Pharmaceutical compositions of testosterone

Abstract

The present invention provides stable pharmaceutical compositions, comprising a pharmaceutically effective amount of testosterone or a pharmaceutically acceptable ester thereof, a pharmaceutically acceptable oil vehicle, and a stabilizing amount of benzyl alcohol, for example, about 1% to 3% weight/volume of benzyl alcohol. The present invention also provides a process for stabilizing testosterone-containing pharmaceutical compositions by ageing them at a temperature of about 20° C. to about 60° C. for at least 48 hours, e.g., prior to secondary packing and labeling. These compositions were stable over the shelf life of the product, without exhibiting crystal formation, even upon storing at temperatures of about 2° C. to about 8° C. Other aspects of the invention relate to methods for making such pharmaceutical compositions, as well as methods of using such pharmaceutical compositions for hormone replacement therapy, e.g., in a male patient having a condition associated with symptoms of deficiency or absence of endogenous testosterone.

Claims

1. An injectable pharmaceutical composition consisting of: about 200 mg/mL testosterone cypionate; (ii) about 15 mg/mL to 30 mg/mL benzyl alcohol; (iii) benzyl benzoate; and (iv) cotton seed oil; wherein the composition is sterilized and suitable for intramuscular injection; and wherein the composition does not exhibit crystal formation for 6 months when stored at a temperature of about 10° C. to about 15° C.

2. A pre-filled syringe consisting of a liquid composition of testosterone cypionate: wherein the composition consists of: about 200 mg/mL testosterone cypionate; (ii) about 15 mg/mL to 30 mg/mL benzyl alcohol; (iii) benzyl benzoate; and (iv) cotton seed oil; wherein the composition is sterilized and suitable for intramuscular injection; and wherein the composition does not exhibit crystal formation for 6 months when stored at a temperature of about 10° C. to about 15° C.

3. An injectable pharmaceutical composition consisting of: about 200 mg/mL testosterone cypionate; (ii) about 15 mg/mL to 30 mg/mL benzyl alcohol; (iii) benzyl benzoate; and (iv) cotton seed oil; wherein the composition is sterilized and suitable for intramuscular injection; and wherein the composition has been aged in containers for at least 48 hours at a temperature of about 20° C. to about 60° C. prior to storage, and wherein the composition does not exhibit crystal formation for 6 months when stored at a temperature of about 2° C. to about 8° C.

4. The injectable pharmaceutical composition according to claim 1, comprising consisting of about 15 mg/mL benzyl alcohol.

5. The injectable pharmaceutical composition according to claim 1, comprising consisting of about 20 mg/mL benzyl alcohol.

6. The injectable pharmaceutical composition according to claim 1, comprising consisting of about 25 mg/mL benzyl alcohol.

7. The injectable pharmaceutical composition according to claim 1, comprising consisting of 30 mg/mL benzyl alcohol.

8. The composition according to claim 2, consisting of about 15 mg/mL benzyl alcohol.

9. The composition according to claim 2, consisting of about 20 mg/mL benzyl alcohol.

10. The composition according to claim 2, consisting of about 25 mg/mL benzyl alcohol.

11. The composition according to claim 2, consisting of 30 mg/mL benzyl alcohol.

12. The composition according to claim 3, consisting of about 15 mg/mL benzyl alcohol.

13. The composition according to claim 3, consisting of about 20 mg/mL benzyl alcohol.

14. The composition according to claim 3, consisting of about 25 mg/mL benzyl alcohol.

15. The composition according to claim 3, consisting of 30 mg/mL benzyl alcohol.

Description

DETAILED DESCRIPTION OF THE INVENTION

(1) As used herein the term “testosterone” refers to the free base or a pharmaceutically acceptable ester, preferably testosterone cypionate (i.e., Testosterone 17β-cyclopentylpropionate). The term “pharmaceutically acceptable ester” may refer to any suitable ester, preferably an ester selected from the group of linear, branched and cyclic C.sub.3-C.sub.16 alkanoates, most preferably testosterone cypionate. By “linear and branched C.sub.3-C.sub.16 alkanoates” is meant aliphatic esters with chain lengths from 3 to 16 carbon atoms. In other aspects, the ester may be selected from the group consisting of acetate, caproate, cyclohexylpropionate, decanoate, enantate benzilic acid hydrazine, furoate, hexahydrobenzoate, hexahydrobenzylcarbonate, hexyloxyphenylpropionate, isobutyrate, isocaproate, ketolaurate, nicotinate, phenylacetate, phenylpropionate, phosphate, valerate, buciclate (20 Aet-1, CDB-1781), polyphloretin phosphate, 17β-(1-((5-(aminosulfonyl)-2-pyridinyl)carbonyl)-L-proline) (EC586), acetate butyrate, acetate propionate, benzoate, butyrate, diacetate, dipropionate, formate, isovalerate, palmitate, phenylbutyrate, stearate and sulfate. In certain preferred embodiments, the ester group is selected from the group consisting of cypionate, enantate, propionate and undecanoate. In certain aspects, the ester group is at the 17β-position of the testosterone molecule.

(2) The term “pharmaceutically acceptable excipient” as used herein means a diluent, carrier, or composition auxiliary, which is non-toxic and inert, which does not have undesirable effects on a subject to whom it is administered and is suitable for delivering a therapeutically active agent to the target site without affecting the therapeutic activity of the said active agent. In certain embodiments, the pharmaceutical compositions may optionally contain pharmaceutically acceptable excipients, including antioxidants, buffers, preservatives, or stabilizing agents.

(3) For purposes of the present invention, a pharmaceutically acceptable solvent is suitable for pharmaceutical use. In several embodiments of the invention, the pharmaceutically acceptable solvent is selected from group consisting of cotton seed oil, castor oil, tea seed oil, sesame oil, linseed oil, peanut oil, olive oil and wheat-germ oil. Within this aspect, the pharmaceutically acceptable solvent is preferably cotton seed oil.

(4) In one embodiment, the present invention relates to stabilized liquid pharmaceutical compositions for intramuscular injection comprising testosterone, a pharmaceutical acceptable solvent, and a solubility enhancer.

(5) In another embodiment, the present invention relates to stabilized liquid pharmaceutical compositions for intramuscular injection comprising (i) testosterone, (ii) a pharmaceutical acceptable solvent selected from group consisting of cotton seed oil, castor oil, tea seed oil, sesame oil, linseed oil, peanut oil, olive oil, wheat-germ oil and mixtures thereof, (iii) a solubility enhancer consisting of benzyl benzoate and (iv) benzyl alcohol.

(6) In an aspect of the invention, the present invention relates to stabilized liquid pharmaceutical compositions for intramuscular injection comprising (i) testosterone, (ii) a pharmaceutical acceptable solvent, preferably cotton seed oil, (iii) a solubility enhancer consisting of benzyl benzoate and (iv) benzyl alcohol.

(7) In an aspect of the invention, the present invention relates to stabilized liquid pharmaceutical compositions for intramuscular injection comprising (i) testosterone, (ii) a pharmaceutical acceptable solvent, preferably cotton seed oil, (iii) a solubility enhancer consisting of benzyl benzoate and (iv) benzyl alcohol, wherein benzyl alcohol is preferably in an amount of about 20 mg/ml to about 25 mg/ml.

(8) In an aspect of the invention, the present invention relates to stabilized liquid pharmaceutical compositions for intramuscular injection comprising (i) testosterone, (ii) a pharmaceutical acceptable solvent, preferably cotton seed oil, (iii) a solubility enhancer consisting of benzyl benzoate and (iv) benzyl alcohol, wherein the benzyl alcohol is in an amount of about 20 mg/ml to about 25 mg/ml and the compositions have been aged for at least about 48 hours at a temperature from about 20° C. to about 60° C.

(9) In certain embodiments, the present application relates to reducing or preventing formation of crystals in the pharmaceutical composition by using an effective amount of benzyl alcohol, and/or ageing the pharmaceutical composition prior to loading. One or both of these techniques may be used, and preferably, both techniques are used together.

(10) As used herein, “ageing” or “holding” compositions refers to a process of storing compositions for at least 48 hours within a controlled temperature range, e.g., a temperature from about 20° C. to about 60° C. The temperature may be kept at a constant single temperature, or may vary slightly within this range. In an aspect of the invention, the present invention relates to aging compositions at a controlled temperature for at least about 48 hours, which will prevent or reduce crystal formation in the said compositions. In some embodiments, the pharmaceutical compositions are aged for an effective length of time, such as between 7-21 days, e.g., 7 days, 15 days, or 3 weeks. In a preferred embodiment, the pharmaceutical compositions are aged for at least 48 hours. The stabilized pharmaceutical compositions can then subsequently be further processed into the finished dosage form, or stored in vials, at suitable temperatures, e.g., 2-8° C. or at 10-15° C. By performing the initial ageing step, it is possible to prevent or reduce the problem of crystal formulation upon storage, e.g., even upon storing at a temperature of about 2° C. to about 8° C.

(11) In another aspect of the invention, the present invention relates to compositions containing stabilizing amount of benzyl alcohol. In certain embodiments, an effective amount of benzyl alcohol is a concentration greater than about 10 mg/mL, and preferably a concentration from about 20 mg/mL to about 25 mg/mL. In certain embodiments, an effective amount of benzyl alcohol is about 15 mg/mL, about 20 mg/mL, about 25 mg/mL or about 30 mg/mL. In certain other embodiments, an effective amount of benzyl alcohol is a concentration greater than about 1% w/v, and preferably a concentration from about 1% w/v to about 3% w/v. When an effective amount of benzyl alcohol is used in the formulation, surprisingly, there is no crystal formation upon storage of the formulation, e.g., such as at room temperature for at least 48 hours or longer durations. By using an effective amount of benzyl alcohol for stabilization according to the invention, the problem with crystal formation can be minimized or avoided.

(12) The pharmaceutical compositions may also comprise one or more pharmaceutically acceptable excipients. For example, the compositions may further comprise a preservative, an antioxidant, and/or a stabilizer. For example, the pharmaceutical compositions of the present invention may contain one or more anti-oxidants, preservatives, complexing agents and chelating agents such as, but are not limited to butylated hydroxyanisole (BHA), butylated hydroxyl toluene (BHT), citric acid, lactic acid, benzoic acid, tocopherol (Vitamin E), monothioglycerol, ascorbic acid, methyl paraben, benzyl alcohol, propyl gallate, surfactants, lipids, thioglycolic acid, niacinamide, nicotinic acid, creatine, cyclodextrins; ethylene diamine tetra acetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), triethanolamine, 8-hydroxyquinoline, tartaric acid, phosphoric acid, gluconic acid, thiodipropionic acid, acetonic dicarboxylic acid, lecithin, di(hydroxyethyl)glycine, sorbitol, methionine, glycerol, propylene glycol, or phenol.

(13) The present invention provides for a composition that may optionally comprise one or more preservatives. The term “preservative” refers to a substance present in a composition that can, at least in part, prevent and/or reduce decomposition of the composition. In some embodiments, the preservative may prevent and/or reduce decomposition of the composition by microbial growth in the composition. Preferably, the preservative is pharmaceutically acceptable.

(14) For purposes of the present invention, preservatives for use in pharmaceutical compositions are known to those skilled in the art and include, but are not limited to, phenol, m-cresol, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, 2-phenoxyethanol, butyl p-hydroxybenzoate, 2-phenylethanol, benzyl alcohol, chlorobutanol, and thiomerosal, or mixtures thereof.

(15) In some embodiments, the preservative may be present in the composition at a concentration of in the range of about 1 to 10 mg/mL, preferably in the range of about 3 and 7 mg/mL, more preferably in the range of about 4 and 5 mg/mL, more preferably at about 4.5 mg/mL.

(16) A pharmaceutically inert gas (e.g., which may be selected from nitrogen or carbon dioxide) may be bubbled into the solution to drive out oxygen. In certain aspects, rubber stoppers may be used for sealing the vials. Certain embodiments additionally relate to sterilizing the finished products, e.g., filtration through a bacterial-retaining filter, aseptic filling, terminal sterilization, incorporation of sterilizing agents, irradiation, and/or heating.

(17) The finished dosage form may be provided in a sealed container (e.g., vials, pre-filled syringes, infusion bags, bottles, ampoules, etc.). In some embodiments, the container may be a single-dose formulation, or a multi-dose formulation. For example, a pre-filled syringe according to the invention may include single use auto injectors, or reusable auto injectors. In some embodiments, the same container may be used for multiple applications of the composition for up to about 10 days after initial use, preferably up to about 15 days, more preferably up to about 30 days, more preferably up to about 45 days, and most preferably up to about 60 days.

(18) The pharmaceutical compositions incorporate a therapeutically effective amount of testosterone or a pharmaceutically acceptable ester thereof. In certain aspects of the invention, the testosterone concentration in the inventive compositions is from about 100 mg/mL to about 200 mg/mL, preferably about 100 mg/mL or about 200 mg/mL.

(19) The present application also relates to certain methods of treatment, comprising administering any of the pharmaceutical compositions described herein to a patient in need thereof, for replacement therapy in a patient having a condition associated with symptoms of deficiency or absence of endogenous testosterone. Examples of diseases and symptoms associated with deficient endogenous levels of testosterone in a man may include primary hypogonadism (congenital or acquired), e.g., testicular failure due to cryptorchidism, bilateral torsion, orchitis, vanishing testis syndrome or orchidectomy, or hypogonadotropic hypogonadism (congenital or acquired), e.g., gonadotropin or LHRH deficiency, or pituitary-hypothalamic injury from tumors, trauma, or radiation. The compositions of the present application can be administered by suitable techniques, such as intramuscular injection deep in the gluteal muscle.

(20) The dosage may be adjusted according to the patient's response and the appearance of adverse reactions. Various dosage regimens have been used to induce pubertal changes in hypogonadal males, e.g., by using lower dosages initially, gradually increasing the dose as puberty progresses, with or without a decrease to maintenance levels, or alternatively by using higher dosages to induce pubertal changes and lower dosages for maintenance after puberty. The chronological and skeletal ages must be taken into consideration, both in determining the initial dose and in adjusting the dose. For example, for replacement in the hypogonadal male, 50-400 mg may be administered every two to four weeks.

(21) Suitable dosage amounts, dosage regimes, administration, and indications can be determined by the skilled artisan, based on the age, sex, and diagnosis of the individual patient, and are also known in the art, e.g., U.S. Pat. No. 7,718,640, which is hereby incorporated in its entirety.

EXAMPLES

(22) The following examples are exemplary and not intended to be limiting. The above disclosure provides many different embodiments for implementing the features of the invention, and the following examples describe certain embodiments. Other modifications and methods known to one of ordinary skill in the art can also be applied to the following experimental procedures, without departing from the scope of the invention.

General HPLC Procedure

(23) Analysis of samples for accelerated and long-term stability studies can be performed using high-performance liquid chromatography (HPLC). For example, the following HPLC procedure can be used to detect and quantify impurities of testosterone, as well as assay calculation. The materials and general conditions are listed below:

(24) TABLE-US-00001 TABLE 1A Chromatographic conditions for assay of testosterone cypionate Chromatographic mode Gradient flow Column Zorbax SB C8, 50 × 4.6 mm, 3.5 μm (part no: 835975-906) Wavelength 254 nm Flow rate 1.5 mL/min Injection volume 20 μL Column oven temperature 40° C. Sample cooler temperature  5° C. Run time About 35 minutes Mobile Phase A 0.77 g of ammonium acetate dissolved in 1000 mL of water and filtered through 0.45 μm PVDF membrane filter Mobile Phase B Acetonitrile

(25) TABLE-US-00002 TABLE 1B Gradient Program Time in % Mobile % Mobile minutes phase A phase B 0 80 20 3 80 20 30 10 90 31 80 20 35 80 20

(26) TABLE-US-00003 TABLE 2A Chromatographic conditions for determining testosterone cypionate related substances Chromatographic mode Gradient flow Column Xterra RP18, 150 × 4.6 mm, 3.5 μm Wavelength 254 nm Flow rate 1.0 mL/minutes Injection volume 10 μL Column oven temperature 40° C. Sample cooler temperature  5° C. Run time 60 minutes Mobile Phase A Add 0.5 mL of trifluoro acetic acid into 1000 mL of water Mobile Phase B Acetonitrile

(27) TABLE-US-00004 TABLE 2B Gradient Program Time in % Mobile % Mobile minutes phase A phase B 0 75 25 3 75 25 35 25 75 54 25 75 55 75 25 60 75 25

Example 1

(28) TABLE-US-00005 TABLE 3 Preparation of Testosterone Cypionate composition, 200 mg/mL Ingredients Qty/mL % (w/v) Qty/30 L batch Testosterone cypionate, USP   200 mg  20.%    6 kg Benzyl benzoate   224 mg 22.4%  6.72 kg Cotton seed oil 542.2 mg 54.2% 16.27 kg Benzyl alcohol   20 mg   2%  0.6 kg Total 986.2 mg 98.6% 29.59 kg

(29) Cotton seed oil and benzyl benzoate were added to the vessel and the mixture was heated to a temperature of about 40° C. to 60° C. Testosterone cypionate was added slowly, in lots, under stirring, for about 90 minutes, maintaining the solution temperature between about 40° C. to 60° C. The mixture was stirred for 60 minutes to ensure complete dissolution of testosterone cypionate. Benzyl alcohol was added slowly to the resultant solution at 40° C.-60° C., and continuously stirred for 30 minutes.

(30) The above-obtained solution was cooled gradually to 23° C. over 3 hours. It was then filtered through a 0.22-micron PVDF filter into a buffer tank. The final solution was filled into USP Type I clear 2 mL glass vial or prefilled syringe with a target 1 mL fill volume. These vials and prefilled syringes were aged at a temperature of about 20° C. to about 30° C. for about 2-weeks, providing stabilized solutions. After aging, the stabilized solutions were then further processed into the final dosage form, e.g., proceeded with packaging and labelling the stabilized solutions.

(31) Some of the above obtained vials and prefilled syringes were stored at a temperature of about 2° C. to about 8° C., and observed every two weeks for crystal formation.

(32) TABLE-US-00006 TABLE 4 Stability observation for crystal formation Time (weeks) 1 mL vial 1 mL prefilled syringe Initial No crystals No crystals 4 No crystals No crystals 12 No crystals No crystals 26 No crystals No crystals

(33) Additional lots of the vials and prefilled syringes from Example 1 were also stored under accelerated (40° C.±2° C./75% RH±5% RH) and long-term (25° C.±2° C./60% RH±5% RH) stability conditions as per ICH stability requirements.

(34) TABLE-US-00007 TABLE 5 Stability observations under accelerated conditions 1 mL vial 1mL prefilled syringe 3 6 3 6 Test Initial Months Months Initial Months Months % assay of 100.4% 99.8%  99.3%  97.9%  100%   101.7%  Testosterone cypionate Total  0.2%  0.23%  0.31%  0.21%  0.22%  0.25% impurities Viscosity  50.5    48.7     50.9     50.3      50.3      50.9     (in cps)

(35) TABLE-US-00008 TABLE 6 Stability observations under long term storage conditions 1 mL vial 1 mL prefilled syringe Test Initial 6 months Initial 6 months % assay of 97.9% 100.1% 100.4% 98.4% Testosterone cypionate Total impurities 0.21%  0.28%  0.21%  0.31% Viscosity (in cps) 50.5 49.8 50.3 53.7

Example 2

(36) TABLE-US-00009 TABLE 7 Preparation of Testosterone Cypionate composition with varying amounts of benzyl alcohol Composition A Composition B Composition C Ingredients Quantity in (mg/mL) Testosterone cypionate 200 200 200 Benzyl benzoate 224 224 224 Cotton seed oil 556 542.20 535.60 Benzyl alcohol 9.45 20 25 Total 989.45 986.2 984.6

(37) Manufacturing Procedure 1. The quantify of Benzyl benzoate shown in the above table was added to cotton seed oil in a glass beaker and stirred for 10 minutes while heating at 40° C.-45° C. to form a solution. 2. Testosterone cypionate was added to the above solution and stirred until the solution was clear. To this solution, benzyl alcohol was added, and stirred continuously for 10 minutes. 3. The solution volume was made up to 100 mL with cotton seed oil which was stirred for 60 minutes. 4. The bulk solution was filled into 10 mL tubular glass vials.

Example 3

(38) Samples from Compositions A, B and C were tested under various conditions (e.g., either stored immediately, or aged at a certain temperature prior to storage), and then observed for crystal formation over time. These results are summarized in Tables 8-14 below.

(39) TABLE-US-00010 TABLE 8 Observation for crystals in the composition A, B and C, when samples stored immediately at 10° C.-15° C. Day Composition A Composition B Composition C 0 (Initial) Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 1 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 2 Needle like Clear pale-yellow Clear pale-yellow crystals were solution solution observed 3 Vials withdrawn Clear pale-yellow Clear pale-yellow (Study solution solution 50 discontinued) Clear pale-yellow Clear pale-yellow solution solution 120 Clear pale-yellow Clear pale-yellow solution solution

(40) Samples from Composition A, B and C were loaded or stored immediately at a temperature range of 10° C.-15° C. after preparation (e.g., without being held at a specific temperature). Composition A shows needle like crystals within 2 days from loading. Composition B and C were free from crystals for 3 months.

(41) TABLE-US-00011 TABLE 9 Observation for crystals in the compositions A, B and C when samples which were initially aged for 7 days at controlled temperature ranges 20° C. to 25° C. and then stored at 10° C.-15° C. Day Composition A Composition B Composition C 0 (Initial) Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 1 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 11 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 21 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 31 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 41 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 44 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 120 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution

(42) Samples from Composition A, B and C were initially held for 7 days at controlled temperature ranges 20° C. to 25° C. and then stored at 10° C.-15° C. Composition A, B and C were free from crystals throughout the entire study.

(43) TABLE-US-00012 TABLE 10 Observation for crystals in the composition A, B and C when samples were initially aged for 15 days at controlled temperature ranges 20° C. to 25° C. and then stored at 10° C.-15° C. Day Composition A Composition B Composition C 0 (Initial) Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 1 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 10 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 21 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 30 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 37 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 120 Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution

(44) Samples from Composition A, B and C were initially aged for 15 days at controlled temperature ranges 20° C. to 25° C. and then stored at 10° C.-15° C. Compositions A, B and C were free from crystals for 3 months.

(45) TABLE-US-00013 TABLE 11 Observation for crystals in the compositions A, B and C when samples were initially aged for 3 weeks at controlled temperature ranges 20° C. to 25° C. and then stored at 2° C. to 8° C. Day Composition A Composition B Composition C 0 (Initial) Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 1 Slight needle like Clear pale-yellow Clear pale-yellow crystals observed solution solution 2 Product freezes Clear pale-yellow Clear pale-yellow solution solution 3 Vials withdrawn (Study Clear pale-yellow Clear pale-yellow discontinued) solution solution 10 Clear pale-yellow Clear pale-yellow solution solution 20 Clear pale-yellow Clear pale-yellow solution solution 29 Clear pale-yellow Clear pale-yellow solution solution 120 Clear pale-yellow Clear pale-yellow solution solution

(46) Samples from Composition A, B and C were initially aged for 3 weeks at controlled temperature ranges 20° C. to 25° C. and then stored at 2° C.-8° C. Composition A shows needle like crystals within one day and the sample was frozen on the second day from loading. Compositions B and C were free from crystals for 3 months.

(47) TABLE-US-00014 TABLE 12 Observation for crystals in the composition A, B and C when samples were initially aged for 1 week at controlled temperature ranges 20° C. to 25° C. and then stored at 2° C. to 8° C. Day Composition A Composition B Composition C 0 (Initial) Clear pale-yellow Clear pale-yellow Clear pale-yellow solution solution solution 1 Needle like crystals Clear pale-yellow Clear pale-yellow observed solution solution 3 Product freezes Clear pale-yellow Clear pale-yellow solution solution 4 Vials withdrawn Clear pale-yellow Clear pale-yellow (Study discontinued) solution solution 10 Clear pale-yellow Clear pale-yellow solution solution 20 Clear pale-yellow Clear pale-yellow solution solution 23 Clear pale-yellow Clear pale-yellow solution solution 120 Clear pale-yellow Clear pale-yellow solution solution

(48) Samples from Compositions A, B and C were initially aged for 1 week at controlled temperature ranges 20° C. to 25° C. and then stored at temperatures between about 2° C. to 8° C. Composition A shows needle like crystals within a day and the sample was frozen on the third day. Composition B and C were free from crystals up to 3 months.

(49) TABLE-US-00015 TABLE 13 Observation for crystals in the composition B and C when samples were not aged, and stored immediately at 2° C. to 8° C. Day Composition B Composition C 0 Clear pale-yellow solution Clear pale-yellow solution (Initial) 1 Clear pale-yellow solution Clear pale-yellow solution 2 Clear pale-yellow solution Clear pale-yellow solution 5 Needle like crystals observed 2-3 weak needle like crystals observed at bottom, not clearly visible 10 Significant growth of crystals Crystal growth observed, but less 14 observed compared to Composition B

(50) Samples from Compositions A, B and C were stored immediately at temperatures between about 2° C. to 8° C. without any ageing. Compositions B and C were observed to have needle like crystals on the fifth day.

(51) TABLE-US-00016 TABLE 14 Observation for crystals in the composition B and C when samples were initially aged for 2 days at controlled temperature ranges 20° C. to 25° C. and then stored at 2° C. to 8° C. Day Composition B Composition C 0 (Initial) Clear pale-yellow solution Clear pale-yellow solution 10 Clear pale-yellow solution Clear pale-yellow solution 13 Clear pale-yellow solution Clear pale-yellow solution 150 Clear pale-yellow solution Clear pale-yellow solution

(52) Samples from Composition B and C were initially aged for 2 days at controlled temperature ranges 20° C. to 25° C. and then stored at temperatures between about 2° C. to 8° C. Compositions B and C were free from crystals for up to 5 months.

(53) Having now fully described this invention, it will be understood by those of ordinary skill in the art that it can be performed within a wide equivalent range of parameters without affecting the scope of the invention or any embodiment thereof. All publications, patent applications and patents disclosed herein are incorporated by reference in their entirety.