FORM 2
THE PATENTS ACT, 1970
(39 of 1970)
&
THE PATENTS RULES, 2003
PROVISIONAL SPECIFICATION
(See section 10 and rule 13)


Title: LIPOSOME COMPOSITION COMPRISING DOCETAXEL
GLENMARK PHARMACEUTICALS LIMITED, An Indian Company registered under The Companies Act, 1956, India
and having its office at
Glenmark House, HDO - Corporate Bldg,
Wing A, B. D. Sawant Marg,
Chakala, Andheri (East),
MUMBAI - 400 099
The following specification describes the invention.


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LIPOSOME COMPOSITION COMPRISING DOCETAXEL
TECHNICAL FIELD OF THE INVENTION The present invention relates to a liposome composition comprising docetaxel, its pharmaceutically acceptable safts, analogs, polymorphs, solvates, single isomers, enantiomers, or mixtures thereof. Particularly, the present invention relates to a liposome composition for parenteral administration comprising docetaxel, processes for preparing the same; and methods of using it. More particularly, the present invention relates to docetaxel liposome compositions to be administered parenterally, as such or, after dilution with an aqueous fluid.
BACKGROUND OF THE INVENTION
Taxanes (also known as "taxoids" or "taxines") like docetaxel are widely used in cancer chemotherapy. Taxanes are often administered via injection or infusion of liquid solutions to better control their blood-borne concentrations. However, due to the aqueous insolubility of taxanes, intravenous injection or infusion of taxanes poses serious problems and challenges for pharmaceutical scientists and physicians. Further such parenteral taxane formulations have the potential for serious side effects in the patients. Various methods for emulsifying, suspending, or encapsulating insoluble taxanes in injectable formulations have been used for decades, but none of those approaches are fully satisfactory for taxanes, and the available commercial formulation of docetaxel still poses serious problems, risks, and drawbacks.
Docetaxel trihydrate (Formula I) is chemically (2R,3S)-N-carboxy- 3-phenylisoserine, N-f-butyl ester, 13-ester with 5(3-20-epoxy-1 ,2α,4,7β,10β, 13α- hexahydroxytax-11 -en-9-one 4-acetate 2-benzoate, trihydrate. It is a white to aimost-white powder with an empirical formula of

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C43H53NO14.3H2O, and a molecular weight of 861,9. It is highly lipophilic and practically insoluble in water.


(Formula I)
Docetaxel is commercially available in the form of an injection concentrate under the brand name TAXOTERE®, marketed by Aventis Pharmaceuticals Inc. It is indicated for the treatment, of neoplasm conditions such as breast cancer, lung cancer and prostate cancer.
TAXOTERE® is sterile, non-pyrogenic, and is available in single-dose vials containing 20 mg (in 0.5 ml) or 80 mg (in 2 ml) docetaxel (expressed on an anhydrous basis). TAXOTERE® injection comprises a two compartment formulation that requires two-step dilution before infusion. The first step involves dilution with the contents of a diluent vial (13 % w/w ethanol in water for injection) without significant foaming, and the second step involves further dilution with infusion fluid for parenteral administration. Thus, the marketed formulation has serious manufacturing, handling and dosing limitations.
The problems associated with TAXOTERE® include, for example, high rates of allergic and/or immune reactions, severe pain at injection sites, serious and potentially permanent damage to blood vessels at or near the site of injection, and the like. Among all side effects, the allergic and/or immune reactions are the most serious and sometimes of fatal risk, and perhaps for

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that reason, the FDA has included a "black box" warning in the approved labels for TAXOTERE®.
However, the severe adverse reactions of TAXOTERE® are believed to be not only due to the docetaxel itself, but may also because of the presence of polysorbate 80, an excipient used in TAXOTERE®, which supposedly causes anaphylaxis and severe hypersensitivity.
U.S. Patent Nos. 5,438,072; 5,670,536; 5,698,582; 5,714,512 and 5,750,561; 6,040,330; 6,071,952; 7,238,369; 7,314,637 describe various taxane/ docetaxel formulations.
U.S. Patent Application Publication Nos. 2005/0232952 and 2006/0292186 and U.S. Patent No. 6,982,282 disclose self-emulsifying delivery systems for poorly soluble drugs. .
U.S. Patent Application Publication No. 2006/0067952 discloses low oil emulsion compositions for delivering taxoids and other insoluble drugs. U.S. Patent Application Publication No. 2007/0082838 describes compositions and methods for preparation of poorly water-soluble drugs with increased stability.
U.S. Patent Application Publication No. 2006/0029658 and 2007/0032438 discloses pharmaceutical compositions comprising taxanes and methods for their preparation.
Without being bound by any theory, the inventors of the present invention believe that formulating docetaxel as liposome composition is likely to reduce hypersensitivity and / or toxicity associated with TAXOTERE® upon parenteral administration in a subject in need thereof.
There is always a need for newer ways and compositions of treating disease conditions, especially those with a high prevalence such as cancer. Hence, it is believed that the liposome compositions comprising docetaxel will be a significant improvement towards fulfilling the unmet medical need for alternative docetaxel compositions with reduced side effects.

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SUMMARY OF THE INVENTION The present invention relates to a liposome composition for parenteral administration comprising
(a) docetaxel in the range of about 0.8 % to about 2 % w/w; and
(b) a lipid mixture in the range of about 10 % to about 20 % w/w; wherein the lipid mixture contains phosphatidylcholine, cholesterol and 2-distearoyl-sn-glycero-3-phosphoethanolamine-N[methoxy(polyethylene glycol)-2000].
In an embodiment, the docetaxel liposome composition for parenteral administration comprises a lipid mixture that includes phosphatidylcholine in the range of about 8 % to about 11 % w/w, or preferably, about 8.5 % to about 10.5 % w/w (based on 100 % total weight of the composition).
In another embodiment, the docetaxel liposome composition for parenteral administration comprises a lipid mixture that includes cholesterol in the range of about 1.5 % to about 3.5 % w/w, or preferably, about 2 % to about 3 % w/w (based on 100 % total weight of the composition).
In yet another embodiment, the docetaxel liposome composition for parenteral administration comprises a lipid mixture that includes 2-distearoyl-sn-glycero-3-phosphoethanolamine-N [methoxy (polyethylene glycol)-2000] in the range of about 2.5 % to about. 4.5 % w/w, or preferably, about 3 % to about 4 % w/w (based on 100 % total weight of the composition).
In an aspect, the weight ratio of docetaxel to lipid in the liposome composition of the present invention ranges from about 1:5 to about 1:25, or preferably, from about 1:10 to about 1:20.
In another aspect, the liposome composition of the present invention further includes sugar and buffer. Preferably, the sugar is sucrose, dextrose, maltose; more preferred being sucrose.

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The docetaxel liposome composition for parenteral administration of the present invention can be in the form of a lyophilized powder or in the form of an aqueous dispersion. Typically, the liposome composition for parenteral administration, when dispersed in an aqueous fluid, exhibits the mean particle size, which is less than about 0.5 urn, or preferably less than about 0.3 urn.
In the context of present invention, the net surface charge on the docetaxel liposome composition for parenteral administration, when dispersed in an aqueous fluid, may be negative, positive or neutral. Preferably, the net surface charge on the dispersed liposome is negative or neutral.
Another embodiment of the present invention provides a process for preparing the liposome composition, said process comprising the steps of:
(a) dissolving the lipid mixture and docetaxel in an organic solvent;
(b) adding the aqueous phase to step (a);
(c) sonicating the composition of step (b) to form liposomes; and
(d) removing the un-entrapped docetaxel by filtration and centrifugation. The process may further include terminal sterilization of the docetaxel
liposome composition by filtration through 0.22 urn membrane, and optionally lyophilization to obtain a powder form.
Yet another embodiment of the present invention provides a method of using the docetaxel liposome composition of the present invention for the treatment of cancer by administering parenteraily a therapeutically effective amount of the liposome composition to a subject in need thereof. Preferably, the subject is a mammal including human.
DETAILED DESCRIPTION OF THE INVENTION The terms used herein are defined as follows. If a definition set forth in the present application and a definition set forth later in a non-provisional application claiming priority from the present provisional application are in

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conflict, the definition in the non-provisional application shall control the meaning of the terms.
The present invention relates to a liposome composition for parenteral administration comprising
(a) docetaxel in the range of about 0.8 % to about 2 % w/w; and
(b) a lipid mixture in the range of about 10 % to about 20 % w/w; wherein the lipid mixture contains phosphatidylcholine, cholesterol and 2-distearoyl-sn-glycero-3-phosphoethanolamine-N[methoxy(polyethylene glyco1)-2000].
The liposome is a closed structure composed of lipid bilayers surrounding an interna! aqueous space. The docetaxel liposome composition for parenteral administration of the present invention can be in the form of a lyophilized powder or in the form of an aqueous dispersion. Typically, the liposome composition for parenteral administration, when dispersed in an aqueous fluid, exhibits the mean particle size, which is less than about 0.5 urn, or preferably less than about 0.3 urn.
In an embodiment, the docetaxel liposome composition for parenteral administration comprises a lipid mixture that includes phosphatidylcholine in the range of about 8 % to about 11 % w/w, or preferably, about 8.5 % to about 10.5 % w/w (based on 100 % total weight of the composition).
In another embodiment, the docetaxel liposome composition for parenteral administration comprises a lipid mixture that includes cholesterol in the range of about 1.5 % to about 3.5 % w/w, or preferably, about 2 % to about 3 % w/w {based on 100 % total weight of the composition).
In yet another embodiment, the docetaxel liposome composition for parenteral administration comprises a lipid mixture that includes 2-distearoyl-sn-glycero-3-phosphoethanolamine-N [methoxy (polyethylene glycol)-2000] in the range of about 2.5 % to about 4.5 % w/w, or preferably, about 3 % to about 4 % w/w (based on 100 % total weight of the composition).

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As used herein, the terms "effective amount" or "therapeutically effective amount" of an docetaxel refers to a non-toxic but sufficient amount to provide the desired effect. The "effective amount" for docetaxel will vary depending on the disease and its severity and the age, weight, physical condition and responsiveness of the mammal to be treated. In one embodiment of the present invention, for parenteral administration, the therapeutically effective amount of liposomal docetaxel to be administered every 3 weeks ranges from about 30 mg/m2 to about 100 mg/m2, or about 40 mg/m2 to about 80 mg/m2. Such an amount can typically be administered over a period of about 1 hour to about 2 hours.
As used herein, the term "subject" includes human and other animals, such as domestic animals (e.g., household pets including cats and dogs) and non-domestic animals (such as wildlife). Preferably, the subject is a human.
The term "treating" or "treatment" as used herein also covers the prophylaxis, mitigation, prevention, amelioration, or suppression of a disorder related to histamine level in a mammal.
The term "active ingredient" (used interchangeably with "active" or "active substance") used herein includes docetaxel or pharmaceutical^ acceptable salts thereof, its pharmaceutically acceptable analogs, polymorphs, solvates, single isomers, enantiomers and mixtures thereof.
By "salts" or "pharmaceutically acceptable salts", it is meant those salts and esters which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, and allergic response, commensurate with a reasonable benefit to risk ratio, and effective for their intended use. Representative acid additions salts include the hydrochloride, hydrobromide, sulphate, and bisulphate. Representative alkali or alkaline earth metal salts include the sodium, calcium, potassium and magnesium salts.

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The term "pharmaceutically acceptable" as used in connection with components includes those components approved by a governmental regulatory agency or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in mammals, such as humans.
The lipid mixture in the context of present invention includes one or more lipids selected from the group consisting of cholesterol (CH), and phospholipids. Phospholipids are triesters of glycerol with two fatty acids and one phosphate ion. Exemplary phospholipids useful in the present invention include, but are not limited to, phosphatidylcholine, lecithin (a mixture of choline ester of phosphorylated diacylglyceride), phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid with about 4 to about 22 carbon atoms, and more generally from about 10 to about 18 carbon atoms and varying degrees of saturation. Non- limiting examples of phospholipids, in the context of the present invention, ajso include natural phospholipids, egg lecithin, soy lecithin, saturated phospholipids, phosphatidyl cholines, phosphatidic acid, sphingomyelins, aminolipids, glycolipids, lysolipids, sterols, cardiolipin and its synthetic derivatives, phospholipids with multifarious functional groups, polymerizable phospholipids, cholesterol, and mixtures thereof. The phospholipid component of the composition can be either a single phospholipid or a mixture of several phospholipids. Phospholipids can be of either natural or synthetic origin. Naturally occurring lecithin can be obtained from a variety of sources including eggs and soybeans. The phospholipids should be acceptable for the chosen route of administration and are generally regarded as safe ("GRAS"). Phospholipids, in the context of present invention, may have a positive charge, a negative charge, or no charge. Accordingly, the phospholipid may be termed as a "cationic phospholipid", "anionic phospholipid" or "neutral phospholipid", respectively.
The lipid mixture of the composition includes one or more lipids selected from phospholipids such as dimyristolphosphatidylcholine (DMPC),

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dipalmitoylphosphatidylcholine (DPPC) and distearoylphasphatidylcholine
(DSPC) and unsaturated phospholipids such as hydrogenated purified soy
bean phosphatidylcholine (HSPC), hydrogenated purified egg yolk
phosphatidylcholine, dilinoleoylphosphatidylcholine (DLPC),
phosphatidylcholine (DOPC), palmitoyloleoyl dioleoylphosphatidylcholine (POPC) and sphingomyelin may be used. Suitable negatively charged lipids such as dioleoylphsophatidylglycerol (DOPG), dioleoyi phosphatidylserine (DOPS), dimyristolphosphatidylglycerol (DMPG) also can be used.
In an aspect, the weight ratio of docetaxel to lipid in the liposome composition of the present invention ranges from about 1:5 to about 1:25, or preferably, from about 1:10 to about 1:20.
In another aspect, the liposome composition of the present invention further includes sugar and buffer. Preferably, the sugar is sucrose, dextrose, maltose; more preferred being sucrose.
The compositions of the present invention may optionally contain pharmaceutically acceptable excipients such as co-solvents or solubilizing agents, antioxidants, pH modifiers and stabilizers, preservatives, suspending and/or viscosity modifying agents, tonicity modifying agents, and other such biocompatible materials or therapeutic agents as are known to a person skilled in the art.
Non-limiting examples of co-solvents, in the context of the present invention, include ethanol, propylene glycol, glycerol, glycofural, polyethylene glycol, diethylene glycol monoethyl ether (TRANSCUTOL®), polyethylene glycol 660 12-hydroxystearate (SOLUTOL®) and the like, and mixtures thereof. "Antioxidants" used in this invention include metal ion chelators and/or reducing agents. Metal ion chelators useful in this invention include, but are not limited to, EDTA (ethylenedinitrilotetraacetic acid), glycine, citric acid and salts thereof. Non-limiting examples of antioxidants also include vitamin E, vitamin E succinate, ascorbic acid, sodium metabisulfite, amino acids,

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flavones, monothioglycerol, L-cysteine, thioglycolic acid and mixtures thereof. Such antioxidants are generally used in concentration ranges of 0.1 to 15% w/w, or 0.5 to 5% w/w. Non-limiting examples of pH modifiers and stabilizers include citric acid, tartaric acid, succinic acid, glutamic acid, ascorbic acid, lactic acid, acetic acid, malic acid, maleic acid, Phosphoric acid, acetic acid, and salts thereof, sodium hydroxide, sodium carbonate, sodium bicarbonate, tris buffer, meglumine, amino acids like histidine and mixtures thereof. Such pH modifiers and stabilizers (synonymously, buffers) maintain a desired pH between about 1 and 8, or between about 2.5 and 5.5 in the composition.
Non-limiting examples of preservatives include parabens such as methyl paraben, propyl paraben; butyl paraben, benzoyl alcohol, cresol and metacrosol, chlorobutanol, phenyl ethanol, thimerosol benzalkonium chloride, and the like, and mixtures thereof.
Non-limiting examples of tonicity modifying agents include sodium chloride, dextrose, mannitol, lactose, propylene glycol, glycerin, and the like, and mixtures thereof.
The docetaxel liposome composition for parenteral administration of the present invention can be in the form of a lyophilized powder or in the form of an aqueous dispersion. In an embodiment, the liposome compositions of the present invention may be diluted with aqueous fluids including water, various buffer solutions having different pH values, parentera| infusion fluids, and other such media. Typically, parenteral infusion fluids include 5 % dextrose solution, 0.9 % sodium chloride solution, Ringer's lactate, mannitol infusion fluid, sucrose infusion fluid, plasma volume expanders, andj mixtures thereof, and dilution typically will produce docetaxel concentrations in the infusion fluid ranging between about 0.05 mg/ml and about 10 mg/ml, or from about 0.06 mg/ml and about 8 mg/ml. Such aqueous fluids can be provided separately, or can be included with a container containing the liposome compositions, in the

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form of a kit. The diluted docetaxel liposome composition can be administered parenterally to a subject in need thereof.
In an aspect, the present invention also relates to a kit for the delivery of docetaxel comprising: (a) a container having the docetaxel liposome composition; and (b) another container having a pharmaceutically acceptable aqueous diluent; provided, upon mixing (a) and (b), a docetaxel concentration ranges from about 0.05 mg/ml and about 10 mg/ml, or between about 0.06 mg/ml and about 8 mg/ml. The diluted docetaxel liposome composition can be administered parenterally to a subject in need thereof.
In an embodiment, the docetaxel liposome composition of the present inventions may be parenterally administered to a subject. Parenteral administration most commonly refers to injections or infusions into blood vessels. In certain embodiments, the mode of administration of the present docetaxel liposome composition is by intravenous, intra-arterial, intrathecal, intraperitoneal, intratumoral, intra-articular, intramuscular or subcutaneous injection, and the like, to be administered as bolus or infusion.
In an aspect of the invention, the docetaxel liposome compositions for parenteral administration of the present invention are both chemically and physically stable. The docetaxel liposome composition is "chemically stable" if the docetaxel in the composition is not substantially chemically degraded during storage under appropriate conditions. The docetaxel liposome composition is "physically stable" if it can be stored under appropriate conditions without significant increases in its average particle size upon dilution, or evidence of phase separation, or aggregation. In an aspect, the docetaxel liposome compositions of the present invention remain stable in their packaging in terms of impurities generated during storage tests at appropriate storage conditions such as at 2°C to 8°C, at 25°C and relative humidity about 60%, and at 40°C and relative humidity about 75%.

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In the context of present invention, the liposome composition for parenteral administration, when dispersed in an aqueous fluid, exhibits the mean particle size, which is less than about 0.5 urn, or preferably less than about 0.3 pm.
As used herein, the term "mean particle size" (or synonymously, "average particle size") refers to the distribution of liposome particles wherein about 50 volume percent of all the particles measured have a size less than the defined average particle size value and about 50 volume percent of all measurable particles measured have a particle size greater than the defined average particle size value. This can be identified by the term "D50" or "d (0.5)"-The particle size can be typically measured using instruments like ZETASIZER® 3000 HS (Malvern® Instruments Ltd., Malvern, Worcestershire, United Kingdom) or NICOMP 388™ ZLS system (PSS-Nicomp Particle Sizing Systems, Santa Barbara, CA.USA).
In the context of present invention, the net surface charge on the docetaxel liposome composition for parenteral administration, when dispersed in an aqueous fluid, may be negative, positive or neutral depending on the ingredients used therein. This charge is often expressed in terms of "zeta potential", which refers to the. electrostatic potential generated by the accumulation of ions at the surface of the particle. Zeta potential can be measured using instruments like ZETASIZER® 3000 HS (Malvern® Instruments Ltd., Malvern, Worcestershire, United Kingdom) or NICOMP 388™ ZLS system (PSS-Nicomp Particle Sizing Systems, Santa Barbara, CA, USA). Preferably, the net surface charge on the dispersed liposome is negative or neutral.
The Docetaxel liposome composition can be formulated by any suitable method. Preferred methods include thin film hydration, solvent injection, freeze-thawing and dehydration-rehydration, removal of surfactant, reverse phase evaporation and ethanol injection. For example, the active ingredient

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and lipid can be dissolved in a suitable solvent, such as methylene chloride, ethanol, methyl acetate, ethyl formate and the like.
Another embodiment of the present invention provides a process for preparing the liposome composition, said process comprising the steps of:
(a) dissolving the lipid mixture and docetaxel in an organic solvent;
(b) adding the aqueous phase to step (a);
(c) sonicating the composition of step (b) to form liposomes; and
(d) removing the un-entrapped docetaxel by filtration and centrifugation. The process may further include terminal sterilization of the docetaxel
liposome composition by filtration through 0.22 pm membrane, and optionally lyophilization to obtain a powder form.
The compositions of the present invention may optionally contain pharmaceutically acceptable excipients such as co-solvents or solubilizing agents, antioxidants, pH modifiers and stabilizers, preservatives, suspending and/or viscosity modifying agents, tonicity modifying agents, and other such biocompatible materials or therapeutic agents as are known to a person skilled in the art.
The liposome composition of the present invention can contain other pharmaceutically acceptable excipients. Examples of these excipients are described in, for example, Howard C. Ansel et. a/., Pharmaceutical Dosage Forms and Drug Delivery Systems, (7th Ed. 1999); Alfonso R. Gennaro et a/., Remington: The Science and Practice of Pharmacy, (20th Ed. 2000); and A. Kibbe, Handbook of Pharmaceutical Excipients, (3rd Ed. 2000), the contents of which are incorporated by reference herein.
Yet another embodiment of the present invention provides a method of using the docetaxel liposome composition of the present invention for the treatment of cancer by administering parenteraliy a therapeutically effective amount of the liposome composition to a subject in need thereof. Preferably, the subject is a mammal including human.

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The docetaxel liposome composition according to the instant invention may be used for the treatment of various cancers (or tumors) like carcinoma, Kaposi's sarcoma, malignancies, uncontrolled tissue or cellular proliferation secondary to tissue injury, and any other disease conditions responsive to docetaxel, and/or prodrugs and derivatives thereof. Among the types of carcinoma, which may be treated particularly effectively with docetaxel are hepatocellular carcinoma and liver metastases, cancers of the gastrointestinal tract, pancreas, prostate and lung, and Kaposi's sarcoma. Generally, the compositions of the present invention, either alone or in combination with other drugs, are useful for treatment of tumors in breast, lung, stomach, head, neck and prostate tissues, esophageal neoplasm, and any other such tumors in mammals.
It will be understood that various modifications may be made to the embodiments disclosed herein. Therefore the above description should not be construed as limiting, but merely as exemplifications of preferred embodiments. Other arrangements and methods may be implemented by those skilled in the art without departing from the scope and spirit of this invention.
The following examples are provided to enable one skilled in the art to practice the invention and are merely illustrative of the invention. The examples should not be read as limiting the scope of the invention.

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EXAMPLES EXAMPLE 1: Docetaxel liposome composition

S.No Ingredients Composition
(% w/w)
1. Docetaxel 1.29
2. Hydrogenated soy phosphatidylcholine (HSPC) 9.53
3. Cholesterol 2.4
4. 1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine-N [methoxy (polyethyleneglycol)-2000] (Ammonium salt) (DSPE-PEG) 3.45
5. Histidine buffer 1.29
6. Sucrose 82.0
Manufacturing process:
1. HSPC, Cholesterol DSPE-PEG and Docetaxel are dissolved in sufficient quantity of chloroform in to 1000 ml round bottom flask.
2. Chloroform is evaporated in rota evaporator at about 40 °C with 300 to 400 m bar pressure so as to obtain a thin film. The dried thin film is heated to a temperature of about 68 °C
3. Thin dried film of step 2 is hydrated with an aqueous phase containing 0.01 M histidine buffer (pH 6.5) and 0.29 M sucrose, which is previously heated up to about 68 °C. The flask is rotated at about 130 RPM to remove the film from glass wall. A viscous gel like liposomal dispersion is formed in the flask.

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4. The volume of liposomal dispersion of step 3 is made up to get required concentration of docetaxel.
5. The diluted docetaxel liposome dispersion of step 4 is homogenized in high pressure homogenizer at about 18,000 pst for 4 cycles.
6. The homogenized sample of step 5 is extruded with 100 nm polycarbonate membrane to get uniform distribution of homogenized liposome and to separate unentrapped docetaxel.
7. The docetaxel liposome dispersion of step 6 is filtered through 0.22 urn syringe filter.
EXAMPLE 2: Docetaxel liposome composition

S.No INGREDIENTS Composition (% w/w)
1. Docetaxel 1.04
2. Hydrogenated soy phosphatidylcholine (HSPC) 9.56
3. Cholesterol 2.41
4. 1,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine-N [methoxy (polyethyleneglycol)-2000] (Ammonium salt) (DSPE-PEG) 3.46
5. Histidine buffer 1.30
6. Sucrose 82.22
Manufacturing process:

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1. HSPC, Cholesterol DSPE-PEG and Docetaxel are dissolved in sufficient quantity of chloroform in to 1000 ml round bottom flask.
2. Chloroform is evaporated in rota evaporator at about 38 °C with 450 m bar pressure so as to obtain a thin film.
3. Again the dried film is dissolved in chloroform, and chloroform is evaporated as in step 2.
4. To the dried film of step 3, chloroform is added to dissolve all the contents of flask.
5. To the solution of step 4 is an aqueous phase containing 0.01 M histidine buffer (pH 6.5) and 0.29 M sucrose, and sonicated 10 min at about 18 °C.
6. After sonication, the chloroform is evaporated slowly in rotary vacuum evaporator to form viscous gel.
7. The chloroform is completely evaporated to form liposome suspension.
8. The docetaxel liposome dispersion of step 7 is homogenized in high pressure homogenizer at about 18,000 psi for 3 cycles.
9. The homogenized sample of step 8 is extruded with 100 nm polycarbonate membrane to get uniform distribution of homogenized liposome and to separate unentrapped docetaxel.
10. The docetaxel liposome dispersion of step 6 is filtered through 0.22 urn syringe filter.

Dated 28 th day of August 2008
I Signed (NameTaranpreet Singh Lamba)
Deputy General Manager-IPM GLENMARK PHARMACEUTICALS LIMITED, Mumbai