WO2014140695A1

WO2014140695A1 - Solid oral formulation of a pyrrolidine substituted flavone compound

Info

Publication number: WO2014140695A1
Application number: PCT/IB2013/052069
Authority: WO
Inventors: Arno Appavoo Enose; Milind VARTAK; Maneesh Nerurkar
Original assignee: Piramal Enterprises Ltd
Current assignee: Piramal Enterprises Ltd
Priority date: 2013-03-15
Filing date: 2013-03-15
Publication date: 2014-09-18
Anticipated expiration: 2015-09-15

Abstract

The present invention relates to a pharmaceutical composition providing a self emulsifying drug delivery system (SEDDS) for oral administration, comprising a compound, (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1- methyl-pyrrolidin-3-yl)-chromen-4-one or a pharmaceutically acceptable salt thereof, along with one or more surfactant and a solubilizer. The present invention also relates to a process for preparation of said pharmaceutical composition and use of the pharmaceutical composition in the treatment of a disease or disorder selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality.

Description

SOLID ORAL FORMULATION OF A PYRROLIDINE SUBSTITUTED FLAVONE

COMPOUND

FIELD OF THE INVENTION

The present invention relates to a pharmaceutical composition comprising (+) trans-2-(2-chloro-phenyl)-57-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl) -chromen-4-one or a pharmaceutically acceptable salt thereof; as the active ingredient and one or more surfactant and a solubilizer; and to methods of preparation of the pharmaceutical composition. The pharmaceutical composition provides for a self emulsifying drug delivery system (SEDDS) for oral administration of the said compound.

BACKGROUND OF THE INVENTION

Metabolism plays an important role in the bioavailability of a certain number of active principles. Poor bioavailability of drugs has been a major limitation in the successful utilization of many therapeutically effective molecules. Many of the new drug candidates tend to be poorly absorbed in the aqueous medium present in the gastrointestinal (Gl) tract. The problem of poor bioavailability is at times further compounded by a faster elimination rate which in turn reduces the efficiency of such molecules being used as a drug of choice.

Approximately, 40 % of new drug candidates have poor solubility and the oral delivery of such drugs is frequently associated with low bioavailability, high intra- and inter- subject variability, and a lack of dose proportionality. For such drugs, absorption is limited to the extent of dissolution. Similarly, there are drugs, which are highly soluble and dissolve rapidly but may be associated with low permeability and bioavailability. However, in these cases it is not the rate of drug dissolution that is usually rate limiting but rather the rate of permeation across the biological membranes.

Parenterally administered drugs are relatively unstable and also generally potent drugs that require strict control of administration to the patient. Moreover, parenteral administration of drugs is relatively painful and causes distress to the patient. Also, parenteral administration of drugs requires strict aseptic procedures, and parenteral dosage forms are costlier than formulations administered by other routes, particularly oral route. The oral route of drug delivery is the major and most desirable route for the treatment of several diseases. Solid oral dosage forms are popular because of the ease of administration, accurate dosage, self-medication, painless administration, and most importantly the patient compliance. Despite the ease of oral route of administration to patients, the drugs used in the treatment of cancers and certain other proliferative disorders were mostly available in parenteral forms until the recent times. A few new oral chemotherapeutic agents have recently been introduced. However, more efforts are required to improve patient compliance by making chemotherapeutic agents or antiproliferative agents available in oral dosage form considering that oral route is convenient and allows administration to take place at home or in non-traditional settings. Also several studies and surveys have revealed that patients prefer oral route of administration.

It is a known fact that bioavailability studies are often an integral part of developing oral agent. The advances in understanding the causes of poor bioavailability have led to significant improvements in the design of technologies to combat these deficiencies. Formulation design is often the route of choice for modifying the oral bioavailability of drugs as it offers a cost effective and relatively simple solution to the problem. US patent no. 7, 271 ,193 discloses flavone derivatives and its tautomeric forms, stereoisomers, optical isomers, pharmaceutically acceptable salts, solvates or polymorphs thereof which are cyclin-dependent kinases (CDK) inhibitors. One of the compounds disclosed in US patent no. 7, 271 ,193 namely (+) trans-2-(2-chloro- phenyl)-5, 7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (herein after referred to as compound A) has been found to be a potent investigational drug through several scientific studies (Mol. Cancer Ther. 2007, 6, 3, 918-925; Mol. Cancer Ther. 2007, 6, 3, 926-934).

Compound A is an effective therapeutic agent for the treatment of an animal or a human suffering from a disease or disorder selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality.

Compound A has been found to be a potent investigational drug particularly in the treatment of cancer amongst other indications specified above but poor bioavailability of the compound may limit its use from the perspective of patient compliance. There exists a need for producing stable oral formulations with improved bioavailability to address the potential problems associated with parenteral forms. The inventors of the present invention have directed their efforts to provide a solution to this problem in the form of a solid oral dosage form which improves the bioavailability of compound A.

The present invention provides the required pharmaceutical composition in the form of an oral solid dosage form and the methods for their preparation.

SUMMARY OF THE INVENTION

In one aspect, the present invention relates to a pharmaceutical composition comprising (+) trans-2-(2-chloro-phenyl)-5, 7-dihydroxy-8-(2-hydroxy methyl-1 - methyl-pyrrolidin-3-yl)-chromen-4-one or a pharmaceutically acceptable salt thereof, as an active ingredient along with one or more surfactant and a solubilizer.

In another aspect, the present invention provides a pharmaceutical composition comprising a compound, (+) trans-2-(2-chloro-phenyl)-5, 7-dihydroxy-8- (2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (referred to herein as compound A), along with one or more surfactant and a solubilizer.

In one aspect, the present invention provides a pharmaceutical composition for oral administration in the form of a capsule dosage form, comprising the compound A. The capsule may be a hard or a soft gelatine capsule. In another aspect, the present invention provides a pharmaceutical composition for oral administration in the form of a tablet dosage form, comprising the compound A. In a further aspect of the present invention, there is provided a method of preparation of a solid oral dosage form comprising the compound A, along with one or more surfactant and a solubilizer.

In another aspect of the present invention, there is provided a method for the treatment of a disease or disorder selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality, comprising administering to the subject therapeutically effective amount of a pharmaceutical composition comprising the compound A, along with one or more surfactant and a solubilizer in the form of a stable oral dosage according to one or more of the embodiments as described herein.

Other aspects and features of the present invention will become apparent from the following detailed description of the invention. DETAILED DESCRIPTION OF THE INVENTION

The general terms used hereinbefore and hereinafter preferably have within the context of this disclosure the following meanings, unless otherwise indicated. Thus, the definitions of the general terms as used in the context of the present invention are provided herein below: The singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise.

The term "pharmaceutically acceptable excipient" refers to a non-therapeutic agent such as diluents, binders, disintegrants, lubricants, glidants, colorants, flavorants, sweeteners and desiccants. The non-therapeutic agents have effects on the physical, chemical and biopharmaceutical properties of the finished dosage form e.g. capsules or tablets.

As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or a diluent that is non-toxic, inert, solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type which is compatible with a subject, preferably a mammal, more preferably a human, and is suitable for delivering a therapeutically active agent to the target site without interfering with the activity of the active pharmaceutical agent.

The term "active pharmaceutical ingredient" (used interchangeably with "active" or "active ingredient") as used herein refers to (+) trans-2-(2-chloro-phenyl)- 5, 7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one or a pharmaceutically acceptable salt thereof. In one embodiment of the invention, the term "active pharmaceutical ingredient" refers to (+) trans-2-(2-chloro-phenyl)-5, 7- dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one

hydrochloride (compound A). The term "therapeutically effective amount", as used herein refers to an amount of active pharmaceutical ingredient effective in producing the desired therapeutic response in a subject in need, for the treatment of a disease or disorder selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality.

The term "cancer" as used herein refers to carcinoma, including that of bladder, breast, colon, kidney, liver, lung, head and neck, ovary, pancreas, stomach, cervix, thyroid, prostate, intestine and skin; hematopoietic tumors of lymphoid lineage, including acute lymphocytic leukemia, B-cell lymphoma, and Burkett's lymphoma; hematopoietic tumors of myeloid lineage, including acute and chronic myelogenous leukemias and promyelocytic leukemia; tumors of mesenchymal origin, including fibrosarcoma and rhabdomysarcoma; and other tumors including melanoma, seminoma, teratocarcinoma, osteosarcoma, neuroblastoma and glioma.

The term "subject" as used herein refers to an animal, preferably a mammal.

The term "mammal" used herein refers to warm-blooded vertebrate animals of the class 'Mammalia', including humans, characterized by a covering of hair on the skin and, in the female, milk-producing mammary glands for nourishing the young. The term mammal includes animals such as cat, dog, rabbit, bear, fox, wolf, monkey, deer, mouse, pig and a human. The preferred mammal is human. As used herein, the terms "treatment", "treat" and "therapy" and the like refer to alleviate, slow the progression, prophylaxis, attenuation or cure of existing disease or condition (e.g., cancer). Treatment also includes treating the symptoms of the disease or condition. The term "bioavailability" refers to the rate and extent to which the active ingredient or active moiety is absorbed from a drug product and becomes available at the site of action (Pharmaceutical Research, 2001 , 18, 12, 1645-1650).

In one aspect, the present invention provides a pharmaceutical composition comprising (+) trans-2-(2-chloro-phenyl)-5, 7-dihydroxy-8-(2-hydroxymethyl-1 - methyl-pyrrolidin-3-yl)-chromen-4-one or a pharmaceutically acceptable salt thereof as an active ingredient and one or more surfactant and a solubilizer.

In one embodiment, the present invention provides a pharmaceutical composition comprising (+) trans-2-(2-chloro-phenyl)-5, 7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A) as an active ingredient and one or more surfactant and a solubilizer.

In an embodiment of the present invention, there is provided a pharmaceutical composition comprising 30 % to 80 % of the active ingredient, 10 % to 30 % of the surfactant and 20 % to 40 % of the solubilizer.

In an embodiment of the present invention, there is provided a pharmaceutical composition comprising 40 % to 70 % of compound A.

In an embodiment of the present invention, the pharmaceutical composition provides for a self emulsifying drug delivery system (SEDDS) for oral administration of (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl- pyrrolidin-3-yl)-chromen-4-one or a pharmaceutically acceptable salt thereof as the active ingredient.

In an embodiment of the present invention, the pharmaceutical composition provides for a self emulsifying drug delivery system (SEDDS) for oral administration of (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl- pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A). Compound A is a representative example of the compounds of formula I as disclosed in US patent no. 7, 271 ,193 and is poorly bioavailable at all physiological pH conditions. Accordingly, an object of the present invention is to increase the bioavailability of the compound A by developing a solid oral dosage form.

In one aspect of the present invention, there is provided a process for the preparation of a capsule dosage form comprising (+) trans-2-(2-chloro-phenyl)-5, 7- dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one or a pharmaceutically acceptable salt thereof as the active ingredient, comprising the steps of: step (a): sifting the active ingredient; step (b): heating a surfactant and a solubilizer at a temperature ranging from 65 to

75 °C to cause melting of the said surfactant and solubilizer to obtain a clear solution; step (c): preparing a blend by adding the sifted active ingredient of step (a) to the solution of step (b) with continuous stirring; step (d): optionally adding pharmaceutically acceptable excipients to the resultant blend; and step (e): filling the resultant blend of step (d) into a hard gelatin capsule to yield the capsule dosage form.

In an embodiment of the invention, a process for the preparation of a capsule dosage form comprising, (+) trans-2-(2-chloro-phenyl)-5, 7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A) as the active ingredient is provided, which process comprises steps (a) to (e) as described above.

In an embodiment of the invention, the pharmaceutically acceptable excipients of step (d) for the preparation of a capsule dosage form are selected from diluents, binders, disintegrants, lubricants, glidants, colorants, flavorants, sweeteners and desiccants, which are described herein below. In an embodiment of the invention, the surfactant contained in the pharmaceutical composition is selected from ionic or non-ionic surfactant.

In one embodiment of the invention, the surfactant include, but are not limited to, non-ionic surfactant selected from Cremophor^® EL, Cremophor^® RH, d-a- tocopherol polyethylene glycol 1000 succinate (vitamin E TPGS or TPGS), polysorbate 20, polysorbate 80, Solutol^® HS 15, sorbitan monooleate, poloxamers, Labrafils^®, Labrasol^®, Gellucire^® 44/14, Softigen^® 767, mono- and di-fatty acid esters of polyethylene glycol or a combination thereof.

In another embodiment of the invention, the surfactant is d-a-tocopherol polyethylene glycol 1000 succinate. This surfactant, d-a-tocopherol polyethylene glycol succinate (Vitamin E TPGS or simply TPGS) is a water-soluble derivative of natural Vitamin E, which is formed by esterification of Vitamin E succinate with polyethylene glycol (PEG).

In an embodiment of the present invention, one or more of the surfactant can act as an emulsifier in the resulting pharmaceutical composition.

In another embodiment of the invention, the solubilizer include, but are not limited to, a group comprising polyethylene glycol (having molecular weight between 300-6000), propylene glycol derivatives, glycerin, Cremophor^®, polysorbates, Lutrol^® Carbitol™, or a combination thereof. In yet another embodiment, the solubilizer is polyethylene glycol (PEG) with molecular weight of 1000, commonly known as PEG 1000.

In an embodiment of the present invention, there is provided a pharmaceutical composition comprising 40 % to 70 % of compound A as the active ingredient, 10 % to 30 % of vitamin E TPGS as the surfactant and 20 % to 40 % of PEG 1000 as the solubilizer.

In one aspect of the present invention, the pharmaceutical composition herein described according to the various embodiments may then be further adapted for oral administration in discrete units such as capsules. In one embodiment of the invention, the capsule may be hard or soft gelatine capsule. In an embodiment, the pharmaceutical composition of the present invention is formulated as hard gelatine capsules. For instance, for oral administration in the form of a capsule, the compositions herein described containing the active ingredient may be utilized as is, or can be further combined with one or more of non-toxic pharmaceutically acceptable carrier such as ethanol, glycerol, glycerin, water, and the like. The pharmaceutical compositions of the invention may be encapsulated in the form of a semisolid or solid matrix or in the form of a liquid in a capsule to yield a solid oral capsule dosage form. Powders may be prepared, for example, by comminuting the composition of the invention, or the active compound, to a suitable fine size and if desired, further mixing with a similarly comminuted pharmaceutically acceptable carrier.

In addition to gelatin, other materials for the capsule sheath or shell include hydroxypropyl methylcellulose (HPMC), cellulose, methylcellulose, starch, or combinations thereof.

Other methods for preparing capsule dosage form (both hard and soft) known in the art may also be utilized.

According to another aspect of the present invention, a process is provided for the preparation of a tablet dosage form comprising (+) trans-2-(2-chloro-phenyl)- 5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one

(compound A) or a pharmaceutically acceptable salt thereof as the active ingredient,, comprising the steps of: step a) sifting the active ingredient and the diluents; step b) mixing the sifted active ingredient with the sifted diluents to obtain a mixture; step c) preparing a binder solution containing a surfactant by dissolving the surfactant and binding agent in purified water; step d) preparing granules by adding binder solution of step (c) to the mixture of step (b), passing the resultant mixture through sieves to obtain wet granules, drying the wet granules at a temperature ranging from 50 °C to 100°C and passing the dried granules through sieves; and step e) sifting glidant and disintegrant through sieves, blending the sifted glidant and the disintegrant with the granules prepared in step (d) to obtain a blend, further adding sifted lubricant to the blend obtained and compressing the final mixture to yield the tablet dosage form. In one embodiment of the invention, a process is provided for the preparation of a tablet dosage form comprising 1 % to 80 % of compound A as the active ingredient.

In an embodiment of the invention, the surfactant used in the preparation of the tablet dosage form is selected from ionic or non-ionic surfactant. In one embodiment of the invention, the surfactant used in the preparation of the tablet dosage form include, but are not limited to, non-ionic surfactant described herein.

In another embodiment of the invention, the surfactant used in the preparation of the tablet dosage form is d-a-tocopherol polyethylene glycol 1000 succinate.

In an embodiment of the invention, the diluents or fillers used in the preparation of the capsule or tablet dosage form include, but are not limited to, cellulose derivatives, such as microcrystalline cellulose or wood cellulose, anhydrous lactose, lactose monohydrate, directly compressible anhydrous lactose, modified lactose monohydrate, sucrose, starch, pregelatinized starch, dextrose, mannitol, fructose, xylitol, sorbitol, corn starch, modified corn starch, calcium carbonate, calcium phosphate, dibasic calcium phosphate dihydrate, dicalcium phosphate, calcium sulfate, dextrins, dextrates, maltodextrin, compressible sugars, or a combination thereof. In another embodiment of the invention, binders used in the preparation of the capsule or tablet dosage form include, but are not limited to, hydroxypropyl cellulose, corn starch, pregelatinized starch, polyvinyl pyrrolidone (PVP), carboxymethylcellulose, lactose, polyethylene glycol, gum acacia, tracaganth, sodium alginate, ethyl cellulose, cellulose acetate, carnauba wax, paraffin, polyethylenes or microcrystalline wax, or a combination thereof. In yet another embodiment of the invention, glidants used in the preparation of the capsule or tablet dosage form include but are not limited to, silicon dioxide, colloidal silicon dioxide, magnesium silicate, magnesium trisilicate, talc, and other forms of silicon dioxide, such as aggregated silicates or hydrated silica, or a combination thereof.

In a further embodiment of the invention, disintegrating agent used in the preparation of the capsule or tablet dosage form include, but are not limited to starch, pregelatinized starch, corn starch, sodium starch glycolate, hydroxypropyl cellulose, agar-agar, calcium carbonate, or sodium carbonate, croscarmellose sodium, crospovidone, or a combination thereof, which can be added to the pharmaceutical composition in order to improve the bioavailability of the medicament when the dosage form is ingested.

In a still further embodiment of the invention, lubricant used in the preparation of the capsule or tablet dosage form include, but are not limited to, magnesium stearate, zinc stearate, calcium stearate, talc, carnauba wax, stearic acid, palmitic acid, sodium oleate, sodium laurel sulfate, glyceryl palmitostearate, palmitic acid, myristic acid, hydrogenated vegetable oils and fats, or a combination thereof.

The pharmaceutical composition of the present invention may optionally also contain a pharmaceutically acceptable antioxidant for stabilization of the dosage form. Examples of such antioxidants include, but not limited to, ascorbic acid, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate and vitamin E.

A preservative, flavoring, desiccant, dispersing or coloring agent can also be added into the pharmaceutical composition of the present invention. In one aspect, the administration of the pharmaceutical composition of the present invention improves oral bioavailability of the compound A in dogs to 30 % to 40 % in comparison to intravenous administration.

According to another aspect of the present invention, there is provided a method for the treatment of a disease or disorder in a subject in need thereof, comprising administering to the subject, the pharmaceutical composition comprising a therapeutically effective amount of the active pharmaceutical ingredient, (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl- pyrrolidin-3-yl)-chromen-4-one, or a pharmaceutically acceptable salt thereof; wherein the disease or disorder selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality.

In an embodiment of the present invention, there is provided a method for the treatment of a disease or disorder in a subject in need thereof, comprising administering to the subject, the pharmaceutical composition comprising a therapeutically effective amount of the active pharmaceutical ingredient, (+) trans-2- (2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)- chromen-4-one, or a pharmaceutically acceptable salt thereof; wherein the disease or disorder is selected from cancer, polycystic kidney disease, radiation induced mucositis or viral infection.

In an embodiment of the present invention, the pharmaceutical composition used in the method described herein above is a solid oral capsule dosage form.

In an embodiment of the present invention, the active pharmaceutical ingredient used in the method described herein above is (+) trans-2-(2-chloro- phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A).

According to another aspect of the present invention, there is provided use of the pharmaceutical composition comprising a therapeutically effective amount of the active pharmaceutical ingredient, (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8- (2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one, or a pharmaceutically acceptable salt thereof; in the treatment of a disease or disorder selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality. In an embodiment of the present invention, there is provided use of the pharmaceutical composition comprising a therapeutically effective amount of the active pharmaceutical ingredient, (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one, or a pharmaceutically acceptable salt thereof; in the treatment of a disease or disorder selected from wherein the disease or disorder is selected from cancer, polycystic kidney disease, radiation induced mucositis or viral infection.

In an embodiment of the present invention, the pharmaceutical composition used herein above is a solid oral capsule dosage form. In an embodiment of the present invention, the active pharmaceutical ingredient used herein above is (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A).

The invention is further described by the following non-limiting examples which further illustrate the invention, and are not intended, nor should they be interpreted to, limit the scope of the invention.

The following abbreviations or terms are used herein:

DMF N, N-dimethylformamide

h Hour

°C Degree centigrade

Rh Relative humidity

CHCIs Chloroform

g Gram

Kg Kilogram

Min Minute(s)

ng Nanogram

mL Millilitre

L Litre

mmol Millimolar

HCI Hydrochloric acid EtOAc Ethyl acetate

Cone. Concentrated

Na₂C0₃ Sodium carbonate

Na₂S0₄ Sodium sulfate

MeOH Methanol

I PA Isopropyl alcohol

TFA Trifluoroacetic acid

A.S.T.M American Society for Testing and Materials

HPLC High Performance Liquid Chromatography

μ Micron

U.S.P United States Pharmacopeia

K2EDTA Dipotassium ethylene diamine tetraacetic acid

AUC(o-t) Area under the curve from the time of dosing to the time of the last measurable concentration

AUC (o Area under the curve from the time of dosing to infinity

Cmax Maximum observed concentration

MRT (o Mean residence time from the time of dosing to infinity

T_max Time of the maximum observed concentration

t-i Terminal half-life

Vz Volume of distribution at terminal phase

F Bioavailability

SD Standard deviation

CL Total body clearance

PO Per os

Examples:

I) Preparation of the compound A, is presented herein as reference example:

Reference example 1 :

(a) Preparation of (+)-frans-2-(2-chlorophenyl)-8-(2-hydroxymethyl-1 -methyl pyrrolidin-3-yl)-5,7-dimethoxy-chromen-4-one Sodium hydride (50 %, 0.54 g, 1 1 .25 mmol) was added in portions to a solution of (-)-frans-1 -[2-hydroxy-3-(2-hydroxymethyl-1 -methyl pyrrolidin-3-yl)-4,6- dimethoxyphenyl)-ethanone (0.7 g, 2.2 mmol) in dry DMF (15 mL) at 0°C, under nitrogen atmosphere and with stirring. After 10 min., methyl 2-chlorobenzoate (1 .15 g., 6.75 mmol) was added. The reaction mixture was stirred at 25 °C for 2 h. Methanol was added carefully below 20 °C. The reaction mixture was poured over crushed ice (300 g), acidified with 1 :1 HCI (pH 2) and extracted using EtOAc (2 x 100 mL). The aqueous layer was basified using a saturated Na₂C0₃ (pH 10) and extracted using CHCI₃ (3 x 200 mL). The organic layer was dried (anhydrous Na₂S0₄) and concentrated. To the residue, cone. HCI (25 mL) was added and stirred at room temperature for 2 h. The reaction mixture was poured over crushed ice (300 g) and made basic using a saturated aqueous Na₂C0₃ solution. The mixture was extracted using CHCI₃ (3 x 200 mL). The organic extract was washed with water, dried (anhydrous Na₂S0₄) and concentrated to obtain the compound, (+)-irans-2-(2-chloro-phenyl)-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-5,7- dimethoxy-chromen-4-one.

Yield: 0.67 g (64 %); mp: 91 - 93°C; [a]_D ²⁵ = + 5.8° (c = 0.7, methanol); IR (KBr): 3431 , 1648, 1598, 1571 cm^"1 ; ¹H NMR (CDCI₃, 300MHz): δ 7.70 (dd, 1 H), 7.52 (m, 1 H), 7.45 (m, 2H), 6.50 (s, 1 H), 6.44 (s, 1 H), 4.17 (m, 1 H), 4.00 (s, 3H), 3.97 (s, 3H), 3.64 (dd, 1 H), 3.40 (d, 1 H), 3.15 (m, 1 H), 2.74 (d, 1 H), 2.52 (m, 1 H), 2.32 (s, 3H), 2.00 (m, 2H); MS (ES+): m/z 430 (M+1 ).

(b) Preparation of (+)-fraA7s-2-(2-chlorophenyl)-5,7-dihydroxy-8-(2-hydroxy methyl-1-methyl-pyrrolidin-3-yl)-chromen-4-one

Molten pyridine hydrochloride (4.1 g, 35.6 mmol) was added to the compound as obtained in part (a) (0.4 g, 0.9 mmol) and heated at 180°C for 1 .5 h. The reaction mixture was cooled to 25 °C, diluted with MeOH (10 mL) and basified using Na₂C0₃ to pH 10. The mixture was filtered and the organic layer was concentrated. The residue was suspended in water (5 mL), stirred for 30 min., filtered and dried to obtain the compound, (+)-frans-2-(2-chloro-phenyl)-5,7- dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one.

Yield: 0.25 g (70 %); IR (KBr): 3422, 3135, 1664, 1623, 1559 cm^"1 ; ¹H NMR (CDCIs, 300MHz): δ 7.56 (d, 1 H), 7.36 (m, 3H), 6.36 (s, 1 H), 6.20 (s, 1 H), 4.02 (m, 1 H), 3.70 (m, 2H), 3.15 (m, 2H), 2.88 (m, 1 H), 2.58 (s, 3H), 2.35 (m, 1 H), 1 .88 (m,

I H) ; MS (ES+): m/z 402 (M+1 ); Analysis: C₂₁ H₂₀CINO₅ C, 62.24 (62.71 ); H, 5.07 (4.97); N, 3.60 (3.48); CI, 9.01 (8.83). (c) Preparation of (+)-fraA7s-2-(2-chlorophenyl)-5,7-dihydroxy-8-(2-hydroxy methyl -1-methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A)

The compound as obtained in part (b) (0.2 g, 0.48 mmol) was suspended in I PA (5 mL) and 3.5 % HCI (25 mL) was added. The suspension was heated to get a clear solution. The solution was cooled and solid filtered to obtain the compound, (+)-irans-2-(2-chlorophenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin- 3-yl)-chromen-4-one hydrochloride or compound A.

Yield: 0.21 g (97 %); mp: 188 - 192°C ; [a]_D ²⁵ = +21 .3° (c = 0. 2, methanol); ¹H NMR (CD₃OD, 300MHz): δ 7.80 (d, 1 H), 7.60 (m, 3H), 6.53 (s, 1 H), 6.37 (s, 1 H), 4.23 (m, 1 H), 3.89 (m, 2H), 3.63 (m, 1 H), 3.59 (dd, 1 H), 3.38 (m, 1 H), 2.90 (s, 3H), 2.45 (m, 1 H), 2.35 (m, 1 H); MS (ES+): m/z 402 (M +1 )( free base).

This compound was subjected to chiral HPLC. Chiral HPLC was done using column Chiralcel OD-H (250 x 4.6 mm) and solvent system hexane:ethanol (92:08) with TFA (0.4 %). The results are recorded at 264 nm with solvent flow rate of 1 ml_/min. Chiral HPLC showed 100 % enantiomeric excess (e.e) of the compound, (+)-irans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxy-methyl-1 -methyl-pyrrolidin- 3-yl)-chromen-4-one hydrochloride.

II) Preparation of dosage form containing compound A Example 1

Preparation of hard gelatin capsule containing compound A

The capsule dosage form was prepared using the ingredients as listed in Table 1 . Table 1 lists the ingredients in % of capsule weight and mg/capsule. Table 1

Procedure:

All the ingredients listed in Table 1 were accurately weighed. Compound A was sifted through sieve # 40 (A.S.T.M, 420 μ). TPGS and PEG 1000 were heated at elevated temperature (for e.g. 70°C) to melt and form a clear solution. To the clear solution obtained, compound A was slowly added with continuous stirring to form a smooth paste. The paste formed was finally filled into hard gelatin capsules. Disintegration time was recorded (according to U.S.P Disintegration test for hard gelatin capsules). The capsule parameters are tabulated in table 1 A and 1 B.

Table 1 A capsule parameters: Size # 2 capsule

Example 2:

Preparation of a tablet by wet granulation technique The tablet dosage form is prepared using the ingredients as listed in Table 2.

Table 2 lists the ingredients in mg/tablet.

Table 2:

Procedure:

All the ingredients (microcrystalline cellulose (MCC), sodium starch glycolate, colloidal silicon dioxide, magnesium stearate) and compound A are accurately weighed. Compound A is sifted through sieve # 20 (A.S.T.M, 850 μ). MCC and sodium starch glycolate are sifted through sieve # 40 (A.S.T.M, 420 μ) and mixed with the sifted compound A in a rapid mixer granulator (RMG). A binder solution is prepared by dissolving Speziol® TPGS pharma and polyethylene glycol 1000 in hot purified water at 70° C. The solution is cooled at room temperature. Mixture of the compound A and other excipients is granulated using the binder solution and the resulting wet granules are passed through sieve # 16 (A.S.T.M, 1204 μ). The granules are then dried at 60 °C in an oven till LOD (loss on drying) is less than 3.5 % w/w at 105°C. The dried granules are then passed through sieve # 30 (A.S.T.M, 600 μ). Colloidal silicon dioxide and sodium starch glycolate are sifted through sieve # 40 (A.S.T.M, 420 μ) and mixed in a polybag with granules as obtained above. Finally this mixture is mixed with magnesium stearate, sifted through sieve # 60 (A.S.T.M, 250 μ) and compressed.

III) Biological activity: Example 3:

In vivo activity of capsule dosage form:

Assay: The assay described herein below was carried out at Medicilon Preclinical

Research (Shanghai) LLC, China.

The pharmacokinetic parameters of compound A were determined in three male beagle dogs by a single bolus intravenous (IV) and oral capsule (PO) administration. The blood samples were collected from the three dogs at pre-dose and post-dose at intervals of 0.25, 0.5, 1 , 2, 4, 6, 8, 10, 12, 24, 48, 72, and 96 h following IV or PO administration. All the dogs underwent a seven or fourteen days wash-out period between IV and PO administration.

Materials and method:

A) Preparation of parenteral formulation of compound A: The formulation of compound A in 5 % dextrose solution at concentration of 2 mg/mL for intravenous administration was prepared at Piramal Enterprises Limited, and was stored at room temperature until picked up for dosing by IV administration. Similarly, the test capsules were provided and dosed to animals.

B) Animal Acquisition and Assignment to Study A total of 4 non-naive male beagle dogs approximately 10 months to 1 year and 4 months of age at transfer, were transferred from the stock colony which were originally received from Beijing Marshall Biotechnology Co., Ltd. and 3 dogs were placed on study. The three dogs were identified by nos. 101 , 102 and 103 during the study. All the treated dogs were fasted overnight prior to oral dosing and food was withheld through the first 4 hours of blood sample collection at each administration. No fasting was done for IV administration.

C) Dose Administration

5 Compound A was administered via cephalic vein as a single bolus intravenous injection in IV administration. For oral administration, the capsules were administered via a single oral dosing. Immediately following oral administration, approximately 10 mL reverse osmosis water was administered. Dose administration information is presented in the following table.

o

Formulation details:

Table 3A: Intravenous formulation

D) Study design:

A total number of 3 male dogs were used in the study according to the following table 4. 0 Table 4:

^**The animals were fasted overnight (12 hours) prior to oral administration.

E) Sample Collection and Bioanalysis

Blood samples (approximately 1 mL) were collected via jugular vein, at pre-dose and post-dose at 0.25, 0.5, 1 , 2, 4, 6, 8, 10, 12, 24, 48, 72, and 96 h following IV or PO administration. Blood samples were placed into tubes containing K2-EDTA and centrifuged at 3,500 rpm for 10 min. at 4°C to separate plasma from the samples. Following centrifugation, the resulting plasma was transferred to clean tubes and stored frozen at -80 °C until bioanalysis.

F) Pharmacokinetic Analysis

The pharmacokinetic (PK) analysis and interpretation of the results were conducted by Medicilon Preclinical Research (Shanghai) LLC. The noncompartmental module of WinNonlin® Professional 5.2 was used to calculate parameters.

The bioavailability was calculated as F (%) = (Doseiv ^χ AUC_orai(o-~)) / (Dose_orai ^χ AUCiv(o-oo)) ^χ 100 %.

Results of PK analysis are represented below in Table 5 and Table 6. IV) Pharmacokinetic parameters of the formulation of compound A in male beagle dogs following intravenous and oral administration:

i) Intravenous administration (2 mg/kg):

Table 5:

ii) Oral administration (10 mg/kg):

Table 6:

Conclusion:

Following intravenous administration of the formulation of compound A according to Table 3A herein above at 2 mg/kg, the mean values of half-life and AUC₍o were 3.19 hours and 717.10 ng/ml_^*h, respectively. The mean values of clearance (CL) and Vz were 2.79 L/h/kg and 12.76 L/kg, respectively.

Following oral administration of the formulation of compound A according to Table 3B herein above at 10 mg/kg, the mean values of half-life and AUC(0-∞) were 7.69 hours and 1 190.32 ng/ml_^*h, respectively. The Cmax and Tmax were 245.16 ng/mL and 0.83 hour, respectively. The mean bioavailability was 33.32 %.

Claims

WE CLAIM:

1 . A pharmaceutical composition comprising a compound, (+) trans-2-(2-chloro- phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one, or a pharmaceutically acceptable salt thereof; as an active pharmaceutical ingredient, along with one or more surfactant and a solubilizer.

2. The pharmaceutical composition according to claim 1 , wherein the active pharmaceutical ingredient is (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A).

3. The pharmaceutical composition according to claim 1 or claim 2, wherein the surfactant is a non-ionic surfactant selected from Cremophor^® EL, Cremophor^® RH, d-a-tocopherol polyethylene glycol 1000 succinate (vitamin E TPGS or TPGS), polysorbate 20, polysorbate 80, Solutol^® HS 15, sorbitan monooleate, poloxamers, Labrafils^®, Labrasol^®, Gellucire^® 44/14, Softigen^® 767, mono- and di-fatty acid esters of polyethylene glycol or a combination thereof.

4. The pharmaceutical composition according to claim 3, wherein the surfactant is d-a-tocopherol polyethylene glycol 1000 succinate (vitamin E TPGS or TPGS).

5. The pharmaceutical composition according to claim 1 or claim 2, wherein the solubilizer is selected from polyethylene glycol (having molecular weight between 300-6000), propylene glycol derivatives, glycerin, Cremophor^®, polysorbates, Lutrol^®, Carbitol™or a combination thereof.

6. The pharmaceutical composition according to claim 5, wherein the solubilizer is polyethylene glycol (PEG) with molecular weight of 1000.

7. The pharmaceutical composition according to claim 2, wherein the active pharmaceutical ingredient comprises 40 % to 70 % of compound A.

8. The pharmaceutical composition according to claim 4, wherein the surfactant comprises 10 % to 30 % of vitamin E TPGS.

9. The pharmaceutical composition according to claim 6, wherein the solubilizer comprises 20 % to 40 % of PEG 1000.

10. The pharmaceutical composition according to claim 1 or claim 2, wherein said composition is encapsulated in a capsule to yield a solid oral capsule dosage form.

1 1 . The capsule dosage form of claim 10, wherein said capsule is a hard or a soft gelatine capsule.

12. A process for the preparation of a solid oral capsule dosage form comprising an active pharmaceutical ingredient, (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one, or a pharmaceutically acceptable salt thereof, comprising the steps of: step (a): sifting the active ingredient; step (b): heating a surfactant and a solubilizer at a temperature ranging from 65 °C to 75 °C to cause melting of the said surfactant and solubilizer to obtain a clear solution; step (c): preparing a blend by adding the sifted active ingredient of step (a) to the solution of step (b) with continuous stirring; step (d): optionally adding pharmaceutically acceptable excipients to the resultant blend; and step (e): filling the resultant blend of step (d) into a hard gelatin capsule to yield the solid oral capsule dosage form.

13. The process for the preparation of a solid oral capsule dosage form according to claim 12, wherein the active pharmaceutical ingredient is (+) trans-2-(2-chloro- phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A).

14. The process according to claim 12 or claim 13, wherein the pharmaceutically acceptable excipients of step (d) are selected from diluents, binders, disintegrants, lubricants, glidants, colorants, flavorants, sweeteners and desiccants.

15. A method of treating a disease or disorder, comprising administering to a patient in need thereof, a pharmaceutical composition according to claim 1 comprising a therapeutically effective amount of the active pharmaceutical ingredient, (+) trans-2- (2-chloro-phenyl)-57-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidi chromen-4-one, or a pharmaceutically acceptable salt thereof, wherein the disease or disorder is selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality.

16. The method according to claim 15, wherein the disease or disorder is selected from cancer, polycystic kidney disease, radiation induced mucositis or viral infection.

17. The method according to claim 15 or claim 16, wherein the pharmaceutical composition is a solid oral capsule dosage form.

18. The method according to any one of the claims 15-17, wherein the active pharmaceutical ingredient is (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A).

19. Use of a pharmaceutical composition according to claim 1 comprising a therapeutically effective amount of the active pharmaceutical ingredient, (+) trans-2- (2-chloro-phenyl)-5,7-dihydroxy-8-(2-hydroxymethyl-1 -methyl-pyrrolidin-3-yl)- chromen-4-one, or a pharmaceutically acceptable salt thereof, for the treatment of a disease or disorder, wherein the disease or disorder selected from cancer, polycystic kidney disease, nephrological disorder, psoriasis, immunological disorder involving unwanted proliferation of leukocytes, restenosis, proliferative smooth muscle disorder, radiation induced mucositis, viral infection, mycotic infection or cardiovascular abnormality.

20. The use according to claim 19, wherein the disease or disorder is selected from cancer, polycystic kidney disease, radiation induced mucositis or viral infection.

21 . The use according to claim 19 or claim 20, wherein the pharmaceutical composition is a solid oral capsule dosage form.

22. The use according to any one of the claims 19-21 , wherein the active pharmaceutical ingredient is (+) trans-2-(2-chloro-phenyl)-5,7-dihydroxy-8-(2- hydroxymethyl-1 -methyl-pyrrolidin-3-yl)-chromen-4-one hydrochloride (compound A).