NOVEL PHARMACEUTICAL COMPOSITIONS OF ROMIDEPSIN

Field of Invention

The present invention relates to the pharmaceutical composition of Romidepsin including its pharmaceutically acceptable salts, solvates, hydrates and polymorphs thereof in lyophilized form. Further the invention also relates to dry fill romidepsin in unit dosage form to be reconstituted for use in injections.

Background of the invention

Romidepsin is a novel compound in a new class of antineoplastic agents known as histone deacetylase (HDAC) inhibitors, which increase acetylation of histones and other proteins. HDAC inhibition is associated with anti-tumor activities, including cell cycle arrest, antiangiogenesis, growth inhibition and apoptosis. Romidepsin is a pan-HDAC inhibitor showing potent inhibition of Class I, II and IV HDACs. Unlike the hydroxamic acid structure common to many of the other HDAC inhibitors in development, romidepsin is a naturally occurring cyclic peptide.

The chemical name of romidepsin is (1 S,4S,7Z,10S,16E,21 R)-7-ethylidene-4,21 - diisopropyl-2-oxa-12, 13-dithia-5,8,20,23-tetrazabicyclo[8.7.6]tricos-16-ene-3,6,9 , 19,22- pentone.

Romidepsin was first reported in the scientific literature in 1994, by a team of researchers from Fujisawa Pharmaceutical Company (now Astellas Pharma) in Tsukuba, Japan, who isolated it in a culture of Chromobacterium violaceum from a soil sample obtained in Yamagata Prefecture. It was found to have little to no antibacterial activity, but was potently cytotoxic against several human cancer cell lines, with no effect on normal cells; studies on mice later found it to have antitumor activity in vivo as well. The first total synthesis of romidepsin was accomplished by Harvard researchers and published in 1996. Its mechanism of action was elucidated in 1998, when researchers from Fujisawa and the University of Tokyo found it to be a histone deacetylase inhibitor with effects similar to those of trichostatin A. Romidepsin is a depsipeptide which contains both amide and ester bonds. In addition to the production of C. violaceum using fermentation, romidepsin can also be prepared by synthetic or semi-synthetic means. The total synthesis of romidepsin reported by Kahn et al. involves 14 steps and yields romidepsin in 18% overall yield. J. Am. Chem. Soc. 1 18:7237-7238, 1996. Romidepsin is approved for the treatment of cutaneous T-cell lymphoma (CTCL) and peripheral T-cell lymphoma (PTCL). Clinical trials are ongoing for other additional indications.

CTCL is a rare, life-altering, and life-threatening form of non-Hodgkin's lymphoma (NHL) which initially presents in the skin. The most common type of CTCL is mycosis fungoides (MF). Skin manifestations of the disease include patches, plaques, tumors, and erythroderma. Disease progression in MF may lead to extracutaneous involvement, including blood, lymph nodes, and viscera. Sezary syndrome (SS), which is a leukemic variant of CTCL, is characterized by pruritic erythroderma, generalized lymphadenopathy, and blood involvement with abnormal circulating T cells known as Sezary cells. Patients with this syndrome often experience intractable pruritus, which is typically the most significant life-altering symptom. Skin lesions may become ulcerative and necrotic with some patients becoming incapacitated due to their skin disease and associated symptoms of pain and pruritus. Infection, including sepsis, can occur due to the breakdown of the cutaneous barrier and compromised immune system. Current treatment regimens for CTCL are associated with a high relapse rate and responses are often short-lived, regardless of disease stage. Patients with Sezary syndrome represent a major therapeutic challenge for physicians treating patients with CTCL as there are currently no therapeutic options that predictably offer meaningful benefit. Peripheral T- cell lymphoma (PTCL) is the overall term for a heterogeneous group of non-Hodgkin's lymphomas arising from clonal proliferation of mature post-thymic lymphocytes. These malignancies tend to be clinically aggressive, respond poorly to chemotherapy, have high relapse rates, and are associated with poor long-term survival. Romidepsin offers an important additional therapeutic option for the treatment of patients with CTCL requiring systemic therapy. The efficacy of romidepsin has been established based on clinically meaningful objective response rates, achievement of clinical complete responses, durability of responses, improvement in all compartments of disease, relief of pruritus, and responses across patient subgroups, including patients with all stages of disease and those with Sezary syndrome. Safety findings associated with romidepsin indicate that the toxicities are manageable and combined with demonstrated clinical benefit in patients with CTCL, suggest a favorable benefit-to-risk ratio.

Commercially, Romidepsin is available under the brand name ISTODAX in United States by Celgene corporation. ISTODAX (romidepsin) for injection is a sterile lyophilized white powder and is supplied in a single-use vial containing 10 mg romidepsin and 20 mg povidone, USP. Diluent for ISTODAX contains 80% (v/v) propylene glycol and 20% (v/v) dehydrated alcohol.

U.S Patent No. 4977138 to Fujisawa Pharmaceutical discloses Romidepsin and novel fermentation method of producing romidepsin. U.S Patent No. 7608280 and 761 1724 discloses crystalline forms of romidepsin and method of producing.

U.S Patent application No. US2012/046442 discloses Romidepsin compositions prepared by lyophilization from a solution of (60:40) (v/v) t-butanol/water or tert-butanol solvent. Romidepsin compositions prepared according to this patent application involve excess process time and may involve additional solvates formation.

Hence there is a need to develop formulations of Romidepsin using alternate solvent or solvent mixtures thereof. Further dry fill formulations of romidepsin powder or a blend of romidepsin and a suitable carrier were found to be stable with desirable characteristics. Moreover romidepsin compositions prepared according to the present invention involves lesser process time with a stable product. Summary of the invention

Aspects of the present invention relates to pharmaceutical compositions of romidepsin including its pharmaceutically acceptable salts, solvates, hydrates and polymorphs thereof, in the form of lyophilized powder for intravenous administration and preparations thereof.

Another aspect of the present invention is to develop manufacturing process to develop pharmaceutical compositions of Romidepsin including its pharmaceutically acceptable salts, solvates, hydrates and polymorphs thereof.

Another aspect of the present invention is to describe lyophilization cycle for pharmaceutical compositions of Romidepsin including its pharmaceutically acceptable salts, solvates, hydrates and polymorphs thereof.

Another aspect of the present invention is to describe suitable solvent or mixture of solvents in a suitable concentration to develop stable romidepsin formulations; wherein the composition is devoid of Tertiary butyl alcohol

Another aspect of the invention is to provide Romidepsin lyophilized composition comprising:

a) Romidepsin or a pharmaceutically acceptable salts, solvates and hydrates thereof; and b) a solvent or mixture of solvents; wherein the composition is devoid of Tertiary butyl alcohol

Another aspect of the invention is to provide Romidepsin lyophilized composition comprising: a) Romidepsin or a pharmaceutically acceptable salts, solvates and hydrates thereof; b) solvent or mixture of solvents; and c) a pharmaceutically acceptable carrier; wherein the composition is devoid of Tertiary butyl alcohol. Another aspect of the invention is to lyophilize romidepsin alone using solvent mixtures and then reconstituted using diluent comprising of polyvinylpyrrolidene (PVP), dehydrated alcohol and propylene glycol.

A further object of the present invention is to provide a dry fill Romidepsin in unit dosage form to be reconstituted for use in injections.

Another aspect of the invention is to describe dry powder filling of romidepsin alone or blend of romidepsin and pharmaceutically acceptable carrier and subsequent gamma radiation sterilization.

Another aspect of the invention is to describe dry powder filling of Romidepsin blend comprising of romidepsin and cyclodextrin.

Detailed description of the invention:

The objective of the present invention is to prepare a stable lyophilized parenteral pharmaceutical compositions of Romidepsin including its pharmaceutically acceptable salts, solvates, hydrates and polymorphs thereof.

Aspects of the present invention relates to pharmaceutical compositions of romidepsin including its pharmaceutically acceptable salts, solvates, hydrates and polymorphs thereof, in the form of lyophilized powder for intravenous administration and preparations thereof.

Another aspect of the present invention is to describe lyophilization cycle for pharmaceutical compositions of Romidepsin including its pharmaceutically acceptable salts, solvates, hydrates and polymorphs thereof by using suitable solvent system in suitable concentrations; wherein the composition is devoid of Tertiary butyl alcohol. Another aspect of the invention is to provide lyophilized Romidepsin composition comprising: a) Romidepsin or a pharmaceutically acceptable salts, solvates and hydrates thereof; and b) a pharmaceutically acceptable carrier; wherein the composition is devoid of Tertiary butyl alcohol

Another aspect of the invention is to lyophilize romidepsin alone using solvent mixtures and then reconstituted using diluent comprising of polyvinylpyrrolidone (PVP), dehydrated alcohol and propylene glycol.

Another aspect of the invention is to describe dry powder filling of romidepsin alone or blend of romidepsin and pharmaceutically acceptable carrier and subsequent gamma radiation sterilization.

Another aspect of the invention is to provide lyophilized Romidepsin formulations comprising:

a) Romidepsin or a pharmaceutically acceptable salts, solvates and hydrates thereof; and

b) a pharmaceutically acceptable carrier such as,

(i) Suitable solvent and/or mixture of solvents thereof

(iii) Bulking agent

(iii) Optionally a tonicity modifier

(iv) Optionally a buffer

(v) Optionally a stabilizer or

(vi) Any suitable pharmaceutically acceptable adjuvant thereof.

A further object of the present invention is to provide a dry fill Romidepsin in unit dosage form to be reconstituted for use in injections.

Another aspect of the invention is to describe dry powder filling of romidepsin alone or blend of romidepsin and pharmaceutically acceptable carrier and subsequent gamma radiation sterilization. Another aspect of the invention is to describe dry powder filling of blend of romidepsin comprising of romidepsin and cyclodextrin.

Lyophilization (also called freeze-drying) refers to a process involving removal of solvent from the formulation using lyophilizer under reduced vaccuum pressure at suitable product temperatures, preferably below 1000 millitorr at suitable product temperature. Lyophilization helps stabilize pharmaceutical formulations by reducing the solvent component or components to levels that no longer support chemical reactions or biological growth. Since drying during lyophilization takes place at a low temperature, chemical decomposition is also reduced. Additionally, freeze dried products have a high specific surface area, which may enhance product dissolution during reconstitution. As used herein, the term "freeze-dried formulation" or "cake" refers to the dried formulation that remains after the solvent has been removed by the process of lyophilization. One goal of lyophilization is to retain the activity of the therapeutic agent while obtaining a pharmaceutically elegant end product.

The injectable formulations of the present invention comprises pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier can be selected from solvent, bulking agent, complexing agents, preservatives, anti-oxidants; stabilizers, tonicity modifiers and any other suitable adjuvant thereof.

As used herein the term "solvent" refers to the liquid component of a formulation that is capable of dissolving or suspending one or more solutes. The term "solvent" can refer to a single solvent or a mixture of solvents. The solvent, as mentioned, can be any liquid in which the material dissolves; the solvent can be a single substance or a mixture of co-solvents Depending on the formulation or the freeze-drying process, it may be desirable to include one or more organic solvents in the liquid formulation. The use of cosolvents is effective technique to enhance the solubility of the drug in the formulation. Suitable solvents include the following, but are not limited to N-methylpyrrolidone (NMP), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), N,N-dimethylacetamide (DMA), tetrahydrofuran (THF), tetrahydropyran, dioxane, trioxane and other cyclic mono-, di- and tri-ethers, lower alkanols (such as Methanol, ethanol, propanol, isopropanol), ethyl acetate, propyl acetate, propylene glycol (PG), polyethylene glycol, glycerine, acetone, acetonitrile, Ethoxyethanol, Toulene, nitromethane, hepatane, Methylcyclohexane, Formic acid, 1 ,2-Dimethoxyethane, 1 ,1 ,2-Trichloroethene, Methylethyl ketone, Cyclohexane, Ethyl formate, Hexane, Tetrahydrofuran, Chloroform, Methyl acetate, 1 ,2-Dichloroethene, tert-Butylmethyl ether, Dichloromethane, Pentane, Ethyl ether, Sulfalone, Ethyleneglycol, Formamide, Tetralin, 1 -pentanol, 3-Methyl-1 - butanol, Anisole, Ν,Ν-dimethylformamide, Methoxyethanol, Cumene, Butyl acetate, 2- Methyl-1 -propanol, Chlorobenzene, Acetic acid, Isobutyl acetate, 1 -Propanol, Pyridine, Methylbutyl ketone, Methyl isobutyl ketone, Xylene, Dimethyl carbonate, Hexafluoroacetone, Chlorobutanol, Dimethylsulfone and carbon tetrachloride or other organic solvents and mixtures of suitable solvents thereof or their equivalents. Optionally water can also be used as a solvent. Mixtures of solvents selected are used in a suitable proportion and suitable quantity to achieve desirable effect.

The solvent or mixture of solvents used in the pharmaceutical composition comprise atleast 10% volume by volume; based on the total weight of the parenteral formulation. Amount of water used in the pharmaceutical composition is in the range of 0.5 to 70 percent; based on the total weight of the parenteral formulation.

The purpose of the bulking agent is to provide bulk to the formulation and enhance cake formation. Suitable bulking agents include the following, but are not limited to raffinose, histidine, Polyvinylpyrrolidone or povidone, sugar alcohols, hydroxylethyl starch, ficoll, sodium chloride, starch, celluloses, gelatin, poloxamers, mannitol, glucose, sucrose, lactose, trehalose, glycine, trehalose, dextrose, maltose, sorbitol, dextran, cyclodextrins or other any suitable saccharides and mixtures thereof. Particularly advantageous oligosaccharides are the cyclodextrins. As the cyclodextrin in the compositions of the invention, there may be used any of the physiologically tolerable substituted or unsubstituted cyclodextrins such as but not limited to α-, β-, γ- and δ cyclodextrins or derivatives, charged cyclodextrins and the like, derivatives wherein one or more of the hydroxy groups are substituted, e.g. by alkyl, hydroxyalkyl, carboxyalkyl, alkylcarbonyl, carboxyalkoxyalkyl, alkylcarbonyloxyalkyl, alkoxycarbonylalkyl or hydroxy-(mono or polyalkoxy)alkyl groups, wherein each alkyl or alkylene moiety preferably contains up to six carbons and other alkylated cyclodextrins. Further examples of substituted cyclodextrins include ethers wherein the hydrogen of one or more cyclodextrin hydroxy groups is replaced by C1 -6 alkyl, hydroxyC1 -6 alkyl, carboxyC1 -6 alkyl or C1 -6 alkoxycarbonyl- C1 -6 alkyl groups or mixed ethers, Sulfated cyclodextrins thereof. E.g. methyl, ethyl, hydroxyethyl, hydroxypropyl beta-cyclodextrin, hydroxy butyl, carboxymethyl or carboxyethyl substituted cyclodextrin, hydroxypropyl dimethyl-β- cyclodextrin, or amino-cyclodextrin, sulfobutylcyclodextrins, sulfobutylether cyclodextrins, sulfobutyl ether beta-cyclodextrin sulfoalkyl ether cyclodextrin (SAE-CD) derivatives and the like. The quantity of cyclodextrin used however will generally be dependent on the quantity of drug and the molar ratio of drug to cyclodextrin lies in the range 1 .0: 0.1 to 1 .0: 50.

Stabilizing agents are typically added to a formulation to improve stability of the protein formulation, for example, by reducing denaturation, aggregation, deamidation and oxidation of the protein during the freeze-drying process as well as during storage. Examples of stabilizing agents include cryoprotectants, lyoprotectants, crystallization inhibitors or any other suitable stabilizer thereof. Suitable stabilizers include the following, but are not limited to Saccharides, including monosaccharides such as glucose, disaccharides such as sucrose (glucose+fructose), lactose (glucose+galactose), maltose (glucose+glucose), and trehalose (alpha-D- glucopyranosyl alpha-D-glucopyranoside), and polysaccharides such as dextran (polysaccharide containing glucose monomers, Crystallization inhibitors such as PVP (polyvinylpyrrolidone), HPC (hydroxypropyl cellulose), or HPMC (hydroxypropylmethylcellulose) and the like can be used. Surfactants also act as suitable stabilizers such as polyoxyethylene sorbitan monolaurate (Tween.TM. 20, Tween.TM. 80), pluronic F-68, Triton. TM. X-100, and sodium dodecyl sulfate (SDS), polysorbate or any other suitable surfactant can be selected. Cyclodextrins are also used as stabilizers. Buffers are typically included in pharmaceutical formulations to maintain the pH of the formulation at a physiologically acceptable pH. The desirable pH for a formulation may also be affected by the active agent. Examples of suitable buffers include buffers derived from an acid such as phosphate, aconitic, citric, glutaric, malic, succinic and carbonic acid, alkali or alkaline earth salt of one of these acids, Tris buffer, histidine buffers, meglumine or any suitable buffer thereof. pH adjusting agents such as, but are not limited to sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium hydroxide, ammonium carbonate, hydrochloric acid, citric acid, lactic acid, phosphoric acid, sodium phosphate, sulfuric acid, and the like can also be used.

Tonicity modifiers are the compounds which make the composition isotonic with blood. Suitable tonicity modifiers include the following, but are not limited to sorbitol, dextrose, glycerol, mannitol, lactose, sucrose or any suitable saccharide thereof, sodium chloride, sodium bicarbonate, calcium chloride, potassium chloride, sodium lactate, Ringer's solution, lactated Ringer's solution, sodium citrate or any suitable salts thereof, amino acids (e.g., arginine, cysteine, histidine, glycine and the like), trehalose and mixtures and salts thereof.

Other excipients may also be added to the lyophilized preparations of the present inventions. Such excipients may include antioxidants, antimicrobials, cryostabilizers, or any other suitable pharmaceutically acceptable adjuvants thereof.

The lyophilized powder or the dry powder or blend is reconstituted with a suitable diluent. Components of the diluent may include the following but are not limited to ethanol, polyethylene glycols or blends containing one or more polyethylene glycols of different grades, propylene glycol, polyvinylpyrrolidone, or agents to adjust solution osmolarity or other parenterally acceptable sugars, polyols, electrolytes or salts e.g., sodium chloride, potassium chloride, sodium citrate and the like) aminoacids (e.g., arginine, cysteine, histidine, glycine), saccharides (e.g., sucrose, glucose, mannitol, dextrose and the like) or any suitable adjuvant thereof. The lyophilized compositions of the present invention can be prepared by the following process; Prepare solvent mixture by mixing required quantity of solvents and optionally water in a glass vessel; Add Romidepsin followed by the excipients (if required) and stir well to get clear solution; Adjust the pH with a pH adjusting agent; Make up the final volume using solvent mixture; Filter the bulk solution followed by filling into vials, and load on to lyophilizer; vials are lyophilized under a vacuum of less than 1000 millitorr and a temperature below 60°C;Run the Lyophilizer as per the pre designed lyophilization cycle; Stopper the lyophilized vials seal them.

Dry fill compositions of the present invention can be prepared by the following process; Add required quantity of romidepsin or bend of romidepsin and excipient in a glass vial. Suitable sterilization techniques are applied to keep the product sterile.

The following examples further describe certain specific aspects and embodiments of the present invention and demonstrate the practice and advantages thereof. It is to be understood that the examples are given by way of illustration only and are not intended to limit the scope of the invention in any manner.

Example:1

Composition:

Manufacturing Process: 1) Solvent mixture was prepared by mixing required quantity of acetonitrile, methanol and water in a glass vessel, maintaining the solution temperature at 5±2°C.

2) Romidepsin was added to 80% of the above solvent mixture in a glass vessel and stirred well to get a clear solution, maintaining the solution temperature at 5±2°C.

3) Polyvinylpyrrolidone was added to the above solution and stirrd well to get clear solution, maintaining the temperature at 5±2°C.

4) pH was adjusted to 3.4 - 4.2 with 0.02N HCI solution.

5) Final volume was made upto to 100% using solvent mixture.

6) The bulk solution was filled into vials and partially stoppered with stoppers and loaded on to Lyophilizer pre-cooled shelves maintained at 2-5 °C.

7) The vials were lyophilized as per the pre designed lyophilization cycle.

Table 1 : Lyophilization Cycle used for manufacturing Romidepsin for Injection:

7 Primary Drying -5 720 H 800 44.9

8 Primary Drying 45 500 R 800 53.2

9 Secondary Drying 45 1790 H 800 83.1

Table 2: Temperature and vacuum profiles for Lyophilization cycle

Example:2

Composition: 2. Polyvinylpyrrolidone (PVP) 20mg

3. Dimethyl sulfoxide (DMSO) 1 .5ml

4. Water for Injection 0.1 ml

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin was taken and dissolved in Dimethyl sulfoxide (DMSO).

2) Polyvinylpyrrolidone dissolved in water was added to the above solution.

3) Solution was filtered and the desired quantity of the solution is filled in the vials.

4) The vials were lyophilized as per the pre designed lyophilization cycle.

Example: 3 Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin was taken and dissolved in Dimethyl sulfoxide (DMSO).

2) Sulfobutylether cyclodextrin was dissolved in the above solution.

3) Solution was filtered and the desired quantity of the solution is filled in the vials.

4) The vials were lyophilized as per the pre designed lyophilization cycle.

Example:4

Composition: S.No Ingredients Qty/unit

1 . Romidepsin 10mg

2. Polyvinylpyrrolidone (PVP) 20mg

3. dimethylacetamide (DMA) 1 .1 ml

4. Water for Injection 0.1 ml

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin was taken and dissolved in dimethylacetamide (DMA).

2) Polyvinylpyrrolidone dissolved in water was added to the above solution.

3) Solution was filtered and the desired quantity of the solution is filled in the vials.

4) The vials were lyophilized as per the pre designed lyophilization cycle.

Example:5

Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin was taken and dissolved in N-methylpyrrolidone (NMP).

2) Polyvinylpyrrolidone dissolved in water was added to the above solution.

3) Solution was filtered and the desired quantity of the solution is filled in the vials.

4) The vials were lyophilized as per the pre designed lyophilization cycle.

Example:6 Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin was taken and dissolved in a mixture of Dimethyl sulfoxide (DMSO) and Acetonitrile solvents.

2) Polyvinylpyrrolidone dissolved in water was added to the above solution.

3) Solution was filtered and the desired quantity of the solution is filled in the vials.

4) The vials were lyophilized as per the pre designed lyophilization cycle.

Example:7

Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin was taken and dissolved in a mixture of Dimethyl sulfoxide (DMSO) and Isopropylalcohol (IPA) solvents.

2) Polyvinylpyrrolidone dissolved in water was added to the above solution.

3) Solution was filtered and the desired quantity of the solution is filled in the vials. 4) The vials were lyophilized as per the pre designed lyophilization cycle.

Example:8

Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin was taken and dissolved in mixture of Dimethyl sulfoxide (DMSO) and Isopropylalcohol (IPA) solvents.

2) Water was added to the above solution.

3) Solution was filtered and the desired quantity of the solution is filled in the vials.

4) The vials were lyophilized as per the pre designed lyophilization cycle.

Example: 9

Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin and PVP were blended and filled in glass vials as a dry powder.

2) The filled vials are sterilized using gamma radiation.

Example: 10 Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin is filled in glass vials and stoppered.

2) The filled vials are sterilized by gamma radiation.

3) Prior to the use the vials are reconstituted with diluent comprising

Propylene glycol and Polyvinylpyrrolidone.

Example: 12 Composition:

Brief manufacturing procedure:

1 ) Required quantity of Romidepsin and Sulfobutylether beta cyclodextrin were blended and filled in glass vials as a dry powder.

2) The filled vials are sterilized using gamma radiation.