CN1925874B

CN1925874B - Compositions and methods of delivery of pharmacological agents

Info

Publication number: CN1925874B
Application number: CN200380109606.9A
Authority: CN
Inventors: N·P·德赛; A·杨; S·X·司; T·德; V·德留; P·宋祥; B·比尔斯·格里姆; 姚强
Original assignee: Abraxis Bioscience LLC
Current assignee: ABRAXIS BIOSCIENCE Inc; Abraxis Bioscience LLC
Priority date: 2002-12-09
Filing date: 2003-12-09
Publication date: 2014-12-17
Anticipated expiration: 2023-12-09
Also published as: ZA200505380B; CN1925874A

Abstract

The present invention relates to a pharmaceutical composition comprising a pharmaceutical agent and a pharmaceutically acceptable carrier, which carrier comprises a protein, for example, human serum albumin and/or deferoxamine. The 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 methods for reducing one or more side effects of administration of the pharmaceutical composition, methods for inhibiting microbial growth and oxidation in the pharmaceutical composition, and methods for enhancing transport and binding of a pharmaceutical agent to a cell.

Description

The method of compositions and delivery of pharmacological agents

To the cross reference of related application

The U.S. Provisional Patent Application 60/432,317 that patent application claims proposes at December in 2002 on the 9th, the U.S. Provisional Patent Application (lawyer's number of putting on record 225519) proposed for 3rd at December in 2003, the U.S. Provisional Patent Application (lawyer's number of putting on record 225549) proposed for 4th at December in 2003 and the interests of U.S. Provisional Patent Application (lawyer's number of putting on record 225585) proposed for 5th at December in 2003.

Invention field

The present invention relates to for parenteral or other inner pharmaceutical compositions using, comprise pharmaceutically active agents, there is when it compares with the similar medicine preparation that can obtain the effect of some undesirable side effect when being reduced in administration.

Background of invention

Generally acknowledge many those of medicine, particularly intravenous administration used for parenteral, cause undesirable side effect as venous stimulation, phlebitis, burn feeling during injection and pain, venous thrombosis, blend the side effect that other administration is relevant outward.Many water insoluble in these medicines, therefore prepare with solubilizing agent, surfactant, solvent and/or emulsifying agent, these solubilizing agents, surfactant, solvent and/or emulsifying agent are irritating, allergic or poisonous (see such as when administered to a patient, Briggs etc., Anesthesis 37,1099 (1982), with Waugh etc., Am.J.Hosp.Pharmacists, 48,1520 (1991)).Often, free drug induction pain or the stimulation upon administration in preparation is present in.Such as, accept in the patient of peripheral vein administration ifosfamide and vinorelbine in the line chemotherapy as advanced Non-small cell lung, 50% in patient observed phlebitis.(see such as Vallejo etc., Am.J.Clin.Oncol., 19 (6), 584-8 (1996)).In addition, vancomycin induction side effect has been shown as phlebitis (see such as, Lopes Rocha etc., Braz.J.Infect.Dis., 6 (4), 196-200 (2002)).Use in patients with solid tumor cisplatin, gemcitabine and SU5416 caused adverse events as DVT formed and phlebitis (see such as, Kuenen etc., J.Clin.Oncol., 20 (6), 1657-67 (2002)).In addition, propofol (a kind of anesthetis) can induction pain when injecting, burn feeling and venous stimulation, particularly when the lipomul administration stable as lecithin (see such as, Tan etc., Anathesia, 53,468-76, (1998)).Medicine of other display administration related side effects comprise such as taxol (paclitaxel) (see such as, the package insert I.V. of taxol), codarone (Amiodarone Hydrochloride) is (see such as, the package insert of Codarone), thyroid hormone T3 or liothyronine (can be used as Triostat to be purchased), phosphinothioylidynetrisaziridine, bleomycin, and diagnostic radiation contrast agent.

Another problem relevant to preparing injectable drug, particularly water-insoluble drug be guarantee aseptic.By before preparation by all components absolute sterilization, then can be completed the aseptic preparation of pharmaceutical emulsion/dispersion by absolutesterility technology in all preparatory phases.But these methods are consuming time and expensive.In addition, at the pH that preparation or lay up period can cause such as reducing by being exposed to air and oxidation of drug formulations, drug degradation, and variable color, make pharmaceutical preparation unstable and/or reduce shelf-life thus.

In order to avoid the problem that pharmaceutical preparation is relevant with the side effect that administration is correlated with, attempt the preparation replaced.Such as about propofol, the method reducing the pain of propofol induction comprises the fat content (such as long chain triglycerides (LCT)) increasing solvent, premedication, use non-steroid drug pretreatment, local anesthesia, opioid, add lignocaine, add cyclodextrin, with microfiltration (see such as, Mayer etc., Anaesthesist, 45 (11), 1082-4 (1996), Davies, Deng Anaesthesia, 57, 557-61 (2002), Doenicke, Deng, Anaesth.Analg., 82, 472-4 (1996), Larsen etc., Anaesthesitis, 50, 842-5 (2001), Lilley etc., Anaesthesia, 51, 815-8 (1996), Bielen etc., Anaesth.Analg., 82 (5), 920-4 (1996), with Knibbe etc., Br.J.Clin.Pharmacol., 47 (6), 653-60 (1999)).But these preparations are induced other side effect (such as cardiovascular complication) or are caused the instability of propofol formulations.

In order to overcome the problem of germ contamination, used antibacterial as EDTA equivalent (as edetate), pentetate or the reagent exploitation propofol formulations containing sulphite, or by their with lower pH preparation (see such as, United States Patent (USP) 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 no WO 99/39696).But because edetate and pentetate are metal ion chelation agents, they have the dangerous potentiality removing essential metal ion from soma.In addition, sulphite is added pharmaceutical preparation and bring potential detrimental effect to pediatric population with to those in the general population of sulfur sensitive.

Therefore, the compositions and the method that reduce or eliminate the side effect relevant with parenteral or vivo medicine-feeding medicine is still needed.Also need the method for aseptic pharmaceutical composition and this compositions of preparation.In addition, need to reduce or eliminate pharmaceutical preparation oxidation with the pharmaceutical composition and the method that prevent medicine instability.

The invention provides these compositionss and method.These and other advantage of the present invention and other inventive features become obvious by from the description of the invention provided here.

Summary of the invention

The invention provides the various embodiments of pharmaceutical composition.One in various embodiment, some or all of performance can find and still belong in the scope of appended claim in the embodiment that the present invention is different.

The invention provides a kind of pharmaceutical composition comprising medicament and pharmaceutical carrier, wherein said pharmaceutical carrier comprises protein as albumin, more preferably human serum albumin, its amount effectively reduces the side effect that pharmaceutical composition is delivered medicine to people by one or more, and wherein said pharmaceutical carrier comprises deferoxamine, it measures the growth of microorganism in effective depressant compositions.The present invention goes back the pharmaceutical composition that providing package contains medicament and pharmaceutical carrier, wherein said pharmaceutical carrier comprises protein as albumin, its amount effectively reduces the side effect that pharmaceutical composition is delivered medicine to people by one or more, and wherein said pharmaceutical carrier comprises deferoxamine, it measures the oxidation in effective depressant compositions.

The invention provides and reduce one or more and method pharmaceutical composition being delivered medicine to the relevant side effect of people, the method comprises (a) comprises medicament and pharmaceutical composition from pharmaceutical carrier to people's administration, and wherein said pharmaceutical carrier comprises albumin and deferoxamine.The present invention is also provided for suppressing growth of microorganism in pharmaceutical composition, or inhibited oxidation, or suppresses the method for growth of microorganism and oxidation.These methods comprise the pharmaceutical composition that preparation comprises medicament and pharmaceutical carrier, and wherein said pharmaceutical carrier comprises deferoxamine with the amount of growth of microorganism in effective depressant compositions or with the amount of effective depressant compositions oxidation.

The present invention is also provided for the method that enhancing medicament is transported to weakness, the method comprises the pharmaceutical composition comprising medicament and pharmaceutical carrier to people's administration, wherein said pharmaceutical carrier comprises albumin, and the ratio of medicament wherein in albumin and pharmaceutical composition is about 18: 1 or less.The present invention further provides the method be combined with external or cells in vivo for strengthening medicament, the method comprises the pharmaceutical composition comprising medicament and pharmaceutical carrier to described external or cells in vivo administration, wherein said pharmaceutical carrier comprises albumin, and the ratio of medicament wherein in albumin and pharmaceutical composition is about 18: 1 or less.

The present invention goes back the pharmaceutical composition of providing package containing medicament and pharmaceutical carrier, and wherein said pharmaceutical carrier comprises albumin with the amount effectively increasing medicine and transport to people's weakness, and wherein the ratio of albumin and medicament is about 18: 1 or less.

The present invention further provides the method by described medicament and protein group incompatible increase medicament being transported to external or cells in vivo, specific cell surface receptors on cell described in wherein said protein bound, the combination of wherein said protein-pharmaceutical agent causes transporting with the combination of described receptor and occurs, and wherein the ratio of albumin and medicament is about 18: 1 or less.

Detailed Description Of The Invention

The invention provides a kind of pharmaceutical composition comprising medicament and pharmaceutical carrier, wherein said pharmaceutical carrier comprises protein as albumin, preferred human serum albumin, its amount effectively reduces the side effect that pharmaceutical composition is delivered medicine to people by one or more, and wherein said pharmaceutical carrier comprises deferoxamine, it measures the growth of microorganism in effective depressant compositions.The present invention goes back the pharmaceutical composition that providing package contains medicament and pharmaceutical carrier, wherein said pharmaceutical carrier comprises protein as albumin, its amount effectively reduces the side effect that pharmaceutical composition is delivered medicine to people by one or more, and wherein said pharmaceutical carrier comprises deferoxamine, it measures the oxidation in effective depressant compositions.

Any suitable medicament may be used in pharmaceutical composition of the present invention.Suitable medicament includes but not limited to anticarcinogen or antineoplastic agent, anti-microtubule agent, immunosuppressant, anesthetis, hormone, for the medicament of cardiovascular disorder, antiarrhythmics, antibiotic, antifungal, antihypertensive, antiasthmatic drug, analgesic, anti-inflammatory agent, arthritis agent, and vasoactive agent.The present invention is also effective with many other medicines classifications.More specifically, suitable medicament includes but not limited to taxanes, (such as taxol

(paclitaxel), and taxotere ^tM(docetaxel)), Epothilones (epothilones), camptothecine, colchicine, amiodarone, thyroid hormones, vasoactive peptide (such as vasoactive intestinal peptide), amphotericin, corticosteroid, propofol, melatonin, cyclosporin, rapamycin (sirolimus), tacrolimus, Mycophenolic Acid, ifosfamide, vinorelbine, vancomycin, gemcitabine, SU5416, phosphinothioylidynetrisaziridine, bleomycin, diagnostic radiation contrast agent, and their derivant.The other medicines that may be used for the present composition are described in the U.S. Patent Application No. 09/446,783 of such as United States Patent (USP) 5,916,596 and common pending trial, and preferably, medicament is propofol, paclitaxel, or docetaxel.More preferably, medicament is propofol or paclitaxel.Most preferably, medicament is propofol.

Taxol

(paclitaxel) (Bristol-Myers Squibb) has activity to ovarian cancer, breast carcinoma, pulmonary carcinoma, the esophageal carcinoma and head and neck cancer.But taxol has shown the induction toxicity relevant with administration, and acute and cumulative toxicity significantly, as bone marrow depression, neutropenic fever, anaphylaxis, and peripheral neuropathy.Because paclitaxel is insoluble in water, typically by polyoxyethylene castor oil (cremophor) as solvent, need large volume infused and special conduit and filter.Polyoxyethylene castor oil is relevant to side effect, these side effect can be serious, comprise anaphylaxis and other anaphylactic reaction, this may require with corticosteroid, antihistamine and the pretreatment of H2 blocker (see such as, Gelderblom etc., Eur.J of Cancer, 37,1590-1598, (2001)).Taxotere ^tM(docetaxel) is used for the treatment of the breast carcinoma of anti-anthracycline, but has also shown the side effect of induced hypersensitivity and fluid retention, and this may be serious.Epothilones (and derivant) also typically administration in polyoxyethylene castor oil, and shown the serious Neutrophilic granulocytopenia of induction, irritated and neuropathy.

Propofol (2,6-Bis(1-methylethyl)phenol) is hydrophobic, water-insoluble oil, is widely used as intravenous anesthetic to induce and keeps general anesthesia and the calmness of humans and animals.Propofol typically directly to deliver medicine in blood flow and passes through blood brain barrier.The pharmaceutical composition comprising propofol must have enough fat-soluble with by this barrier and the related mechanism suppressing brain.Propofol is 1.0+/-0.02 μM (see such as, Tonner etc., Anesthesiology, 77,926-931 (1992)) at 22.5 DEG C of maxima solubilities had in water.Therefore, propofol usually used as the Emulsion preparation containing solubilizing agent, surfactant, solvent, or as oil-in-water emulsion preparation (see such as, U.S. patent 6,150,423,6,326,406 and 6,362,234).Except active agents, compositions of the present invention also comprises pharmaceutical carrier or excipient.The selection of carrier is not necessarily crucial, and any carrier known in the art may be used in compositions.The selection of carrier is preferably partially by the concrete position of pharmaceutical composition administration with determine for the concrete grammar of administration medicine compositions.Preferably, pharmaceutical carrier comprises protein.Any suitable protein can be used.The example of suitable protein includes but not limited to albumin, comprises the immunoglobulin of IgA, lipoprotein, apolipoprotein B, β-2-macroglobulin, Elityran etc.Most preferably, pharmaceutical carrier comprises albumin, most preferably human serum albumin.Be suitable for of the present invention comprise albuminised protein can be natural origin or synthesis preparation.

Human serum albumin (HSA) is M _rthe globulin of the high soluble of 65K, is made up of 585 aminoacid.HSA is most rich in protein and account for the 70-80% of human plasma colloid osmotic pressure in blood plasma.The aminoacid sequence of HSA comprises 17 disulphide bridgeses altogether, a free mercaptan (Cys 34), and single tryptophan (Trp 214).Vein uses HSA solution to show for prevention and therapy hypovolemic shock (see such as, Tullis, JAMA, 237,355-360,460-463, (1977)) and Houser etc., Surgery, Gynecology and Obstetrics, 150,811-816 (1980)) and exchange transfusion be used for the treatment of together blood of neonate hyperbilirubinemia (see such as, Finlayson, Seminars in Thrombosisand Hemostasis, 6,85-120, (1980)).

Human serum albumin (HSA) has multiple hydrophobic binding site, and (8 for fatty acid altogether, the endogenic ligand of HSA) and in conjunction with various medicine, particularly neutral or electronegative hydrophobic compound (Goodman etc., The Pharmacological Basis of Therapeutics, 9th edition, McGraw-Hill New York (1996)).Two high affinity combined sites have been proposed at subdomain IIA and IIIA of HSA, they are the hydrophobic capsules highly extended, at near surface, there is charged lysine and arginine residues, as the junction point of polar ligand features (see such as, Fehske etc., Biochem.Pharmcol., 30, 687-92 (1981), Vorum, Dan.Med.Bull., 46, 379-99 (1999), Kragh-Hansen, Dan.Med.Bull., 1441, 131-40 (1990), Curry etc., Nat.Struct.Biol., 5, 827-35 (1998), Sugio etc., Protein.Eng., 12, 439-46 (1999), He etc., Nature, 358, 209-15 (1992), with Carter etc., Adv.Protein.Chem., 45, 153-203 (1994)).Paclitaxel and propofol have shown in conjunction with HSA (see such as, Paal etc., Eur.J.Biochem., 268 (7), 2187-91 (2001), Purcell etc., Biochim.Biophys.Acta, 1478 (1), 61-8 (2000), Altmayer etc., Arzneimittelforschung, 45,1053-6 (1995), and Garrido etc., Rev.Esp.Anestestiol.Reanim., 41,308-12 (1994)).In addition, docetaxel has shown in conjunction with human plasma protein fraction (see such as, Urien etc., Invest.NewDrugs, 14 (2), 147-51 (1996)).Therefore, although it is specifically theoretical not wish to be bound by any one, but think that, by protein as caused the reduction of the side effect relevant with administration medicine compositions in albumin is included in pharmaceutical composition of the present invention, this is due to human serum albumin and the combination being present in any free drug in compositions at least partly.

The albuminised amount in pharmaceutical composition of the present invention of being included in changes according to the approach of pharmaceutically active agents, other excipient and expection administration and site.Ideally, the albuminised amount comprised in the composition is the amount owing to effectively reducing the side effect of one or more active agents to people's administration pharmaceutical composition of the present invention.Typically, pharmaceutical composition is prepared into liquid form, then albumin is added in solution.Preferably, the pharmaceutical composition of liquid form comprise about 0.1 % by weight to about 25 % by weight albumin of (such as about 0.5 % by weight, about 5 % by weight, about 10 % by weight, about 15 % by weight, or about 20 % by weight).Most preferably, the pharmaceutical composition of liquid form comprises the albumin of about 0.5 % by weight to about 5 % by weight.Pharmaceutical composition such as can pass through lyophilizing, spraying dry, fluid bed drying, wet-granulation method and other proper method dehydration known in the art.When in solid form, during as prepared compositions by wet-granulation method, fluid bed drying and other method well known by persons skilled in the art, preferably albumin is coated on active agents and other excipient (if existence) as solution.HSA solution preferably has an appointment 0.1 % by weight to about 25 % by weight albumin of (about 0.5 % by weight, about 5 % by weight, about 10 % by weight, about 15 % by weight, or about 20 % by weight).

Except albumin, compositions of the present invention preferably also comprises deferoxamine.Deferoxamine is separated the natural product from Streptomyces pilous, can form iron complex.Deferoxamine mesylate such as injecting USP is approved as iron chelating agent by Food and Drug Administration and is available for intramuscular, subcutaneous and intravenous administration.Deferoxamine mesylate USP is that white is to linen powder.It is soluble in water and its molecular weight is 656.79.The chemical name of deferoxamine mesylate is N-[5-[3-[(5-Aminopentyl)-Hydroxycarboamoyl]-propyl-acylamino-] amyl group]-3 [[5-((N-hydroxyacetamido) amyl group]-carbamoyl] propionyl hydroxamic acid one mesylate (salt), its structural formula is C ₂₅h ₄₈n ₆o ₈.CH ₃sO ₃h.As described embodiments, the growth of microorganism in deferoxamine or its analog, derivant or salt (such as mesylate) depressant compositions and oxidation, think free drug in its binding compositions.Deferoxamine also shows in conjunction with phenolic compound (see such as, Juven etc., J.Appl.Bacteriol., 76 (6), 626-31 (1994)).Paclitaxel, docetaxel, propofol etc. or similar phenol or there is phenol or phenyl substituent.Therefore, think that deferoxamine can in conjunction with the free drug in pharmaceutical composition of the present invention or the amount reducing the free drug in pharmaceutical composition of the present invention, stimulation when also reducing thus or alleviate injection or pain.

The amount comprising deferoxamine in the composition or its preferred salt (i.e. the mesylate of deferoxamine) will depend on active agents and other excipient.Ideally, the amount of deferoxamine, its salt and analog thereof effectively suppresses the amount of growth of microorganism and/or inhibited oxidation in the composition.As mentioned above, typically pharmaceutical composition is prepared in liquid form, then deferoxamine and analog thereof is added in the form of a solution.Preferably, the pharmaceutical composition of liquid form comprise about 0.0001 % by weight to about 0.5 % by weight deferoxamine, its salt or its analog of (such as, about 0.005 % by weight, about 0.1 % by weight, or about 0.25 % by weight).More preferably, the compositions of liquid form comprises the preferred de-iron amine salt of similar quantity, deferoxamine mesylate.More preferably, the pharmaceutical composition of liquid form comprises the deferoxamine mesylate of about 0.1 % by weight.When in solid form, as mentioned above, during as prepared compositions by wet-granulation method, fluid bed drying and other method well known by persons skilled in the art, preferably deferoxamine mesylate is added active agents and other excipient (if existence) as solution.Deferoxamine mesylate solution preferably has an appointment 0.0001 % by weight to about 0.5 % by weight deferoxamine of (such as, about 0.005 % by weight, about 0.1 % by weight, or about 0.25 % by weight).

Consistent with the present invention, pharmaceutical composition can comprise other medicament, and excipient or stabilizing agent are to improve the performance of compositions.Such as, in order to the negative zeta potential dripped by increase nanoparticle or nanometer increases stability, some electronegative component can be added.These electronegative components include but not limited to the bile salts of bile acid, described bile acid is made up of the following: glycocholic acid, cholic acid, chenodeoxy cholic acid, taurocholic acid, sweet chenodeoxy cholic acid (glycochenodeoxycholic acid), cattle sulphur chenodeoxy cholic acid (taurochenodeoxycholic acid), lithocholic acid, ursodesoxycholic acid (ursodeoxycholic acid), dehydrocholic acid and other; Phospholipid; comprise lecithin (egg yolk) base phospholipid; this lecithin (egg yolk) base phospholipid comprises following phosphatidylcholine: POPC; the sub-oleoyl phosphatidylcholine of palmityl; the sub-oleoyl phosphatidylcholine of stearoyl; stearoyl oleoyl phosphatidylcholine, stearoyl palmitoylphosphatidylcholine, and dipalmitoyl phosphatidyl choline.Other phospholipid comprises L-α-dimyristoyl phosphatidyl choline (DMPC), DOPC (DOPC), distearoyl phosphatidylcholine (DSPC), HSPC (HSPC), D-α-phosphatidylcholine, β-acetyl group-γ-O-cetyl, L-α-phosphatidylcholine, β-acetyl group-γ-O-cetyl, DL-α-phosphatidylcholine, β-acetyl group-γ-O-cetyl, L-α-phosphatidylcholine, β-acetyl group-γ-O-octadecyl, L-α-phosphatidylcholine, β-arachidonic acyl-γ-O-cetyl, L-α-phosphatidylcholine, β-acetyl group-γ-O-(Linolenic Acid-cis-thiazolinyl), D-α-phosphatidylcholine, β-arachidonic acyl-γ-O-palmityl, 3-sn-phosphatidylcholine, 2-arachidonic acyl-1-stearoyl, L-α-phosphatidylcholine, β-arachidonic acyl-γ-stearoyl, L-α-phosphatidylcholine, two arachidonic acyls, L-α-phosphatidylcholine, two (behenolyls), L-α-phosphatidylcholine, β-(cis-8, 11, 14-20 carbon triolefin acyl)-γ-O-cetyl, L-α-phosphatidylcholine, beta-oil acyl-γ-myristoyl, L-α-phosphatidylcholine, β-(pyrene-1-base) capryl-γ-palmityl, 3-sn-phosphatidyl-N, N-dimethylethanolamine, 1, 2-bis-palmityl, L-α-PHOSPHATIDYL ETHANOLAMINE, two (heptadecanoyls), 3-sn-PHOSPHATIDYL ETHANOLAMINE, 1, 2-bis-lauroyl, 3-sn-PHOSPHATIDYL ETHANOLAMINE, 1, 2-bis-myristoyl, 3-sn-PHOSPHATIDYL ETHANOLAMINE, 1, 2-bis-oleoyl, 3-sn-PHOSPHATIDYL ETHANOLAMINE, 1, 2-bis-palmityl, L-α-PHOSPHATIDYL ETHANOLAMINE, two palmityls, L-α-PHOSPHATIDYL ETHANOLAMINE, two palmityls, N-dansyl, L-α-PHOSPHATIDYL ETHANOLAMINE, two palmityls, N, N-dimethyl, L-α-GLYCEROL,DIMYRISTOYL PHOSPHATIDYL (sodium salt) (DMPG), DPPG (sodium salt) (DPPG), DSPG (sodium salt) (DSPG), N-(carbonyl-methoxy poly (ethylene glycol) 2000)-1, 2-distearyl-sn-glycerol-3-phosphate ethanolamine sodium (MPEG-DSPE), L-α-phosphatidic acid, two capryl sodium salts, L-α-phosphatidic acid, two (17 phosphinylidyne) sodium salt, 3-sn-phosphatidic acid, 1, 2-bis-myristoyl sodium salt, L-α-phosphatidic acid, two caprylyl sodium salts, L-α-phosphatidic acid, two oleoyl sodium salts, L-α-phosphatidic acid, two palmityl sodium salts, L-α-phosphatidyl-DL-glycerol, two myristoyl sodium salts, L-α-phosphatidyl-DL-glycerol, two oleoyl sodium salts, L-α-phosphatidyl-DL-glycerol, two palmityl ammonium salts, L-α-phosphatidyl-DL-glycerol, distearyl ammonium salt, L-α-phosphatidyl-DL-glycerol, beta-oil acyl-γ-palmityl ammonium salt, L-α-phosphatidylinositols ammonium salt, L-α-phosphatidyl-4 sodium alkoxide, L-α-phosphatidyl-Serine, two oleoyl sodium salts, L-α-phosphatidyl-Serine, with two palmityl sodium salts.The surfactant of electronegative emulsifying agent is also suitable as additive, such as sodium cholesteryl sulfate etc.

Medicament (such as propofol) can be used alone or is dissolved in the immiscible solvent of water.The immiscible solvent of various water can be used as soybean oil, safflower oil, Oleum Gossypii semen, Semen Maydis oil, Oleum helianthi, Oleum Arachidis hypogaeae semen, Oleum Ricini, olive oil.Preferred oil is vegetable oil, wherein most preferably soybean oil.Soybean oil can use within the scope of the 1%-10% of composition weight.Preferred soybean oil is present in compositions with the amount of about 3 % by weight.

Pharmaceutical composition of the present invention can use medicinal surfactants stabilize.Term used herein " surfactant " refers to amphiphilic surface active groups.Surfactant can be anionic, cationic, nonionic and amphoteric ion type.Any suitable surfactant can be included in pharmaceutical composition of the present invention.Suitable surfactant comprises nonionic surfactant as phospholipid, polyoxyethylene sorbitan ester, and tocopheryl polyethylene glycol succinate.Preferred surfactant is lecithin, Tween 80 and vitamin E-t d-alpha-tocopherol polyoxyethylene-1000 succinate (TPGS).For the preparation containing soybean oil, preferably lecithin and be no more than 1.2 % by weight of compositions for containing the preparation of 3% soybean oil, preferably 1.1 % by weight.For the preparation not containing soybean oil, Tween 80 or D-ALPHA-tocopheryl polyethylene glycol 1000 succinate are preferred surfactants.Typical case, the Tween 80 of 0.1-1.5 % by weight or the D-ALPHA-tocopheryl polyethylene glycol 1000 succinate of 0.5-4 % by weight are suitable.Preferably, the Tween 80 of 1.5 % by weight or the D-ALPHA-tocopheryl polyethylene glycol 1000 succinate of 1 % by weight is used.The example of other suitable surfactant is described in such as Becher, Emulsions:Theory and Practice, Robert E.KriegerPublishing, in Malabar, Fla. (1965).

There is the suitable preparation (see such as, U.S. patent 5,916,596) of various pharmaceutical composition of the present invention.Lower series preparation and method are only exemplary and not in any limiting sense.The preparation being suitable for oral administration can be made up of the following: (a) liquid solution, as the compound be dissolved in diluent of effective dose, described diluent is as water, saline or orange juice, (b) capsule, sachet or tablet, the active component as solid or granule separately containing scheduled volume, (c) suspensoid in appropriate liquid, and the Emulsion that (d) is suitable.Tablet form can comprise one or more lactose, mannitol, corn starch, potato starch, microcrystalline Cellulose, arabic gum, gelatin, silica sol, cross-linking sodium carboxymethyl cellulose, Talcum, magnesium stearate, stearic acid, and other excipient, coloring agent, diluent, buffer agent, wetting agent, antiseptic, flavoring agent, and pharmaceutically-compatible excipient.Lozenge form can comprise active component in flavoring agent, described flavoring agent is generally sucrose and arabic gum or Tragacanth, and in inert base, comprise the lozenge of active component, described inert base is as gelatin and glycerol, or sucrose and arabic gum, Emulsion, gel etc. besides the active ingredients also containing all excipient as known in the art.

The preparation being suitable for parenteral comprises aqueous and nonaqueous isotonic sterile injection liquid, it can contain antioxidant, buffer agent, antibacterial, with make preparation and expect the solute that the blood of receiver is isotonic, and suspending agent, solubilizing agent, thickening agent, the aqueous of stabilizing agent and antiseptic and nonaqueous sterile suspensions can be comprised.Preparation can provide with the container of unit dose or multiple dose sealing, and as peace is cutd open and bottle, and under can being stored in lyophilization (lyophilizing) condition, this only requires and adds sterile liquid excipient before at once using, and such as water is to inject.Extemporaneous injection solutions and suspensoid can be prepared from the sterilized powder of aforesaid kind, granule and tablet.Preferred ejection preparation.

The preparation being suitable for Aerosol administration comprises pharmaceutical composition of the present invention, it comprises the aqueous and nonaqueous isotonic sterile solution that can contain antioxidant, buffer agent, antibacterial and solute, and suspending agent, solubilizing agent, thickening agent, the aqueous of stabilizing agent and antiseptic and non-aqueous sterile suspensions can be comprised, these components separately or with other suitable combination of components, aerosol preparations can be prepared into pass through inhalation.These aerosol preparations can put into pressurization acceptable propellant, described propellant as dichlorodifluoromethane, propane, nitrogen etc.They can also be mixed with the medicine for non-pressured preparations, as in aerosol apparatus or nebulizer.

Other suitable preparation is possible, such as, can prepare suppository by using various substrate (base) such as emulsifying base or water-insoluble substrate.The preparation being suitable for vagina administration can be provided as vaginal suppository, tampon, ointment, gel, paste, foam, or spray formu, and it is besides the active ingredients also containing all suitable carriers as known in the art.

In a preferred embodiment of the invention, pharmaceutical composition is mixed with the pH with 4.5-9.0, more preferably the pH of 5.0-8.0.Can also by add suitable Osmolyte regulator as glycerol make pharmaceutical composition and blood isotonic.In addition, pharmaceutical carrier preferably also contains not containing water or the water for injection of pyrogen, USP.Preferably, pharmaceutical composition of the present invention is prepared to sterile aqueous formulation, nanoparticle, oil-in-water emulsion, or water-in-oil emulsion.Most preferably, pharmaceutical composition is oil-in-water emulsion.

According to the present invention, for the pharmaceutical composition comprising propofol, by being dissolved in separately by propofol in the immiscible solvent of water and the aqueous phase of preparation containing albumin, deferoxamine, surfactant and other water soluble ingredient, oil-in-water emulsion can be prepared by mixed with water for oil.Coarse emulsion under the pressure of 10,000 to 25,000psi high pressure homogenization a recirculation 5-20 cycle to form desirable Emulsion.Preferred pressure is 15,000 to 20,000psi, more preferably 10,000psi.Coarse emulsion can a recirculation 7-15 cycle and preferred 15 cycles of recirculation.Alternatively, point open walk by homogenizer can be used.

Preferably, pharmaceutical composition of the present invention can be less than granule or the drop of about 200 nanometers (nm) containing size.Such as, at paclitaxel, docetaxel, rapamycin, cyclosporin, in propofol and other situation, the average-size of these dispersions is less than 200nm.

The present invention provides in addition and reduces one or more and the method to the relevant side effect of people's administration medicine compositions.The method comprises the pharmaceutical composition comprising medicament and pharmaceutical carrier to people's administration, and wherein said pharmaceutical carrier comprises albumin and deferoxamine.Those identical aspects of the inventive method are more than also applicable in conjunction with the description of the pharmaceutical composition of pharmaceutical composition of the present invention statement, medicament and pharmaceutical carrier and composition thereof.

In the context of the present invention, the pharmaceutical composition of the present invention dosage that delivers medicine to people changes along with concrete pharmaceutical composition, medication and the concrete position that is treated.This dosage should be enough to realize expected response within the expected time, as to the treatment of specified disease or prophylactic response, or when medicament is that anesthetis is as anesthesia reaction during propofol.

Although any mode to people's administration medicine compositions suitably may be used in the context of the invention, preferably pharmaceutical composition of the present invention is delivered medicine to people by intravenous administration, intraarterial delivery, feeding drug into pulmones, oral administration, suction, intravesical administration, intramuscular administration, intrarterial, subcutaneous administration, eye drops, intrathecal drug delivery or transdermal administration.Such as, pharmaceutical composition of the present invention can by inhalation to treat respiratory passage diseases.The side effect relevant with sucking pharmaceutical composition of the present invention is minimum, because albumin is the natural component of respiratory tract internal layer and secretions.The present composition may be used for treatment respiratory passage diseases as pulmonary fibrosis, bronchiolitis obliterans, pulmonary carcinoma, bronchovesicular cancer etc.

Method of the present invention causes and the reduction to relevant one or more side effect of people's administration medicine compositions.These side effect comprise such as, bone marrow depression, neurotoxicity, irritated, inflammation, venous stimulation, phlebitis, pain, skin irritation, and combination.But these side effect are only exemplary, new compositions and the method for the application of the invention can reduce or avoid other side effect relevant with various medicament or side effect to combine.

The present invention provides the method for growth of microorganism in depressant compositions in addition." suppression growth of microorganism " refers to and eliminate microorganism completely from pharmaceutical composition, or reduces amount or the speed of growth of microorganism in pharmaceutical composition.The method comprises the pharmaceutical composition that preparation comprises medicament and pharmaceutical carrier, and wherein said pharmaceutical carrier comprises deferoxamine, its salt, its analog, and combination, the growth of microorganism in their the effective depressant compositions of amount.In addition, the invention provides the method for depressant compositions oxidation.The method comprises the pharmaceutical composition that preparation comprises medicament and pharmaceutical carrier, and wherein said pharmaceutical carrier comprises deferoxamine, its salt, its analog, and combination, the oxidation of their the effective depressant compositions of amount.The description of the pharmaceutical composition of more than illustrating in conjunction with pharmaceutical composition of the present invention, medicament and pharmaceutical carrier and component thereof is also applicable to those identical aspects of the inventive method.

Comprise deferoxamine in the composition, or the amount of its preferred salt (mesylate of deferoxamine) will depend on active agents and other excipient.Ideally, deferoxamine in compositions, its salt, the amount of its analog effectively suppresses the amount of growth of microorganism and/or inhibited oxidation.As mentioned above, typically, pharmaceutical composition is prepared in liquid form, then by deferoxamine, and its salt, and its analog is added in solution.Preferably, the pharmaceutical composition of liquid form comprises the deferoxamine of about 0.0001 % by weight to about 0.5 % by weight (such as about 0.005 % by weight, about 0.1 % by weight or about 0.25 % by weight), its salt, or its analog.More preferably, the compositions of liquid form comprises the preferred de-iron amine salt of similar quantity, deferoxamine mesylate.Most preferably, the pharmaceutical composition of liquid form comprises the deferoxamine mesylate of about 0.5 % by weight.When compositions is prepared in solid form, as mentioned above, during as prepared compositions by wet-granulation method, fluid bed drying and other method well known by persons skilled in the art, preferably deferoxamine mesylate is coated to active agents as solution, and other excipient (if existence).Deferoxamine mesylate solution preferably has the deferoxamine of about 0.0001 % by weight to about 0.5 % by weight (such as about 0.005 % by weight, about 0.1 % by weight or about 0.25 % by weight).

The present invention is also provided for the method that enhancing medicament is transported to weakness, the method comprises the pharmaceutical composition comprising medicament and pharmaceutical carrier to people's administration, wherein said pharmaceutical carrier comprises albumin, and wherein in pharmaceutical composition the ratio of albumin and medicament be about 18: 1 or less.The present invention further provides the method be combined with external or cells in vivo for strengthening medicament, the method comprises the pharmaceutical composition comprising medicament and pharmaceutical carrier to described external or cells in vivo administration, wherein said pharmaceutical carrier comprises albumin, and wherein in pharmaceutical composition the ratio of albumin and medicament be about 18: 1 or less.The description of pharmaceutical composition, medicament, pharmaceutical carrier, route of administration and the component thereof more than illustrated in conjunction with pharmaceutical composition of the present invention and the inventive method is also applicable to transport those the identical aspects with associated methods.

For by enhancing transport to weakness or be used for strengthening in the method for medicament and Cell binding, pharmaceutical carrier preferably comprises albumin, most preferably human serum albumin.Be not bound by any one specifically theoretical, the ratio of the medicament thought in protein such as human serum albumin and pharmaceutical composition affects ability and the cytotropic transhipment of medicament of medicament and Cell binding.In this, the higher rate of protein and medicament is usually with the Cell binding of the medicament of difference with transport relevant, and this may be the result that receptor is competed on cell surface.The ratio of protein such as albumin and active agents must be make enough pharmaceutical agent and Cell binding or be transported to cell.The exemplary scope of protein-drug preparations is protein and drug ratio (w/w) is 0.01: 1 to about 100: 1.More preferably, ratio is 0.02: 1 to about 40: 1.Although the ratio of protein and medicament must to different protein and pharmaceutical agent combinations optimization, usually the ratio of protein such as albumin and medicament be about 18: 1 or less (such as about 15: 1, about 10: 1, about 5: 1, or about 3: 1).More preferably, ratio is about 0.2: 1 to about 12: 1.Most preferably, ratio is about 1: 1 to about 9: 1.Preferably, preparation is substantially free of polyoxyethylene castor oil, more preferably not containing polyoxyethylene castor oil EL

(BASF).Polyoxyethylene castor oil EL

being nonionic emulsifier, is as Oleum Ricini and polyoxyethylated polyethers.As mentioned above, polyoxyethylene castor oil is typically used as the solvent of paclitaxel, and with may be serious side effect relevant (see such as, Gelderblom etc., above).

Medicament can be any suitable medicament described herein (such as, propofol, paclitaxel, or docetaxel).In addition, medicament can be nucleotide sequence, most preferably DNA sequence.This respect, pharmaceutical composition of the present invention can be transported gene delivery to cell by receptor-mediated/cave, chamber (caveolar)/capsule.In order to by DNA sequence as gene or other hereditary material, include but not limited to plasmid or c-DNA, in transporte to cells (such as endotheliocyte or tumor cell), can prepare and comprise the pharmaceutical composition of albumin in conjunction with hereditary material.Because have at the tumor cell of inflammation part and other cell, the height of protein is absorbed, preferably hereditary material to be sucked in these cell types and can be incorporated in the hereditary material of cell to reach effective therapeutic effect.Protein such as human serum the use of albumen please work the risk of the non-virus carrier doing to transmit hereditary material and the not disease relevant to virus or side effect.Such as, the pharmaceutical composition comprising coding beta-galactosidase or green fluorescent protein (GFP) and albuminised nucleotide sequence can be prepared, and with derive from human umbilical vein or the microvascular contact endothelial cells of people's lung, to promote that nucleotide sequence is incorporated in endotheliocyte.The integration of nucleotide sequence can use methods known in the art to detect, as such as fluorescence or dyeing.

In the present invention in the method that strengthens medicament and transport to weakness, weakness can be any suitable disease or disease.Preferably, weakness is cancer, cardiovascular disease, or arthritis.

In the present invention in the method that strengthens medicament and be combined with external or cells in vivo, pharmaceutical composition is delivered medicine to external or cells in vivo.Ideally, cell is zooblast.More preferably, cell is mammalian cell, and most preferably, cell is people's cell.Preferably pharmaceutical composition is delivered medicine to cells in vivo.Cell can be any suitable cell of desired target target as administration medicine compositions.Such as, cell can be positioned at or derive from digestive system tissue, comprises such as esophagus, stomach, small intestinal, colon, rectum, anus, liver, gallbladder, and pancreas.Cell can also be positioned at or derive from respiratory system tissue, comprises such as larynx, lung, and bronchus.Cell can be positioned at or derive from the cervix uteri such as forming sex reproductive system, body of uterus, ovary hole, vagina, prostate, testis, and penis, and the bladder of composition urinary system, kidney, renal pelvis, and ureter.Cell can be positioned at or derive from the tissue of cardiovascular system, comprises such as endotheliocyte and myocardial cell.Cell can also be positioned at or derive from lymphoid tissue (such as lymphocyte), nervous system (such as neuron or neurogliocyte), and hormonal system (such as thyroid cell).Preferably, cell is positioned at or derives from the tissue of cardiovascular system.Most preferably, cell is endotheliocyte.In the present invention for strengthening in the content of the method for transhipment and medicament and Cell binding, pharmaceutical composition contacts ideally more than a kind of cell.

In another aspect of this invention, the present invention may be used for treating tumor cell for the method strengthening transhipment and enhancing medicament and Cell binding.Compared with normal cell, the protein uptake that tumor cell display strengthens, this protein comprises albumin and transferrins.Because tumor cell divides fast, so compared with normal cell, they require more source of nutrition.The tumor research that the present invention contains the pharmaceutical composition of paclitaxel and human serum albumin shows that tumor is absorbed the height of albumin-paclitaxel.Have been found that this is the phenomenon due to the albumin-drug transport by glycoprotein 60 (" gp60 ") receptor do not recognized in the past, this glycoprotein 60 receptor is specific for albumin.

Therefore, according to another aspect of the present invention, other protein transport receptors that the specific gp60 receptor of albumin and tumor cell exist can be used as the target of Tumor suppression growth.By using the antibody of anti-gp60 receptor or combination, other large or micromolecular compound of blocking-up or inactivation gp60 blocks gp60 receptor, with blocking-up other protein transport receptors on tumor cell or tumor endothelial cell, can blocking protein to the transhipment of these cells, reduce their growth rate thus and cause cell death.Therefore, the blocking-up of this mechanism causes the treatment of the patient (such as people) suffering from cancer or other disease.Qualification specific protein receptor blocking-up/combinations be by screen any amount of for be separated gp60 or other receptor as the compound of gp16 or gp30 or by use full cell preparation come.In addition, suitable animal model also may be used for this object, as such as contained coding gp60 or cFLIP or " knocking out " for the gene transporting other special albumen the mice suddenlyd change.Therefore, qualification blocks or method in conjunction with the compound of gp60, gp16, gp30 or other protein receptor is within the scope of the present invention.

In addition, to block or compound in conjunction with gp60 receptor or other protein receptor may be used for treating the several conditions comprising cancer.About the treatment of cancer, blocking-up or binding compounds can be used as single medicament or be combined with other standard chemotherapy or multiple chemotherapy.Such as, it and conventional chemotherapy or be used for the treatment of cancer with albumin-drug pharmaceutical compositions of the present invention (it shows high accumulation in tumor), the compound then transported to tumor cell with blocking protein.Blocking compound can before other chemotherapy or anticarcinogen or in conjunction with administration.Therefore, anyly can to block or compound in conjunction with gp60 receptor or other protein receptor is within the scope of the present invention.

Albumin-drug compositions of the present invention, as such as albumin-paclitaxel, albumin-docetaxel, albumin-Epothilones, albumin-camptothecine, or albumin-rapamycin and other, may be used for disease therapy.Think that these pharmaceutical compositions are effectively because of the receptor-mediated transhipment of the protein-drug composition increased to desired area (such as tumor).Do not wish to be bound by any concrete theory, be considered to such as albumin-paclitaxel compositions to transporting mechanism through lung of tumor and albumin-paclitaxel and albumin-rapamycin transhipment by the transhipment of the protein-drug composition causing the receptor-mediated transhipment of therapeutic effect.Transhipment is by the impact of the existence of gp60, gp16 or gp30 in these tissues.Therefore, to disease such as inflammation (such as arthritis) or tumor sites transhipment and gp60, gp16 or gp30 receptor is correlated with and cause the medicine of therapeutic effect and protein-drug composition to be considered as compositions of the present invention.

According to another aspect of the present invention, endotheliocyte can with the co-culture of cells with specific function.Component needed for endotheliocyte and other cell type incubation as islet cells, hepatocyte, neuroendocrine cell and other cell allows is if protein and other Hubeiwan are to the transfer of these cells.Endotheliocyte provides these components to the transfer of cultured cells type so that condition in analogue body, namely wherein these cell types are usually close with endothelial cell tight and will depend on endotheliocyte and transport nutrition, somatomedin, hormone signal etc., and these normal functions for them are needs.When endotheliocyte does not exist, previously can not cultivate these different cell types fully and obtain physiological property.External or islets of langerhans ex vivo, the differentiation of hepatocyte or neuroendocrine tissue and suitable function is allowed in the existence with endotheliocyte in the culture expecting cell type.Therefore find with lack cultivate under endotheliocyte those compared with, the Dual culture of endotheliocyte and islets of langerhans causes the islets of langerhans of the physiological property (ability of such as excreting insulin) with improvement.Then this tissue can use in vitro or be transplanted in body to treat disease (the such as diabetes in the situation of islet cells caused owing to lacking suitable cell function, hepatic dysfunction in hepatocyte situation, and neuroendocrine disturbance in the situation of neuroendocrine cell or pain relief).The cell (as mentioned above) deriving from other tissue and organ also can with endotheliocyte Dual culture to provide identical benefit.In addition, Dual culture may be used for hereditary material to be incorporated in target cell type.Find that albuminised existence is highly profitable in these cultures.

The following example illustrates the present invention further, but certainly should not be considered to limit its scope by any way.

Embodiment 1

Example illustrates that the preparation containing paclitaxel and albuminised pharmaceutical composition.The preparation of Paclitaxel-albumin compositions is described in United States Patent (USP) 5,439,686 and 5,916, and in 596, they are intactly incorporated into this by reference.Particularly, 30mg paclitaxel is dissolved in 3.0ml dichloromethane.This solution is added in 27.0ml human serum albumin solution (2%w/v).Add deferoxamine as required.This mixture homogenize 5 minutes (Vitris homogenizer, model TempestI.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to rotary evaporator, 40 DEG C, fast remove dichloromethane 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the paclitaxel particles of acquisition is 50-220nm (Z-meansigma methods, Malvem Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.

Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When compared with the toxicity of the paclitaxel be dissolved in cremophor formulations, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 2

Example illustrates that the preparation containing amiodarone and albuminised pharmaceutical composition.30mg amiodarone is dissolved in 3.0ml dichloromethane.Solution is added in 27.0ml human serum albumin solution (1%w/v).Add deferoxamine as required.Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to rotary evaporator, 40 DEG C, fast remove dichloromethane 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the amiodarone granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.

Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When compared with the toxicity of the amiodarone be dissolved in Tween formulation, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 3

Example illustrates that the preparation of the pharmaceutical composition containing liothyronine and albumin compositions.Liothyronine (or suitable salt) is dissolved in water-alcoholic solutions or alkaline solution with the concentration of 0.5-50mg/ml.Alcohol (or alkalescence) solution to be added in albumin soln (0.1-25%w/v) and to stir.Stirring uses the low sheraing of agitator or uses the high shear of ultrasonoscope or homogenizer.Under the liothyronine of low concentration, obtain (5-1000 μ g/ml) settled solution.When concentration increases, stable emulsus suspensoid is obtained.These solution or suspensoid are filtered by sterile filters.Organic solvent is removed by evaporation or other proper method.

Embodiment 4

Example illustrates that the preparation containing rapamycin and albuminised pharmaceutical composition.30mg rapamycin is dissolved in 2ml chloroform/ethanol.Solution is added in 27.0ml human serum albumin solution (3%w/v).Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.

Embodiment 5

Example illustrates that the preparation containing epothilone B and albuminised pharmaceutical composition.30mg epothilone B is dissolved in 2ml chloroform/ethanol.Solution is added in 27.0ml human serum albumin solution (3%w/v).Add deferoxamine as required.Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When compared with the toxicity of the epothilone B be dissolved in cremophor formulations, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 6

Example illustrates that the preparation containing colchicine dimer and albuminised pharmaceutical composition.30mg colchicine-dimer is dissolved in 2ml chloroform/ethanol.Solution is added in 27.0ml human serum albumin solution (3%w/v).Add deferoxamine as required.Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When compared with the dimeric toxicity of the colchicine be dissolved in tween, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 7

Example illustrates that the preparation containing docetaxel and albuminised pharmaceutical composition.30mg docetaxel is dissolved in 2ml chloroform/ethanol.Solution is added in 27.0ml human serum albumin solution (3%w/v).Add deferoxamine as required.Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When with the standard solvent being dissolved in this medicine--time compared with the toxicity of the docetaxel in tween/ethanol, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 8

Example illustrates that the preparation containing docetaxel and albuminised pharmaceutical composition.150mg docetaxel is dissolved in 1ml ethyl acetate and butyl acetate and 0.5ml oil such as soybean oil or vitamin E oil.Use solvent and the oil of other ratio, these compositionss are also considered a part of the present invention.Also optionally add a small amount of electronegative component, such as benzoic acid (0.001%-0.5%).Then solution is added in 27.0ml human serum albumin solution (5%w/v).Add deferoxamine as required.Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, MalvernZetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When with the standard solvent being dissolved in this medicine--time compared with the toxicity of the docetaxel in tween/ethanol, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 9

Example illustrates that the preparation containing taxane IDN5390 and albuminised pharmaceutical composition.150mg IDN5390 is dissolved in 1ml ethyl acetate and butyl acetate and 0.5ml oil such as soybean oil or vitamin E oil.Use solvent and the oil of other ratio, these compositionss are also considered a part of the present invention.Also optionally add a small amount of electronegative component, such as benzoic acid (0.001%-0.5%).Then solution is added in 27.0ml human serum albumin solution (5%w/v).Add deferoxamine as required.Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When compared with the toxicity of the IDN5390 be dissolved in tween, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 10

Example illustrates that the preparation containing taxane IDN5109 and albuminised pharmaceutical composition.150mg IDN5109 is dissolved in 2ml chloroform/ethanol.Use solvent and the oil of other ratio, these compositionss are also considered a part of the present invention.Also optionally add a small amount of electronegative component, such as benzoic acid (0.001%-0.5%).Then solution is added in 27.0ml human serum albumin solution (5%w/v).Add deferoxamine as required.Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.When compared with the toxicity of the IDN5109 be dissolved in tween, the present invention contains the significantly lower toxicity of albuminised pharmaceutical composition display.

Embodiment 11

Example illustrates that the preparation containing 10-hydroxycamptothecine (10HC) and albuminised pharmaceutical composition.30mg 10HC is dissolved in 2.0ml DMF/ dichloromethane/soybean oil.Then solution is added in 27.0ml human serum albumin solution (3%w/v).Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.

Embodiment 12

Example illustrates that the preparation containing cyclosporin and albuminised pharmaceutical composition.30mg cyclosporin is dissolved in 3.0ml dichloromethane.Then solution is added in 27.0ml human serum albumin solution (1%w/v).Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to Rotavap, under decompression (30mm Hg), removes solvent 20-30 minute at 40 DEG C fast.The dispersion obtained is translucent, and the typical average diameter of the granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.

Embodiment 13

Example illustrates that containing oil and the preparation comprising cyclosporin and albuminised pharmaceutical composition.30mg cyclosporin is dissolved in the suitable oil (Oleum sesami containing 10% orange oil) of 3.0ml.Then solution is added in 27.0ml human serum albumin solution (1%w/v).Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The typical average diameter of the granule obtained is 50-220nm (Z-meansigma methods, Malvern Zetasizer).Direct use dispersion or by optionally adding suitable antifreezing agent lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.

Embodiment 14

Example illustrates that the preparation containing amphotericin and albuminised pharmaceutical composition.30mg amphotericin is dissolved in 3.0ml methyl pyrrolidone/dichloromethane.Solution is incorporated in 27.0ml human serum albumin solution (1%w/v).Mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to rotary evaporator, 40 DEG C, fast remove solvent 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the amphotericin granule of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.Will be appreciated that for the amount of the medicine of the present embodiment, solvent, protein, type and ratio by the restriction of any mode.Add other component such as lipid, cholate etc. and also produce suitable preparation.

Embodiment 15

The present embodiment illustrates preclinical pharmacokinetics and the pharmacodynamics of the pharmaceutical composition containing albumin and paclitaxel.

Carry out several Preclinical metabolism and pharmacokinetics study in Mouse and rat to assess the possible advantage of albumin-paclitaxel pharmaceutical composition relative to cremophor-paclitaxel (taxol) pharmaceutical composition.These study proof: the pharmacokinetics of (1) albumin-paclitaxel is in rats linear, and the pharmacokinetics of taxol is nonlinear relative to dosage, (2) comprise the pharmaceutical composition display lower plasma A UC and C of albumin and paclitaxel _max, point out compared with taxol albumin-paclitaxel compositions to the distribution faster (excretion is similar) of tissue, (3) comprise the lower C of the pharmaceutical composition display of albumin and paclitaxel _maxthe toxicity of the reduction that this possible explanation is correlated with blood peak levels compared with taxol, (4) compared with taxol, the half-life comprising the pharmaceutical composition display of albumin and paclitaxel in rats high about 2-doubly and in the mice of carrying tumor high 4-doubly, and (5) comprise the metabolism of metabolism lower than the paclitaxel in taxol pharmaceutical composition of paclitaxel in the pharmaceutical composition of albumin and paclitaxel.Inject latter 24 hours in rats, for the pharmaceutical composition comprising albumin and paclitaxel, the gross activity of 44% is still relevant to paclitaxel, compares down for taxol only 22%.For the final effect of the above-mentioned pharmacokinetics of the pharmaceutical composition display containing albumin and paclitaxel, namely the cellular uptake strengthened, the half-life extended and lower metabolism to cause compared with taxol tumor AUC in the mice of carrying tumor high 1.7 times, tumor C _maxhigh 1.2 times, and long 1.7 times of tumor half-life.

Embodiment 16

Example illustrates that the side effect of reduction relevant with albuminised pharmaceutical composition to comprising paclitaxel and the toxicity of reduction.

Owing to there is no the peculiar property comprising paclitaxel and albuminised pharmaceutical composition under polyoxyethylene castor oil, comprising the toxicity of paclitaxel and albuminised pharmaceutical composition significantly lower than taxol.In preclinical study in Mouse and rat, the display of single dose studies on acute toxicity in mice for containing paclitaxel and albuminised pharmaceutical composition, its LD ₅₀about 59 times of dose ratio taxol.In multiple dose toxicity research in mice, for containing paclitaxel and albuminised pharmaceutical composition, its LD ₅₀about 10 times of dose ratio taxol.Further research assessment is by myelosuppressive degree in the rat containing paclitaxel and albuminised pharmaceutical composition and taxol process.Under result such as to be presented at the dosage, comprise paclitaxel and albuminised pharmaceutical composition produces significantly less bone marrow depression than taxol in rats.In studies on acute toxicity in rats, in the animal accepting 9mg/kg taxol, observe cerebral cortical necrosis or serious neurotoxicity, but in acceptance up to not observing comprising in the animal of paclitaxel and albuminised pharmaceutical composition of 120mg/kg dosage.Therefore, compared with the standard pharmaceutical composition comprising paclitaxel, comprise the remarkable reduction that albuminised existence in the pharmaceutical composition of paclitaxel causes side effect and toxicity.

Embodiment 17

Example illustrates that and comprise paclitaxel and the clinical effectiveness of albuminised pharmaceutical composition in people.

Be provide containing paclitaxel and the pharmaceutical composition of albumin (" albumin-paclitaxel ") evidence supporting the Side effect of reduction compared with cremophor-paclitaxel compositions (taxol) more than the clinical research in 500 people patients so far.In the I phase of 19 patients is studied, every maximum tolerated dose of albumin-paclitaxel that provides for 3 weeks is 300mg/m ².This is significantly higher than the dosage of the cremophor-paclitaxel of usual administration, namely within every 3 weeks, provides a 175mg/m ².In these patients, hematotoxicity is gentle, does not have allergy, mild neuropathies, with the side effect not having administration relevant as venous stimulation etc.

In another was studied the I phase of 27 patients, the maximum tolerated dose of the albumin-paclitaxel provided according to weekly program is 125-150mg/m ².This is significantly higher than the dosage of the cremophor-paclitaxel of usual administration, namely according to the 80mg/m that weekly program provides ².In these patients, hematotoxicity is gentle, does not have allergy, mild neuropathies, with the side effect not having administration relevant as venous stimulation etc.

Within every 3 weeks respectively in 43 and 63 patients, provide 175 or 300mg/m ²two II phases of albumin-paclitaxel study, hematotoxicity is low, at 175mg/m ²and 300mg/m ²lower only have 7% and 24% patient ANC < 500/mm respectively ³.At 175mg/m ²and 300mg/m ²lower have the patient of 0% and 14% that serious neuropathy occurs respectively.Serious allergy does not occur, the relevant side effect of administration does not occur as venous stimulation, injection pain etc., these side effect are significantly lower than using taxol to experience.

In the III phase of being compared with taxol (containing cremophor-paclitaxel) by albumin-paclitaxel compositions ABI-007 tests, the dosage significantly higher (260mg/m of ABI-007 ², compare the 175mg/m of taxol ²), show that it is tolerated better.When compared with cremophor-paclitaxel, albumin-paclitaxel compositions also shows the neutrophilic granulocytopenia significantly reduced.

Embodiment 18

Example illustrates that the preclinical efficacy of the enhancing using the pharmaceutical composition containing albumin and paclitaxel.

The relatively vitro cytotoxicity research display of the effect of albumin-paclitaxel and Taxol on cervical squamous cell carcinoma A431: albumin-paclitaxel cytotoxic activity increase about 4 times, the IC of albumin-paclitaxel and taxol ₅₀be respectively 0.0038 and 0.012 μ g/ml.

Five different people Xenograft Tumor Models (MX-1 breast in athymic mouse, NCI-H522 lung, SK-OV-3 ovary, PC-3 prostate, with HT-29 colon) in, the MTD of ABI-007 or etc. malicious dose ratio taxol height 1.5-3.4 doubly, in all tumors except lung tumor (p=0.15), cause the improvement (p < 0.05) of statistically significant in tumor growth delay.

In MX 1 breast model, the animals survived of the absolutely albumin-paclitaxel treatment of (100%) 103 days, 20-40% survival in the group for the treatment of with the taxol of DE of comparing down.

Embodiment 19

Example illustrates that the clinical efficacy using the pharmaceutical composition comprising albumin and paclitaxel of intraarterial delivery to strengthen.

During the I/II phase comprising the pharmaceutical composition of albumin and paclitaxel at intraarterial delivery studies, as described herein, recruit head and neck cancer patient (N=31) and cancer of anal canal patient (N=12).By percutaneous super-selective endoarterial infusion, the dosage of dosage administration in 30 minutes is from 120mg/m ²progressively be increased to 300mg/m ², q 3-4 week.Head and neck cancer patient shows the response rate of 76% (N=29), and cancer of anal canal patient shows the response rate of 64% (N=11).

Embodiment 20

Example illustrates that containing 3% oil and the preparation comprising propofol and albuminised pharmaceutical composition.

Following preparation contains the oil-in-water emulsion of the propofol of 1 % by weight.Also stir until aqueous phase is prepared in dissolving by glycerol (2.25 % by weight) and human serum albumin's (0.5 % by weight) being added in water for injection.By aqueous phase by filter (0.2um filter).By at about 50 DEG C-60 DEG C, lecithin (0.4 % by weight) and propofol (1 % by weight) are dissolved in soybean oil (3 % by weight) and also stir until oil phase is prepared in dissolving.Oil phase to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization in 15 cycles of 5 DEG C of recirculatioies under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.The pharmaceutical composition obtained comprises the component (be weight %) of following usual scope: propofol 0.5-5%; Human serum albumin 0.5-3%; Soybean oil 0.5-3.0%; Lecithin 0.12-1.2%; Glycerol 2.25%; Water for injection in right amount to 100; PH 5-8.Optionally add suitable chelating agen, such as deferoxamine (0.001-0.1%).

Embodiment 21

Example illustrates that containing 5% oil and the preparation comprising propofol and albuminised pharmaceutical composition.

Following preparation contains the oil-in-water emulsion of the propofol of 1 % by weight.Also stir until aqueous phase is prepared in dissolving by glycerol (2.25 % by weight) and human serum albumin's (0.5 % by weight) being added in water for injection.By aqueous phase by filter (0.2um filter).By at about 50 DEG C-60 DEG C, lecithin (0.8 % by weight) and propofol (1 % by weight) are dissolved in soybean oil (5 % by weight) and also stir until oil phase is prepared in dissolving.Oil phase to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization in 15 cycles of 5 DEG C of recirculatioies under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.The pharmaceutical composition obtained comprises the component (be weight %) of following usual scope: propofol 0.5-5%; Human serum albumin 0.5-3%; Soybean oil 0.5-10.0%; Lecithin 0.12-1.2%; Glycerol 2.25%; Water for injection in right amount to 100; PH 5-8.Optionally add suitable chelating agen, such as deferoxamine (0.001-0.1%).

Embodiment 22

Example illustrates that the preparation of the pharmaceutical composition containing propofol and albumin, not oil-containing.

Use is similar to method described in embodiment 18, and preparation contains the propofol composition of albumin and Tween 80.By by glycerol (2.25 % by weight), human serum albumin's (0.5 % by weight), Tween 80 (1.5 % by weight) and deferoxamine mesylate (0.1 % by weight) to be added in water for injection and to stir until dissolve and prepare aqueous phase.By aqueous phase by filter (0.2 μm of filter).Propofol (1 % by weight) to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization in 15 cycles of 5 DEG C of recirculatioies under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.The pharmaceutical composition obtained comprises the component (be weight %) of following usual scope: 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%; Water for injection in right amount to 100; PH 5-8.

Embodiment 23

Example illustrates that the preparation of the pharmaceutical composition containing propofol, albumin and D-ALPHA-tocopheryl polyethylene glycol 1000 succinate, not oil-containing.

Use is similar to method described in embodiment 19, and preparation contains the propofol composition of albumin and D-ALPHA-tocopheryl polyethylene glycol 1000 succinate.By by glycerol (2.25 % by weight), human serum albumin's (0.5 % by weight), D-ALPHA-tocopheryl polyethylene glycol 1000 succinate (1 % by weight) and deferoxamine mesylate (0.1 % by weight) to be added in water for injection and to stir until dissolve and prepare aqueous phase.By aqueous phase by filter (0.2 μm of filter).Propofol (1 % by weight) to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization in 15 cycles of 5 DEG C of recirculatioies under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.The pharmaceutical composition obtained comprises the component (be weight %) of following usual scope: propofol 0.5-5; Human serum albumin 0.5-3%; D-ALPHA-tocopheryl polyethylene glycol 1000 succinate 0.5-4.0%; Optionally deferoxamine mesylate 0.0001-0.1%; Glycerol 2.25%; Water for injection in right amount to 100; PH 5-8.

Embodiment 24

Example illustrates that the preparation of the pharmaceutical composition containing propofol, albumin, D-ALPHA-tocopheryl polyethylene glycol 1000 succinate and 1% oil.

The Emulsion of the propofol of 1 % by weight is contained by following method preparation.Also stir until aqueous phase is prepared in dissolving by glycerol (2.25 % by weight) and human serum albumin's (0.5 % by weight) being added in water for injection.By aqueous phase by filter (0.2 μm of filter).Surfactant such as D-ALPHA-tocopheryl polyethylene glycol 1000 succinate (0.5%) is added in aqueous phase.Oil phase is made up of propofol (1 % by weight) and 1% soybean oil.Oil phase to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization 5 DEG C of recirculation maximum 15 cycles under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.

The pharmaceutical composition obtained comprises the component (be weight %) of following usual scope: propofol 0.5-5%; Human serum albumin 0.01-3%; D-ALPHA-tocopheryl polyethylene glycol 1000 succinate 0.1-2%; Soybean oil or other oil (0.1%-5%); Glycerol 2.25%; Water for injection in right amount to 100; PH 5-8.Optionally add deferoxamine (0.001 % by weight-0.1 % by weight).

Embodiment 25

Example illustrates that the preparation of the pharmaceutical composition containing propofol, albumin, D-ALPHA-tocopheryl polyethylene glycol 1000 succinate, 1% oil and electronegative component.

The Emulsion of the propofol of 1 % by weight is contained by following method preparation.Also stir until aqueous phase is prepared in dissolving by glycerol (2.25 % by weight) and human serum albumin's (0.5 % by weight) being added in water for injection.By aqueous phase by filter (0.2 μm of filter).Surfactant such as D-ALPHA-tocopheryl polyethylene glycol 1000 succinate (0.5%) is added in aqueous phase.Oil phase is made up of propofol (1 % by weight) and 1% soybean oil.Add a small amount of electronegative component (0.001%-1%), such as phospholipid or cholate.Oil phase to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization 5 DEG C of recirculation maximum 15 cycles under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.

Embodiment 26

Example illustrates that the preparation of the pharmaceutical composition containing propofol, albumin, D-ALPHA-tocopheryl polyethylene glycol 1000 succinate, 1% oil and electronegative component (NaTDC).

The Emulsion of the propofol of 1 % by weight is contained by following method preparation.Also stir until aqueous phase is prepared in dissolving by glycerol (2.25 % by weight) and human serum albumin's (0.5 % by weight) being added in water for injection.By aqueous phase by filter (0.2 μm of filter).Surfactant such as D-ALPHA-tocopheryl polyethylene glycol 1000 succinate (0.5%) is added in aqueous phase.Oil phase is made up of propofol (1 % by weight) and 1% soybean oil.Add a small amount of electronegative component (0.001%-1%), such as NaTDC.Oil phase to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization 5 DEG C of recirculation maximum 15 cycles under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.

Embodiment 27

Example illustrates that the preparation of the pharmaceutical composition containing propofol, albumin, D-ALPHA-tocopheryl polyethylene glycol 1000 succinate, 1% oil and electronegative component (phospholipid, cholate, polyamino acid etc.).

The Emulsion of the propofol of 1 % by weight is contained by following method preparation.Also stir until aqueous phase is prepared in dissolving by glycerol (2.25 % by weight) and human serum albumin's (0.5 % by weight) being added in water for injection.By aqueous phase by filter (0.2 μm of filter).Surfactant such as D-ALPHA-tocopheryl polyethylene glycol 1000 succinate (0.5%) is added in aqueous phase.Oil phase is made up of propofol (1 % by weight) and 1% soybean oil.Add a small amount of electronegative component (0.001%-1%), such as phosphatidylcholine.Oil phase to be added in aqueous phase and 10,000RPM homogenize 5 minutes.By thick Emulsion high pressure homogenization 5 DEG C of recirculation maximum 15 cycles under 20,000psi.Alternatively, the split tunnel by homogenizer is used.Final Emulsion is filtered (0.2 μm of filter) and stores under nitrogen.

Embodiment 28

Example illustrates that propofol and albuminised combination.

Following mensuration propofol and albuminised combination.The dissolubility of propofol is tested in water and in containing albuminised solution.250 μ L propofol to be added in 10mL water or albumin soln and to stir 2 hours in scintillation vial.Then solution to be transferred in 15mL polyethylene centrifuge tube and remain on 40 DEG C about 16 hours.Measure the propofol in water and albumin soln sample.The dissolubility of propofol in water is measured as 0.12mg/ml.The dissolubility of propofol in albumin soln depends on albuminised concentration, is increased to 0.44mg/ml when albuminised concentration is 2% (20mg/ml).By solution ultrafiltration by 30kD MWCO filter and the propofol measured in filtrate.Find for propofol/aqueous solution, the propofol of 61% can reclaim in filtrate, and for propofol/albumin soln, only 14% is recovered in filtrate, display propofol and albuminised sizable combination.Based on these results, because albumin is combined with propofol, albumin is added the minimizing that the pharmaceutical composition comprising propofol causes the amount of free propofol.

Embodiment 29

Example illustrates that the minimizing by filtration/film contact free propofol in pharmaceutical composition.

As observed in experiment described in embodiment 28, comprise the minimizing that the filtration of the pharmaceutical composition of propofol or ultrafiltration cause the amount of free propofol.Disoprivan and according to the present invention prepare containing albuminised pharmaceutical composition, separately containing 1% propofol (10mg/ml), use 30kD membrane ultrafiltration.HPLC is used to measure the amount of free propofol in filtrate.Concentration for Disoprivan free propofol in filtrate is about 17 μ g/ml, and is about 7 μ g/ml for the concentration of the free propofol of pharmaceutical composition of the present invention in filtrate.Effective minimizing that result corresponds to for the free propofol comprising propofol and albuminised pharmaceutical composition is greater than 2 times.

Embodiment 30

Example illustrates that to people's administration and comprise propofol and albuminised pharmaceutical composition.

Enter row stochastic double blind, comprise propofol and albuminised pharmaceutical composition and propofol formulations--the disadvantageous dermal sensation of Disoprivan be purchased to compare.Carry out testing and obtaining Informed Consent Form from experimenter according to good clinical practice (Good Clinical Practices).The Adult human subjects of any sex can participate in, and prerequisite is that they have unbroken, the normal the back of the hand surface skin in surface.

Originally the preparation be stored in refrigerator is brought in room temperature environment, then 10 μ L preparations is slowly placed on simultaneously the back of the hand of experimenter's two handss.The hands recording them is for the general reaction of preparation and sensation.The result of this research is listed in Table 1.

Table 1

Embodiment 31

Example illustrates that the application as the deferoxamine of antioxidant in the pharmaceutical composition comprising propofol.

Comprise propofol and deferoxamine mesylate, and the pharmaceutical composition containing tween or TPGS is stored in 4 DEG C, 25 DEG C or 40 DEG C, to detect the effect of deferoxamine mesylate in preventing propofol to be oxidized.The concentration these preparations being measured in time to propofol with, measure the antioxidant activity of deferoxamine.These data are reported as the effect % relative to time zero in following table 2 and 3.

Table 2. albumin/Tween formulation

Table 3. albumin/TPGS preparation

Under these conditions, deferoxamine is enough to the oxidation level reducing propofol.This effect is more remarkable under higher temperature.Remarkable oxidation is there is not at 4 DEG C.Use and do not carried out this research by inert material or teflon-coating stopper.

Embodiment 32

Example illustrates that the interior transmission of lung containing paclitaxel and albuminised pharmaceutical composition (ABI-007).

The object of this research be measure [ ³h] ABI-007 time-histories in blood and select tissue after instiling in SpragueDawley rat trachea.

Based on the dose volume of every kg body weight 1.5mL, calculate the target volume of the intratracheal dose formulation to animals administer.Penn-Century micro-sprayer (the model 1A-1B that dosing apparatus is connected by the luer-lock syringe airtight with 1-mL; Penn-Century, Inc., Philadelphia, PA; Purchased from DeLong Distributors, Long Branch, NJ).By the dosage particles inhalation dose doser of proper volume, weigh the device of filling and record weight.Conduit is put into the trachea of dopey animal, the micro-sprayer part of dosing apparatus is placed on tracheal strips by conduit, dosage.After dosed administration, again weigh empty dosing apparatus, the weight difference as dosing apparatus before and after dosed administration calculates the dosage of administration.Be 4.7738 ± 0.0060 (CV 1.5059) mg paclitaxel/kg body weight for all animal mean doses.

Moment after following predetermined close administration collects the blood sample of about 250 μ L from the jugular vein sleeve pipe of the indwelling of JVC rat: 1,5,10,15,30, and 45 minutes (min), and Isosorbide-5-Nitrae, and 8, and 24 hours (h).24 h blood samples, and the blood sample collected from the animal of putting to death 10 minutes, 45 minutes and 2 hours is collected from the rat of anesthesia by heart puncture when putting to death.The all blood samples being used for gross activity analysis are distributed in preweighted sample cell, sample cell is weighed again, by the weight of each sample of subtraction.Measure total tritium level of the blood aliquot of the blood sample collected from jugular vein and the 250-μ L collected from each animal when putting to death.

For all rats, in blood, in the Cmax of tritium 5 minutes (0.0833 hour) times after dosed administration, are observed.The elimination half-life of the tritium measured in the interval from 4h to 24h is 19.73h to 43.02h.It should be noted that this interval only comprises three data points, it can explain the variability of this parameter.Tritium is about 0.04L/h from the apparent clearance rate blood.The result of these experiments is listed in following table 4.

Table 4. intratracheal instillation [ ³h] after ABI-007 in rats blood tritium concentration (mg-eq/L) for the non-compartmental analysis of time graph

As the Functional Analysis of time deriving from after administration in rat vein [ ³h] radioactive mean blood concentration of ABI-007 so that assessment derive from intrarterial [ ³h] bioavailability of tritium of ABI-007.This analysis causes the 24-hour AUC (AUC eventually) of 6.1354mg-eq hr/L.Based on these data, derive from intrarterial [ ³h] radioactivity of ABI-007 is high bioavailability.These analyses are based on gross activity.

Derive from [ ³h] tritium of ABI-007 is rapidly absorbed after intratracheal instillation.Intrarterial [ ³h] after ABI-007 in blood tritium average absorption and eliminate the half-life (being respectively k01 half-life and k10 half-life) and be respectively 0.0155+/-0.0058 hour and 4.738+/-0.366 hour.Tritium is 0.1235+/-0.0180L/ hour (see above table 4) from the average apparent clearance rate blood.

Derive from [ ³h] tritium of ABI-007 absorbed and distributed after intrarterial.Tritium time-histories is in blood described by two compartment models well, and average absorption and elimination half-life are respectively 0.0155 and 4.738 hour.Within 10 minutes after intrarterial, in lung, reclaim the dosage of about 28%.Except gastrointestinal tract, when all inspections, in other tissue, be recovered to the maximum dosage being less than 1%.

Based on from previously using [ ³h] Capxol ^tMthe intravenous dose result of study of carrying out, for three animals of this dosage group, the bioavailability deriving from the tritium of intrarterial is 1.229 ± 0.268 (meansigma methods ± SD).But, it should be noted that this estimation for bioavailability is based on gross activity.Surprisingly, the paclitaxel that the lung approach that the application of the invention contains albuminised compositions transmits fast can biological utilisation, shows the fabulous transhipment striding across lung endothelium.Not to be noted toxicity in animal, this is wonderful, since it is known lung transfer sell endotoxin causes pulmonary toxicity.

Latter 24 hours of dosed administration (intrarterial of 27%), the radioactivity of a great deal of was present in gastrointestinal tract (comprising inclusions).In the gastrointestinal tract the existence of tritium may be due to bile excretion or tritium from respiratory tract by mucociliary clearance and the removing of swallowing subsequently.

Embodiment 33

Example illustrates that the Aerotech II and the Pari aerosol apparatus that comprise paclitaxel and albuminised pharmaceutical composition for pulmonary's transmission.

This research uses Paclitaxel-albumin pharmaceutical composition A BI-007 to carry out under the following conditions: room temperature (20-23 DEG C), relative humidity (48-54%), ambient pressure (629mmHg), nebulizer flowrate (is 10L/min for Aerotech II; Be 7L/min for Pari), total flow rate (28.3L/min), it (is 23lb/in for Aerotech II that nebulizer pressure declines ²; Be 32lb/in for Pari ²), running time (15-60 second), sample volume (1.5mL), ABI-007 paclitaxel concentration (5,10,15 and 20mg/mL).

When being reconstructed with the concentration of 5-15mg/mL by ABI-007, Aerotech II and Pari aerosol apparatus both provide acceptable aggregate efficiency (30%-60%).Pari nebulizer efficiency has the efficiency higher than Aerotech II aerosol apparatus.Pari nebulizer efficiency increases along with ABI-007 concentration and has and decline a little.Observe fabulous fine fraction (74%-96%).Aerotech II aerosol apparatus has the fine fraction higher than Pari aerosol apparatus.Fine fraction and concentration have nothing to do.

Use the ABI-007 solution of 15mg/mL, transmit 100mg paclitaxel being less than Pari aerosol apparatus in 30 minutes.Use the ABI-007 solution of 10mg/mL or 15mg/mL, in about 65 minutes, Aerotech II aerosol apparatus transmits 100mg paclitaxel.For Aerotech II and Pari aerosol apparatus detection perform stability.Until medicine drains, the aerosol concentration of two kinds of aerosol apparatus and efficiency are stable.In the twice that the speed of 15mg/mL, Pari aerosol apparatus consumption medicine is Aerotech II aerosol apparatus, produce the aerosol concentration higher than Aerotech II aerosol apparatus.

In a word, when by lung administration, the nanoparticle/albumin formulation (ABI-007) of paclitaxel shows fabulous bioavailability in rats.Under dosage, there is no the visible marking of early stage toxicity.The lung transmission of Nanoparticle paclitaxel (ABI-007) can use conventional nebulizers to realize.

Embodiment 34

Present embodiment describes the interior transmission of lung of the pharmaceutical composition comprising albumin and rapamycin.The object of this research is that the lung being determined at the later rapamycin in blood of instillation in Sprague Dawley rat trachea absorbs, and compares with intravenous drip.

Dose volume based on every kg body weight 1mL calculates the target volume to the intratracheal dose formulation of animals administer.Penn-Century micro-sprayer (the model 1A-1B that dosing apparatus is connected by the luer-lock syringe airtight with 1-mL; Penn-Century, Inc., Philadelphia, PA; Purchased from DeLong Distributors, Long Branch, NJ).By the dosage particles inhalation dose doser of proper volume, weigh the device of filling and record weight.Conduit is put into the trachea of dopey animal, the micro-sprayer part of dosing apparatus is placed on tracheal strips by conduit, dosage.After dosed administration, again weigh empty dosing apparatus, the weight difference as dosing apparatus before and after dosed administration calculates the dosage of administration.

From the jugular vein sleeve pipe of the indwelling of rat, the blood sample of about 250 μ L is collected: 1,5,10,15,30 in the following predetermined close administration moment, and 45 minutes (min), and Isosorbide-5-Nitrae, 8, and 24 hours (h).The blood sample of all analyses is assigned in the sample cell of weighing in advance, sample cell is weighed again, by the weight of each sample of subtraction.LC/MS/MS is used to measure total rapamycin concentrations of the blood sample collected.

Surprisingly, result is presented at by not having significant difference between lung approach and the blood concentration of rapamycin transmitted by intravenous.Use comprises albuminised pharmaceutical composition, and the bioavailability of the rapamycin transmitted by lung approach is calculated as 109%, shows the fabulous transhipment striding across lung endothelium.

Embodiment 35

The present embodiment illustrate feeding drug into pulmones prepare according to the present invention comprise rapamycin and albuminised pharmaceutical composition after, the tissue distribution of albumin-rapamycin.The object of this research is determined at rapamycin lung absorption in the tissue after instillation in Sprague Dawley rat trachea, and compared with intravenous drip.

In each moment of 10 minutes, 45 minutes, 2 hours and 24 hours from the brain often organizing 3 rats, in lung regulating liver-QI, collect sample.Collect sample and use LC/MS/MS analyzing total rapamycin concentrations.Result display when being transmitted by lung compared with intravenous transmission rapamycin concentrations larger in lung tissue.But compared with transmitting with intravenous (IV), when being transmitted by tracheal strips (IT), the total concentration of rapamycin in brain is lower.In liver, no matter IT or IV transmits, and the concentration of rapamycin seems do not have difference.Based on these results, transmit rapamycin in lung and go for treatment disease (i.e. lung transplantation), wherein the rapamycin of high local concentrations will be useful.

Embodiment 36

Example illustrates that oral delivery contains paclitaxel and albuminised pharmaceutical composition (ABI-007).

The ABI-007 of tritiate is used for the oral administration biaavailability measuring paclitaxel in rats after oral garage.After overnight fast, 5 rats are given the paclitaxel of 5.5mg/kg in ABI-007 (group A) and another 5 rats (group B) use cyclosporin (5.0mg/kg) pretreatment, then use the taxol treatment of 5.6mg/kg in ABI-007.After measuring the radioactivity in blood sample by burning, carry out 0.5,1,2,3,4,5,6,8,12, and the pharmacokinetic analysis of the blood sample extracted for 24 hours.By measuring oral administration biaavailability compared with the intravenous data previously obtained.Result is listed in following table 5.

Table 5. after oral administration 3H-paclitaxel source radioactive average A UC 0-24, C _max, T _maxabsorb with %

Group treatment dosage/approach AUC0-24 absorbs Cmax (mg/kg) Tmax

mg/kg (μg eq×hr/mL) (％) (μg×eq/mL) (hr)

The 5.5/PO of A in normal saline (P) 2.92 44.3 0.245 1

ABI-007

B at the raw 5/PO (C) containing CsA, 8.02 121.1 0.565 0.5

5.6/PO (P) in reason saline

ABI-007

By AUC 0-24IV (6.06 μ g × hr./mL) and IV dosage (5.1mg/kg) for calculating PA (data based on the IV dosage of ABI-007).

Independent ABI-007 is observed to the oral administration biaavailability of 44%.This is significantly higher than for other formulation for paclitaxel viewed.When processing animal with cyclosporin (CsA), bioavailability is increased to 121%.This expects because CsA is the known inhibitor of p-glycoprotein pump, its usually by Inhibitor as paclitaxel absorbs from GI road.The bioavailability being greater than 100% can by explaining the absorption again after paclitaxel bile excretion to GI road.Other known absorption inhibitor or reinforcing agent also may be used for this object.

Embodiment 37

Example illustrates that the paclitaxel that improves after administration comprises paclitaxel and albuminised pharmaceutical composition penetrating to erythrocyte and tumor cell.

By people MX-1 breast tumor fragments subcutaneous transplantation in athymic mouse.With ³the preparation of H paclitaxel paclitaxel comprises paclitaxel and albuminised pharmaceutical composition (" Paclitaxel-albumin ") and taxol as above, and specific activity is 25 μ Ci/mg paclitaxels.When gross tumor volume reaches about 500mm ³time, the administration in saline medium-sized vein by radiolabeled for 20mg/kg Paclitaxel-albumin or taxol.By the sampling of blood plasma, blood and tumor tissues and upon administration 5,15, and 30 minutes and 1,3,8, and 24 hours display analysis radioactivity.WinNonlin, Pharsight, USA is used to analyze Tumor pharmacokinetic (AUC and absorption constant).

Paclitaxel-albumin display to the fast allocation of erythrocyte (RBC), as blood plasma after intravenous administration medicine/blood radioactivity ratio be reduced to fast consistent shown in.Within 1 hour after administration Paclitaxel-albumin, just there occurs to the distribution completely in RBC.By contrast, the paclitaxel being formulated as taxol is many lentamente to the distribution of RBC, and until does not also complete more than 8 hours.

Paclitaxel-albumin shows the fast allocation to tumor tissues, absorption constant (K _a) 3.3X larger than taxol.K _a0.43hr is respectively for Paclitaxel-albumin and taxol ^-1and 0.13hr ^-1.The quick picked-up of paclitaxel causes the tumor AUC of Paclitaxel-albumin higher than taxol by 33%.AUC is respectively 3632nCi*hr/g and 2739nCi*hr/g for Paclitaxel-albumin and taxol.

Embodiment 38

Example illustrates that the safety containing paclitaxel and albuminised pharmaceutical composition to mice administration.

Every day uses the Paclitaxel-albumin or the taxol process athymic mouse that increase dosage gradually, continuous 5 days.By survival rate to plotted versus dosage to measure LD ₅₀.Compared with taxol, Paclitaxel-albumin greatly adds survival rate (p=0.017, ANOVA).For q1d × 5 program, the LD of Paclitaxel-albumin and taxol ₅₀be calculated as 47mg/kg/ days and 30mg/kg/ days respectively.Under the dosage level of 13.4mg/kg/ days, Paclitaxel-albumin and taxol are all tolerated well, and mortality rate is respectively 1% (in 72 mices 1 death) and 4% (in 47 mices 2 death).Under the dosage level of 20mg/kg/ days, the mortality rate of Paclitaxel-albumin is 1% (in 72 mices 1 death), and taxol mortality rate is 17% (in 47 mices 8 death) (p=0.0025).Under the dosage level of 30mg/kg/ days, the mortality rate of Paclitaxel-albumin is 4% (in 72 mices 3 death), and taxol mortality rate is 49% (in 47 mices 23 death) (p < 0.0001).

Embodiment 39

Example illustrates that for Paclitaxel-albumin compositions, stride across the IDN5111 IDN 5111 transporting mechanism of microvascular endothelial cells (EC).

Due to the EPR effect that " seepage " vascular in tumor causes, nanoparticle and albumin-paclitaxel compositions can be accumulated in tumor tissues.Albumin, by the dysuria with lower abdominal colic of the receptor in the caveolae at cell surface place, is transported through EC by the special gp60 receptor (albondin) of albumin.This dysuria with lower abdominal colic mechanism allows albumin-paclitaxel to be transported in the intercellular space of bottom.On the contrary, the polyoxyethylene castor oil in taxol suppresses paclitaxel and albuminised combination, significantly reduces the transhipment of paclitaxel to tumor.In addition, gp16 and gp30 receptor also relates to the albuminised intracellular transport of the modification containing the paclitaxel combined, and cause the combination of paclitaxel and the endotheliocyte increased, blood vessel formation against function is larger compared with taxol.

Embodiment 40

Example illustrates that compared with taxol, the increase of the endothelium dysuria with lower abdominal colic containing paclitaxel and albuminised pharmaceutical composition.

People's lung microvascular endothelial cells (HLMVEC) is cultured on transwell and converges.Be that 20 μ g/mL the present invention contain paclitaxel and albuminised pharmaceutical composition or taxol and are added in upper transwell room by concentration.

Exometer continuous monitoring paclitaxel is used to be transported to transhipment in lower room from upper room by dysuria with lower abdominal colic.Also use only containing Flutax and without albuminised contrast.Contrast containing Flutax does not show transhipment, proves the integrity converging HLMVEC monolayer.Under the existence of 5%HSA (physiological concentration), the transhipment from the paclitaxel of albumin-paclitaxel compositions is more faster than the paclitaxel from taxol.Transport velocity constant (K _t) 1.396hr is respectively for albumin-paclitaxel compositions and taxol ^-1and 0.03hr ^-1.The total amount that the transhipment of albumin-paclitaxel compositions strides across the paclitaxel of monolayer strides across the paclitaxel of monolayer total amount than the transhipment of taxol is high 3 times.

Embodiment 41

Example illustrates that compared with taxol, the endotheliocyte (EC) improved containing paclitaxel and albuminised pharmaceutical composition combines.

Human umbilical vein endothelial cells (HUVEC) is cultivated on the microtitration plate of 96-hole.In an experiment, under the existence of polyoxyethylene castor oil EL/EtOH (carrier of taxol) increasing concentration, HUVEC and paclitaxel (paclitaxel of Flutax-Oregon Green labelling) are reacted.In another experiment, the pharmaceutical composition containing albumin and Flutax and Taxol-Flutax composition react with HUVEC under various final concentration.The combination of paclitaxel and cell is subject to the suppression of polyoxyethylene castor oil.By the IC of 0.02% of polyoxyethylene castor oil EL/EtOH ₅₀display suppresses.The polyoxyethylene castor oil of this concentration has been presented at taxol chemotherapeutic period and has continued at least 24 hours.Therefore, it is correlated process in body.Under all detectable concentrations, the paclitaxel from albumin-paclitaxel compositions of significant quantity and Cell binding.By contrast, taxol observed seldom or do not observe combination.

Embodiment 42

Example illustrates that compared with taxol, the albumin of the improvement containing paclitaxel and albuminised pharmaceutical composition combines.

Human serum albumin (HSA) is fixed on plastic ELISA plate.Paclitaxel (paclitaxel of Flutax-Oregon Green labelling) and fixing HSA are reacted under the existence of polyoxyethylene castor oil EL/EtOH increasing concentration.In another experiment, albumin-paclitaxel-Flutax compositions and Taxol-Flutax composition react under the final concentration of 20 μ g paclitaxel/mL with the HSA fixed.Paclitaxel and albuminised combination are subject to the suppression of polyoxyethylene castor oil.By the IC of 0.003% of polyoxyethylene castor oil EL/EtOH ₅₀display suppresses.The polyoxyethylene castor oil of this concentration has been presented at taxol chemotherapeutic period and has continued at least 24 hours.Therefore, it is correlated process in body.Under relevant pharmacologic paclitaxel concentration (20 μ g/mL), the paclitaxel from albumin-paclitaxel compositions of significant quantity is combined with fixing HSA.By contrast, combination is not observed for taxol.

Embodiment 43

Example illustrates that compared with taxol, for the paclitaxel increased containing paclitaxel and albuminised pharmaceutical composition to albuminised transhipment.

By taxol-Flutax and albumin-paclitaxel-Flutax compositions with 20 μ g/mL, 40 μ g/ml, and 80 μ g/ml are with the 5%HSA in Hanks buffer or mix with serum.Separating mixture in natural 3-14% polyacrylamide gel immediately, measures the amount of the paclitaxel be combined with albumin by scanning fluorimeter.Compared with taxol, for albumin-paclitaxel compositions, paclitaxel is faster to the transhipment of HSA.When by serum or 5%HSA and albumin-paclitaxel-Flutax compositions or Taxol-Flutax composition incubation, more paclitaxel and HSA common-battery are swum.Compared with Taxol-Flutax composition, 20 μ g/ml, 40 μ g/ml, with the albumin-paclitaxel-Flutax compositions of 80 μ g/ml after contacting 5%HSA, have 45%, 60% respectively, and 33% more paclitaxel is transported to HSA.For 260mg/m ²aBI-007, C _maxfor about 20 μ g/mL, therefore this is important physiological disposition.

Embodiment 44

Example illustrates that glycoprotein receptor gp60 is responsible for combination and the dysuria with lower abdominal colic of albumin-paclitaxel.

Fluorescently-labeled paclitaxel (Flutax) albumin compositions contacts with the microvascular endothelial cells in culture.Examine under a microscope fluorescence staining, evidence is speckle regions, it is speculated that the gp60 receptor be combined with albumin-paclitaxel.This confirms by using the albumin of rhodamine labelling, and the albumin of rhodamine labelling is identical with the punctuate fluorescence position of paclitaxel.

Embodiment 45

Example illustrates that the albumin of recruitment can be competed with the combination of paclitaxel.

Albumin is fixed on microtitration plate.Fluorescent paclitaxel is added in hand-hole, use scanning fluorimeter to measure the combination of paclitaxel.The albumin of recruitment is added the suppression level also measuring the paclitaxel be combined with fixing albumin in hand-hole.Data display increases along with the albuminised amount added, and observes the corresponding reduction of combination.Similar effect is observed for the combination with endotheliocyte.The combination of the albumin concentration suppression paclitaxel that this display is higher.Preferred the present invention has the albuminised compositions of relatively low amount.

Embodiment 46

Example illustrates that the albumin of relatively low amount in pharmaceutical composition of the present invention causes stable compositions.

In order in seminar's compound, whether the albumin of relatively low amount affects the stability of pharmaceutical composition of the present invention, preparation is containing the albuminised albumin-paclitaxel compositions of low amounts.When check under different temperatures (2-8 DEG C, 25 DEG C and 40 DEG C) formation of potency of paclitaxel, impurity, particle diameter, pH and other stability canonical parameter time, find that these compositionss are equally stable with the albuminised compositions with more a large amount.Therefore this preferably there is the albuminised compositions of relatively low amount, because can greatly reduce costs and allow the combination of increase and cytotropic transhipment.

Embodiment 47

Example illustrates that the pharmaceutical composition containing albumin and paclitaxel, it has high albumin and paclitaxel ratio.

30mg paclitaxel is dissolved in 3.0ml dichloromethane.Solution is added to (ratio being equivalent to albumin and paclitaxel is 27) in 27.0ml human serum albumin solution (3%w/v).Add deferoxamine as required.This mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to rotary evaporator, 40 DEG C, fast remove dichloromethane 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the paclitaxel particles of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.

Embodiment 48

Example illustrates that the pharmaceutical composition containing albumin and paclitaxel, it has low albumin and paclitaxel ratio.

Particularly, 300mg paclitaxel is dissolved in 3.0ml dichloromethane.Solution is added to (ratio being equivalent to albumin and paclitaxel is 4.5) in 27ml human serum albumin solution (5%w/v).Add deferoxamine as required.This mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to rotary evaporator, 40 DEG C, fast remove dichloromethane 20-30 minute under decompression (30mm Hg).The dispersion obtained is translucent, and the typical average diameter of the paclitaxel particles of acquisition is 50-220nm (Z-meansigma methods, Malvern Zetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.

Embodiment 49

Example illustrates that the pharmaceutical composition containing albumin and paclitaxel, it has medium albumin and paclitaxel ratio.

Particularly, 135mg paclitaxel is dissolved in 3.0ml dichloromethane.Solution is added in 27ml human serum albumin solution (5%w/v).Add deferoxamine as required.This mixture homogenize 5 minutes (Vitris homogenizer, model Tempest I.Q.) under low RPM, to form thick Emulsion, is then transferred in high-pressure homogenizer (Avestin).Emulsifying is carried out, simultaneously by least 5 cycles of Emulsion recirculation under 9000-40,000psi.The system of acquisition is transferred to rotary evaporator, under decompression (30mm Hg), removes dichloromethane 20-30 minute at 40 DEG C fast.The dispersion obtained is translucent, and the typical average diameter of the paclitaxel particles of acquisition is 50-220nm (Z-meansigma methods, MalvernZetasizer).By further for this dispersion lyophilizing 48 hours.By adding sterilized water or saline, the cake block of acquisition can easily be reconstructed into original dispersion.Particle diameter is after reconstitution identical with before lyophilizing.In the compositions of the present invention, the calculating ratio (w/w) of albumin and paclitaxel is about 10.

Embodiment 50

Example illustrates that by the rheumatoid arthritis in albumin-paclitaxel compositions treatment animal model.

The arthritis model that collagen protein in Louvain rat is induced is used for detecting albumin-paclitaxel compositions to arthritic therapeutic effect.Monitor that the pawl size of laboratory animal is to assess arthritic seriousness.

After arthritis develops completely (usually about 9-10 days after collagen injections), laboratory animal is divided into different groups and accepts albumin-paclitaxel 1mg/kg q.o.d with intraperitoneal, or albumin-paclitaxel 0.5mg/kg+ prednisone 0.2mg/kg q.o.d. (therapeutic alliance) reaches 6 dosage, then individual dosage reaches three weeks on every Mondays.The measuring claw size when treating beginning (the 0th day) and per injection medicine.Only accept one group of normal saline in contrast.At the end of experiment, the group pawl size accepting albumin-paclitaxel realizes the reduction of 42%, and therapeutic alliance group display pawl size reduces 33%, and when matched group pawl size starts with treatment compared with increase about 20%.

In a word, albumin-paclitaxel compositions shows therapeutic effect to arthritis.The combination of albumin-paclitaxel can be positioned the position of arthritic lesions by the transhipment of receptor-mediated mechanism as gp60.

Embodiment 51

Example illustrates that the application of albumin-paclitaxel compositions in treatment cardiovascular restenosis.

In animal, paclitaxel outflow support (Paclitaxel eluting stents) causes incomplete healing, in some situation, causes lacking the lasting suppression to neointima growth in the artery.This research examines new systemic delivery albumin-paclitaxel present composition for the effect reducing in-stent restenosis.

The albumin-paclitaxel of saline reconstruct is detected in 38 New Zealand white rabbit accepting bilateral iliac artery support.Using the endoarterial infusion administration of the dosage (1.0-5.0mg/kg dose of paclitaxel) of albumin-paclitaxel as 10-minute; Control animal accepts carrier (0.9% normal saline).

In ensuing chronic experiment, 5.0mg/kg albumin-paclitaxel is supplied to support, at the 28th day intravenous repeated doses administration 3.5-mg/kg albumin-paclitaxel; These researchs stopped at 3 months.At the 28th day, reduce average neointimal thickness (p <=0.02) by doses of albumin-paclitaxel >=2.5mg/kg, evidence is the healing extended.But effect of the 5.0mg/kg albumin-paclitaxel of single dose was lost before 90 days.On the contrary, after upper bracket, 28 days second time repeat administration 3.5mg/kg albumin-paclitaxel cause (comparing with contrast with single dose 5.0mg/kg albumin-paclitaxel in the lasting suppression of 90 days stylish inner film thickness, p <=0.009), neointima almost heals completely.

Although the albumin-paclitaxel of system reduced neointima growth at 28 days, sustained neointimal is suppressed to require single repeat dose.Therefore, the present composition is applicable to Cardiovarscular as restenosis.The present invention comprises medicament (the such as rapamycin except paclitaxel, other taxanes, Epothilones etc.) compositions be all applicable to treat blood vessel or artificial blood vessel grafts as in the patient needing hemodialysis for artery-vein entrance those in restenosis.

Claims

1. one kind comprises the pharmaceutical composition of medicament and pharmaceutical carrier, wherein said pharmaceutical carrier comprises albumin, and wherein said pharmaceutical carrier comprises the deferoxamine effectively measured for the growth of microorganism suppressed in described pharmaceutical composition, wherein said medicament is paclitaxel or propofol, wherein said pharmaceutical composition comprises: the deferoxamine of 0.0001 % by weight to 0.5 % by weight, the albumin of 0.1 % by weight to 25 % by weight, the propofol of 0.1 % by weight to 5 % by weight or the paclitaxel of 0.1 % by weight to 1 % by weight, and after this pharmaceutical composition lyophilizing and dehydration, there is the particle diameter of 50-220nm.

2. the pharmaceutical composition of claim 1, wherein said pharmaceutical composition comprises the albumin of 0.5 % by weight to 5 % by weight.

3. the pharmaceutical composition of claim 1, wherein said albumin is human serum albumin.

4. the pharmaceutical composition of claim 1, wherein said medicament is propofol.

5. the pharmaceutical composition of claim 1, wherein said pharmaceutical composition comprises the deferoxamine of 0.1 % by weight.

6. the pharmaceutical composition of claim 1, wherein said pharmaceutical composition comprises deferoxamine mesylate.

7. the pharmaceutical composition of claim 1, wherein said pharmaceutical composition is oil-in-water emulsion.

8. the pharmaceutical composition of claim 6, wherein said medicament is propofol, and described deferoxamine mesylate is with the amount existence of 0.0001 % by weight to 0.5 % by weight.

9. the pharmaceutical composition of claim 1, wherein said pharmaceutical composition is unit dose.

10. the pharmaceutical composition of claim 1, wherein said pharmaceutical composition is multiple dose.

The pharmaceutical composition of 11. claim 1, wherein said pharmaceutical composition is contained in the container of sealing.

Pharmaceutical composition in 12. claim 1, wherein said medicament is paclitaxel.

The pharmaceutical composition of 13. claim 1, wherein said pharmaceutical composition there is no polyoxyethylene castor oil.

The pharmaceutical composition of 14. claim 1, wherein said pharmaceutical composition does not have polyoxyethylene castor oil.