WO2008042841A2

WO2008042841A2 - Docetaxel compositions

Info

Publication number: WO2008042841A2
Application number: PCT/US2007/080063
Authority: WO
Inventors: Kocherlakota Chandrasekhar; Tarun Singh; Bejgum Santosha; Banda Nagaraju; Ramesh Mullangi; Ravi Kumar Trivedi
Original assignee: Dr. Reddy's Laboratories Limited; Dr. Reddy's Laboratories, Inc.
Priority date: 2006-10-02
Filing date: 2007-10-01
Publication date: 2008-04-10
Also published as: WO2008042841A3

Abstract

Emulsion-preconcentrate compositions comprising docetaxel, processes for preparing the same, and their methods of use. The docetaxel compositions are suitable for parenteral administration to treat neoplasm conditions, upon dilution with aqueous fluids.

Description

DOCETAXEL COMPOSITIONS

INTRODUCTION TO THE INVENTION

The present invention relates to pharmaceutical compositions comprising docetaxel, pharmaceutically acceptable salts thereof, its pharmaceutically acceptable analogs, polymorphs, solvates, single isomers, enantiomers and mixtures thereof, and processes for preparing the same. Further, the present invention relates to emulsion-preconcentrate compositions comprising docetaxel, processes for preparing the same, and their methods of use. More particularly, the present invention relates to docetaxel compositions for parenteral administration after dilution with aqueous fluids.

Taxanes (also known as "taxoids" or "taxines") are widely used in cancer chemotherapy. Because they tend to be highly toxic, 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 these drugs poses serious problems and challenges for pharmaceutical scientists and physicians as well as the potential for serious side effects in the patients. Various methods for emulsifying, suspending, or encapsulating insoluble drugs in injectable formulations have been used for decades, but none of those approaches are fully satisfactory for taxanes, and the best available formulations of taxanes pose serious problems, risks, and drawbacks. Such problems 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 for that reason, the FDA has requested the manufacturers of taxanes viz. paclitaxel (TAXOL^®) and docetaxel (TAXOTERE^®) to include a "black box" warning in the approved labels for these products.

However, the severe adverse reactions of these two important drugs are not due to the drugs themselves, but to the excipient ingredients used in their formulations. For TAXOL^®, anaphylaxis and severe hypersensitivity are caused by the presence of the excipients Cremophor^® EL (polyoxyethylated castor oil), and for TAXOTERE^®, polysorbate 80. Docetaxel is an antineoplastic agent belonging to the taxoid family. It is indicated for the treatment of neoplasm conditions such as breast cancer, lung cancer and prostate cancer. A chemical name for docetaxel is (2R,3S)-N-carboxy- 3-phenylisoserine, N-f-butyl ester, 13-ester with 5β-20-epoxy-1 ,2α,4,7β,10β,13α- hexahydroxytax-11-en-9-one 4-acetate 2-benzoate, trihydrate, and docetaxel is represented by structural Formula 1.

Formula 1

Docetaxel is a white to almost-white powder with an empirical formula of 0₄₃H₅₃NOi₄-SH₂O, and a molecular weight of 861.9. It is highly lipophilic and practically insoluble in water. Docetaxel is commercially available in the form of an injection concentrate under the brand name TAXOTERE^®, marketed by Aventis Pharmaceuticals Inc. 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.

U.S. Patent Nos. 5,438,072, 5,698,582, 5,714,512 and 5,750,561 describe docetaxel formulations.

U.S. Patent No. 5,670,536 discloses a docetaxel composition prepared by homogenization and having an average particle size less than 200 nm. U.S. Patent No. 6,040,330 describes a formulation comprising taxane and N-methyl pyrrolidone to be diluted with parenteral delivery vehicle before use.

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, yielding emulsions upon dilution with aqueous phase.

U.S. Patent No. 6,071 ,952 describes a stabilized injectable composition comprising a taxoid.

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. 2007/0032438 discloses pharmaceutical compositions comprising taxanes and methods for their preparation.

The current approaches for reducing the side effects of the commercial product are mainly focused on developing formulations that do not contain polyoxyethylated castor oils and polysorbates. Several attempts have been made to deliver paclitaxel using alternative systems, such as nanoparticles described by R. Cavalli et al., European Journal of Pharmaceutical Sciences, 2000, 10:305- 309, liposomes described by P. Crosasso et al., Journal of Controlled Release, 2000, 63:19-30, and A. Sharma et al., Pharmaceutical Research, 1994, 11 :889- 896). However, success at this time is still limited.

A further aspect that remains to be addressed in the use of compositions for the delivery of docetaxel includes the ability to modify the delivery characteristics of the vehicle using the composition to modify the delivery to different organs in the body. To date there are no report of emulsion compositions of docetaxel with enhanced volume of distribution. An enhanced volume of distribution correlates with a higher tissue distribution and lower amount of drug in the central compartment (blood compartment). Since docetaxel is indicated for the treatment of solid tumors located in the organs such as the breast, lung and prostate (among other tumors) it is reasonable that a composition for the delivery of docetaxel which has a higher volume of distribution when compared with the currently available solution composition would preferentially deliver drug to the tissue compartment with a higher probability of delivery to the target organs. Conflicting research articles demonstrate uncertainty in correlating the charge on dispersed particles (for example, emulsion droplets, liposomes, nanoparticles, and the like) and their tissue distribution patterns as well as potentials for targeting (such as preferential accumulation of particles in specific tumor tissues). For example, R. R. Patlolla and V. Venkateswarlu, Journal of Pharmaceutical Sciences, 2005, 94 (2), 437-445, investigated the tissue distribution patterns of parenteral emulsions of etoposide. They observed that the volume of distribution at steady state ("Vss") for an etoposide parenteral emulsion was higher than that of etoposide solution when administered to mice. The same authors in another research publication Journal of Drug Targeting, 2005, 13 (10), 543-553 demonstrated improved area under the curve ("AUC") and reduced clearance ("CL") for an etoposide parenteral emulsion, as compared to an etoposide solution, when administered to rats.

A review article by Yang and Benita, Drug Development Research, 2000, 50, 476-486, teaches that the negatively charged droplets of parenteral emulsions are "certainly problematic for drug delivery to non-RES tissues" (such as lungs) as observed in tissue distribution studies in mice. A report of a cationic parenteral nano-emulsion technology (NOVASORB^®, developed by Novagali Pharma SA) suggests that cationic emulsions have biological advantages over anionic emulsions, since cell walls are anionic (L. Rabinovich-Guilatt and G. Lambert, Drug Delivery Report, Autumn/Winter 2005, pages 71-72).

Campbell et al., Cancer Research, 2002, 62, 6831-6836, studied the effect of cationic charge on the distribution of liposomes in mice. They reported that although the cationic liposome charge influenced the extent of liposome uptake by organs such as lungs and spleen, the overall tumor uptake was not affected by liposome charge in biodistribution studies.

The present invention provides docetaxel compositions in the form of emulsion-preconcentrates that readily disperse, upon dilution, in aqueous infusion fluids, thus addressing an unmet need of easy-to-prepare and easy-to-use formulations of docetaxel. Pharmaceutical compositions of the present invention require a single step dilution of an entire vial contents with infusion fluid for parenteral administration, unlike the current TAXOTERE^® injection concentrate. There is always a need for newer ways and compositions of treating disease conditions, especially ones with a high prevalence such as cancer. Hence, emulsion-preconcentrate compositions comprising taxanes like docetaxel will be a significant improvement towards fulfilling the unmet medical need for alternative docetaxel compositions with reduced side effects.

These and other such needs are addressed by the instant invention.

SUMMARY OF THE INVENTION

An aspect of the present invention provides an emulsion-preconcentrate composition comprising docetaxel and an anionic unsaturated phospholipid, wherein said composition, upon dilution with an aqueous fluid, forms an 'oil-in- water' type emulsion.

Another aspect of the present invention provides an emulsion- preconcentrate composition comprising docetaxel, oil, and a mixture of anionic unsaturated phospholipid and neutral phospholipid.

Further aspect of the present invention provides an emulsion preconcentrate comprising docetaxel, an oil, a phospholipid or a mixture of phospholipids, and optionally a co-solvent, wherein at least one phospholipid is an anionic phospholipid

In an embodiment, an emulsion-preconcentrate composition comprises docetaxel in a dispersed or dissolved state, and an 'oil-in-water' type emulsion readily formed upon dilution of the composition with an aqueous fluid can be administered parenterally to a mammal in need thereof.

In certain embodiments, an average droplet size of 'oil-in-water' type emulsion resulting from dilution of an emulsion-preconcentrate composition with an aqueous fluid is less than about 400 nm. In another embodiment, droplets of 'oil-in-water' type emulsion resulting from dilution of an emulsion-preconcentrate composition with an aqueous fluid have a net negative charge.

In a further aspect, emulsion-preconcentrate compositions of the present invention remain stable for commercially acceptable periods under storage conditions such as about 2 ⁰C to 8 ⁰C, about 25 ⁰C and relative humidity about 60 %, and about 40 ⁰C and relative humidity about 75 %.

In a still further aspect, emulsion-preconcentrate compositions of the present invention exhibit reduced mortality as compared to presently marketed compositions when, following dilution, they are parenterally administered to a mammal.

Another aspect of the present invention provides injectable oil-in-water emulsions comprising a therapeutically effective amount of docetaxel and at least one phospholipid, wherein said emulsions exhibit an improved volume of distribution than is obtained with reconstituted presently marketed compositions, upon parenteral administration to a mammal.

A further aspect of the present invention provides injectable oil-in-water emulsions comprising a therapeutically effective amount of docetaxel and at least one phospholipid, wherein the emulsions exhibit improved systemic exposure than is obtained with reconstituted presently marketed compositions, upon parenteral administration to a mammal at the same doses.

A still further aspect of the present invention provides injectable oil-in-water emulsions comprising a therapeutically effective amount of docetaxel and at least one phospholipid, wherein the emulsions exhibit reduced toxicity than is obtained with reconstituted presently marketed compositions, upon parenteral administration to a mammal at the same doses.

A yet further embodiment of the invention provides an emulsion preconcentrate comprising docetaxel, an oil, a phospholipid or a mixture of phospholipids, and optionally a co-solvent, wherein at least one phospholipid is an anionic phospholipid imparting a net negative charge about -10 mV to about -70 mV to emulsion droplets formed upon dilution of the preconcentrate with an aqueous fluid. DETAILED DESCRIPTION

The present invention relates to pharmaceutical compositions of docetaxel, pharmaceutically acceptable salts thereof, its pharmaceutically acceptable analogs, polymorphs, solvates, single isomers, enantiomers and mixtures thereof, and processes for preparing the same. Further, the present invention relates to emulsion-preconcentrate compositions comprising docetaxel, processes for preparing the same, and their methods of use. More particularly, the present invention relates to docetaxel compositions for parenteral administration upon dilution with aqueous fluids. Emulsion-preconcentrate compositions (or "emulsion preconcentrate", also called self-emulsifying drug delivery systems ("SEDDS"), self-nanoemulsifying drug delivery systems ("SNEDDS"), or self-microemulsifying drug delivery systems ("SMEDDS")) are mixtures comprising drug(s), oil(s) and surfactant(s), ideally isotropic, and sometimes containing cosolvents, which emulsify spontaneously to produce fine oil-in-water emulsions when introduced into aqueous diluents under gentle agitation. SEDDS or SNEDDS typically produce emulsions with average droplet sizes in the range of about 100-400 nm (also called nanoemulsions), while SMEDDS, upon dilution with aqueous diluents, produce transparent microemulsions, which are thermodynamically stable isotropic "solutions" with average droplet sizes less than about 100 nm. SeIf- emulsification has been shown to be specific to the nature of the oil/surfactant pair, the surfactant concentration and oil/surfactant ratio, and the temperature at which self-emulsification occurs. In support of these factors, it has also been demonstrated that only very specific pharmaceutical excipient combinations could lead to efficient self-emulsifying systems. It is the appropriate choice of oil and surfactant (and cosurfactant) together with their proper concentrations, which provides an optimum, custom-designed, self-emulsifying formulation. The mechanism by which self-emulsification occurs is not yet well understood and there are different schools of thought. Various theories including interfacial or mixed film theories, solubilization theories and thermodynamic theories have been proposed to explain spontaneous emulsion formation. Further such emulsion- preconcentrate compositions require minimal energy input in terms of simple mixing for dispersion in aqueous fluids, without involving complex unit operations like rigorous dispersion and costly equipment like homogenizers. "Therapeutically effective amount" (used interchangeably with "pharmaceutically effective amount") refers to the quantity of a drug (e.g., docetaxel) that is effective to treat a disease or disorder (e.g., cancer), at a reasonable benefit/risk ratio applicable to any medical treatment. The term "therapy" or "treatment" as used herein refers to management and care of a patient for the purpose of combating a condition, such as a disease or a disorder. The term is intended to include the full spectrum of treatments for a given condition from which the patient is suffering, such as administration of docetaxel to alleviate the symptoms or complications, to delay the progression of the disease, disorder or condition, to alleviate or relief the symptoms and complications, and/or to cure or eliminate the disease, disorder or condition as well as to prevent the condition. The patient to be treated is preferably a mammal, in particular a human, and may also include animals such as rats, mice, dogs, cats, cows, sheep and pigs. An "oil-in-water emulsion" refers to a dispersion system in which an oil is dispersed as small droplets (the "discrete phase," also referred to as "the oil phase") in an aqueous medium (the "continuous phase," also referred to as "the aqueous phase").

"Marketed composition" in the context of present invention refers to a conventional two-compartment TAXOTERE^® formulation that requires two step dilution before parenteral administration. TAXOTERE is marketed by Aventis Pharmaceuticals Inc.

An aspect of the present invention provides emulsion-preconcentrate compositions comprising docetaxel and an anionic unsaturated phospholipid, wherein said composition, upon dilution with an aqueous diluent, forms an oil-in- water-type emulsion.

In an embodiment, an emulsion-preconcentrate composition comprises docetaxel, oil, and a phospholipid, optionally with other pharmaceutically acceptable additives suitable for parenteral use, wherein docetaxel is in a dispersed or dissolved state, and such composition readily disperses, upon dilution, in aqueous infusion fluids to yield an oil-in-water-type emulsion of defined average droplet size. In one aspect of this embodiment, compositions of the present invention may require gentle shaking after dilution so as to form the oil-in- water emulsion dispersed uniformly throughout the bulk of aqueous fluid. In another embodiment, an emulsion-preconcentrate composition comprises docetaxel in a dispersed or dissolved state, and an 'oil-in-water' type emulsion is readily formed upon dilution of said composition with an aqueous diluent and can be administered parenterally to a mammal in need thereof. The term "oil" is used herein in a general sense to identify hydrocarbon derivatives, carbohydrate derivatives, or similar water-insoluble organic compounds that are frequently liquid at body temperatures, e.g., about 37 ⁰C, and are generally regarded as safe ("GRAS"). It includes glycerides or non-glycerides. The term "oil component" or "oil phase" refers to an oil, or a combination of multiple oils. In certain embodiments, an oil component of the present invention comprises a monoglyceride, a diglyceride, a triglyceride, or a mixture thereof. In certain embodiments, an oil component comprises an ester formed between one or more fatty acids and an alcohol other than glycerol. The oil component is frequently a lipophilic material that is triglyceride or propylene glycol diester oil, or a combination of both, either natural or synthetic in origin.

Oils include vegetable oils and medium chain triglycerides ("MCTs"). Vegetable oil refers to oils derived from plant seeds or nuts. Exemplary vegetable oils include, but are not limited to, almond oil, borage oil, black currant seed oil, corn oil, safflower oil, soybean oil, sesame oil, cottonseed oil, peanut oil, olive oil, coconut oil, palm oil, canola oil, etc. Vegetable oils are typically "long-chain triglycerides," formed when three fatty acid molecules (usually about 14 to about 22 carbons in length, with unsaturated bonds in varying numbers and locations, depending on the source of the oil) form ester bonds with the three hydroxyl groups on glycerol. In certain embodiments, vegetable oils of highly purified grade (also called "super refined") are generally used to ensure safety and stability of oil- in-water emulsions. In certain embodiments, hydrogenated vegetable oils, which are produced by controlled hydrogenation of the vegetable oil, may be used in the present invention. MCTs can be either naturally derived or synthetic. MCTs are made from fatty acids that are usually about 8 to about 12 carbons in length. Like vegetable oils, MCTs have been used extensively in emulsions designed for injection as a source of calories, for patients requiring parenteral nutrition. Representative oils are commercially available as MIGLYOL^® 840 (medium chain diesters of propylene glycols) and MIGLYOL^® 812 (medium chain triglyceride) from SASOL^® GmbH, Germany, CRODAMOL^® GTCC-PN (medium chain triglyceride) from Croda^® Inc. of Parsippany, NJ. U.S.A., or NEOBEE^® M-5 oil from PVO International^®, Inc., of Boonton, N.J. U.S.A. Other low-melting medium chain oils may also be used in the present invention.

Examples of suitable oils for purposes of this invention include triesters of glycerol with fatty acids having 6 to 14 carbon atoms. Suitable oils further include propylene glycol esters of capric and caprylic acids having from 19 to 23 carbon atoms. The triglycerides may be further subdivided into saturated, mono- unsaturated and polyunsaturated triglycerides, depending on whether the fatty acyl moieties of the triglyceride contain zero, one, or more than one double carbon-carbon bond. Mono or polyunsaturated long chain triglycerides, short chain and medium chain triglycerides, particularly short or medium chain, and more particularly medium chain triglycerides are useful for the present invention. In certain embodiments, combinations of vegetable oil and MCTs are used in the present invention. Such combinations generally have a long history of safe use in combination and provide superior stability for the emulsions of this invention. The specific type of vegetable oil used (i.e., soybean oil, corn oil, safflower oil, etc.) is not critical, so long as it is safe, well tolerated, pharmaceutically acceptable, chemically stable and provides dispersions having a desired average droplet size range. In the context of the present invention, "phospholipids" are compounds that prevent the separation of emulsion into individual oil and aqueous phases. Typical amphiphilic phospholipids useful in the present invention generally are (1 ) compatible with the other ingredients of the oil-in-water emulsions of the present invention, (2) do not interfere with the stability or efficacy of the docetaxel in the emulsions, (3) are stable and do not deteriorate in the preparation, and (4) are non-toxic.

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, phosphatidyl chlorine, 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, also include amphiphiles having hydrophilic-lipophilic balance (HLB) values in the range of about 2 to 40, or about 4 to 20, such as phospholipids comprising 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. Without being bound by any theory, the charge of phospholipid present in the emulsion-preconcentrate composition can determine the net charge on the surface of droplets generated upon dilution with aqueous fluid, and may further affect the performance characteristics of such droplets in biological milieu. Thus, when an anionic phospholipid is present to a larger extent in the composition, then a net negative charge could be expected on the droplets. The total charge can be varied through the use of anionic, neutral or cationic phospholipids individually or can be modified through the use of mixtures of phospholipids. Anionic materials such as sodium lauryl sulfate, sodium ethylhexyl sulfate, sodium lauryl ether sulfate, di-octyl sulphosuccinate and sodium soaps of coconut, palm and rice bran oils and the like can also be used with phospholipids to impart a net negative charge to the emulsion droplets.

Particularly, anionic phospholipids such as egg phosphatidyl glycerol ("EPG"), 1 ,2-dioleoyl-sn-glycero-3-[phospho-rac-(1 -glycerol)] ("DOPG"), 1 ,2- dipalmitoyl-sn-glycero-3-phosphoglycerol ("DPPG"), 1 ,2-dimyristoyl-sn-glycero-3- phosphoglycerol ("DMPG"), 1 ,2-Dioleoyl-sn-glycero-3-(phospho-L-serine) ("DOPS"), 1 ,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (¹DPPS"), 1 ,2- Dimyristoyl-sn-glycero-3-(phospho-L-serine) ("DMPS"), 1 ,2-Dioleoyl-sn-glycero-3- phosphate, monosodium salt ("DOPA"), 1 ^-Dipalmitoyl-sn-glycero-S-phosphate, monosodium salt ("DPPA"), i ^-Dimyristoyl-sn-glycero-S-phosphate, monosodium salt ("DMPA") and the like, and their pharmaceutically acceptable salts and mixtures thereof; and neutral phospholipids such as egg phosphatidyl choline ("EPC"), 1 ^-Dilinoleoyl-sn-glycero-S-phosphocholine ("DLPC"), 1 ,2-Dimyristoyl- sn-glycero-3-phosphocholine ("DMPC"), 1 ,2-Dipalmitoyl-sn-glycero-3- phosphocholine ("DPPC"), 1 ,2-Distearoyl-sn-glycero-3-phosphocholine ("DSPC"), 1 ^-Dioleoyl-sn-glycero-S-phosphocholine ('DOPC"), 1 ,2-Dimyristoyl-sn-glycero-3- phosphoethanolamine ("DMPE"), 1 ,2-Dipalmitoyl-sn-glycero-3- phosphoethanolamine ("DPPE"), 1 ^-Dioleoyl-sn-glycero-S-phosphoethanolamine ("DOPE"), 1-Oleoyl-2-hydroxy-sn-glycero-3-phosphocholine ("LOPC") and the like and mixtures thereof; and combinations of such anionic and neutral phospholipids; have been found to be useful in the context of the present invention. In certain embodiments, the anionic phospholipids containing one or more double bond (i.e. unsaturated anionic phospholipids) such as EPG, DOPG, DOPA, DOPS and their pharmaceutically acceptable salts and mixtures thereof, have been observed to provide desired properties.

In an embodiment an emulsion-preconcentrate composition of the present invention comprises docetaxel, phospholipid, oil, and co-solvent. Typically, an emulsion-preconcentrate composition comprises the following concentrations (% w/w) of the ingredients: a) Docetaxel about 1 % to about 6 %, or about 2 % to about 4 %. b) Phopsholipid(s) about 10% to about 30%, or about 15% to about 25%. c) Oil(s) about 4% to about 10 %, or about 5 % to about 8 %. d) Co-solvent(s) about 55 % to about 85 %, or about 65 % to about 75 %. In some embodiments of the present invention, anionic phospholipid contents in the emulsion-preconcentrate compositions in the range of about 10 % to about 40 % w/w, or about 15 % to about 30 % w/w, of the total phospholipids have been observed to be useful.

An aspect of the present invention relates to emulsion-preconcentrate compositions comprising docetaxel, oil, and a mixture of anionic unsaturated phospholipid and neutral phospholipid.

In an embodiment of the present invention, the emulsion-preconcentrate contains the weight ratio of the anionic phospholipid to the neutral phospholipid ranges between about 2:1 to about 1 :9, or about 1 :2 to about 1 :7, respectively. In certain embodiments, the phospholipids useful in the context of present invention include unsaturated anionic phospholipids and neutral phospholipids. Typical unsaturated anionic phospholipids include EPG, DOPG, DPPG, DMPG, DOPS, DPPS, DMPS, DOPA, and the like and mixtures thereof, and neutral

5 phospholipids comprise DLPC, DMPC, DPPC, DSPC, DOPC, DMPE, DPPE, and the like and mixtures thereof.

In an embodiment, an emulsion-preconcentrate composition of the present invention comprises docetaxel, EPG, DOPG, DMPC, DOPC, LOPC, soybean oil and MIGLYOL® 840, and pharmaceutical acceptable excipients.

IO In certain embodiments, weight ratios of DMPC to DOPC in the emulsion- preconcentrate composition ranging between about 9:1 and about 5:5, or about 8:2 and about 6:4, have been found to be useful. Also the LOPC content in the range of at least about 5 %, or about 10 %, by weight of the total phospholipids in the emulsion-preconcentrate composition was found to be useful.

15 In certain embodiments, the presence of DOPG in an emulsion- preconcentrate composition in the range of about 1% to about 20 % w/w, or about 5% to about 15 % w/w, of the total phospholipids has been observed to yield good quality emulsions upon dilution with aqueous fluids.

In an embodiment, an emulsion-preconcentrate composition of the present

>0 invention comprises weight ratios of docetaxel to phospholipid in the range of about 1 :1 to about 1 :50, or about 1 :5 to about 1 :30, or about 1 :10 to about 1 :20. In another embodiment, an emulsion-preconcentrate composition of the present invention comprises weight ratios of oil(s) to phospholipid in the range of about 1 :1 to about 1 :20, or about 1 :2 to about 1 :15, or about 1 :2 to about 1 :10.

.5 Following are different embodiments within the context of the present invention wherein various weight ratios (% w/w) of the ingredients in the emulsion- preconcentrate compositions are provided: a) In the phospholipids, ratios of anionic phospholipid to neutral phospholipid ranging between about 1 :1 and about 1 :10, or about 1 :2 to

50 about 1 :5. b) In the oils, ratios of soybean oil to Miglyol 840 ranging between about 1 : 1 to about 1 : 10, or about 6:4 to about 2:8. c) In the co-solvents, ratios of propylene glycol to ethanol ranging between about 1 :100 to about 2:98, or about 5:95 to about 8:92. The emulsion-preconcentrate compositions comprising docetaxel and phospholipid of the present invention have viscosities in the range of about 1 cP to about 25 cP, or about 1.5 cP to about 20 cP, at ambient temperature.

The emulsion-preconcentrate compositions comprising docetaxel and phospholipid of the present invention have surface tensions in the range of about 5 dyne/cm² to about 50 dyne/cm², or about 10 dyne/cm² to about 40 dyne/cm², at ambient temperature.

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.

An aspect of the present invention provides use of a co-solvent or solubilizing agent in the compositions to solubilize other components of the system. 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. A metal ion chelator functions as an antioxidant by binding to metal ions and thereby reduces the catalytic effect of metal ions on oxidation reactions of the drug, oil and/or phospholipid components. Metal 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, 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, and salts thereof, sodium hydroxide, sodium carbonate, sodium bicarbonate, tris buffer, meglumine, amino acids and mixtures thereof. Such pH modifiers and stabilizers 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 suspending and/or viscosity modifying agents include polymers like polyvinylpyrrolidone, Hydroxypropylmethyl cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, 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.

In an aspect of the invention, the compositions are both chemically and physically stable. An emulsion-preconcentrate composition is "chemically stable" if the docetaxel in the composition is not substantially chemically degraded during storage under appropriate conditions. An emulsion composition is "physically stable" if it can be stored under appropriate conditions without significant increases in its average droplet size upon dilution, or evidence of phase separation, creaming, or droplet aggregation. In an aspect, the emulsion-preconcentrate 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%.

In an embodiment, a process for preparation of an emulsion- preconcentrate composition of the present invention comprises: a) dissolving oil, cosolvent, and phospholipid, optionally with other pharmaceutically acceptable excipients, by mixing and/or warming to about 40 ⁰C to 50 ⁰C under sonication and vortexing; b) dissolving docetaxel in the solution of step a) by mixing and/or warming to about 40 ⁰C to 50 ⁰C under sonication and vortexing; c) aseptically filtering the solution of step b); and d) filling the filtrate into a vial with inert gas purging and closing it. Although generally the sequence of mixing the ingredients is not very critical in preparation of solutions, typically emulsion-preconcentrate compositions are prepared by first mixing oil, cosolvent and phospholipid to form a solution and then dissolving or dispersing active ingredient.

The emulsion preconcentrate composition of the present invention may be packaged in various materials, including pre-filled syringes, glass containers, polymer coated glass containers and polymeric containers.

The emulsion-preconcentrate compositions of the present invention may be stored at about 2°C to 8°C, or at ambient conditions, prior to dilution.

In an embodiment, emulsion-preconcentrate compositions of the present invention may be diluted with aqueous fluids including water, various buffer solutions having different pH values, parenteral 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, and mixtures thereof, and dilution typically will produce docetaxel concentrations in the infusion fluid ranging between about 0.01 mg/ml and about 10 mg/ml, or between about 0.1 mg/ml and about 1 mg/ml. Such aqueous fluids can be provided separately, or can be included with a container containing an emulsion preconcentrate, in the form of a kit.

In an aspect, the present invention also relates to a kit for the delivery of docetaxel comprising: (a) a container having emulsion preconcentrate; and (b) a pharmaceutically acceptable aqueous diluent; providing, upon mixing (a) and (b), a docetaxel concentration about 0.01 mg/ml to about 10 mg/ml.

As used herein, the term "average droplet size" (or synonymously, "mean droplet size") refers to the distribution of emulsion droplets wherein about 50 volume percent of all the droplets measured have a size less than the defined average droplet size value and about 50 volume percent of all measurable droplets measured have a droplet size greater than the defined average droplet size value. This can be identified by the term "D₅₀." The droplet size can be typically measured using instruments like the Zetasizer^® 3000 HS, from Malvern^® Instruments Ltd., Malvern, Worcestershire, United Kingdom. In certain embodiments, the average droplet size of 'oil-in-water' type emulsions resulting from dilution of an emulsion-preconcentrate composition with an aqueous fluid is less than about 500 nm, or less than about 400 nm, or less than about 300 nm, or less than about 200 nm, and such emulsion may be called a "nanoemulsion" or "mini-emulsion" in the context of the present invention. Typically, an average droplet size of emulsions of the present invention ranges between about 50 nm and about 400 nm, or about 100 nm and about 300 nm. The droplet sizes generally will be greater than about 25 nm.

The net charge on the oil droplets generated after dilution of an emulsion- preconcentrate composition with 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 droplet. Zeta potential can be measured using instruments like the Zetasizer^® 3000 HS, from Malvern^® Instruments, Ltd.

In some embodiments, droplets of "oil-in-water" types of emulsions resulting from dilution of an emulsion-preconcentrate composition with an aqueous fluid have a negative charge. Typically the magnitude of net droplet charge (or zeta potential) ranges between about - 10 mV and about - 70 mV, or about - 20 mV and about - 60 mV, or about - 30 mV and about - 50 mV.

In some embodiments, the emulsions of the present inventions may be parenterally administered to a subject. "Parenteral" includes any mode of administration that does not go through the digestive tract, but excludes transmembrane delivery such as skin patches. Parenteral administration most commonly refers to injections or infusions into blood vessels. In certain embodiments, the mode of administration of the present emulsions is by intravenous, intra-arterial, intrathecal, intraperitoneal, intratumoral, intra-articular, intramuscular or subcutaneous injection or infusion, and the like.

In vivo performance of oil-in-water emulsions of the present invention can be evaluated by pharmacokinetic studies in animal models, which is one of the widely accepted tools. Typically, such pharmacokinetic studies involve administration of the composition to pre-conditioned subjects, and then monitoring the course of the active in the body with respect to time. Such monitoring comprises collection of samples of biological fluids such as blood, urine, sputum, and the like at periodic intervals post dosing, and determining the active ingredient content in the samples using suitable analytical techniques.

Determination of any active ingredient (analyte) content in biological fluids generally involves two steps: sample preparation and determination of analyte concentration in the sample using a suitable analytical technique. Sample preparation comprises collection of biological fluid, optional pretreatments such as freezing and thawing, and extraction of analyte in suitable solvent system for further estimation.

When required, a blood sample can be analyzed for analyte concentration using a suitable analytical technique such as liquid chromatography ("LC"), gas chromatography ("GC"), thin layer chromatography ("TLC"), supercritical fluid chromatography ("SFC") and the like. In many instances, a high performance liquid chromatography ("HPLC") technique is suitable in the context of the present invention. As is known to a person skilled in the art, various detector systems can be employed along with the above-mentioned analytical techniques. A few examples of suitable detectors include ultraviolet ("UV") detectors, fluorescence detectors, refractive index ("Rl") detectors, radiation detectors (for detecting radio-labeled compounds), and mass spectrometer ("MS") detectors. In an embodiment of the present invention, an emulsion-preconcentrate composition is suitably diluted using parenteral infusion fluid so as to get a desired drug concentration. Upon such dilution, and optionally with mild shaking, an oil-in- water emulsion is readily formed. Such formed emulsion is then administered to animal models via a parenteral route. In a particular embodiment, an emulsion-preconcentrate composition comprising docetaxel was diluted with 5 % dextrose infusion fluid to produce a final concentration of 2 mg/ml. This emulsion, when injected intravenously into rats at a dose of 20 mg/kg, resulted in improved pharmacokinetic parameters.

Various pharmacokinetic parameters that demonstrate bioavailability of an active ingredient include maximum concentration of drug in plasma, ("C₀" in case of intravenous administration and "C_max" for other routes of administration), and the area under the plasma concentration vs. time curve ("AUC"). Parameters such as volume of distribution (calculated by dividing the total amount of drug in the body by the drug blood concentration) provide useful insight about the distribution of active inside the body.

"Volume of distribution" also known as apparent volume of distribution, is a pharmacological term used to quantify the distribution of a medication throughout the body after oral or parenteral dosing. It is defined as the volume in which the amount of drug would need to be uniformly distributed in to produce the observed blood concentration. The units for volume of distribution are typically ml or liters per kilogram (kg) of body weight.

In an aspect, the present invention provides injectable oil-in-water emulsions comprising therapeutically effective amounts of docetaxel and at least one phospholipid, wherein said emulsion exhibits improved volume of distribution over that obtained with reconstituted presently marketed compositions, upon parenteral administration to a mammal.

The term "systemic exposure" is used synonymously with "area under the curve" in the context of the present invention. In the field of pharmacokinetics, the area under the curve ("AUC") is the area under the curve in a plot of concentration of drug in plasma against time, after administration.

In another aspect, the present invention provides injectable oil-in-water emulsions comprising therapeutically effective amounts of docetaxel and at least one phospholipid, wherein said emulsions exhibit improved systemic exposure over that obtained with reconstituted presently marketed compositions, upon parenteral administration to a mammal at the same doses.

Toxicity is a measure of the degree to which something is toxic or poisonous. In the context of the present invention, toxicity of the formulation is tested in terms of mortality of subject animals subsequent to parenteral administration of an emulsion composition, and determination of the number of animals that die during or after completion of a designed study, out of the total number of animals involved in such study. Toxicity can also be quantified by other parameters such as maximum tolerated dose ("MTD"), hematological counts, occurrence of side effects, and the like. A further aspect of the present invention provides injectable oil-in-water emulsions comprising therapeutically effective amounts of docetaxel and at least one phospholipid, wherein said emulsion exhibits reduced toxicity than is obtained with reconstituted presently marketed compositions, upon parenteral administration to a mammal at the same doses. In a still further aspect, emulsion-preconcentrate compositions of the present invention exhibit reduced mortality as compared to presently marketed compositions when they are, after dilution, parenterally administered to a mammal at the same doses. In certain embodiments, the emulsion preconcentrate compositions of the present invention exhibit improvement in MTD as compared to presently marketed compositions when they are, after dilution, parenterally administered to a mammal at the same doses. The emulsion-preconcentrate compositions according to the instant invention, and the oil-in-water emulsions obtained upon dilution of these emulsion- preconcentrate compositions, may be used for the treatment of various disease states like cancers, tumors, Kaposi's sarcoma, malignancies, uncontrolled tissue or cellular proliferation secondary to tissue injury, and any other disease conditions responsive to toxoids such as docetaxel, and/or prodrugs and derivatives thereof. Among the types of carcinoma, which may be treated particularly effectively with docetaxel, other taxoids, and their prodrugs and derivatives, 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.

The following examples will further illustrate certain specific aspects and embodiments of the invention, are being provided solely for purposes of illustration, and are not intended to limit the scope of the invention in any manner.

Table 1 : Abbreviations of terms used

EXAMPLES 1-3: Emulsion-preconcentrate compositions of docetaxel with varying phospholipids.

Manufacturing processes: Example 1 :

1. Vitamin E succinate and citric acid were dissolved in the mixture of ethanol and propylene glycol in a glass vial.

2. Lipids other than DC cholesterol or cholesterol were dissolved in the above mixture by warming at 40⁰C to 50 ⁰C and mixing under sonication till the solution was clear.

3. DC cholesterol or cholesterol was dissolved by vortex mixing, warming at 40⁰C to 50 ⁰C and by sonicating in the solution of step 2.

4. Docetaxel trihydrate was dissolved completely by sonication and vortex mixing.

5. The vial was purged with nitrogen gas and stoppered with an ETFE coated stopper, followed by sealing with a tamper-evident seal.

6. The composition of step 5 was stored at about 2 ⁰C to 8 ⁰C. Manufacturing process for Example 2 and Example 3 were similar to that described in Example 1 , with the required components.

Method of dilution to obtain emulsion:

Emulsion-preconcentrate of docetaxel was added to 5% dextrose solution such that a concentration range of docetaxel between 0.1 mg/ml and 0.8 mg/ml was obtained before parenteral administration.

Properties of emulsions were obtained after dilution (measured using a Zetasizer^® 3000 HS, Malvern Instruments).

EXAMPLE 4: Comparative tumor volume reduction data for Example 1 composition, TAXOTERE^®, adriamycin, and control in nude mice having MCF-7 tumors.

* Total dose of 43 mg/kg by i.v. administration in 3 doses Q4D (every four days) * 3.

^** Total dose of 50 mg/kg by i.v. administration in 3 doses Q4D * 3.

* Total dose of 15 mg/kg by i.v. administration in 3 doses Q4D * 3. ** Untreated group.

From the above data it is inferred that relative tumor volumes in the animals treated with present invention and TAXOTERE are smaller compared to adriamycin treated subjects and untreated subjects.

EXAMPLES 5-6: Emulsion-preconcentrate compositions of docetaxel.

Manufacturing process:

1. Ethanol and propylene glycol were mixed and vitamin-E-succinate and citric acid were dissolved in it.

2. To solution of step 1 , soybean oil and phospholipids were added and mixed.

3. Mixture of step 2 was warmed to 40-50 ⁰C to dissolve its content with stirring.

4. Solution of step 3 was cooled and docetaxel was dissolved in it by mixing.

Properties of emulsions were obtained after dilution (measured using a

Zetasizer^® 3000 HS, Malvern Instruments).

EXAMPLES 7-10: Emulsion-preconcentrate compositions of docetaxel with different phospholipids.

Manufacturing process for Examples 7-10:

1. Ethanol and propylene glycol were mixed with Miglyol and soybean oil (as present in a composition).

2. Various phospholipids (as present) were dissolved in the solution of step 1.

3. All other the ingredients except docetaxel were mixed and warmed to 40-50 0C to dissolve its content with stirring.

4. Docetaxel was added to the mixture of step 3, and dissolved by sonication and vortex mixing.

5. The solution of step 4 was filled into stoppered vials with nitrogen gas purging followed by sealing.

The emulsion preconcentrate compositions were diluted with 5 % dextrose solution in a stepwise manner. First a stock dispersion of 1 mg/ml strength was prepared, which was subsequently diluted 100 times with 5 % dextrose solution to get a concentration of 10 μg/ml. The diluted sample (oil-in-water emulsion) was used for determination of droplet size and its charge in terms of zeta potential. Properties of emulsions were obtained after dilution of emulsion preconcentrate compositions (measured using a Zetasizer^® 3000 HS, Malvern Instruments).

EXAMPLES 11-14: Emulsion-preconcentrate compositions of docetaxel and their pharmacokinetic parameters in rats.

Manufacturing process was similar to that described for Example 7. Properties of emulsions were obtained after dilution of emulsion preconcentrate compositions with aqueous fluid similar to that described for Example 7 (measured using a Zetasizer^® 3000 HS, Malvern Instruments).

Comparative pharmacokinetic evaluation data for Examples 11 through 14 and TAXOTERE®, in rats (dose single i.v. bolus, 20 mg/kg, n = 4). Study Protocol:

Animal and gender: rats; male. Animal weight range: 24 g to 34 g. Pre-experimental condition of animals: fed state.

Formulation strength: 2 mg/ml. Dose and Schedule: 20 mg per kg, single dosing. Route of administration: intravenous.

The determination of docetaxel content in plasma after intravenous administration of the emulsion compositions produced after suitable dilution of Examples 11-14 emulsion-preconcentrate compositions was carried out using a HPLC analytical technique with following set of conditions: Column: INERTSIL® ODS (4.6x100 mm; 3 μm). Mobile phase: Ammonium acetate (pH 6.0):acetonitrile (15:85, v/v). Internal standard: Paclitaxel (10 μl_ of 5 μg/ml solution).

Detector: Mass spectrophotometer ("MS"). Injection volume: 30 μL Flow rate: 0.6 mL/minute. Retention time: 2.40 minutes (approximately). Linearity range: 0.005 to 10 μg/mL. Data Treatment: The pharmacokinetic data were processed using WINNONLIN® statistical software. Further, the volume of distribution was calculated using Sigma Plot® analysis.

Parameter Example 11 Example 12 Example 13 Example 14 TAXOTERE

C₀ (μg/ml) 18.53 ± 3 .57 9.55 ± 1. 94 9.37 ± 1. 27 5.13 ± 1. 04 6 03 ± 0. 93

AUC (o-t)

4.70 ± 0. 81 5.22 ± 1. 02 5.59 ± 0. 72 4.61 ± 0. 66 3 10 ± 0. 69 (μg hour/ml)

Volume of distribution 62.73±40 .24 NA^* NA* NA^* 24 .35 ± 5 .92 (Vz) (UWg)

^*Not analyzed.

EXAMPLE 15: Toxicity study for emulsion-preconcentrate compositions of docetaxel.

Formulations tested:

Emulsion-preconcentrate compositions comprising docetaxel of Example 1 1 , Example 12 and Example 14 vis-a-vis TAXOTERE^® with Dextrose saline solution as a control and one placebo formulation according to Example 1 1 , but omitting docetaxel. Administration:

Intra-venous administration of 3 doses: 20 mg/kg (Dose 1 ); 40 mg/kg (Dose 2); and 80 mg/kg (Dose 3, split into 2 equal portions and spaced two hours apart). Volume of injection: 20 ml/kg. Rate of injection: 0.4 ml/minute. Schedule: day 0, day 4 and day 8 (Q4D^χ3). Study Duration: 13 days. Animal species: Athymic mice (males).

Grouping: 6 mice for TAXOTERE * 3 doses = 18 mice. 6 mice for Example 1 1 * 3 doses = 18 mice. 6 mice for Example 12 * 3 doses = 18 mice. 6 mice for Example 14 x 3 doses = 18 mice. 6 mice for 5% dextrose saline solution = 6 mice.

6 mice for placebo (for Example 1 1 ) = 6 mice.

mg per kg.

From these observations, it is apparent that the emulsion-preconcentrate compositions of Example 11 and Example 12 exhibit reduced toxicity in terms of mortality at 20 mpk and 40 mpk dose when compared against TAXOTERE.

Hematological toxicity study was conducted with nude mice administered TAXOTERE and the compositions of Example 11 , 12 and 14, and blood samples were taken for analysis on the 13^th day.

^* mg per kg.

^** White blood cell. No significant difference in hematological toxicity was observed for the tested parameters between the compositions of Example 11 , 12, 14, and TAXOTERE at the dose of 20 mg per kg.

Example 11 composition and TAXOTERE at the dose of 40 mg per kg did not show significant differences in the tested parameters for hematological toxicity.

EXAMPLE 16: Emulsion-preconcentrate composition comprising docetaxel and exhibiting stability on storage.

Manufacturing process:

1. All the ingredients except docetaxel were mixed and warmed to 40-50 ⁰C to dissolve with stirring.

2. Docetaxel was added to the mixture of step 1 , and dissolved by sonication and vortex mixing for about 10 minutes. 3. The solution of step 2 was aseptically filtered and filled into stoppered vials

(equivalent to about 40 mg docetaxel per vial) with nitrogen gas purging followed by sealing. 4. The product of step 3 was stored between 2 ⁰C and 8 ⁰C.

Properties of emulsions were obtained after dilution of emulsion preconcentrate compositions with aqueous fluid similarly to the procedure described for Example 7 (measured using a Zetasizer^® 3000 HS, Malvern^ Instruments).

Stability study of the composition of Example 16.

Storage conditions: 2 ⁰C - 8 ⁰C, 25 °C/60 % relative humidity (RH), and 40 °C/75 % RH.

Sampling points: initial, 1 month.

Stability packaging: USP type I glass vials with ETFE coated rubber stoppers. HPLC analytical method for stability study:

Mobile phase: Acetate buffer (0.02M) pH 4.5:acetonitrile (60:40 v/v).

Buffer: Acetate buffer (0.02M) pH 4.5.

Column: LICHROSPHERE® RP 18e, 250><4.6mm, 5 μm.

Flow rate: 1.5 ml / minute.

Wave length: 230 nm.

Temperature: 25°C.

Load: 20 μl.

Run time: 30 minutes (For diluted standard preparation).

Run time: 80 minutes (For blank, placebo, and sample preparation).

* Stability of emulsion produced after dilution with 5 % Dextrose solution a determined by visual observation. Emulsion with no separation or sedimentation was considered to be physically stable. Manufacturing processes for Examples 17-46 were similar to that described in Example 16.

EXAMPLE 17: Emulsion-preconcentrate composition comprising docetaxel, which on dilution with aqueous fluid yields anionic emulsion.

Properties of emulsion obtained after dilution of emulsion preconcentrate compositions with aqueous fluid similar to that described for Example 7 (measured using a Zetasizer^® 3000 HS, Malvern Instruments).

Stability study of composition of Example 17.

Storage condition: 2 ⁰C - 8 ⁰C.

Sampling points: initial, after 30 days and after 75 days.

Stability packaging: USP Type 1 glass vials with ETFE coated stoppers and a tamper-evident seal.

* Stability of emulsion produced after dilution with 5 % dextrose solution as determined by visual observation. Emulsion with no separation or sedimentation was considered to be physically stable.

EXAMPLES 18-21 : Emulsion-preconcentrate compositions comprising docetaxel and varying amounts of EPG, DOPG, and LOPC, and their properties.

From these observations, it can be concluded that with increasing EPG and DOPG concentrations, the mean droplet size remains fairly constant. However, during storage of the Example 21 composition at 2-8 °C, a precipitation potential was observed.

EXAMPLES 22-24: Emulsion-preconcentrate compositions comprising docetaxel and varying amounts of DMPC, DOPC and LOPC, and their properties.

From these observations, it can be concluded that when DMPC, DOPC and LOPC content is varied, the mean droplet size remains fairly constant. EXAMPLES 25-28: Emulsion-preconcentrate compositions comprising docetaxel and varying amounts of EPG and LOPC, and their properties.

From these observations, it can be concluded that when EPG and LOPC content is varied, the mean droplet size remains fairly constant. Absence of DOPG in these compositions does not significantly affect the mean droplet sizes.

EXAMPLES 29-32: Emulsion-preconcentrate compositions comprising docetaxel and varying amounts of DMPC, DOPC and LOPC, and their properties.

From these observations, it can be concluded that in compositions devoid of DOPG, increasing DMPC and DOPC, and correspondingly decreasing LOPC, causes variations in the mean droplet sizes.

EXAMPLES 33-36: Emulsion-preconcentrate compositions comprising docetaxel and varying amounts of DMPC, DOPC and LOPC, and their properties.

From these observations, it can be concluded that in the presence of DOPG, compositions with varying DMPC and DOPC contents yield satisfactory mean droplet sizes. EXAMPLES 37-42: Emulsion-preconcentrate compositions comprising docetaxel and varying amounts of soybean oil and Miglyol, and their properties.

From these observations, it can be concluded that varying ratios of soybean oil and Miglyol yields compositions, which upon dilution produce oil-in- water emu ^■ls- ^"ions w -iⁱt^lh mean droplet ^*. sizes about 200 nm.

EXAMPLES 43-46: Emulsion-preconcentrate compositions comprising docetaxel and varying amounts of propylene glycol and ethanol, and their properties.

From these observations, it can be concluded that varying amounts of ethanol and propylene glycol yields compositions, which upon dilution produce oil- in-water emulsions with mean droplet sizes less than 200 nm.

Claims

CLAIMS:

1. An emulsion preconcentrate comprising docetaxel, an oil, a phospholipid or a mixture of phospholipids, and optionally a co-solvent, wherein at least one phospholipid is an anionic phospholipid imparting a net negative charge about -10 mV to about -70 mV to emulsion droplets formed upon dilution of the preconcentrate with an aqueous fluid.

2. The emulsion preconcentrate of claim 1 , wherein a net negative charge about -20 mV to about -60 mV is imparted to emulsion droplets.

3. The emulsion preconcentrate of claim 1 , wherein a net negative charge about -30 mV to about -50 mV is imparted to emulsion droplets.

4. The emulsion preconcentrate of claim 1 , wherein droplets formed upon dilution have mean sizes less than about 500 nm.

5. The emulsion preconcentrate of claim 1 , wherein droplets formed upon dilution have mean sizes less than about 400 nm.

6. The emulsion preconcentrate of claim 1 , wherein droplets formed upon dilution have mean sizes less than about 300 nm.

7. The emulsion preconcentrate of claim 1 , wherein droplets formed upon dilution have mean sizes less than about 200 nm.

8. The emulsion preconcentrate of claim 1 , wherein an anionic phospholipid comprises: egg phosphatidyl glycerol; 1 ,2-dioleoyl-sn-glycero-3-[phospho-rac-(1- glycerol)]; 1 ^-dipalmitoyl-sn-glycero-S-phosphoglycerol; 1 ,2-dimyristoyl-sn- glycero-3-phosphoglycerol; 1 ,2-dioleoyl-sn-glycero-3-(phospho-L-sehne); 1 ,2- dipalmitoyl-sn-glycero-3-phospho-L-sehne; 1 ,2-dimyristoyl-sn-glycero-3-(phospho- L-serine); 1 ,2-dioleoyl-sn-glycero-3-phosphate, monosodium salt; 1 ,2-dipalmitoyl- sn-glycero-3-phosphate, monosodium salt; or 1 ,2-dimyristoyl-sn-glycero-3- phosphate, monosodium salt; and any combinations of two or more.

9. The emulsion preconcentrate of claim 1 , wherein an anionic phospholipid content is about 10 % w/w to about 40 % w/w.

10. The emulsion preconcentrate of claim 1 , wherein an anionic phospholipid content is about 15 % w/w to about 30 % w/w.

11. The emulsion preconcentrate of claim 1 , wherein a mixture of phospholipids comprises an anionic phospholipid and a neutral phospholipid.

12. The emulsion preconcentrate of claim 11 , wherein a weight ratio of anionic phospholipid to neutral phospholipid is between about 2:1 and about 1 :9.

13. The emulsion preconcentrate of claim 11 , wherein a weight ratio of anionic phospholipid to neutral phospholipid is between about 1 :2 and about 1 :7.

14. The emulsion preconcentrate of claim 1 , having a docetaxel content about

1 % w/w to about 6 % w/w.

15. The emulsion preconcentrate of claim 1 , having a docetaxel content about

2 % w/w to about 4 % w/w.

16. The emulsion preconcentrate of claim 1 , wherein an oil content is about 4 % w/w to about 10 % w/w.

17. The emulsion preconcentrate of claim 1 , wherein an oil content is about 5 % w/w to about 8 % w/w.

18. The emulsion preconcentrate of claim 1 , wherein a weight ratio of docetaxel to phospholipid is about 1 :1 to about 1 :50.

19. The emulsion preconcentrate of claim 1 , wherein a weight ratio of docetaxel to phospholipid is about 1 :5 to about 1 :30.

20. The emulsion preconcentrate of claim 1 , wherein a weight ratio of oil to phospholipid is about 1 :1 to about 1 :20.

21. The emulsion preconcentrate of claim 1 , wherein a weight ratio of oil to phospholipid is about 1 :2 to about 1 :15.

22. An oil-in-water emulsion produced upon dilution of the emulsion preconcentrate of claim 1 with an aqueous fluid, the emulsion containing about 0.01 mg/ml to about 10 mg/ml docetaxel and having mean droplet sizes less than about 500 nm.

23. The oil-in-water emulsion of claim 22, wherein a net charge on emulsion droplets is about -20 mV to about -60 mV.

24. The oil-in-water emulsion of claim 22, wherein a net charge on emulsion droplets is about -20 mV to about -60 mV.

25. The oil-in-water emulsion of claim 22, having mean droplet sizes less than about 400 nm.

26. The oil-in-water emulsion of claim 22, having mean droplet sizes less than about 300 nm.

27. The oil-in-water emulsion of claim 22, having mean droplet sizes less than about 200 nm.

28. A method of treating neoplasm conditions, comprising administering a therapeutically effective amount of an oil-in-water emulsion of claim 22.

29. A kit for the delivery of docetaxel comprising: (a) a container having the emulsion preconcentrate of claim 1 ; and (b) a container having a pharmaceutically acceptable aqueous diluent; and providing, upon mixing the contents of (a) and (b), an emulsion having docetaxel concentrations about 0.01 mg/ml to about 10 mg/ml and mean droplet sizes less than about 500 nm.