The present invention is directed to anti-progestational agents of compound of general formula (I) and process for the preparation thereof. The present invention is further directed to pharmaceutical composition thereof and method of treatment using the same.

BACKGROUND OF THE INVENTION

Steroids with antihormonal activity have been explored in past few decades with some success. However, there are various road blocks that a steroid chemist faces when developing anti-progestational agents.

Anti-progestational agents have been explored with great interest due to its vast ranging therapeutic activities. Anti-progestational agents have been explored in treatment of utrine fibroids, endometritis and as contraceptive agents.

Various anti-progestational agents have been discovered having therapeutic potential. Mifepristone is the first steroid of this type known for potential as contraceptive agent. Ulipristal acetate is a synthetic selective progesterone receptor modulator, is the first selective progesterone modulator known for therapeutic potential for uterine fibroids. It was initially approved for the preoperative treatment of moderate to severe uterine fibroid symptoms in women of reproductive age.

Despite clinical success, many anti-progestational agents have been lately known to cause severe toxicities like hepatotoxicity amongst other side effects which have raised serious questions regarding safety of these agents. Therefore, there is a need to explore novel anti- progestational agents having desired safety and dosing profile.

Dose reduction could provide reduction of side effects of the individual therapeutic compounds. Dose reduction could also improve patient compliance and enhance psychological impact. Therefore, there is a need to explore reduced dose novel compositions of prior art compounds.

The present inventors have surprisingly found novel anti-progestational agents of compound of general formula (I) which meets unmet medical needs of prior art compounds. SUMMARY OF THE INVENTION

In one aspect the present invention relates to anti-progestational agents of compound of general formula (I) and the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof comprising:

wherein, R is selected from the group comprising hydrogen, alkyl or optionally substituted aryl, heteroaryl, cycloalkyl.

In one aspect the present invention relates to anti-progestational agents of compound of general formula (I) and the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof.

wherein, R is selected from group comprising of: alkyl substituted with aryl, alkyl with

substituted heteroaryl or or wherein m and p are independently selected from 0 to 3, n refers to degree of polymerization and Z is alkyl or amine.

In another aspect the present invention relates to process for the preparation compound of general formula (I) comprising reacting compound of formula (II) with hydroxylamine salt of compound of formula (III).

wherein, R has same meaning as defined above and X represents counter-ion used in preparation of salt of hydroxylamine.

In one aspect the present invention relates to anti-progestational agents of compound of general formula (I-a) and the geometric isomers, pharmaceutically acceptable salts and solvates thereof.

In another aspect the present invention relates to process for the preparation compounds of general formula (I-a) comprising reacting compound of formula (Il-a) with hydroxylamine salt of compound of formula (III- a). wherein, X represents counter-ion used in preparation of salt of hydroxylamine.

In one aspect the present invention relates to pharmaceutical composition comprising anti-progestational agents of compounds of general formula (I) or anti-progestational agents of compound of formula (I- a) and pharmaceutically acceptable excipients.

In one aspect the present invention relates to method of treating a disorder or a condition selected from group comprising of: utrine fibroids, endometritis, uterine leiomyoma, meningioma, dysmenorrhea, metastatic breast cancer, contraception, emergency postcoital contraceptive, inducement of cervical ripening in a mammal, including a human, comprising administering to a mammal in need of such treatment an amount of a compound of general formula (I) or compound of formula (I-a) or the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof, that is effective in treating such disorder or condition.

In another aspect the present invention relates to use of anti-progestational agents of compound of general formula (I) or anti-progestational agents of compound of formula (I-a) in contraception, emergency postcoital contraceptive, inducement of cervical ripening, inducement of labor.

In another aspect the present invention relates to administration of anti-progestational agents of compound of general formula (I) or anti-progestational agents of compound of formula (I-a) of the present invention through an intrauterine device.

In another aspect the present invention relates to treatment of utrine fibroids, menses regulation, endometritis, uterine leiomyoma, dysmenorrhea by administration of the anti- progestational agents of compound of general formula (I) or compound of formula (I-a) of the present invention, leuprolide, elagolix, ulipristal acetate, vilaprisan, relugolix or fezolinetant through an intrauterine device. In another aspect the present invention relates to method of administration of the reduced dose of leuprolide, elagolix, ulipristal acetate, vilaprisan, relugolix or fezolinetant through an intrauterine device.

DETAILED DESCRIPTION OF THE INVENTION

It is to be understood that the invention is not limited in its application to the details set forth in the following description or exemplified by the examples. The invention is capable of other embodiments or of being practiced or carried out in various ways. Also, it is to be understood that the phraseology and terminology employed herein is for the purpose of description and should not be regarded as limiting.

The term “salt” or “pharmaceutically acceptable salt” as used herein refers to pharmaceutically acceptable acid addition salts formed with organic or inorganic acids. Exemplary of such organic salts are those but not limited to maleic, fumaric, benzoic, ascorbic, succinic, oxalic, bis-methylenesalicylic, methanesulfonic, ethanedisulfonic, acetic, propionic, tartaric, salicylic, citric, gluconic, lactic, malic, mandelic, cinnamic, citraconic, aspartic, stearic, palmitic, itaconic, glycolic, p-toulenesulfonic acid, p-aminobenzoic, glutamic, benzenesulfonic, theophylline acetic acids and the like. Exemplary of such inorganic salts are those but not limited to hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric acids and the like. Preferred acid addition salt is hydrochloride salt.

The term“alkyl,” as used herein refers to a straight or branched chain, or cyclic hydrocarbon radical, or combination thereof, which may be fully saturated, mono- or polyunsaturated and can include di- and multivalent radicals, having the number of carbon atoms designated (i.e. Ci-Ciomeans one to ten carbons). Examples of saturated hydrocarbon radicals include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, homologs and isomers of, for example, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group is one having one or more double bonds or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4- pentadienyl, 3-(l,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs and isomers.

The term“optionally substituted aryl” as used herein refers to aryl compounds having zero, one, two, three or four substituents, and a substituted aryl includes aryl compounds having one, two, three or four substituents, wherein the substituents include groups such as, for example, alkyl, halo, amino substituents or the like.

The term “optionally substituted heteroaryl” as used herein refers to a heteroaryl compounds having zero, one, two, three or four substituents, and a substituted heteroaryl includes heteroaryl compounds having one, two, three or four substituents, wherein the substituents include groups such as, for example, alkyl, halo, amino substituents or the like.

The term “optionally substituted cycloalkyl” as used herein refers to a cycloalkyl compounds having zero, one, two, three or four substituents, and a substituted cycloalkyl includes cycloalkyl compounds having one, two, three or four substituents, wherein the substituents include groups such as, for example, alkyl, halo, amino substituents or the like.

The term“pharmaceutically acceptable excipient” as used herein includes vehicles, adjuvants, or diluents or other auxiliary substances, such as those conventional in the art, which are readily available to the public. For example, pharmaceutically acceptable excipients include pH adjusting and buffering agents, tonicity adjusting agents, stabilizers, wetting agents and the like.

The term“intrauterine device” or“IUD” as used herein includes a device comprising a substantially C-, omega-, or triangular shaped device which is adapted to lie in the uterine cavity.

The term“reduced dose” as used herein refers to a dose of the respective drug that is lower, e.g. by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% (or any range in between these doses) lower than the usual approved dose of the same drug administered in the same or similar setting, i.e. in the same or similar patient groups with the same or similar treatment(s). The usual dose may be the dose that has frequently been used in the past and/or is mainly used in the same or similar setting, i.e. in the same or similar patient groups with the same or similar treatment(s).

The term "base" as used herein includes but not limited to inorganic base such as ammonia or hydroxide, carbonate, or bicarbonate of a metal cation or ammonia or organic bases such as organic primary, secondary, or tertiary amine. The base may be chosen as appropriate depending on various reaction conditions known to those skilled in the art.

The term "conventional techniques" as used herein includes but not limited to distillation, distillation under reduced pressure or vacuum, evaporation, solvent, anti-solvent, spray drying, lyophilization or freeze drying. The term "organic solvent" or "solvent" or“anti -solvent” as used herein includes but not limited to polar protic and aprotic solvents as well as non-polar solvents selected from water, hydrocarbons, ketones, alcohols, ethers, esters, halogenated solvents, dimethyl sulfoxide (DMSO) and dimethylformamide (DMF), pyridine, phenol, DMA, carbon disulphide, acetic acid, acetonitrile and mixtures thereof. Hydrocarbons include but not limited to such as benzene, toluene, xylene, pentane, hexane, heptane, cyclo hexane and tetraline. Ketones include but not limited to such as acetone, methyl ethyl ketone, cyclohexanone and methyl isobutyl ketone. Alcohols include but not limited to such as methanol, ethanol, propanol, butanol, octanol, ethanediol, 1, 2-propane diol and S (+)-l, 2-propane diol. Ethers include but not limited to such as diethyl ether, di isopropyl ether, di butyl ether, methyl tert-butyl ether, 1 ,4-dioxane, tetrahydrofuran and cyclo pentyl methyl ether. Halogenated solvents include but not limited to such as chloroform, carbon tetrachloride, methylene chloride and l,2-dichloro ethane. Esters include but not limited to such as methyl acetate, ethyl acetate, isopropyl acetate, n-butyl acetate and n-propyl acetate. The term“solvate” as used herein refers to a compound which contains a stoichiometric or non- stoichiometric amount of solvent bound by non-covalent intermolecular forces. When the solvent is water, hydrate is formed. The term "hydrate" as used herein refers to a compound which is formed by the union of water with the parent compound.

The starting material, compound of formula (II) and compound of formula (Il-a), used for the preparation of compound of general formula (I) and compound of formula (I-a) respectively, was prepared according to process disclosed in prior-art and known to person having ordinary skills in the art.

In one embodiment, the present invention is directed to anti-progestational agents of compound of general formula (I) and the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof.

wherein, R is selected from group comprising hydrogen, alkyl or optionally substituted aryl, heteroaryl, cycloalkyl.

In one embodiment, the present invention is directed to anti-progestational agents of compound of general formula (I) and the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof.

wherein, R is selected from group comprising of: alkyl substituted with aryl, alkyl with

substituted heteroaryl or or o wherein m and p are independently selected from 0 to 3, n refers to degree of polymerization and Z is alkyl or amine.

In another embodiment, the present invention is directed to anti-progestational agents and the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof comprising:

10 In another embodiment, the present invention is directed to process for the preparation compound of general formula (I) comprising reacting compound of formula (II) with hydroxylamine salt of compound of formula (III).

wherein, R has same meaning as defined above and X represents counter-ion used in preparation of salt of hydroxylamine.

In one embodiment, the present invention is directed to anti-progestational agents of compound of general formula (I-a) and the geometric isomers, pharmaceutically acceptable salts and solvates thereof.

In another embodiment, the present invention is directed to anti-progestational agents of compound of general formula (I-a) and the geometric isomers, pharmaceutically acceptable salts and solvates thereof wherein geometric isomers can be separated by techniques, such as high- performance liquid chromatography (HPLC), upercritical fluid chromatography, capillary electrochromatography (CEC), chiral chromatography or other alternative techniques known to person having ordinary skill in the art.

In another embodiment, the present invention is directed to process for the preparation compound of general formula (I-a) comprising reacting compound of formula (Il-a) with hydroxylamine salt of compound of formula (Ill-a).

wherein, X represents counter-ion used in preparation of salt of hydroxylamine.

In another embodiment, the process for the preparation of compound of general formula (I) or compound of formula (I-a) as referred herein can be carried out in presence of a base, selected from group comprising of sodium acetate, sodium hydroxide, potassium hydroxide or sodium methoxide. One preferred base is sodium acetate. In another embodiment, the process for the preparation of compound of general formula

(I) or compound of formula (I-a) or pharmaceutically acceptable salt as referred herein further comprised isolation of title compounds by conventional techniques.

In one embodiment, the present invention is directed to pharmaceutical composition comprising compound of general formula (I) or compound of formula (I-a) and optionally pharmaceutically acceptable excipients.

In one embodiment, the present invention is directed to method of treating a disorder or a condition selected from group comprising of: utrine fibroids, endometritis, uterine leiomyoma, meningioma, dysmenorrhea, metastatic breast cancer, contraception, emergency postcoital contraceptive, inducement of cervical ripening in a mammal, including a human, comprising administering to a mammal in need of such treatment an amount of a compound of general formula (I) or compound of formula (I-a) or the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof, that is effective in treating such disorder or condition

In one embodiment, the present invention is directed to use of compound of general formula (I) or compound of formula (I-a) in contraception, emergency postcoital contraceptive, inducement of cervical ripening, inducement of labor. In one embodiment, the present invention is directed to a method of administration of compound of general formula (I) or compound of formula (I-a) or the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof to reduce at least one toxicity associated with compound of formula (II) or compound of formula

(P- a).

In one embodiment, the present invention is directed to a method of administration of a compound of general formula (I) or compound of formula (I-a) or the geometric isomers, enantiomers, diastereomers, racemates, pharmaceutically acceptable salts and solvates thereof which is at least lower by 10% than dosage of compound of formula (II) or compound of formula (P- a).

The compound of general formula (I) or compound of formula (I-a) of the present invention may be administered by a variety of methods. Thus, compound of general formula (I) or compound of formula (I-a) are active by the oral route and may be administered in solutions, suspensions, emulsions, tablets, including sublingual and intrabuccal tablets, soft gelatin capsules, including solutions used in soft gelatin capsules, aqueous or oil suspensions, emulsions, pills, lozenges, troches, tablets, syrups or elixirs and the like. The compound of general formula (I) or compound of formula (I-a) are active by parenteral administration and may be administered by depot injection, implants including biodegradable implants, intramuscular and intravenous injections.

Compositions may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents. Tablets containing the active ingredient in admixture with nontoxic pharmaceutically acceptable excipients which are suitable for manufacture of tablets are acceptable. These excipients may be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as maize starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or may be coated by known techniques to delay disintegration and adsorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate alone or with a wax may be employed. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the invention contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include a suspending agent, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing or wetting agents such as a naturally occurring phosphatide (e.g., lecithin), a condensation product of an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate), a condensation product of ethylene oxide with a long chain aliphatic alcohol (e.g., heptadecaethylene oxycetanol), a condensation product of ethylene oxide with a partial ester derived from a fatty acid and a hexitol (e.g., polyoxyethylene sorbitol mono-oleate), or a condensation product of ethylene oxide with a partial ester derived from fatty acid and a hexitol anhydride (e.g., polyoxyethylene sorbitan mono-oleate). The aqueous suspension may also contain one or more preservatives such as ethyl or npropyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, such as sucrose, aspartame or saccharin. Ophthalmic formulations, as is known in the art, will be adjusted for osmotic pressure.

Oil suspensions may be formulated by suspending the active incredient in a vegetable oil, such as arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oil suspensions may contain a thickening agent, such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an antioxidant such as ascorbic acid.

Dispersible powders and granules of the invention suitable for preparation of an aqueous suspension by the addition of water may be formulated from the active ingredients in admixture with a dispersing, suspending and/or wetting agent, and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those disclosed above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present. The pharmaceutical compositions of the invention may also be in the form of oil-in- water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, a mineral oil, such as liquid paraffin, or a mixture of these. Suitable emulsifying agents include naturally- occurring gums, such as gum acacia and gum tragacanth, naturally occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan mono-oleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan mono-oleate. The emulsion may also contain sweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, such as glycerol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, such as a solution of l,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water and Ringer's solution, an isotonic sodium chloride. In addition, sterile fixed oils may conventionally be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may likewise be used in the preparation of injectables.

The compound of general formula (I) or compound of formula (I-a) of the present invention may also be administered in the form of suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable nonirritating excipient which is solid at ordinary temperatures but liquid at the rectal temperatures and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

The compound of general formula (I) or compound of formula (I-a) of the present invention may be administered by intranasal, intraocular, intravaginal, and intrarectal routes including suppositories, insufflation, powders and aerosol formulations.

The compound of general formula (I) or compound of formula (I-a) of the present invention may be administered by the topical route and may be administered as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, paints, powders, and aerosols. The compound of general formula (I) or compound of formula (I-a) of the present invention possess antiprogestational and/or antiglucocorticoid activity. Products having anti glucocorticoid activity are of particular value in pathological conditions characterized by excess endogenous glucocorticoid such as Cushing's syndrome, hirsutism and in particular when associated with the adrenogenital syndrome, ocular conditions associated with glucocorticoid excess such as glaucoma, stress symptoms associated with excess glucocorticoid secretion and the like. Products having progestational activity are of particular value as progestational agents, ovulation inhibitors, menses regulators, contraceptive agents, agents for synchronization of fertile periods in cattle, endometriosis, and the like. When used for contraceptive purposes, they may conveniently be admixed with estrogenic agents, such as for example as ethynylestradiol or estradiol esters. Products having anti-progestational activity are characterized by antagonizing the effects of progesterone. As such, they are of particular value in control of hormonal irregularities in the menstrual cycle and for synchronization of fertile periods in cattle.

The compound of general formula (I) or compound of formula (I-a) of the present invention may be used for control of fertility during the whole of the reproductive cycle. They are of particular value as postcoital contraceptives, for rendering the uterus inimical to implantation, and as "once a month" contraceptive agents. They may be used in conjunction with prostaglandins, oxytocics and the like.

A further important use for the compound of general formula (I) or compound of formula (I-a) of the present invention is in treatment of utrine fibroids. Utirine fibroids (leiomyomas or myomas) are extremely common benign neoplasms of uterus having prevalence exceeding 50% in premonopasusal women.

Another important use for the compound of general formula (I) or compound of formula (I-a) of the present invention lies in their ability to slow down growth of hormone -dependent cancers. Such cancers include kidney, breast, endometrial, ovarian cancers, and prostate cancer which are characterized by possessing progesterone receptors and may be expected to respond to the products of this invention. Other utilities of anti-progestational agents include treatment of fibrocystic disease of the breast. Certain cancers and in particular melanomas may respond favorably to corticoid/anticorticoid therapy.

The compound of general formula (I) or compound of formula (I-a) of the present invention may be administered to any warm-blooded mammal such as humans, domestic pets, and farm animals. The amount of active ingredient that may be combined with a carrier material to produce a single dosage form will vary depending upon the disease treated, the mammalian species, and the particular mode of administration. For example, a unit dose of the steroid may preferably contain between 0.1 milligram and 1 gram of the active ingredient. A more preferred unit dose is between 0.001 and 0.5 grams. It will be understood, however, that the specific dose level for any particular patient will depend on a variety of factors including the activity of the specific compound employed; the age, body weight, general health, sex and diet of the individual being treated; the time and route of administration; the rate of excretion; other drugs which have previously been administered; and the severity of the particular disease undergoing therapy, as is well understood by those of skill in the art.

The compound of general formula (I) or compound of formula (I-a) of the present invention can be administered through an intrauterine device (IUD). Administration through IUD can be advantageous in treatment of utrine fibroids, menses regulation, endometritis, uterine leiomyoma, dysmenorrhea amongst others. Administration through IUD can also provide symptomatic relief in utrine fibroids due to likelihood of less menstrual blood loss.

The intrauterine device as hereinbefore defined comprises, an inserter for the insertion of the IUD in the fundal region of the uterine cavity, an external slidable flange on the hollow tube extends substantially transversely thereof and drug reservoir amongst other parts.

The intrauterine device as hereinbefore defined can also be used in treatment of utrine fibroids, menses regulation, endometritis, uterine leiomyoma, dysmenorrhea by administration of the compound of general formula (I) or compound of formula (I-a) of the present invention, leuprolide, elagolix, ulipristal acetate, vilaprisan, relugolix or fezolinetant.

In another aspect the present invention relates to method of administration of the reduced dose of leuprolide, elagolix, ulipristal acetate, vilaprisan, relugolix or fezolinetant through an intrauterine device.

According to a still further aspect of the invention there is provided a method of insertion of an intrauterine device as hereinbefore defined using an inserter as hereinbefore defined, the method comprising straightening the device and inserting it into the hollow tube and then inserting the plunger into the tube, inserting the tube through the cervical canal into the uterus until the slidable flange rests against the cervical os, fully inserting the plunger into the tube to transfer the device into the uterine cavity, removing the plunger, drawing the ends of the device together, by exerting traction on a bead which connects the threads to the device, separating the bead from the threads, and then withdrawing the inserter and trimming the threads at the os cervix. Examples

The following examples are provided here to enable one skilled in the art to practice the invention and merely illustrate the process of the present invention. However, it is not intended in any way to limit the scope of the present invention. l-H NMR spectra are recorded at 400 MHz on a Brucker Avance-III HD. Dimethyl sulfoxide-d6 is used as solvent, and tetramethylsilane (TMS) is used as internal reference standard.

Mass spectra was recorded on a Waters SQD mass spectrometer equipped with an electrospray interface (LC-MS) connected with Acquity H-class system.

HPLC system (Agilent-l260 series (manufactured by Agilent Technologies) LC system, equipped with PDA detector, quaternary gradient pump, auto injector and Agilent OpenLab CDS (EZChrom Edition) Version A.04.07 and Build 04.07.28 (designed by Agilent Technologies) was used to determine purity of compounds.

Chromatographic Parameters:

Column: YMC TRIAT C18 (150*4.6) mm, 5m

Detector wavelength: 290 nm

Flow rate: 1.0 mL /min

Column oven temperature: 25 °C

Sample cooler temperature: Ambient

Injection volume: 10 pL

Run time: 14 min

Mobile Phase: (A) 0.1% Formic Acid in Water

(B) 0.1 % Formic Acid in Acetonitrile

Diluent: Water: Acetonitrile (1:1 v/v)

Elution: Gradient

Needle was I: Acetonitrile Example-1: Preparation of compound of formula (I-a)

Compound of formula (Il-a) (0.30 g, 0.00063 lmol, 1.0 equiv) was dissolved in acetic acid (3 ml) at room temperature under nitrogen atmosphere. Sodium acetate (0.160 g, 0.001956 mol, 3.1 equiv) and hydroxylamine-HCl (formula III-a, X= Cl ) (0.132 g, 0.0018923 mmol, 3.0 equiv) were added to the reaction mass at room temperature to get white suspension. The reaction mass was stirred at room temperature for 45 minutes. After completion of the reaction, the suspension was poured in to water (15 ml, 50 vol) and the suspension was stirred for 1 hour. The solid product was filtered, washed with water (5 ml x 2) and dried under vacuum at 40°C for 30 minutes to afford title compound (I-a) (0.135 g) as off-white solid (Purity: 99.18% by HPLC). Mass (m/z): 491.5 [M+H] 1H NMR (400 MHz, DMSO) d 10.650 (s, 0.58H), 10.363 (s, 0.31H), 7.015-6.994 (d, 2H), 6.646-6.624 (d, 2H), 6.392 (s, 0.33H), 5.83 (s, 0.0036H), 4.325-4.308 (d, 1H), 2.819 (s, 6H), 2.684-2.508 (s, 3H), 2.196-1.638 (m, 16H), 1.390-1.238 (m, 2H), 0.203- 0.190 (d, 3H).

Yield: 43.8%

Separation of mixture of E/Z isomer of compound of formula (I-a)

Compound of formula (I-a) (mixture of E and Z isomer) were separated using preparative HPLC. The fractions obtained were lyophilized to isolate the product.

Fraction- 1 was identified as Z-isomer and fraction-2 was identified as E-isomer.

PREP HPLC system (Schimadzu LC-20AP (manufactured by Schimadzu Corporation) LC system, equipped with UV detector (SPD-20A), binary gradient pump, manual injector and Lab solution software was used.

Chromatographic Parameters:

Column: SUNFIRE C18, (250 x 19 mm), 5 m

Detector wavelength: 202 and 285 nm

Flow rate: 14 mL /min

Column oven temperature: N/A

Auto sampler temperature: N/A

(Manual injector)

Injection volume: ~2 ml Run time: 70 minutes

Mobile Phase:

A) 0.1% FA + lOmM Ammonium acetate in water

B) 100% ACN

Elution: Gradient

Needle wash: Acetonitrile

Fraction-1 (Z-isomer):- (0.135 g, 36%)

LCMS: 97.48% m/z: 491.36 [M+l] ⁺

HPLC: 95.68%

¾ NMR (400 MHz, DMSO): d 10.374 (s, 1H), 7.013 (d, 2H), 6.647 (d, 2H), 6.401 (s, 1H), 4.33l(d, 1H), 2.845 (s, 5H), 2.691 - 2.754 (m, 3H), 2.171 - 2.272 (m, 2H), 2.119 (s, 4H), 1.847 - 2.009 (m, 7H), 1.677 (t, 2H),l.329(q, 2H), 0.218 (s, 3H).

Fraction-2 (E-isomer):- (0.209 g, 83.6%)

LCMS: 97.34% m/z: 491.41 [M+l] ⁺

HPLC: 96.80%

*H NMR (400 MHz, DMSO): d 10.668 (s, 1H), 7.018 (d, 2H), 6.649 (d, 2H), 5.847 (s, 1H), 4.329(d, 1H), 2.844 (s, 5H), 2.689 - 2.767 (m, 3H), 2.522 (s, 2H), 2.118 - 2.184 (m, 5H), 1.961 - 2.011 (m, 4H), 1.841 - 1.892 (m, 1H), 1.631 - 1.772 (m, 4H), 1.265 - 1.386 (m, 2H), 0.205 (s, 3H)

Chemical stability of E/Z isomer of compound of formula (I-a)

Study protocol:

Test item requirement: 100m L of 10hM DMSO stock

No. of replicates: One (Single)

Incubation time: 0, 15, 30, 60 and 120 Min

Incubation temperature: 37 °C

Test concentration: 5 mM

Buffer system: pH 1.2- Simulated Gastric Fluid (SGF) without enzyme

QC reference compound: ketoconazole

Analysis method: LCMS-MS (single analysis)

Deliverables: % parent compound remaining at each point and half life Study Results:

Example-2: Preparation of compound of formula (I-a)

Compound of formula (Il-a) (0.30 g) was dissolved in acetic acid (3 ml) at room temperature under nitrogen atmosphere. Sodium acetate (0.160 g, 3.1) and hydroxylamine-HCl (formula III- a, X= Cl ) (0.132 g) were added to the reaction mass at room temperature to get white suspension. The reaction mass was stirred at room temperature for 45 minutes. After completion of the reaction, the suspension was poured in to water (15 ml) and the suspension was stirred for 1 hour. The solid product was filtered, washed with water (5 ml x 2) and dried under vacuum at

40°C for 30 minutes to afford title compound (I-a) (0.156 g) as off-white solid.

Yield: 50.6%

Example-3: Preparation of compound of formula (I-a)

Compound of formula (Il-a) (0.30 g) was dissolved in methanol (5 ml) at room temperature under nitrogen atmosphere. Sodium methoxide (0.105 g) and hydroxylamine-HCl (formula Ill-a, X= Cl ) (0.132 g) were added to the reaction mass at room temperature to get white suspension. The reaction mass was stirred at room temperature for 45 minutes. After completion of the reaction, the suspension was poured in to water (15 ml) and the suspension was stirred for 1 hour. The solid product was filtered, washed with water (5 ml x 2) and dried under vacuum at 40°C for 30 minutes to afford title compound (I-a) (0.171 g) as off-white solid.

Yield: 55.5%

Example-4: Preparation of compound of formula (I-b)

Compound of formula (II) (0.35 g) was dissolved in acetic acid (5 ml) at room temperature under nitrogen atmosphere. Sodium acetate (0.185 g) and methylhydroxyl-amine-HCl (formula Ill-b, X= Cl ) (0.182 g) were added to the reaction mass at room temperature to get white suspension. The reaction mass was stirred at room temperature for 45 minutes. After completion of the reaction, the suspension was poured in to water (25 ml) and the suspension was stirred for 1 hour. The solid product was filtered, washed with water (5 ml x 2) and dried under vacuum at 40°C for 30 minutes to afford title compound (I-b) (0.169 g) as off-white solid.

Yield: 45.92%

Example-5: Preparation of compound of formula (I-c)

Compound of formula (II) (0.40 g) was dissolved in acetic acid (5 ml) at room temperature under nitrogen atmosphere. Sodium acetate (0.213 g) and benzylhydroxylamine-amine-HCl (formula III-c, X= Cl ) (0.402 g) were added to the reaction mass at room temperature to get white suspension. The reaction mass was stirred at room temperature for 45 minutes. After completion of the reaction, the suspension was poured in to water (30 ml) and the suspension was stirred for 1 hour. The solid product was filtered, washed with water (5 ml x 2) and dried under vacuum at 40°C for 30 minutes to afford title compound (I-c) (0.203 g) as off-white solid.

Yield: 41.6%