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Patent US5616341 - High drug:lipid formulations of liposomal antineoplastic agents - Google Patents
A method for encapsulation of antineoplastic agents in liposomes is provided, having preferably a high drug:lipid ratio. Liposomes may be made by a process that loads the drug by an active mechanism using a transmembrane ion gradient, preferably a transmembrane pH gradient. Using this technique, trapping...http://www.google.com/patents/US5616341?utm_source=gb-gplus-sharePatent US5616341 - High drug:lipid formulations of liposomal antineoplastic agents
Publication number US5616341 A
Application number US 08/112,875
Also published as US5744158, US5795589, US6083530
Publication number 08112875, 112875, US 5616341 A, US 5616341A, US-A-5616341, US5616341 A, US5616341A
Patent Citations (42), Non-Patent Citations (109), Referenced by (59), Classifications (16), Legal Events (7)
High drug:lipid formulations of liposomal antineoplastic agents
US 5616341 A
(i) liposomes which comprise an ionizable antineoplastic agent selected from the group consisting of doxorubicin and daunorubicin, a release-inhibiting aqueous buffer comprising citric acid and a bilayer comprising a lipid which comprises a phospholipid; and
(ii) an aqueous solution external to the liposomes which is basic with respect to the release-inhibiting buffer, wherein
the weight ratio of antineoplastic agent to lipid in the liposomes is from at least about 0.1:1 to about 3:1.
2. The composition of claim 1, wherein the liposome has an average diameter of from about 60 nm to about 300 nm and wherein the liposome is unilamellar.
15. A liposomal antineoplastic agent treatment system which comprises:
(a) a liposome comprising a release-inhibiting aqueous buffer comprising citric acid and a bilayer comprising a lipid which comprises a phospholipid;
(b) an aqueous solution which is basic with respect to the release-inhibiting buffer; and
(c) an ionizable antineoplastic agent selected from the group consisting of doxorubicin and daunorubicin,
wherein the liposome and aqueous solution are combined so as to establish a pH gradient across the bilayer,
whereby the antineoplastic agent is loaded into the liposomes so as to form the liposome composition of claim 1.
16. A dehydrated liposome which comprises an ionizable antineoplastic agent selected from the group consisting of doxorubicin and daunorubicin, a buffering agent and a bilayer comprising a lipid which comprises a phospholipid, wherein the buffering agent comprises citric acid and wherein the weight ratio of drug to lipid in the liposome is at least about 0.1:1.
FIG. 2 is a graph of release characteristics of liposomal-doxorubicin (EPC:cholesterol, 55:45 mol:mol, 29±2/100 drug/lipid wt./wt.) containing 300 mm citrate, dialyzed against buffer at 37° C. of pH 4.0 (open circles) and pH 7.5 (closed circles) at 37° C.
FIG. 3 is a graph of a citrate-doxorubicin interaction resulting from mixing experiments at varying citrate pH values. The mM doxorubicin remaining in solution following centrifugation is plotted as a function of citrate pH: 4 mM doxorubicin, mixed at 60° C. then cooled to 25° C. (closed squares); 4 mM doxorubicin mixed at 25° C. (open squares); 20 mM doxorubicin mixed at 60° C. then cooled to 25° C. (closed circles); and 4 mM doxorubicin mixed in 20 mM/HEPES, 150 mM NaCl, at 25° C. for comparison (open circle).
FIG. 7 is a graph demonstrating the effect of temperature on uptake of 5-fluorouracil ("FU"). The delta T reflects a temperature increase from 21° C. to 60° C.
FIG. 8 is a graph depicting the effect of external buffer on FU release at 37° C.
Doxorubicin retention in EPC/cholesterol (55:45) vesicles exhibiting a pH gradient can be increased by employing citrate/carbonate buffer systems such that less than about 5% drug release is observed over 24 h at 37° C. This vesicle-entrapped doxorubicin also appears stable to serum components; less than 5% doxorubicin is released over 24 hours for vesicles incubated at 37° C. in 95% fresh human serum. In association assays, where doxorubicin was incubated with HEPES buffer at pH 7.5, and citrate buffers (sodium citrate) at pH ranging from about 4.0-7.5, citrate interacts with doxorubicin and precipitates, whereas HEPES buffer does not. Such a buffer combination, that is, citrate/carbonate, acts to reduce the rate of release of the drug from the liposomes. Other release-reducing buffer combinations can be used such as oxalic acid/potassium phosphate or succinic acid/sodium bicarbonate, with citric acid/sodium carbonate or citric acid/sodium bis phosphate preferred.
In order to determine whether an ionizable antineoplastic agent will load into liposomes in response to a transmembrane pH gradient, EPC-containing liposomes are made (about 1.0 mM EPC) with a 3 H-DPPC tracer and with a relatively acidic or basic internal medium such as 300 mM citric acid at about pH 4.0. These liposomes are extruded about 10 times according to the LUVET procedure through 2 100 nm filters, followed by adjustment of the external pH to a relatively basic or acidic pH, for example, sodium carbonate, at about pH 11.0. Following the formation of the pH gradient, the agent to be loaded, spiked with a radioactive isotope of the agent, is admixed with the liposomes to about 200 uM (per 1.0 mM lipid used). The liposomes are separated from free, unentrapped agent on G50-M Sephadex minicolumns at 500×g for 3 minutes into 13×100 mm tubes, and radioactivity counted in a scintillation counter. Uptake of the drug inn moles per umole of lipid is then plotted over incubation time. One hundred percent of the available doxorubicin is taken up into liposomes under these conditions.
In the case of doxorubicin, commercially available forms, such as powdered, solid, and methylparaben-containing forms (Adriamycin R. D. F., Adria Laboratories, Inc., Columbus, Ohio) may be used in the invention. When the methylparaben-containing form is employed, an aqueous solution such as saline may be added to that form, thereby dissolving it, followed by the admixing of this suspension with the liposomes which have the transmembrane pH gradient across their bilayers. Such admixing at 60° C. for about 10 minutes results in more than about 98% encapsulation of the doxorubicin.
Lipids which can be used in the liposome formulations of the present invention include phospholipids such as phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidic acid (PA), phosphatidylinositol (PI), sphingomyelin (SPM), and the like, alone or in combination. The phospholipids can be synthetic or derived from natural sources such as egg or soy. The phospholipids dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylglycerol (DMPG) may also be used. In the preferred embodiments, egg phosphatidylcholine (EPC), and cholesterol are used in preferably a 55:45 mole ratio. In other embodiments, distearoylphosphatidyl choline (DSPC), dipalmitoylphosphatidylcholine (DPPC), or hydrogenated soy phosphatidylcholine (HSPC) may be used in a mole ratio of 55:45 with cholesterol. Dimyristoylphosphatidylcholine (DMPC) and diarachidonoyl phosphatidylcholine (DAPC) may similarly be used. Due to the elevated transition temperatures (Tc) of lipids such as DSPC (Tc of about 65° C.), DPPC (Tc of about 45° C.) and DAPC (Tc of about 85° C.), such lipids are preferably heated to about their Tc or temperatures slightly higher (e.g., up to about 5° C. higher) than the Tc in order to make these liposomes.
The liposomes formed by the procedures of the present invention may be lyophilized or dehydrated at various stages of formation. For example, the lipid film may be lyophilized after removing the solvent and prior to adding the drug. Alternatively, the lipid-drug film may be lyophilized prior to hydrating the liposomes. Such dehydration may be carried out by exposure of the lipid or liposome to reduced pressure thereby removing all suspending solvent. The liposomes may be dehydrated in the presence of a hydrophilic agent according to the procedures of Bally et al., PCT Publication No. 86/01102, published Feb. 27, 1986, entitled "Encapsulation of Antineoplastic Agents in Liposomes", and Janoff et al., PCT Publication No. 86/01103, published Feb. 27, 1986, entitled "Dehydrated Liposomes", or Schneider et al., in U.S. Pat. No. 4,229,360, issued Oct. 29, 1980. Alternatively or additionally, the hydrated liposome preparation may also be dehydrated by placing it in surrounding medium in liquid nitrogen and freezing it prior to the dehydration step. Dehydration with prior freezing may be performed in the presence of one or more protective agents, such as sugars in the preparation according, to the techniques of Bally, et al., PCT Application No. 86/01103published Feb. 27, 1986, relevant portions of which are hereby incorporated by reference. Such techniques enhance the long-term storage and stability of the preparations. For example, the liposomal-antineoplastic agent can be mixed with a sugar solution in a sugar: lipid w/w ratio of about 0.5:1 to about 50:1, and preferably about 20:1. Upon rehydration, such liposomes retain essentially all the antineoplastic agent previously loaded, for such liposomes sized through 100 and 200 nm pore size filters. In a preferred embodiment, the sugar is mannitol, or mannitol:glucose:lactose in a 2:1:1 w/w/w ratio. Following rehydration in distilled water, the preparation is preferably heated for ten minutes at an elevated temperature, for example 60° C. Other suitable methods may be used in the dehydration of the above-disclosed liposome preparations. The liposomes may also be dehydrated without prior freezing.
Liposomes that passively entrapped doxorubicin were made using the materials as above, by suspending doxorubicin in buffer (20 mM HEPES, 150 mM NaCl, pH 7.5) to 2.0 mM doxorubicin, prior to the lipid hydration step. The liposomes were frozen and thawed and extruded as above. Active entrapment of doxorubicin was accomplished by preparing vesicles in buffer at pH 4.0, increasing the exterior pH to 7.5 with 1.0M Na2 CO3, and incubating the vesicles (20 mM lipid) with doxorubicin (10 mg lipid/ml) at 60° C. for 5 minutes.
The materials and procedures of Example 6 were employed to determine the in vitro stability of the liposome-doxorubicin preparations. Release experiments were performed as follows: 10-fold dilute liposome samples were dialyzed for 24 hours against 1000 volumes of 20 mM HEPES, 150 mM NaCl (pH 7.5) at 37° C. At 1, 2, 4, 8, 10, and 24 hours post-preparation, a 150 ul aliquot was removed and the entrapped doxorubicin was determined.
After completion of the entrapment procedure according to Example 6, 20 ul of the doxorubicin-liposomes were diluted to 200 ul with 20 mM HEPES, 150 mM NaCl (pH 7.5). An aliquot of this diluted sample (20 ul) was assayed for lipid phosphate by the procedure of Bartlett, J. Biol. Chem. 1959, 234:466-468. A second 20 ul sample of the diluted preparation was removed and placed in a glass test tube, to which Triton X-100 (1.0 ml of 1% w/w) was added. The sample was heated in a water bath at 40° C. for 2 minutes and vortically mixed. Absorbance of the sample was read at 480 nm in a spectrophotometer. Sample readings were compared to a standard curve of doxorubicin samples containing known amounts of the agent which have been diluted with 1.0 ml of Triton X-100.
Sephadex G-50 (medium grade) columns were prepared at 1.0 ml capacity that had been pre-swollen with gel in 20 mM HEPES, 150 mM NaCl (pH 7.5). Columns were centrifuged at 500×g for 3 minutes followed by a repeat spin, to pack columns. Doxorubicin-liposome samples were applied (150 ul of the 10×diluted samples) to the columns, followed by application of 50 ul of buffer, and centrifuged at 3000 rpm for 5 minutes. The eluant was vortically mixed until homogenous. Aliquots (25 ul) were removed and analyzed for phosphate and doxorubicin as described above.
EPC/cholesterol (2.1:1 wt. ratio) was dispersed in 150 mM citric acid (pH 4.0) to yield 200 mg total lipid/ml buffer. The resulting MLVs were frozen and thawed 7 times with vortical mixing prior to each freezing step. The resulting FATMLVs were extruded 5 times through 2 stacked 0.2 um pore size filters to make VET200 s. The liposomes were then diluted 2 fold with unbuffered saline and the pH brought to 7.5 with 1N NaOH. The equivalent of 1.0 ml of liposomes before pH adjustment were added to 133 mg of doxorubicin/lactose and 3.7 mg Na2 CO3 contained in a sealed vial (20 ml capacity). Both the liposomes and the doxorubicin-containing vial were heated to 60° C. for 5 minutes prior to admining. After admixing, the liposomes were heated at 60° C. for 5 minutes with vortical mixing every minute. The sample was then cooled to room temperature. An aliquot of the sample (50 ul) was removed and diluted to 0.5 ml with 20 mM HEPES, 150 mM NaCl (pH 7.5). An aliquot of this sample (150 ul) was applied to a 1.0 ml Sephadex G-50 column as described previously. Phosphate and doxorubicin were quantitated as described previously, in the eluant and the original samples.
EPC/cholesterol (55/45 mole ratio) was dried from chloroform to a thin film on a 500 ml capacity round bottom flask (400 mg total lipid). The film was hydrated with 4.0 ml of 300 mM citric acid at pH 4.0, forming MLVs. These MLVs were extruded through 2 stacked 0.22 um Nucleopore membrafil filters followed by extrusion 10 times through a 0.1 um Nucleopore membrafil (tortuous path) filter. To 1.0 ml of the resulting filtrate sample as added 275 ul of 1M Na2 CO3, which raised the exterior pH to 8.3. An aliquot (0.6 ml) was heated for 3 minutes at 60° C., as was a 10 mg sample of doxorubicin. The liposome aliquot was added to the 10 mg doxorubicin and heated at 60° C. for 5 minutes. The sample was divided into 2 parts. Part 1 was diluted 10 times with 30 mM HEPES, 150 mM NaCl, at pH 7.5. Part 2 was diluted 10 times with 300 mM citric acid at pH 4.0. Both samples were placed into dialysis bags and dialyzed at 37° C. against 1000 volumes of their respective buffers. At 1 hour, a 150 ul aliquot was removed and analyzed for doxorubicin and lipid phosphate as previously described, after passage down a 1.0 ml Sephadex column equilibrated in the respective buffer.
Results are graphed in FIG. 3, a graph of a citrate-doxorubicin interaction resulting from mixing experiments at varying citrate pH values. The mM doxorubicin remaining in solution following centrifugation is plotted as a function of titrate pH: 4 mM doxorubicin, mixed at 60° C. then cooled to 25° C. (closed squares); 4 mM doxorubicin mixed at 25° C. (open squares); 20 mM doxorubicin mixed at 60° C. then cooled to 25° C. (closed circles); and 4 mM doxorubicin mixed in 20 mM/HEPES, 150 mM NaCl, at 25° C. for comparison (open circle).
Liposomes were made according to the procedures of Example 2. Where the P388 leukemia model was employed, tumor cells (1×105 cells in 0.1 ml, were injected i.p into female CDF-1 mice. One day after tumor inoculation, the mice were treated with liposomal doxorubicin (5 mg/kg dose) via tail vein injection. Dosage was calculated according to the mean weight of each group, and weights were determined on day 0 (day of tumor injection) and day 5. Deaths were recorded on a daily basis.
Male shinogi mice (25-40g, 9 per group) were injected subcutaneously with 1×105 SC-115 cells obtained from a primary tumor in previously inoculated mice. Tumor growth was monitored by palpation and tumor measurements with a vernier caliper. Upon growth of the tumor to 0.5-2.0 g (tumor weight=[width2 ×length]/2, measurements in mm), mice were administered liposomal doxorubicin dose of 13 mg/kg i.v. at seven day intervals (3 injections of the indicated dose). Tumor growth was monitored 3 times weekly for 50 days post first treatment or until the tumor weight exceeded 9 g at which time the animal was sacrificed. Treatment doses were based on the initial animal weights prior to tumor inoculation.
Liposomes were prepared by hydrating a film of DSPC/cholesterol (55:45 molar ratio) in 300 mM citric acid buffer (pH 4.0) with vortex mixing. These MLVs (100 mg total lipid/ml buffer) were extruded 10 times through a 200 nm pore size polycarbonate filters in a thermojacket LUVET heated to 60° C. Liposomes were added to a solution of 1 mg/ml vincristine sulfate (Oncovin, available from Eli Lilly and Co., Indianapolis, Ind.) to achieve a drug to total lipid weight ratio of approximately 0.17:1. To this was added a sufficient amount of 1.0M Na2 HPO4 to bring the pH of the solution to about 7.0. The samples were then heated at 60° C. for 10 minutes at which time the drug was encapsulated inside the liposomes at a trapping efficiency in excess of 98%.
Egg phosphatidylcholine (15 mg) was dispersed in 2 ml of 300 mM citric acid., p H 4.0 and the resulting MLVs frozen in liquid nitrogen and thawed in warm water (approximately 35° C.) a total of five times. The lipid was then extruded 10 times through two stacked 100 nm pore size polycarbonate filters using the LUVET procedure. A proton gradient was created by passage of the vesicles over a Sephadex G-50 (fine) column (1.5 cm×10 cm) preequilibrated with 300 mM NaCl, 20 mM HEPES, p H 7.5. An aliquot of the large unilamellar vesicles eluted from the column was diluted in 300 mM NaCl, mM HEPES, p H 7.5 to a lipid concentration of 0.75 mgml-1 in a total volume of 2 ml and then daunorubicin (113 ug) added from a stock solution (5.64 mgml-1) in distilled water. The mixture was incubated at room temperature (25° C.) and at intervals of 2, 10, 20, 30, 60 and 120 minutes, 100 ul aliquots were centrifuged through 1 ml "minicolumns" of Sephadex G-50 (fine) to remove any unencapsulated daunorubicin from the vesicles. The concentration of entrapped daunorubicin was determined from its absorbance at 500 nm in a Shimadzu UV-265 spectrophotometer following solubilization of the vesicles in 1% Triton X-100. Lipid was quantified by liquid scintillation counting using tracer levels of 3 H-DPPC. In excess of 98% of the daunorubicin was encapsulated by the vesicles giving a drug to lipid molar ratio of 1:5.
TABLE 1______________________________________TRAPPING EFFICIENCIES OF VARIOUSLIPOSOMAL VINCRISTINE PREPARATIONS  TEM-  PERA-                        TRAPPINGSAM-   TURE     VINC      DRUG:LIPID                               EFFI-PLE    (°C.)           SOURCE    (wt:wt)   CIENCY______________________________________EPC/   60       SIGMA     0.24:1     95.0CHOLEPC/   60       ONCOVIN   0.29:1     88.0CHOLHSPC/  21       SIGMA     0.20:1     15.0CHOLHSPC/  60       SIGMA     0.20:1    100.0CHOLDSPC/  60       ONCOVIN   0.24:1    100.0CHOL______________________________________
1 Bangham, et al.; "Diffusion of Univalent Iona Across the Lamellae of Swollen Phospholipids", 1965; J. Mol. Biol., 13:238-252.
2 * Bangham, et al.; Diffusion of Univalent Iona Across the Lamellae of Swollen Phospholipids , 1965; J. Mol. Biol., 13:238 252.
5 Casey et al.; "Active Proton Uptake by Chromaffin Granules: Observation by Amine Distribution and Phosphorus-31 Nuclear Magnetic Resonance Techniques" 1977, Biochemistry 16(5), pp. 972-976.
6 * Casey et al.; Active Proton Uptake by Chromaffin Granules: Observation by Amine Distribution and Phosphorus 31 Nuclear Magnetic Resonance Techniques 1977, Biochemistry 16(5), pp. 972 976.
9 Crommelin et al; Chem. Abs. vol. 99, 1983, Abs #128259c.
10 * Crommelin et al; Chem. Abs. vol. 99, 1983, Abs 128259c.
11 Crommelin, et al., "Preparation and characteriation of doxorubicin-containing liposomes: I. Influence of liposome charge and pH of hydration medium on loading capacity and particle size", Int. J. Pharms, 16, (1983), 79-92.
12 Crommelin, et al., "Preparation and characterization of doxorubicin-containing liposomes, II. Loading capacity, long-term stability and doxorubicin-bilayer interaction mechanism", Chemical Abstracts, vol. 100, 1984, Abs. 109032w.
13 * Crommelin, et al., Preparation and characteriation of doxorubicin containing liposomes: I. Influence of liposome charge and pH of hydration medium on loading capacity and particle size , Int. J. Pharms, 16, (1983), 79 92.
14 * Crommelin, et al., Preparation and characterization of doxorubicin containing liposomes, II. Loading capacity, long term stability and doxorubicin bilayer interaction mechanism , Chemical Abstracts, vol. 100, 1984, Abs. 109032w.
15 Deamer, et al., "The response to Fluorescent Amine to pH Gradients Across Liposome Membranes", 1972, Biochim. Biophys, Acta, 274, pp. 323-335.
16 * Deamer, et al., The response to Fluorescent Amine to pH Gradients Across Liposome Membranes , 1972, Biochim. Biophys, Acta, 274, pp. 323 335.
17 Forssen, et al., "Improved Therapeutic Benefits of Doxorubicin by Entrapment in Anionic Liposomes", 1983; Cancer Res. 43:546-550.
18 * Forssen, et al., Improved Therapeutic Benefits of Doxorubicin by Entrapment in Anionic Liposomes , 1983; Cancer Res. 43:546 550.
19 Gabizon, et al., "Enhancement of Adriamycin Delivery to Liver Metastatic Cells with Increased Tumoricidal Effect Using Liposomes as Drug Carriers", 1983; Cancer Res. 43:4730-4735.
20 * Gabizon, et al., Enhancement of Adriamycin Delivery to Liver Metastatic Cells with Increased Tumoricidal Effect Using Liposomes as Drug Carriers , 1983; Cancer Res. 43:4730 4735.
21 Gabizon, et al.; "Liposomes as In Vivo Carriers of Adriamycin: Reduced Cardiac Uptake and Preserved Antitumor Activity in Mice", 1982; Cancer Res. 42:4734-4739.
22 * Gabizon, et al.; Liposomes as In Vivo Carriers of Adriamycin: Reduced Cardiac Uptake and Preserved Antitumor Activity in Mice , 1982; Cancer Res. 42:4734 4739.
23 Gannphati, et al., "Effect of cholesterol content of liposomes on the encapsulation, efflux and toxicity of adriamycin", 1984; Biochem. Pharmacol. 33:698-700.
24 * Gannphati, et al., Effect of cholesterol content of liposomes on the encapsulation, efflux and toxicity of adriamycin , 1984; Biochem. Pharmacol. 33:698 700.
25 Garcia, et al.; "Mechanism of lactose translocation in proteoliposomes reconstituted with lac carrier protein prified from Escherishia coli", Biol. Abs. vol. 77(7):6013, 1984, Abs. 54638.
26 * Garcia, et al.; Mechanism of lactose translocation in proteoliposomes reconstituted with lac carrier protein prified from Escherishia coli , Biol. Abs. vol. 77(7):6013, 1984, Abs. 54638.
27 Gregoriadis, "Targeting of Drugs: Implications of Medicine", 1981;The Lancet, 241-247.
28 * Gregoriadis, Targeting of Drugs: Implications of Medicine , 1981;The Lancet, 241 247.
29 Groom, et al., "Liposomes", Chemical Abstracts, vol. 102, 1985, Abs #67398d.
30 * Groom, et al., Liposomes , Chemical Abstracts, vol. 102, 1985, Abs 67398d.
31 Herman, et al., "Prevention of Chronic Doxorubicin Cardiotoxicity in Beadles by Liposomal Encapsulation", 1983; Cancer Res. 43:5427-5432.
32 * Herman, et al., Prevention of Chronic Doxorubicin Cardiotoxicity in Beadles by Liposomal Encapsulation , 1983; Cancer Res. 43:5427 5432.
33 Kano and Fendler, "Pyranine as a Sensitive pH Probe for Liposome Interiors and Surfaces", Biochim. Biophys. Acta, 1978, 509, pp. 289-299.
34 * Kano and Fendler, Pyranine as a Sensitive pH Probe for Liposome Interiors and Surfaces , Biochim. Biophys. Acta, 1978, 509, pp. 289 299.
35 Kirby and Gregoriadis, "The Effect of Lipid Composition of Small Unilamellar Liposomes Containing Melphalan and Vincristine on Drug Clearance After Injection into Mice" Abstract of Biochem. Pharmacol. 1983, 32(4), pp. 609-615.
36 * Kirby and Gregoriadis, The Effect of Lipid Composition of Small Unilamellar Liposomes Containing Melphalan and Vincristine on Drug Clearance After Injection into Mice Abstract of Biochem. Pharmacol. 1983, 32(4), pp. 609 615.
37 Kirby, et al., "Dehydration-rehydration vesicles: a simple method for high yield drug entrapment in liposomes", Chemical Abstracts,. vol. 102, 1985, Abs #84326w.
38 Kirby, et al., "The Effect of Lipid Composition of Small Unilamellar Liposomes Containing Melphalan and Vincristine on Drug Clearance After Injection into Mice", 1983; Biochem,. Pharma., vol. 32(4) pp. 609-615.
39 * Kirby, et al., Dehydration rehydration vesicles: a simple method for high yield drug entrapment in liposomes , Chemical Abstracts,. vol. 102, 1985, Abs 84326w.
40 * Kirby, et al., The Effect of Lipid Composition of Small Unilamellar Liposomes Containing Melphalan and Vincristine on Drug Clearance After Injection into Mice , 1983; Biochem,. Pharma., vol. 32(4) pp. 609 615.
41 Kornberg, et al., "Measurement of Transmembrane Potentials in Phospholipid Vesicles", 1972, Proc. Nat. Aca. Sci. USA 69(6), pp. 1508-1513.
42 * Kornberg, et al., Measurement of Transmembrane Potentials in Phospholipid Vesicles , 1972, Proc. Nat. Aca. Sci. USA 69(6), pp. 1508 1513.
43 Layton, et al.; "A Comparison of the Therapeutic Effects of Free and Liposomally Encapsulated Vincristine in Leukemic Mice", 1980; Europe. J. Cancer., vol. 16, 945-950.
44 * Layton, et al.; A Comparison of the Therapeutic Effects of Free and Liposomally Encapsulated Vincristine in Leukemic Mice , 1980; Europe. J. Cancer., vol. 16, 945 950.
45 * Lopez Berestein, et al., Liposomal Amphotericin B For the Treatment of Systemic Fungal Infections in Patients with Cancer: A Preliminary Study , 1985; J. Infect. Dis., 151:704 710.
46 Lopez-Berestein, et al., "Liposomal Amphotericin B For the Treatment of Systemic Fungal Infections in Patients with Cancer: A Preliminary Study", 1985; J. Infect. Dis., 151:704-710.
47 Mayer et al.: "Influence of Vesicle Size, Lipid Composition, and Drug-to-Lipid Ratio on the Biological Activity of Liposomal Doxorubicin in Mice", Cancer Res. 49:5922-5930 (1989).
48 * Mayer et al.: Influence of Vesicle Size, Lipid Composition, and Drug to Lipid Ratio on the Biological Activity of Liposomal Doxorubicin in Mice , Cancer Res. 49:5922 5930 (1989).
49 Mayer, et al., "Uptake of adriamycin into large unilamellar vesicles in response to a pH gradient", 1986; Biochim. Biophys. Acta., 857:123.
50 Mayer, et al., "Uptake of antineoplastic agents into large unilamellar vesicles in response to a membrane potential", 1985; Biochem. Biophys. Acta., 816:294-302.
51 * Mayer, et al., Uptake of adriamycin into large unilamellar vesicles in response to a pH gradient , 1986; Biochim. Biophys. Acta., 857:123.
52 * Mayer, et al., Uptake of antineoplastic agents into large unilamellar vesicles in response to a membrane potential , 1985; Biochem. Biophys. Acta., 816:294 302.
53 Mayer, et al.; "Solute distributions an trapping efficiencies observed in freeze-thawed multilamellar vesicles", 1985; Biochim. Biophys. Acta., 187:193-196.
54 * Mayer, et al.; Solute distributions an trapping efficiencies observed in freeze thawed multilamellar vesicles , 1985; Biochim. Biophys. Acta., 187:193 196.
55 Minow, et al., "Adriamycin (NSC-123127) Cardiomyophathy--An Overview With Determination of Risk Factors", 1975; Cancer Chemother. Rep. 6: 195-201.
56 * Minow, et al., Adriamycin (NSC 123127) Cardiomyophathy An Overview With Determination of Risk Factors , 1975; Cancer Chemother. Rep. 6: 195 201.
57 Moro, et al., "Purification of Liposome Suspensions", Chemical. Abstracts,. vol. 94, 1981, p. 372, Abs. 52931g.
58 * Moro, et al., Purification of Liposome Suspensions , Chemical. Abstracts,. vol. 94, 1981, p. 372, Abs. 52931g.
59 Nichols and Deamer, "Catecholamine Uptake and Concentration by Liposomes Maintaining pH Gradients" 1976, BBA 455, pp. 269-271.
60 * Nichols and Deamer, Catecholamine Uptake and Concentration by Liposomes Maintaining pH Gradients 1976, BBA 455, pp. 269 271.
61 Olson, et al., "Characterization, Toxicity and Therapeutic Efficacy of Adriamycin Encapsulated in Liposomes", 1982; Br. J. Cancer Clin. Oncol., 18-167.
62 * Olson, et al., Characterization, Toxicity and Therapeutic Efficacy of Adriamycin Encapsulated in Liposomes , 1982; Br. J. Cancer Clin. Oncol., 18 167.
63 Papahadjopoulos, et al., "Phospholipd Model Membranes", 1967: Biochim. Biophys. Acta., 135:624-638.
64 * Papahadjopoulos, et al., Phospholipd Model Membranes , 1967: Biochim. Biophys. Acta., 135:624 638.
65 Rahman, et al., "Doxorubicin-induced Chronic Cardiotoxicity and its Protection by Liposomal Administration", 1982; Cancer Res. 42:1817.
66 Rahman, et al., "Liposomal Protection of Adriamycin-induced Cardiotoxicity in Mice", 1980; Cancer Res. 40:1532-1536.
67 Rahman, et al., "Pharmacological, Toxicological, and Therapeutic Evaluation in Mice of Doxorubicin Entrapped in Cardiolipid Liposomes", 1985; Cancer Res. 45:796-803.
68 * Rahman, et al., Doxorubicin induced Chronic Cardiotoxicity and its Protection by Liposomal Administration , 1982; Cancer Res. 42:1817.
69 * Rahman, et al., Liposomal Protection of Adriamycin induced Cardiotoxicity in Mice , 1980; Cancer Res. 40:1532 1536.
70 * Rahman, et al., Pharmacological, Toxicological, and Therapeutic Evaluation in Mice of Doxorubicin Entrapped in Cardiolipid Liposomes , 1985; Cancer Res. 45:796 803.
71 Richardson, et al., "Tissue Distribution and Tumour Localization of 99m-Technetium-Labeled Liposomes in Cancer Patients", 1979; Br. J. Cancer 40:35-43.
72 * Richardson, et al., Tissue Distribution and Tumour Localization of 99m Technetium Labeled Liposomes in Cancer Patients , 1979; Br. J. Cancer 40:35 43.
73 Rosa, et al., in Transport in Biomembranes: Model Systems and Reconstitution, R. Antolini ed., "Liposomes Containing Doxorubicin: An Example of Drug Targeting", 1982; pp. 243-256.
74 * Rosa, et al., in Transport in Biomembranes: Model Systems and Reconstitution, R. Antolini ed., Liposomes Containing Doxorubicin: An Example of Drug Targeting , 1982; pp. 243 256.
75 Rosa, et al.; "Absorption and Tissue Distribution of Doxorubicin Entrapped in Liposomes following Intravenous or Intraperitoneal administration:", 1983; Pharmacol. 25:221-229.
76 * Rosa, et al.; Absorption and Tissue Distribution of Doxorubicin Entrapped in Liposomes following Intravenous or Intraperitoneal administration: , 1983; Pharmacol. 25:221 229.
77 Ryman, et al., "Liposomes--Further Considerations of Their Possible Role as Carriers of Therapeutic Agents", 1983; Targeting of Drugs, pp. 235-248.
78 * Ryman, et al., Liposomes Further Considerations of Their Possible Role as Carriers of Therapeutic Agents , 1983; Targeting of Drugs, pp. 235 248.
79 Shakov, et al., "Reconstitution of highly purified proton-translocating pyrophosphatase from Rhodospirillum rubrum", Biol. Abs. 77(12):10459, 1984, Abs. 94506.
80 * Shakov, et al., Reconstitution of highly purified proton translocating pyrophosphatase from Rhodospirillum rubrum , Biol. Abs. 77(12):10459, 1984, Abs. 94506.
81 Shinozawa, et al., "Tissue Distribution and Antitumor Effect of Liposome-Entrapped Doxorubicin (Adriamycin) in Ehrlich Solid Tumor-Bearing Mouse", 1981; Acta. Med. Okayama, 35:395-405.
82 * Shinozawa, et al., Tissue Distribution and Antitumor Effect of Liposome Entrapped Doxorubicin (Adriamycin) in Ehrlich Solid Tumor Bearing Mouse , 1981; Acta. Med. Okayama, 35:395 405.
83 Szoka, Jr., et al., "Comparative Properties and Methods of Preparation of Lipid Vesicles (Liposomes)", 1980; Ann. Rev. Biophys. Bioeng. 9:467-509.
84 * Szoka, Jr., et al., Comparative Properties and Methods of Preparation of Lipid Vesicles (Liposomes) , 1980; Ann. Rev. Biophys. Bioeng. 9:467 509.
85 * U.S. application No. 122,354, Forssen, filed Nov. 18, 1987.
86 * U.S. application No. 122,613, Bally et al. filed Nov. 17, 1987 Pending.
87 * U.S. application No. 161,141, Popescu et al., filed, Feb. 25, 1988 Pending.
88 * U.S. application No. 164,580, Janoff et al., filed Mar. 7, 1988 Pending.
89 * U.S. application No. 22,157, Janoff et al., filed Mar. 5, 1987 Abandoned.
90 * U.S. application No. 220,388, Mehlhorn, filed Jul. 12, 1988.
91 * U.S. application No. 236,700, Janoff, et al., filed Aug. 25, 1988.
92 * U.S. application No. 284,751, Bally, et al., filed Dec. 12, 1988.
93 * U.S. application No. 310,495, Cullis et al., filed Feb. 13, 1989 Pending.
94 * U.S. application No. 360,964, Janoff, et al., filed Jun. 26, 1989.
95 * U.S. application No. 4,762, Cullis et al., filed Jan. 7, 1987 Pending.
96 * U.S. application No. 61,837, Hope et al., filed Jun. 12, 1987 Pending.
97 * U.S. application No. 622,502, Cullis et al., filed Jun. 20, 1984 Abandoned.
98 * U.S. application No. 622,690, Cullis et al., filed Jun. 20, 1984 Abandoned.
99 * U.S. application No. 638,809, Janoff, et al., filed Aug. 8, 1984.
100 * U.S. application No. 660,573, Lenk et al., filed Oct. 12, 1984 Pending.
101 * U.S. application No. 69,908, Janoff et al., filed Jul. 6, 1987 Abandoned.
102 * U.S. application No. 749,161, Bally, et al., filed Jun. 26, 1985.
103 * U.S. application No. 752,423, Bally et al., filed Jul. 5, 1985 Abandoned.
104 * U.S. application No. 759,419, Janoff, et al., filed Jul. 26, 1985.
105 * U.S. application No. 788,017, Cullis et al., filed Oct. 16, 1985 Abandoned.
106 * U.S. application No. 800,545, Cullis et al., filed Nov. 21, 1985 Abandoned.
107 * U.S. application No. 874,575, Hope et al., filed Jun. 16, 1986 Abandoned.
108 Van Hoesel, et al., "Reduced cardiotoxicity and nephrotoxicity with preservation of anittumor activity of Doxorubicin entrapped in stable liposomes in the LOU/M", Chemical. Abstracts,. vol. 101, 1984, 163307y.
109 * Van Hoesel, et al., Reduced cardiotoxicity and nephrotoxicity with preservation of anittumor activity of Doxorubicin entrapped in stable liposomes in the LOU/M , Chemical. Abstracts,. vol. 101, 1984, 163307y.
US6475517 * Jul 5, 2001 Nov 5, 2002 Mitsubishi Chemical Corporation Method for preparing closed vesicles
US7048944 Sep 25, 2002 May 23, 2006 Mitsubishi Chemical Corporation Method for preparing closed vesicles
US8241663 * Mar 25, 2005 Aug 14, 2012 Terumo Kabushiki Kaisha Liposome preparation
US8349360 Oct 6, 2005 Jan 8, 2013 Bc Cancer Agency Liposomes with improved drug retention for treatment of cancer
US8709474 Dec 9, 2012 Apr 29, 2014 Bc Cancer Agency Liposomes with improved drug retention for treatment of cancer
US9737485 Jul 21, 2016 Aug 22, 2017 Zoneone Pharma, Inc. Remote loading of sparingly water-soluble drugs into liposomes
US20030022379 * Sep 25, 2002 Jan 30, 2003 Toshiaki Tagawa Method for preparing closed vesicles
US20080279916 * Mar 25, 2005 Nov 13, 2008 Terumo Kabushiki Kaisha Liposome Preparation
US20090324698 * Oct 14, 2005 Dec 31, 2009 Polymun Scientific Immunbiologische Forschung Gmbh Liposomal composition comprising an active ingredient for relaxing smooth muscle, the production of this composition and the therapeutic use thereof
CN101229127B Nov 26, 2003 Oct 10, 2012 吉里德科学公司 Liposome preparation
EP0834309A2 * Sep 18, 1997 Apr 8, 1998 Artur Herzog Dr. Mesmer Use of a liposome solution for enhancing the activity and/or reducing the toxicity of drugs
EP0834309A3 * Sep 18, 1997 Jan 19, 2000 Artur Herzog Dr. Mesmer Use of a liposome solution for enhancing the activity and/or reducing the toxicity of drugs
EP1233754A2 * Nov 30, 2000 Aug 28, 2002 The Regents of The University of California Targeted drug delivery with a cd44 receptor ligand
EP1233754A4 * Nov 30, 2000 Jun 7, 2006 Univ California Targeted drug delivery with a cd44 receptor ligand
EP2682106A1 Jul 3, 2012 Jan 8, 2014 Phares Pharmaceutical Research N.V. Method of solubilizing biologically active compounds
WO1999049716A2 * Mar 24, 1999 Oct 7, 1999 Clemens Unger Method for producing liposomal formulations of active agents by high-pressure homogenisation
WO1999049716A3 * Mar 24, 1999 Jan 13, 2000 Clemens Unger Method for producing liposomal formulations of active agents by high-pressure homogenisation
WO2002080883A3 * Mar 26, 2002 Dec 11, 2003 Mathew Louis Steven Leigh Method and composition for solubilising a biologically active compound with low water solubility
WO2007111720A2 * Dec 1, 2006 Oct 4, 2007 Rigel Pharmaceuticals, Inc. Formulation of insoluble small molecule therapeutics in lipid-based carriers
WO2007111720A3 * Dec 1, 2006 Dec 27, 2007 Rigel Pharmaceuticals Inc Formulation of insoluble small molecule therapeutics in lipid-based carriers
WO2008127358A2 * Oct 10, 2007 Oct 23, 2008 Jina Pharmaceuticals, Inc. Aqueous systems for the preparation of lipid-based pharmaceutical compounds; compositions, methods, and uses thereof
WO2008127358A3 * Oct 10, 2007 Dec 24, 2008 Jina Pharmaceuticals Inc Aqueous systems for the preparation of lipid-based pharmaceutical compounds; compositions, methods, and uses thereof
U.S. Classification 424/450, 436/164, 436/829, 264/4.3, 514/908, 424/1.21