Liposomes useful for drug delivery

• US 20070116753A1
• Filed: 05/02/2005
• Published: 05/24/2007
• Est. Priority Date: 05/03/2004
• Status: Active Grant

• Current Assignee: IPSEN Biopharm Limited (Ipsen Sa)
• Original Assignee: Hermes Biosciences Inc. (Merrimack Pharmaceuticals, Inc.)
• Inventors: Hong, Keelung, Kirpotin, Dmitri, Drummond, Daryl
• Granted Patent: US 8,147,867 B2
• Time in Patent Office: Days
• US Class Current: 424/450
• CPC Class Codes: A61K 31/337 having four-membered rings, e.g. taxol
• CPC Class Codes: A61K 31/4375 the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
• CPC Class Codes: A61K 31/4745 condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines yohimbine derivatives, vinblastine A61K31/475; ergoline derivatives A61K31/48
• CPC Class Codes: A61K 31/475 having an indole ring, e.g. yohimbine, reserpine, strychnine, vinblastine vincamine A61K31/4375
• CPC Class Codes: A61K 31/704 attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin digitoxin A61K31/7048
• CPC Class Codes: A61K 47/10 Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
• CPC Class Codes: A61K 47/24 containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
• CPC Class Codes: A61K 47/26 Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
• CPC Class Codes: A61K 47/28 Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61K 47/6913 the liposome being modified on its surface by an antibody
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/127 Liposomes
• CPC Class Codes: A61K 9/1271 Non-conventional liposomes, e.g. PEGylated liposomes, liposomes coated with polymers liposome as conjugate A61K47/6911
• CPC Class Codes: A61K 9/1277 Processes for preparing; Proliposomes
• CPC Class Codes: A61K 9/1278 Post-loading, e.g. by ion or pH gradient
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07H 13/12 : by acids having the group -X-C(=X)-X-, or halides thereof, in which each X means nitrogen, oxygen, sulfur, selenium or tellurium, e.g. carbonic acid, carbamic acid
• CPC Class Codes: Y10S 977/773 : Nanoparticle, i.e. structure having three dimensions of 100 nm or less
• CPC Class Codes: Y10S 977/906 : Drug delivery
• CPC Class Codes: Y10S 977/907 : Liposome

Abstract

The present invention provides liposome compositions containing substituted ammonium and/or polyanion, and optionally with a desired therapeutic or imaging entity. The present invention also provide methods of making the liposome compositions provided by the present invention.

Claims

1-12. -12. (canceled)

13. A composition comprising a liposome in a medium, said liposome having an interior space separated from the medium by a membrane comprising one or more lipids, wherein said interior space comprises an anion selected from the group consisting of a polyanionized polyol or polyanionized sugar, and wherein said liposome comprises a transmembrane gradient effective for retention of an entity within said liposome.

14. The composition of claim 13 wherein said transmembrane gradient is an ion gradient, an ammonium ion gradient, a pH gradient, an electrochemical potential gradient, a solubility gradient, or a gradient of a substituted ammonium ion comprising at least one C—

N bond.

15. The composition of claim 14 wherein said anion comprises

(i) a polyol moiety selected from the group consisting of a monosaccharide, a disaccharide, a linear polyhydroxylated compound, a cyclic polyhydroxylated compound, an aliphatic alcohol having more than one hydroxyl group, an alicyclic alcohol having more than one hydroxyl group, or a heterocyclic alcohol having more than one hydroxyl group, and

(ii) linked to said polyol moiety, at least two strongly anionic functional groups.

16. The composition of claim 15 wherein at least one of said strongly anionic functional groups is a sulfuric acid ester, a phosphoric acid ester, a boric acid ester, a sulfonic acid group, a phosphonic acid group, a thiocarbonic acid group, a dithiocarbonic acid group, a derivative thereof, or an analog thereof.

17. The composition of claim 15 wherein said polyol moiety is selected from the group consisting of arabinose, ribose, xylose, glucose, galactose, sorbose, fructose, maltose, sucrose, lactose, trehalose, ethylene glycol, prolylene glycol, glycerol, treitol, erythritol, pentaerythritol, ribitol, arabitol, sorbitol, mannitol, lactitol, maltitol, fructitol, glucitol, xylitol, or inositol.

18. The composition of claim 15 wherein the anion is polysulfated sucrose having from 3 to 8 sulfate groups per molecule.

19. The composition of claim 15 wherein said polysulfated sucrose is sucrose octasulfate.

20. The composition of claim 15 wherein the polysulfated sucrose is sucrose octasulfate.

21. The composition of claim 15 wherein the concentration of said anion in said liposome interior is at least about 0.05, 0.1, 0.2, 0.5, 0.6, 0.7, or 1.0 gram-equivalent/L.

22. The composition of claim 15 wherein the concentration of said anion in said liposome interior is about 0.65 gram-equivalent/L or about 1.0 gram-equivalent/L.

24. The composition of claim 15 wherein the interior space of the liposome further comprises a globally cationic entity, a therapeutic entity, or a detectable marker, wherein the globally cationic entity is other than an ammonium ion or substituted ammonium ion.

25. The composition of claim 24 wherein said entity is present in said interior space at a concentration which exceeds the concentration of said entity in said medium.

26. The composition of claim 24 wherein said medium is essentially free from said entity.

27. The composition of claim 24 wherein molar ratio of said entity to the totality of said lipids is at least about 0.05, at least about 0.1, at least about 0.2, at least about 0.3, at least about 0.5, at least about 0.7, or at least about 1.0.

28. The composition of claim 24 wherein said therapeutic entity is an antimicrobial therapeutic, antiviral therapeutic, or an anti-neoplastic therapeutic.

29. The composition of claim 24 wherein said therapeutic entity is an aminoglycoside antibiotic or a fluoroquinolone derivative.

30. The composition of claim 28 wherein said therapeutic entity is selected from a group consisting of:

a topoisomerase inhibitor, a farnesyltransferase inhibitor, a tyrosine kinase inhibitor, a cyclin-dependent kinase inhibitor, a phosphatase inhibitor, an aurora kinase inhibitor, a microtubule depolymerizing agent, a microtubule stabilizing agent, an alkylating agent, an enzyme, an enzyme inhibitor, a histone deacetylase inhibitor, an antimetabolite, a receptor-binding agent, a hormone, a hormone antagonist, a nucleotide, a polynucleotide, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

31. The composition of claim 28 wherein said anti-neoplastic therapeutic is selected from a group consisting of an anthracycline compound, a camptothecin compound, a vinca alkaloid, an ellipticine compound, a taxane compound, a wortmannin compound, a pyrazolopyrimidine compound, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

32. The composition of claim 24 wherein said therapeutic entity is selected from a group consisting of:

doxorubicin, daunorubicin, mitomycin C, epirubicin, pirarubicin, rubidomycin, carcinomycin, N-acetyladriamycin, rubidazone, 5-imidodaunomycin, N-acetyldaunomycin, daunoryline, mitoxanthrone, camptothecin, 9-aminocamptothecin, 7-ethylcamptothecin, 10-hydroxycamptothecin, 9-nitrocamptothecin, 10,11-methylenedioxycamptothecin, 9-amino-10,11-methylenedioxycamptothecin, 9-chloro-10,11-methylenedioxycamptothecin, irinotecan, topotecan, lurtotecan, silatecan, (7-(4-methylpiperazinomethylene)-10,11-ethylenedioxy-20(S)-camptothecin, 7-(4-methylpiperazinomethylene)-10,11-methylenedioxy-20(S)-camptothecin, 7-(2-N-isopropylamino)ethyl)-(20S)-camptothecin, vincristine, vinblastine, vinorelbine, vinflunine, vinpocetine, vindesine, ellipticine, 6-3-aminopropyl-ellipticine, 2-diethylaminoethyl-ellipticinium and salts thereof, datelliptium, retelliptine, paclitaxel, docetaxel, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

33. The composition of claim 24 wherein said therapeutic entity is selected from a group consisting of:

an antihistamine ethylenediamine derivative, bromphenifamine, diphenhydramine, an anti-protozoal drug, quinolone, iodoquinol, an amidine compound, pentamidine, an antihelmintic compound, pyrantel, an anti-schistosomal drug, oxaminiquine, an antifungal triazole derivative, fliconazole, itraconazole, ketoconazole, miconazole, an antimicrobial cephalosporin, cefazolin, cefonicid, cefotaxime, ceftazimide, cefuoxime, an antimicrobial beta-lactam derivative, aztreopam, cefmetazole, cefoxitin, an antimicrobial of erythromycine group, erythromycin, azithromycin, clarithromycin, oleandomycin, a penicillin compound, benzylpenicillin, phenoxymethylpenicillin, cloxacillin, methicillin, nafcillin, oxacillin, carbenicillin, a tetracycline compound, novobiocin, spectinomycin, vancomycin;

an antimycobacterial drug, aminosalicyclc acid, capreomycin, ethambutol, isoniazid, pyrazinamide, rifabutin, rifampin, clofazimine, an antiviral adamantane compound, amantadine, rimantadine, a quinidine compound, quinine, quinacrine, chloroquine, hydroxychloroquine, primaquine, amodiaquine, mefloquine, an antimicrobial qionolone, ciprofloxacin, enoxacin, lomefloxacin, nalidixic acid, norfloxacin, ofloxacin, a sulfonamide;

a urinary tract antimicrobial, nitrofurantoin, trimetoprim;

a nitroimidazoles derivative, metronidazole, a cholinergic quaternary ammonium compound, ambethinium, neostigmine, physostigmine, an anti-Alzheimer aminoacridine, tacrine, an anti-Parkinsonal drug, benztropine, biperiden, procyclidine, trihexylhenidyl, an anti-muscarinic agent, atropine, hyoscyamine, scopolamine, propantheline, an adrenergic compound, dopamine, serotonin, albuterol, dobutamine, ephedrine, epinephrine, norepinephrine, isoproterenol, metaproperenol, salmetrol, terbutaline, a serotonin reuptake inhibitor, an ergotamine derivative, a myorelaxant of a curare series, a central action myorelaxant, baclophen, cyclobenzepine, dentrolene, nicotine, a nicotine receptor antagonist, a beta-adrenoblocker, acebutil, amiodarone, a benzodiazepine compound, ditiazem, an antiarrhythmic drug, diisopyramide, encaidine, a local anesthetic compound, procaine, procainamide, lidocaine, flecaimide, quinidine;

an ACE inhibitor, captopril, enelaprilat, fosinoprol, quinapril, ramipril;

an opiate derivative, codeine, meperidine, methadone, morphine, an antilipidemic, fluvastatin, gemfibrosil, an HMG-coA inhibitor, pravastatin, a hypotensive drug, clonidine, guanabenz, prazocin, guanethidine, granadril, hydralazine, a non-coronary vasodilator, dipyridamole, an acetylcholine esterase inhibitor, pilocarpine, an alkaloid, physostigmine, neostigmine, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

34. The composition of claim 24 wherein at least 90% of said entity remains encapsulated in the liposome after 6 months at 2-8°

C.

35. The composition of claim 24 wherein at least 80% of said entity remains encapsulated in the liposome after 2 years at 2-8°

C.

36. The composition of claim 24 wherein the entity is contained in the liposomes of the composition at a first entity-to-lipid ratio, and wherein 24 hours following administration of said composition into the bloodstream of a mammal, the entity remains contained in the liposomes at a second entity-to-lipid ratio wherein the second entity-to-lipid ratio is over 50%, at least 60%, or at least 70% of the first substance-to-lipid ratio.

37. The composition of claim 36 wherein the entity is a vinca alkaloid, an analog thereof, or a derivative thereof.

38. The composition of claim 36 wherein the entity is vinorelbine.

39. The composition of claim 36 wherein said mammal is a rat.

105. The composition of claim 24 wherein the liposome comprises a targeting moiety.

106. The composition of claim 105 wherein said targeting moiety is a protein, a peptide, a polysaccharide, a polynucleotide, natural small molecule, a synthetic small molecule, a combination thereof, or a derivative thereof.

107. The composition of claim 105 wherein said targeting moiety is a naturally, synthetically, or recombinantly produced protein comprising an antigen binding sequence of an antibody.

108. The composition of claim 105 wherein said targeting moiety is an antibody, an antigen-binding fragment thereof, a single-chain protein comprising the antigen-binding polypeptide sequences of an antibody, a single-domain antibody, an analog of any of the foregoing, or a derivative of any of the foregoing.

109. The composition of claim 105 wherein said targeting moiety is linked to said liposome membrane and exposed to said medium.

110. The composition of claim 109 wherein said linked targeting moiety comprises a hydrophilic polymer that links said liposome membrane to said moiety.

111. The composition of claim 110 wherein said hydrophilic polymer is poly(ethylene glycol).

112. The composition of claim 111 wherein said poly(ethylene glycol) has molecular weight from about 250 to about 30,000.

113. The composition of claim 105 wherein the targeting moiety effects internalization of said liposome in a cell.

114. The composition of claim 105 wherein the targeting moiety selectively binds to a receptor tyrosine kinase, a growth factor receptor, an angiogenic factor receptor, a transferrin receptor, a cell adhesion molecule, or a vitamin receptor.

115. The composition of claim 114 wherein said tyrosine kinase receptor is a growth factor receptor

116. The composition of claim 115 wherein said tyrosine kinase growth factor receptor is an EGFR, ErbB-2 (HER-2), ErbB-3 (HER3), or ErbB-4 (HER4).

117. The composition of claim 114 wherein said angiogenic factor receptor is bFGF receptor, or VEGF receptor.

118. The composition of claim 114 wherein said cell adhesion molecule is an integrin.

119. The composition of claim 113 wherein the cell is a malignant cell.

120. A method of encapsulating an entity into a liposome comprising a step of contacting the composition of claim 15 with the entity for the time sufficient for the substance to become encapsulated into the liposome.

121. The method of claim 120 wherein at least a portion of said entity enters the liposome interior space.

122. The method of claim 120 wherein at least 90% of said entity enters the liposome interior space.

123. The method of claim 120 wherein the proportion of said entity that becomes encapsulated into the liposomes is at least 80%, at least 90%, or at least 95%.

124. The method of claim 120 wherein the entity is a globally cationic entity, a therapeutic entity, or a detectable marker.

125. The method of claim 120 wherein molar ratio of said entity to the totality of said lipids is at least about 0.05, about 0.1, about 0.2, or about 0.3.

126. The method of claim 120 wherein said therapeutic entity is an antimicrobial therapeutic, antiviral therapeutic, or an anti-neoplastic therapeutic.

127. The method of claim 120 wherein said therapeutic entity is an aminoglycoside antibiotic or a fluoroquinolone derivative.

128. The method of claim 124 wherein said therapeutic entity is selected from a group consisting of:

a topoisomerase inhibitor, a farnesyltransferase inhibitor, a tyrosine kinase inhibitor, a cyclin-dependent kinase inhibitor, a phosphatase inhibitor, an aurora kinase inhibitor, a microtubule depolymerizing agent, a microtubule stabilizing agent, an alkylating agent, an enzyme, an enzyme inhibitor, a histone deacetylase inhibitor, an antimetabolite, a receptor-binding agent, a hormone, a hormone antagonist, a nucleotide, a polynucleotide, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

129. The method of claim 124 wherein said therapeutic entity is selected from a group consisting of an anthracycline compound, a camptothecin compound, a vinca alkaloid, an ellipticine compound, a taxane compound, a wortmannin compound, a pyrazolopyrimidine compound, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

130. The method of claim 124 wherein said therapeutic entity is selected from a group consisting of:

doxorubicin, daunorubicin, mitomycin C, epirubicin, pirarubicin, rubidomycin, carcinomycin, N-acetyladriamycin, rubidazone, 5-imidodaunomycin, N-acetyldaunomycin, daunoryline, mitoxanthrone, camptothecin, 9-aminocamptothecin, 7-ethylcamptothecin, 10-hydroxycamptothecin, 9-nitrocamptothecin, 10,11-methylenedioxycamptothecin, 9-amino-10,11-methylenedioxycamptothecin, 9-chloro-10,11-methylenedioxycamptothecin, irinotecan, topotecan, lurtotecan, silatecan, (7-(4-methylpiperazinomethylene)-10,11-ethylenedioxy-20(S)-camptothecin, 7-(4-methylpiperazinomethylene)-10,11-methylenedioxy-20(S)-camptothecin, 7-(2-N-isopropylamino)ethyl)-(20S)-camptothecin, vincristine, vinblastine, vinorelbine, vinflunine, vinpocetine, vindesine, ellipticine, 6-3-aminopropyl-ellipticine, 2-diethylaminoethyl-ellipticinium and salts thereof, datelliptium, retelliptine, paclitaxel, docetaxel, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

131. The method of claim 124 wherein said therapeutic entity is selected from a group consisting of:

an antihistamine ethylenediamine derivative, bromphenifamine, diphenhydramine, an anti-protozoal drug, quinolone, iodoquinol, an amidine compound, pentamidine, an antihelmintic compound, pyrantel, an anti-schistosomal drug, oxaminiquine, an antifungal triazole derivative, fliconazole, itraconazole, ketoconazole, miconazole, an antimicrobial cephalosporin, cefazolin, cefonicid, cefotaxime, ceftazimide, cefuoxime, an antimicrobial beta-lactam derivative, aztreopam, cefmetazole, cefoxitin, an antimicrobial of erythromycine group, erythromycin, azithromycin, clarithromycin, oleandomycin, a penicillin compound, benzylpenicillin, phenoxymethylpenicillin, cloxacillin, methicillin, nafcillin, oxacillin, carbenicillin, a tetracycline compound, novobiocin, spectinomycin, vancomycin;

an antimycobacterial drug, aminosalicyclc acid, capreomycin, ethambutol, isoniazid, pyrazinamide, rifabutin, rifampin, clofazimine, an antiviral adamantane compound, amantadine, rimantadine, a quinidine compound, quinine, quinacrine, chloroquine, hydroxychloroquine, primaquine, amodiaquine, mefloquine, an antimicrobial qionolone, ciprofloxacin, enoxacin, lomefloxacin, nalidixic acid, norfloxacin, ofloxacin, a sulfonamide;

a urinary tract antimicrobial, nitrofurantoin, trimetoprim;

a nitroimidazoles derivative, metronidazole, a cholinergic quaternary ammonium compound, ambethinium, neostigmine, physostigmine, an anti-Alzheimer aminoacridine, tacrine, an anti-Parkinsonal drug, benztropine, biperiden, procyclidine, trihexylhenidyl, an anti-muscarinic agent, atropine, hyoscyamine, scopolamine, propantheline, an adrenergic compound, dopamine, serotonin, albuterol, dobutamine, ephedrine, epinephrine, norepinephrine, isoproterenol, metaproperenol, salmetrol, terbutaline, a serotonin reuptake inhibitor, an ergotamine derivative, a myorelaxant of a curare series, a central action myorelaxant, baclophen, cyclobenzepine, dentrolene, nicotine, a nicotine receptor antagonist, a beta-adrenoblocker, acebutil, amiodarone, a benzodiazepine compound, ditiazem, an antiarrhythmic drug, diisopyramide, encaidine, a local anesthetic compound, procaine, procainamide, lidocaine, flecaimide, quinidine;

an ACE inhibitor, captopril, enelaprilat, fosinoprol, quinapril, ramipril;

an opiate derivative, codeine, meperidine, methadone, morphine, an antilipidemic, fluvastatin, gemfibrosil, an HMG-coA inhibitor, pravastatin, a hypotensive drug, clonidine, guanabenz, prazocin, guanethidine, granadril, hydralazine, a non-coronary vasodilator, dipyridamole, an acetylcholine esterase inhibitor, pilocarpine, an alkaloid, physostigmine, neostigmine, a derivative of any of the foregoing, a pro-drug of any of the foregoing, and an analog of any of the foregoing.

132. The method of claim 120 wherein the contacting is in an aqueous solution.

133. The method of claim 132 wherein said aqueous solution has pH in the range of about 4 to about 7.

134. The method of claim 132 wherein said aqueous solution has ionic strength equivalent or less than that of 50 mM sodium chloride.

135. The method of claim 132 wherein the aqueous solution has ionic strength equivalent or less than that of 50 mM sodium chloride.

136. The method of claim 132, wherein the aqueous solution has pH between about 6.0 and about 7.0, and the entity is a vinca alkaloid, an analog thereof, or a derivative thereof.

137. The method of claim 136 wherein the pH of the aqueous solution of about 6.5, and the entity is vinorelbine.

138. The method of claim 135 wherein the pH of the aqueous solution is between about 5 and about 7, and the substance is a camptothecin derivative.

139. The method of claim 138 wherein the pH of the aqueous solution is between about 5.0 and about 6.5, and the substance is Topotecan or Irinotecan.

140. The method of claim 134, wherein following the contacting, the ionic strength of the aqueous solution is increased to more than that of 50 mM sodium chloride.

141. The method of claim 140 wherein said ionic strength value is increased to at least that of 100 mM sodium chloride.

142. The method of claim 141 wherein said ionic strength value is increased to at least that of 150 mM sodium chloride.

156. A kit for providing a liposomally encapsulated entity, comprising the composition of claim 15, an instruction for using the composition to encapsulate an entity, and optionally, in a separate container, the entity.

157. A kit of claim 156 wherein the entity is a globally cationic substance, a therapeutic entity, or a detectable marker.

158. A composition of claim 15 wherein the liposome comprises a substituted ammonium compound having in a aqueous solution at ambient temperature a pKa value of at least 8.0, at least 8.5, at least 9.0, at least 9.5, or at least 10.0.

159. A method of claim 120 wherein the composition contacted with the entity comprises a liposome, said liposome comprises a substituted ammonium compound having in a aqueous solution at ambient temperature a pKa value of at least 8.0, at least 8.5, at least 9.0, at least 9.5, or at least 10.0.

162. The composition of claim 158 wherein the substituted ammonium is pharmaceutically inert.

164. The composition of claim 158 wherein the concentration of the substituted ammonium in the interior space is at least about a concentration of 0.05 M, 0.1 M, 0.2 M, 0.5 M, 0.6 M, or 0.7 M.

23. (canceled)

40. A composition comprising a liposome comprising one or more lipids and an anti-neoplastic therapeutic entity encapsulated therein at an entity-to lipid molar ratio of at least about 0.10,

wherein said liposome composition has in vivo anti-neoplastic activity at least a four-fold higher than the anti-neoplastic activity of the entity in a free non-liposomal form, and

wherein the toxicity of said liposome composition administered to a mammal is equal or less than the toxicity of said entity administered to said mammal in a free non-liposomal form.

41. The composition of claim 40 wherein the toxicity of said liposome composition administered to a mammal is at least two times less or at least three times less than the toxicity of said entity administered to said mammal in a free non-liposomal form.

42. The composition of claim 40 wherein the toxicity of said liposome composition administered to a mammal is at least four times less than the toxicity of said entity administered to said mammal in a free non-liposomal form.

43. The composition of claim 40 wherein the entity is a pro-drug.

44. The composition of claim 40 wherein said entity is a camptothecin topoisomerase I inhibitor, a camptothecin prodrug, an analog thereof, or a derivative thereof.

45. The composition of claim 44 wherein said camptothecin prodrug is irinotecan.

46. The composition of claim 44 wherein said entity administered into the bloodstream of a mammal has a half-release time from said liposomes of at least 10 hours.

47. The composition of claim 44 wherein said entity administered into the bloodstream of a mammal has a half-release time from said liposomes of at least 24 hours.

48. The composition of claim 44 wherein said liposome comprises a polyanion selected from a group consisting of a polyanionized sugar and polyanionized polyol.

49. The composition of claim 48 wherein said polyanion is sucrose octasulfate, or inositol hexaphosphate.

50. The composition of claim 44 wherein said liposome comprises a biodegradable polyanionic polymer.

51. The composition of claim 50 wherein the biodegradable polyanionic polymer is polyphosphate.

52. The composition of claim 44 wherein said liposome contains a substituted ammonium compound having a formula

R1-(R2-)N+(—

R3)-R4

wherein N is an ammonium nitrogen atom, each of R1, R2, R3, R4 is independently a hydrogen atom or an organic group having each independently not more than 8 carbon atoms, and in totality not more than 18 carbon atoms inclusive, wherein at least one of R1, R2, R3, R4 is an organic group;

wherein the organic group is independently alkyl, alkylidene, heterocyclic alkyl, cycloalkyl, aryl, alkenyl, cycloalkenyl, or a hydroxy-substituted derivative thereof, optionally including a S, O, or N atoms forming an ether, ester, thioether, amine, or amide bond; and

wherein at least three of R1, R2, R3, R4 are the organic groups;

or at least one of the organic groups has a secondary or tertiary carbon atom directly linked to the ammonium nitrogen atom.

53. The composition of any of the claim 44 wherein the topoisomerase inhibitor is entrapped at a concentration of at least about 0.30 mole drug per mole lipid.

54. The composition of claim 53 wherein the topoisomerase inhibitor is entrapped at a concentration of at least about 0.40 mole drug per mole lipid and about 1.7 mole drug per mole lipid.

55. The composition of claim 53 wherein the mammal is a mouse.

56. The composition of claim 53 wherein the anti-tumor activity is determined in vivo in an HT-29 tumor model or a BT-474 tumor model.

57. The composition of claim 46 wherein the mammal is a rat.

161. The composition of claim 44 wherein the liposome comprises a substituted ammonium ion having at least one C—

N bond and selected from a primary ammonium, a secondary ammonium, a tertiary ammonium, and a quaternary ammonium, and having in a aqueous solution at ambient temperature a pKa value of at least 8.0, at least 8.5, at least 9.0, at least 9.5, or at least 10.

58. A composition for administering a vinca alkaloid drug, comprising liposomes in a medium, said liposomes having an interior space and a membrane separating said interior from said medium, said membrane comprising one or more lipids, said liposomes comprising a vinca alkaloid drug entrapped in the liposomes at a first drug/lipid ratio, wherein 24 hours following administration of said composition into the bloodstream of a mammal, the vinca alkaloid drug remains entrapped in aid liposomes at a second drug/lipid ratio which is over about 50%, at least about 60%, or at least about 70% of said first drug/lipid ratio.

61. The composition of claim 58 wherein the first drug/lipid molar ratio is at least 0.05, at least 0.1, or at least 0.2.

62. The composition of claim 61 wherein said first drug/lipid mass ratio is at least about 0.05 mg/mmol, at least about 0.1 mg/mmol, or at least about 0.3 mg/mmol.

63. The composition of claim 58 wherein the liposomes comprise an inside/outside transmembrane ion gradient effective for retaining of the drug within the liposomes.

64. The composition of claim 58 wherein the interior space comprises a polyanion selected from the group of polymeric polyanion, polyanionized polyol, or polyanionized sugar.

65. The composition of claim 64 wherein the interior space comprises ammonium ion or a substituted ammonium ion.

66. The composition of claim 65 wherein the substituted ammonium ion is a primary ammonium, a secondary ammonium, a tertiary ammonium, or a quaternary ammonium ion.

68. The composition of claim 65 wherein said polyanion is comprised in said interior space at a concentration of at least 0.05 gram-equivalent/L, at least 0.2 gram-equivalent/L, at least 0.5 gram-equivalent per liter, or at least 0.6 gram-equivalent/L.

69. The composition of claim 68 wherein the vinca alkaloid drug is vincristine, vinblastine, vinorelbine, vinflunine, vindesine, vinpocetine, an analog thereof, or a derivative thereof.

70. The composition of claim 69 wherein said vesicle-forming lipid comprises phosphatidylcholine and cholesterol.

71. The composition of claim 70 wherein said phosphatidylcholine is selected from the group consisting of a natural lecithin, a hydrogenated natural lecithin, a synthetic lecithin, 1,2-distearoyl-lecithin, 1,2-dipalmitoyl-lecithin, 1,2-dimyristoyl-lecithin, 1,2-dioleoyl-lecithin, 1-stearoyl-2-oleoyl-lecithin, and 1-palmitoyl-2-oleoyl lecithin.

72. The composition of claim 70 wherein said phosphatidylcholine and said cholesterol are contained in molar ratio of about 3:

2.

73. The composition of claim 58 wherein said lipids comprises a neutral PEG-lipid derivative or an anionic PEG-lipid derivative.

74. The composition of claim 73 wherein said neutral PEG-lipid derivative is from about 0.1 mol. % to about 10 mol. % of the totality of said lipids.

75. The composition of claim 73 wherein said neutral PEG lipid derivative is PEG-ceramide or PEG-diacylglycerol.

76. The composition of claim 73 wherein said anionic PEG-lipid derivative is contained at less than 1 mol % of the total lipid.

77. The composition of claim 76 wherein said anionic PEG-lipid derivative is N-(PEG)-diacyl-phosphatidylethanolamine.

78. The composition of claim 58 wherein the mammal is a rat.

160. The composition of claim 66 wherein the liposome comprises a substituted ammonium ion having in a aqueous solution at ambient temperature a pKa value of at least 8.0, at least 8.5, at least 9.0, at least 9.5, or at least 10.

163. The composition of claim 160 wherein the substituted ammonium is pharmaceutically inert.

59-60. -60. (canceled)

67. (canceled)

79. A composition for administering a vinca alkaloid drug, comprising liposomes in a medium, said liposomes having an interior space and a membrane separating said interior from said medium, said membrane comprising lipids,

wherein said lipids comprise lecithin and cholesterol in the molar ratio of about 3:

2; and

wherein said vinca alkaloid drug is contained in said liposomes at the drug/lipid ratio from about 0.15 mg/mmol lecithin to about 0.55 mg/mmol lecithin; and

wherein said interior space also contains a biodegradable polyanionic polymer, a polyanionized polyol, or a polyanionized sugar; and

wherein said polymer or sugar is contained in said interior space of the liposomes at a concentration from about 0.5 gram-equivalent/L to about 1.0 gram-equivalent/L; and

wherein said vinca alkaloid drug is vinorelbine, vincristine, or vinblastine.

80. The composition of claim 79 wherein said polyanionized sugar is sucrose octasulfate.

81. The composition of claim 80 also comprising an ionically neutral poly(ethylene glycol)-lipid derivative in the amount from about 0.1 mol. % to about 10 mol. % of the total lipid.

82. The composition of claim 81 wherein said poly(ethylene glycol)-lipid derivative is poly(ethylene glycol)-dialkylglycerol, poly(ethylene glycol)-diacylglycerol, or poly(ethylene glycol)-ceramide, wherein molecular weight of poly(ethylene glycol) portion of said poly(ethylene glycol)-lipid derivative is from about 250 to about 10,000.

83. The composition of claim 80 also comprising an anionic poly(ethylene glycol)-lipid derivative in the amount from about 0.1 mol. % to about 0.9 mol. % of the total lipid.

84. The composition of claim 83 wherein said anionic poly(ethylene glycol)-lipid derivative is PEG-phosphatidylethianolamine, wherein molecular weight of the poly(ethylene glycol) portion of said poly(ethylene glycol)-lipid derivative is from about 250 to about 10,000.

85. The composition of claim 79 wherein half-release time of said vinca alkaloid drug from said liposomes in the blood circulation of a mammal is over 24 hours.

86. The composition of claim 85 wherein the mammal is a rat.

87. A composition comprising a liposome having an interior space and comprising a taxane compound wherein said taxane compound is contained essentially within the interior space of said liposome.

88. The composition of claim 87 wherein said taxane is ionically neutral molecule.

89. The composition of claim 88 wherein the molecular structure of said taxane does not comprise a hydrophilic polymer moiety.

90. The composition of claim 89 wherein the amount of the encapsulated taxane is at least 0.05 mole per mole of said lipid.

91. The composition of claim 89 wherein the amount of said encapsulated taxane is at least 0.1 mole per mole of said lipid.

92. The composition of claim 89 wherein the interior space is essentially free from a solubilizing aid selected from a micelle-forming surfactant compound and a cyclodextrin compound.

93. The composition of claim 92 wherein the taxane is paclitaxel or docetaxel.

94. The composition of claim 94 wherein the liposome comprises one or more lipids, and the lipids comprise a lipid derivatized with a hydrophilic polymer.

95. The composition of claim 94 wherein said hydrophilic polymer-derivatized lipid comprises up to 20 mole % of the totality of said lipids.

96. The composition of claim 94 wherein said hydrophilic polymer-derivatized lipid comprises less than 1 mole % of the totality of said lipids.

97. The composition of claim 94 wherein said liposome has blood circulation longevity in a mammal of less than two times higher than the circulation longevity of an identically composed liposome except for the absence of said hydrophilic polymer-derivatized lipid.

98. The composition of claim 97 wherein said mammal is a rat.

99. The composition of claim 96 wherein said hydrophilic polymer-derivatized lipid comprises from about 0.1 mole % to about 0.9 mole % of the totality of said lipids.

100. The composition of claim 97 wherein the hydrophilic polymer is poly(ethylene glycol), or a derivative thereof.

101. The composition of claim 100 wherein the hydrophilic polymer-derivatized lipid is a poly(ethylene glycol)-derivatized phospholipid, a poly(ethylene glycol)-derivatized diacylglycerol, a poly(ethylene glycol)-derivatized dialkylglycerol, a poly(ethylene glycol)-derivatized ceramide, a poly(ethylene glycol)-derivatized fatty acid, a poly(ethylene glycol)-derivatized fatty alcohol, or a poly(ethylene glycol)-derivatized sterol.

102. The composition of claim 97 wherein the hydrophilic polymer is poly(ethylene glycol) having at least three ethylene glycol units.

103. The composition of claim 102 wherein the hydrophilic polymer is poly(ethylene glycol) having molecular weight from about 200 to about 10,000.

104. The composition of claim 102 wherein the hydrophilic polymer is poly(ethylene glycol) having molecular weight from about 500 to about 5,000.

143. A method for preparing a liposomes containing an encapsulated entity, said process comprising the steps of:

(a) providing a pre-entity of said entity;

(b) encapsulating said pre-entity into a liposome having an interior space; and

(c) providing a condition inside the liposome to convert said encapsulated pre-entity into said entity within the interior space of said liposome to obtain said entity in a liposomally encapsulated form

wherein said entity is an organic compound or a compound comprising a coordination complex of a metal of the platinum group.

144. The method of claim 143 wherein said encapsulating step comprises contacting the pre-entity with said liposome having a transmembrane gradient for the time sufficient for encapsulation of said derivative into said liposome.

145. The method of claim 144 wherein said transmembrane gradient is an ion gradient, a pH gradient, an electrochemical potential gradient, or a solubility gradient.

146. The method of claim 144 wherein said ion gradient is a gradient of an ion selected from the group of ammonium ion, and a substituted form of ammonium ion comprising at least one C—

N bond.

147. The method of claim 144 wherein said pre-entity is a globally cationic derivative of the entity.

148. The method claim 143 wherein said compound is a pharmaceutical.

149. The method of claim 148 wherein said organic compound is a taxane compound

150. The method of claim 149 wherein said taxane compound is paclitaxel or docetaxel.

151. The method of claim 149 wherein said pre-entity comprises an ester of the hydroxyl group in any of the positions 2′

or 7′

of the taxane molecule wherein said ester comprises a titratable amine.

152. The method of claim 151 wherein said pre-entity is 2′

-(2-(N,N′

-diethylamino)propionyl)-paclitaxel or 2′

-(2-(N,N′

-diethylamino)propionyl)-docetaxel.

153. The method of claim 143 wherein said condition is a change in pH.

154. The method of claim 143 wherein said condition is an enzymatic cleavage of a labile bond.

155. The method of claim 143 wherein said condition is the process of hydrolysis, photolysis, radiolysis, thiolysis, ammonolysis, reduction, substitution, oxidation, or elimination.