flow rate ( 10 L / min for Aerotech II; 32 lb / in ^{2 for} Pari, operating time (15 to 60 seconds), sample volume (1.5 mL), ABI-007 paclitaxel concentration (5, 10, 15, and 20 mg / mL).

When ABI-007 was reconstituted to a concentration range of 5-15 mg / mL, both Aerotech II and Pari nebulizers provided suitable overall efficiencies (30-60%). The efficiency of the Pari Nebulizer was higher than that of the Aerotech II Nebulizer. Pari nebulizer efficiency decreased slightly with increasing ABI-007 concentration. Excellent fine particle fractions were observed (74% -96%). Aerotech II nebulizers had a finer particle fraction than Pari nebulizers. Fine particle fractions were not concentration dependent.

Pari nebulizer delivered 100 mg of paclitaxel in less than 30 minutes using a 15 mg / mL ABI-007 solution. Aerotech II Nebulizer delivered 100 mg of paclitaxel after about 65 minutes using either 10 mg / mL or 15 mg / mL of ABI-007 solution. Performance stability was tested for both Aerotech II and Pari nebulizers. Both aerosol concentrations and efficiencies were stable until the drug was exhausted. At 15 mg / mL, the Pari nebulizer consumed the drug at twice the rate of the Aerotech II nebulizer, producing a higher aerosol concentration than that of the Aerotech II nebulizer.

In conclusion, nanoparticle / albumin preparations of paclitaxel (ABI-007) show excellent bioavailability in rats when administered by intrapulmonary route. There was no obvious signal for early toxicity at the doses administered. Intrapulmonary delivery of nanoparticle paclitaxel (ABI-007) can be accomplished using conventional nebulizers.

Example  34

This example describes intrapulmonary administration of a pharmaceutical composition comprising albumin and rapamycin. The purpose of this study was to perform intratracheal instillation in Sprague Dawley rats and then to determine pulmonary uptake of rapamycin in the blood compared to intravenous instillation.

The target volume of the endotracheal formulation to be administered to the animals was calculated based on the dose volume of 1 mL per kg of body weight. The endotracheal administration device consisted of a Penn-Century microspray attached to a 1-mL gas-tight luer-lock syringe (Model 1A-1B; Penn-Century, Inc., Philadelphia, PA; purchased from DeLong Distributors, Long Branch, NJ). The appropriate volume of the dosage formulation was drawn into the dosing device and the filled device was weighed to record the weight. The catheter is placed in the trachea of the anesthetized animal, the microspray portion of the administration device is placed in the trachea via the catheter and then dosed once. After administration of a single dose, the empty dose device was reweighed to calculate the dose administered as the weight difference before and after dose administration.

About 250 μl of blood samples were collected from the rat's intrinsic jugular vein cannula at the following previously determined dose-time points: 1, 5, 10, 15, 30, and 45 minutes (min), and 1, 4, 8, And 24 hours (h). All blood samples were dispensed into previously weighed sample tubes, and the sample tubes were weighed again and the weight of each sample was calculated by subtraction. Collected blood samples were assayed for total rapamycin concentration using LC / MS / MS.

Surprisingly, the results showed no significant difference in blood levels of rapamycin administered intravenously and rapamycin administered intravenously. The bioavailability of rapamycin administered by the intrapulmonary route using a pharmaceutical composition comprising albumin was calculated to be 109%, indicating a good transport of the drug through the pulmonary endothelial.

Example  35

This example shows the tissue distribution of albumin-rapamycin after intrapulmonary administration of a pharmaceutical composition comprising rapamycin and albumin prepared according to the present invention. The purpose of this study was to determine the uptake of rapamycin in the tissues after intratracheal instillation in Sprague-Dawley rats compared to intravenous instillation.

The target volume of the endotracheal formulation to be administered to the animals was calculated based on the dose volume of 1 mL per kg of body weight. The dosing device consisted of a Penn-Century microspray attached to a 1-mL gas-tight luer-lock syringe (Model 1A-1B; Penn-Century, Inc., Philadelphia, PA; Purchased from DeLong Distributors, Long Branch, NJ). The appropriate volume of the dosage formulation was drawn into the dosing device and the filled device was weighed to record the weight. The catheter is placed in the trachea of the anesthetized animal, the microspray portion of the administration device is placed in the trachea via the catheter and then dosed once. After administration of a single dose, the empty dose device was reweighed to calculate the dose administered as the weight difference before and after dose administration.

Samples were collected from three rats per brain, lung, and liver at 10, 45, 2, and 24 hour points. The sample was collected and analyzed for total rapamycin concentration using LC / MS / MS. The results showed that the concentration of rapamycin in the lung tissue was higher than intravenous delivery when delivered into the lung. However, the total concentration in the brain was lower at intratracheal administration (IT) compared to intravenous administration (IV). In the liver, IT or IV delivery showed no difference in rapamycin concentrations. Based on these results, intrapulmonary administration of rapamycin would be appropriate for the treatment of diseases where high local concentrations of rapamycin are beneficial (eg lung transplantation).

Example  36

This example illustrates oral administration of a pharmaceutical composition (ABI-007) comprising paclitaxel and albumin.

Rats were subjected to oral gabage using tritiated ABI-007 and then determined oral bioavailability of paclitaxel. After fasting overnight, 5 rats group A) received 5.5 mg / kg of paclitaxel in ABI-007 and another 5 rats (group B) were pretreated with cyclosporine (5.0 mg / kg) and then ABI 007 mg / kg paclitaxel was administered. Pharmacokinetic analysis of blood samples taken at 0.5, 1, 2, 3, 4, 5, 6, 8, 12, and 24 hours was performed after radioactivity was determined by burning in blood samples. Oral bioavailability was determined by comparing the intravenous data already obtained. The results are shown in Table 5 below.

It was used to calculate _{AUC 0 -24 IV (6.06 ㎍ x} hr./mL) and IV doses (5.1 gmkg) (data based on the IV dose of ABI-0007) percent absorption.

ABI-007 alone showed 44% oral bioavailability. This is significantly larger than other paclitaxel preparations. Bioavailability increased to 121% when the animals were treated with cyclosporin (CsA). This is expected to be because CsA is a known inhibitor of the p-glycoprotein pump that normally inhibits the absorption of compounds such as paclitaxel from the gastrointestinal tract. Bioavailability greater than 100% can be explained by reuptake after paclitaxel's bile excretion into the gastrointestinal tract. Other known inhibitors or absorption enhancers can also be used for this purpose.

Example  37

This example shows an improvement in the infiltration of paclitaxel into erythrocytes and tumor cells upon administration of a pharmaceutical composition comprising paclitaxel and albumin.

Human MX-1 breast cancer fragments were implanted subcutaneously into athymic mice. Pharmaceutical compositions comprising paclitaxel and albumin (“paclitaxel-albumin”) and Taxol as described above were prepared to have ³ H paclitaxel with a specific activity of 25 μCi / mg paclitaxel. When the tumor volume reached about 500 mm ³ , paclitaxel-albumin or taxol labeled with 20 mg / kg of the radioisotope was administered intravenously in saline. Plasma, blood, and tumor tissues were sampled and analyzed for radioactivity at 5, 15, and 30 minutes and 1, 3, 8, and 24 hours post dose. Tumor pharmacokinetic constants (AUC and absorption constant) were analyzed using WiNonlinn, Pharsight, USA.

Paclitaxel-albumin showed rapid distribution to red blood cells (RBCs) as indicated by the rapid decrease in the plasma / blood radioactivity ratio after intravenous administration of the drug. Full distribution to RBCs appeared early about 1 hour after paclitaxel-albumin administration. In contrast, the distribution of paclitaxel formulated with taxol was much slower and not complete after more than 8 hours.

Paclitaxel-albumin showed rapid distribution to tumor tissue with an absorption constant (K _a ) 3.3 times greater than Taxol. K _a is paclitaxel - it was ^1-2 albumin and Taxol respectively 0.43 hr ^-1 and 0.13 hr. Rapid uptake of paclitaxel resulted in tumor AUC 33% higher for paclitaxel-albumin than taxol. The AUC was 3632 nCi * hr / g and 2739 nCi * hr / g for paclitaxel-albumin and taxol, respectively.

Example  38

This example shows the stability of pharmaceutical compositions comprising paclitaxel and albumin administered to mice.

Athymic mice were treated with paclitaxel-albumin and taxol with increasing doses for 5 consecutive days. Survival was indicated for dose to determine LD ₅₀ . Survival was significantly improved for paclitaxel-albumin compared to Taxol (p = 0.017, ANOVA). LD ₅₀ of paclitaxel-albumin and taxol was calculated to be 47 mg / kg / day and 30 mg / kg / day, respectively, on the q1d × 5 schedule. At the dose level of 13.4 mg / kg / day, both paclitaxel-albumin and Taxol were well tolerated with 1% lethality (1 death in 72 mice) and 4% (2 deaths in 47 mice), respectively. (well tolerated). At the dose level of 20 mg / kg / day, paclitaxel-albumin mortality was 1% (1 death in 72 mice) compared to 17% (8 deaths in 47 mice) compared to Taxol. (p = 0.0025). At a dose level of 30 mg / kg / day, paclitaxel-albumin mortality was 4% (3 deaths in 72 mice) compared to 49% (23 deaths in 47 mice) compared to Taxol. (p <0.0001).

Example  39

This example shows a novel paclitaxel transport mechanism through microtubule endothelial cells (EC) of the paclitaxel-albumin composition.

Nanoparticles and albumin-paclitaxel compositions can accumulate in tumor tissue by EPR effects resulting from 'leaking' blood vessels in tumors. Albumin specific gp60 receptors (albondines) transport albumin through the EC by transcytosis of the receptors in the caveolae of the cell surface. This transcytosis mechanism allows transport to the interstitial space just below albumin-paclitaxel. In contrast, cremophores in Taxol inhibit paclitaxel binding to albumin, significantly reducing the transport of paclitaxel to tumors. In addition, the gp16 and gp30 receptors are involved in the transport of altered albumin-containing binding paclitaxel, which increases the binding of paclitaxel to endothelial cells, resulting in a significant antiangiogenic effect compared to taxol.

Example  40

This example shows an increase in endothelial transcytosis of pharmaceutical compositions comprising paclitaxel and albumin compared to Taxol.

Human Lung Microvessel Cells (HLMVEC) were grown to confluence in clear wells. The pharmaceutical composition of the invention comprising paclitaxel and albumin or Fultax containing fluorescent paclitaxel was added to the upper transparent well chamber at a concentration of 20 μg / mL.

The transport of paclitaxel by transcytosis from the upper chamber to the lower chamber was continuously monitored using a fluorometer. A control containing only Flutax without albumin was also used. Controls with Flutax showed no transport at all and demonstrate completeness of the confluent HLMVEC monolayer. In the presence of 5% HSA (physiological concentration), the transport of paclitaxel from the albumin-paclitaxel composition was much faster than paclitaxel from Taxol. Albumin-transport rate constant (K _t) of a paclitaxel composition and Taxol are each 1.396 hr ^-1 and 0.03 ^hr-1. The total amount of paclitaxel transported through the monolayer was three times greater for albumin-paclitaxel than for taxol.

Example  41

This example shows that pharmaceutical compositions comprising paclitaxel and albumin improve endothelial cell (EC) binding as compared to taxol.

Human Umbilical Vein Endothelial Cells (HUVECs) were grown on 96-well microtiter plates. In one experiment, pataxtaxel (Flutax-Oregon Geen labeled paclitaxel) was reacted with HUVEC with increasing concentrations of Taxol's medium, Cremophor EL-EtOH. In another experiment, pharmaceutical compositions comprising albumin and Flutax and Taxol-Flutax compositions were reacted with HUVEC at various final concentrations. Binding of paclitaxel to cells was inhibited by Cremophor. Inhibition showed an IC ₅₀ of Cremophor EL / EtOH of 0.02%. This concentration of Cremophor has been shown to persist for more than 24 hours of Taxol treatment. Therefore, it is an in vivo related process. At all concentrations tested, significant amounts of paclitaxel from the albumin-paclitaxel composition were bound to the cells. In contrast, in the case of Taxol, little or no binding was observed.

Example  42

This example shows that pharmaceutical compositions comprising paclitaxel and albumin improve albumin binding over taxol.

Human serum albumin (HSA) was immobilized on an ELISA plate. Fataxtaxel (Flutax-Oregon Geen labeled paclitaxel) was reacted with fixed HSA with increasing concentration of Cremophor EL-EtOH. In another experiment, albumin-paclitaxel-Flutax composition and Taxol-Flutax composition were reacted with fixed HSA at a final concentration of 20 μg paclitaxel / mL. The binding of paclitaxel to albumin was inhibited by Cremophor. Inhibition showed an IC ₅₀ of Cremophor EL / EtOH of 0.003%. This concentration of Cremophor has been shown to persist for more than 24 hours of Taxol treatment. Therefore, it is an in vivo related process. At the relevant pharmacological paclitaxel concentration (20 μg / mL), a significant amount of paclitaxel was bound to immobilized HSA from the albumin-paclitaxel composition. In contrast, in the case of Taxol, little or no binding was observed.

Example  43

This example shows that pharmaceutical compositions comprising paclitaxel and albumin improve paclitaxel transport to albumin over taxol.

Taxol-Flutax and Albumin-Paclitaxel-Flutax compositions were mixed with one of 5% HSA or serum in Hanks buffer at concentrations of 20 μg / mL, 40 μg / mL, and 80 μg / mL. The mixture was separated on a natural 3-14% polyacrylamide gel and the amount of paclitaxel bound to albumin was determined by scanning fluorometer. The transport of paclitaxel to HSA is faster for albumin-paclitaxel compositions compared to Taxol. When serum or 5% HSA was incubated with albumin-paclitaxel-Flutax composition or Taxol-Flutax composition, more paclitaxel was electrophoresed with HSA. Upon exposure to 5% HSA, the albumin-paclitaxel-Flutax composition was 45%, 60%, and 33% at 20 μg / mL, 40 μg / mL, and 80 μg / mL, respectively, compared to the taxol-Flutax composition. More paclitaxel was transported to HSA. Upon exposure to human serum, the albumin-paclitaxel-Flutax composition was 121%, 31%, and 83% more at 20 μg / mL, 40 μg / mL, and 80 μg / mL, respectively, than the taxol-Flutax composition. Many paclitaxel was transported to HSA. The C _max of ABI-007 at 260 mg / m ² is about 20 μg / mL, therefore this is an important in vivo process.

Example  44

This example shows that the glycoprotein receptor gp60 is responsible for the binding and transcytosis of albumin-paclitaxel.

Fluorescent label paclitaxel albumin composition was contacted with microvascular endothelial cells in culture. Fluorescent staining was observed under the microscope, which was considered the gp60 receptor binding albumin-paclitaxel. This was confirmed by using rhodamine-labeled albumin, which is localized with highlighted fluorescence of paclitaxel.

Example  45

This example shows that increasing the amount of albumin can compete with the binding of paclitaxel.

Albumin was immobilized on microtiter plates. Fluorescent paclitaxel was added to the wells and binding of paclitaxel was measured using a scanning fluorometer. Increasing the amount of albumin was added to the wells, and the extent to which paclitaxel binding to immobilized albumin was inhibited was measured. The data showed that as the amount of added albumin increased, binding decreased. Similar effects were observed in binding to endothelial cells. This indicates that high concentrations of albumin inhibit the binding of paclitaxel. Therefore, the composition of the present invention is preferably a composition having a lower amount of albumin.

Example  46

This example shows that a lower amount of albumin in the pharmaceutical composition of the present invention results in a stable composition.

To investigate whether a smaller amount of albumin in the composition affects the stability of the pharmaceutical composition of the present invention, an albumin-paclitaxel composition with a small amount of albumin was prepared. Months of investigation of the titer of paclitaxel, impurity production, particle size, pH, and other typical stability parameters have been found to be as stable as compositions with higher amounts of albumin. Thus, compositions with lower amounts of albumin are preferred because they not only increase transport and binding to cells, but also significantly reduce cost.

Example  47

This example shows a composition comprising albumin and paclitaxel having a high ratio of albumin to paclitaxel.

30 mg of paclitaxel was dissolved in 3.0 mL of methylene chloride. The solution was added to 27.0 mL of human serum albumin (3% w / v) (the ratio of albumin to paclitaxel was 27). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest I.Q.) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the methylene chloride was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting paclitaxel particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization.

It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of paclitaxel dissolved in the cremophore formulation, the pharmaceutical compositions of the invention containing albumin exhibited substantially lower toxicity.

Example  48

This example shows a composition comprising albumin and paclitaxel having a low ratio of albumin to paclitaxel.

Specifically, 30 mg of paclitaxel was dissolved in 3.0 mL of methylene chloride. The solution was added to 27.0 ml of human serum albumin solution (5% w / v) (the ratio of albumin to paclitaxel was 4.5). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest I.Q.) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the methylene chloride was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting paclitaxel particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization.

It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of paclitaxel dissolved in the cremophore formulation, the pharmaceutical compositions of the invention containing albumin exhibited substantially lower toxicity.

Example  49

This example shows a composition comprising albumin and paclitaxel having a medium ratio of albumin to paclitaxel.

Specifically, 135 mg of paclitaxel was dissolved in 3.0 mL of methylene chloride. The solution was added to 27.0 ml of human serum albumin solution (5% w / v). Deferoxamine was added essentially. The mixture was homogenized for 5 minutes at low RPM (Vitris homogenizer, model Tempest I.Q.) to form crude, and then transferred to high pressure homogenizer (Avestin). The emulsification was carried out at 9000-40,000 psi while recycling the emulsion for at least 5 cycles. The resulting system was then transferred to a rotary evaporator and the methylene chloride was quickly removed at 40 ° C. under reduced pressure (30 mm Hg) for 20-30 minutes. The resulting dispersion was translucent and the typical average diameter of the resulting paclitaxel particles was in the range of 50-220 nm (Z-mean, Malvern Zetasizer). The dispersion was further lyophilized for 48 hours. The resulting cake was easily reconstituted by adding sterile water or saline to give the original dispersion. The particle size after reconstitution was the same as before lyophilization. The calculated ratio (w / w) of albumin to paclitaxel in the compositions of the present invention is about 10.

It should be understood that the amounts, types, and ratios of drugs, solvents, and proteins used in these examples are not intended to be limiting in any way. Compared to the toxicity of paclitaxel dissolved in the cremophore formulation, the pharmaceutical compositions of the invention containing albumin exhibited substantially lower toxicity.

Example  50

This example shows the treatment of rheumatoid arthritis with an albumin-paclitaxel composition in an animal model.

A collagen-induced arthritis model in Louvain rats was used to test the therapeutic effect of albumin-paclitaxel composition on arthritis. Foot size in experimental animals was monitored to monitor the severity of arthritis.

After sufficient arthritis has developed (usually ˜9-10 days after collagen injection), the experimental animals are intraperitoneally injected 6 times with albumin-paclitaxel 1 mg / kg qod, or albumin-paclitaxel 0.5 mg / kg + prednisone 0.2 mg / kg qod (Complex therapy) was divided into different groups that were administered once weekly for 3 weeks following administration. Foot size was measured at the start of treatment (day 0) and each time a drug was injected. Only one group received normal saline as a control. At the end of the experiment, the group receiving albumin-paclitaxel had a 42% reduction in foot size compared to when treatment was initiated, and the combination treatment group had a 33% reduction in foot size, whereas the control group increased about 20% in foot size. It was.

In conclusion, albumin-paclitaxel compositions demonstrated a therapeutic effect against arthritis. Albumin-paclitaxel combinations seem to localize arthritis lesion sites through receptor-mediated mechanisms such as gp60.

Example  51

This example shows the use of albumin-paclitaxel compositions for treating cardiovascular restenosis.

Stents secreting paclitaxel in animals indicate incomplete treatment and in some cases result in a lack of sustained inhibition of neointimal proliferation in the arteries. In this study, we tested the efficacy of a new system-delivered albumin-paclitaxel composition to reduce in-stent restenosis.

Saline-reconstituted albumin-paclitaxel was tested in 38 New Zealand White rabbits receiving symmetrical iliac artery stents. A dose of albumin-paclitaxel (a paclitaxel dose of 1.0-5.0 mg / kg) was administered as an intraarterial infusion for 10 minutes and control animals received medium (0.9% saline).

In the follow-up long-term experiment, 5.0 mg / kg of albumin-paclitaxel was given to stents with or without 3.5 mg / kg repeat albumin-paclitaxel intravenously on day 28; This study was terminated at 3 months. On day 28, neointimal thickness was reduced by administration of albumin-paclitaxel> = 2.5 mg / kg with evidence of delayed treatment (p <= 0.02). However, the efficacy of albumin-paclitaxel 5.0 mg / kg single dose was lost at 90 days. In contrast, repeated administration of 3.5 mg / kg of second albumin-paclitaxel administered after 28 days resulted in a sustained inhibition of neointimal thickness with nearly complete neointimal healing on day 90 (p <= 0.009 vs. albumin-). Paclitaxel 5.0 mg / kg single dose and control).

Although system albumin-paclitaxel reduced neointimal proliferation on day 28, a single repeated dose was required for sustained neoendothelial inhibition. Thus, the compositions of the present invention are suitable for the treatment of cardiovascular diseases such as restenosis.

Compositions of the present invention comprising paclitaxel other than pharmaceutical substances, such as rapamycin, other taxanes, epotyrones and the like, may be used for arterial-vein access in patients requiring stenosis or hemodialysis in blood vessels. It is suitable for the treatment of restenosis in artificial vascular grafts as used.

Claims (46)

A pharmaceutical composition comprising a pharmaceutical substance and a pharmaceutically acceptable carrier, wherein the pharmaceutically acceptable carrier comprises albumin, wherein the ratio of albumin to pharmaceutical agent (w / w) in the pharmaceutical composition is A pharmaceutical composition that is 18: 1 or less. The pharmaceutical composition of claim 1, wherein the ratio of albumin to pharmaceutical agent (w / w) in the pharmaceutical composition is 12: 1 or less. The pharmaceutical composition of claim 2, wherein the ratio (w / w) of albumin to the pharmaceutical substance in the pharmaceutical composition is 9: 1 or less. The pharmaceutical composition of claim 3, wherein the ratio of albumin to pharmaceutical agent in the pharmaceutical composition is 1: 1 to 9: 1. The pharmaceutical composition of claim 4, wherein the ratio of albumin to pharmaceutical agent in the pharmaceutical composition is 9: 1. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises 0.1% to 25% by weight of albumin. The pharmaceutical composition of claim 6, wherein the pharmaceutical composition comprises 0.5% to 5% by weight of albumin. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises nanoparticles comprising albumin and a pharmaceutical substance. The pharmaceutical composition of claim 8, wherein the nanoparticles in the composition have a size of less than 200 nm. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition comprises nanoparticles comprising albumin and a pharmaceutical substance. The pharmaceutical composition of claim 10, wherein the nanoparticles in the composition have a size of less than 200 nm. The pharmaceutical composition of claim 1, wherein the pharmaceutical substance is taxane. The pharmaceutical composition of claim 12, wherein the taxane is paclitaxel. The pharmaceutical composition of claim 12, wherein the taxane is doxytaxel. The pharmaceutical composition of claim 11, wherein the taxane is paclitaxel. The pharmaceutical composition of claim 9, wherein the albumin is human serum albumin. The pharmaceutical composition of claim 15, wherein the albumin is human serum albumin. The pharmaceutical composition of claim 17, wherein the ratio (w / w) of albumin to paclitaxel in the pharmaceutical composition is 9: 1. 10. The pharmaceutical composition of claim 9, wherein the pharmaceutical composition does not contain polyethers of castor oil and ethylene oxide. The pharmaceutical composition of claim 15, wherein the pharmaceutical composition does not contain polyethers of castor oil and ethylene oxide. The pharmaceutical composition of claim 1, wherein the pharmaceutical agent is rapamycin. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is contained in a sealed container. The pharmaceutical composition of claim 1, wherein the composition is a unit dose. The pharmaceutical composition of claim 1, wherein the composition is in multiple doses. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is sterile. The pharmaceutical composition of claim 1, wherein the composition is dehydrated. The pharmaceutical composition of claim 1, wherein the composition is lyophilized. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is a liquid composition. The pharmaceutical composition of claim 28, wherein the pharmaceutical composition further comprises saline. The pharmaceutical composition of claim 28, wherein the pH of the composition is from 5.0 to 8.0. 31. The pharmaceutical composition of any one of claims 1-30, wherein the pharmaceutical composition is formulated for use in the treatment of a disease. The pharmaceutical composition of claim 31, wherein the disease is cancer. The pharmaceutical composition of claim 32, wherein the cancer is breast cancer. The pharmaceutical composition of claim 32, wherein the cancer is lung cancer. The pharmaceutical composition of claim 32, wherein the cancer is pancreatic cancer. The pharmaceutical composition of claim 32, wherein the cancer is ovarian cancer. 31. The pharmaceutical composition of any one of claims 1-30, wherein the pharmaceutical composition is formulated for use in enhancing the transport of the pharmaceutical substance to cells. 31. The pharmaceutical composition of any one of the preceding claims, wherein the pharmaceutical composition is formulated for use in enhancing binding of the pharmaceutical substance to cells. The pharmaceutical composition of claim 37, wherein the cell is an endothelial cell. The pharmaceutical composition of claim 38, wherein the cell is an endothelial cell. 31. The pharmaceutical composition of any one of claims 1-30, wherein the pharmaceutical composition is formulated for use in reducing one or more side effects associated with administering the pharmaceutical substance to a human. The pharmaceutical composition of any one of claims 1 to 30, wherein the pharmaceutical composition is intravenous, intraarterial, pulmonary, oral, inhaled, alveolar, intramuscular, intratracheal, subcutaneous, intraocular, intrathecal, Or pharmaceutical compositions for use in transdermal administration. The pharmaceutical composition of claim 42, wherein the pharmaceutical composition is for use in intravenous administration. 31. The pharmaceutical composition of any one of claims 1-30, wherein the pharmaceutical composition is for use in humans. 31. The pharmaceutical composition of any one of claims 1-30, wherein the pharmaceutical composition is for injection. A method of making the pharmaceutical composition of claim 1, comprising combining the pharmaceutical agent such that the ratio (w / w) of the albumin to the pharmaceutical agent is 18: 1 or less.