FIELD OF INVENTION
This invention relates to novel (1E,6E)-1-(4-(2-([1,1'-biphenyl]-4-yl)-2-oxoethoxy)-3-methoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione, its synthesis and uses thereof, covering anti-polycystic ovary syndrome (PCOS).
BACKGROUND AND PRIOR ART
Poly-Cystic Ovarian Syndrome (PCOS) is common amongst adolescent and pre-menopausal women, affecting 4-20% of reproductive women globally [Deswal R, Narwal V, Dang A, Pundir CS. The prevalence of polycystic ovary syndrome: a brief systematic review. Journal of Human Reproductive Sciences. 2020 Oct;13(4):261.]. Luteinizing hormone (LH)/Follicle-stimulating hormone (FSH) imbalance due to abnormally frequent pulsatile stimulation of the hypothalamus, and consequent surge in androgen levels due to over-secretion of gonadotropin-releasing hormone (GnRH) in the women’s triggers polycystic ovary syndrome (PCOS). Hirsutism, acne, insulin resistance, irregular menses, and obesity are the characteristic symptoms of PCOS [Balen A. The pathophysiology of polycystic ovary syndrome: trying to understand PCOS and its endocrinology. Best practice & research clinical obstetrics & gynaecology. 2004 Oct 1;18(5):685-706.]. Biochemically, PCOS is typically characterized by elevated testosterone levels, accompanied by diminished blood progesterone levels. The current mainstay treatment for PCOS and its complications include oral contraceptives (estrogen and progesterone-based), antihyperglycemic drugs (metformin and gliptins), temporary symptomatic relief by androgen inhibitors (finasteride, spironolactone), eflornithine cream (for hirsutism) and GnRH agonists to restore LH-FSH balance. However, none of these treatments can be long-term, due to their severe adverse effects, including impotency, alopecia, gynecomastia, etc. Also, steroidal treatment has severe hormonal adverse effects [Witchel SF, Oberfield SE, Peña AS. Polycystic ovary syndrome: pathophysiology, presentation, and treatment with emphasis on adolescent girls. Journal of the Endocrine Society. 2019 Aug;3(8):1545-73.; Poirier D. Inhibitors of 17ß-hydroxysteroid dehydrogenases. Current medicinal chemistry. 2003 Mar 1;10(6):453-77.]. Thus, medicinal chemistry research in this field is consistently behind the search for newer chemotherapeutic agents.
It is evident from the literature that (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione, also named as (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione, possesses beneficial properties for the treatment of the polycystic ovarian syndrome. (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione in its nano-particulate form has proven to be a remedial treatment against PCOS [Raja MA, Maldonado M, Chen J, Zhong Y, Gu J. Development and evaluation of curcumin encapsulated self-assembled nanoparticles as potential remedial treatment for PCOS in a female rat model. International Journal of Nanomedicine. 2021;16:6231.]. Also, (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione has positive effects on various diseases associated with PCOS such as increased body weight, hyperglycaemia and hyperlipidaemia. (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione is experimentally proven to reduce increased body weight in PCOS patients. Also, PCOS-associated hyperglycaemia and hyperlipidaemia are found to be readily mitigated after the treatment by (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione [Jamilian M, Foroozanfard F, Kavossian E, Aghadavod E, Shafabakhsh R, Hoseini A, Asemi Z. Effects of curcumin on body weight, glycemic control and serum lipids in women with polycystic ovary syndrome: A randomized, double-blind, placebo-controlled trial. Clinical nutrition ESPEN. 2020 Apr 1;36:128-33.].
Other than PCOS, (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione is also beneficial in various types of cancer, including hormone-dependent cancer such as breast and ovarian cancer. (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione is investigated and successfully established to be a potent Nuclear Factor-kB inhibitor, to quench the progression of ovarian cancer, and associated angiogenesis [Lin YG, Kunnumakkara AB, Nair A, Merritt WM, Han LY, Armaiz-Pena GN, Kamat AA, Spannuth WA, Gershenson DM, Lutgendorf SK, Aggarwal BB, Sood AK. Curcumin InhibitsTumor Growth and Angiogenesis in Ovarian Carcinoma by Targeting the Nuclear Factor-KB Pathway. Clinical Cancer Research. 2007 Jun 1; 13;11: 3423-3430]. Another mechanism through which (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione acts as an anticancer agent is via the arrest of the G2/M phase of the cell cycle in tumour cells. It modulates the AkT and p38 MAPK proteins to elicit its anticancer activity [Weir NM, Selvendiran K, Kutala VK, Tong L, Vishwanath S, Rajaram M, Tridandapani S, Anant S, Kuppusamy P. Curcumin induces G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by modulating Akt and p38 MAPK. Cancer biology & therapy. 2007 Feb 1;6(2):178-84.].
Thus, the versatile therapeutic utility of (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione is well-established from past research. However, it still possesses few challenges from the pharmacokinetic point of view due to its poor bioavailability, that is by the virtue of its inability to readily cross the biological membrane [Dei Cas M, Ghidoni R. Dietary curcumin: correlation between bioavailability and health potential. Nutrients. 2019 Sep 8;11(9):2147.]. Hence, the development of (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione by the application of synthetic and medicinal chemistry to enhance its pharmacokinetic properties, thereby increasing lipophilicity and thus, bioavailability can help tackle this problem. The introduction of biphenyl moiety through a linker on (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione has helped us to increase its therapeutic potential against polycystic ovarian syndrome as well as cancer.
Therefore, it is an objective of the invention to synthesize novel (1E,6E)-1-(4-(2-([1,1'-biphenyl]-4-yl)-2-oxoethoxy)-3-methoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione as anti-PCOS agent, which has activity in the low micromolar ranges and uses thereof.
SUMMARY OF THE INVENTION
Compounds having formula S, its method of synthesis and uses thereof are described herein.

(S)
wherein;
Unsubstituted/substituted biphenyl moiety is linked to hydroxyl group of (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione, with an ethereal linkage bridged with ethanone linker;
R1 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R2 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R3 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
X1 and X2, Y1 and Y2, Z1 and Z2 independently represent hydrogen or together form a chemical bond.
The method of synthesis of compounds of formula (S) comprises at least one of the following reactions;
i. Synthesis of 1-(1, 1’-biphenyl]-4-yl)-2-chloroethan-1-one (III) by condensation of biphenyl (I) with chloroacetyl chloride (II), in presence of anhydrous aluminium chloride in carbon disulfide (step i, Scheme 1)
ii. Synthesis of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) by condensation of (III) with (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (IV), in the presence of base anhydrous K2CO3 in solvent acetone (step ii, Scheme 1)
The process is shown in Scheme 1 below:

In another aspect, the compound of formula (S) of the present invention comprise;
1. (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa)
DESCRIPTION OF FIGURES
Fig 1 depicts histomicrograph of the ovary showing (a) Ovaries in control group (CON); (b) Polycystic ovaries (PCOS); (c) Effect on polycystic ovaries on treatment with Sa (PCOS+CA); and effect on polycystic ovaries on treatment with nanoconjugates of Sa on PCOS induced rats (PCOS+NCA). In PCOS condition, number of cysts increases confirming the pathology of the condition. 1E,6E)-1-(4-(2-([1,1'-biphenyl]-4-yl)-2-oxoethoxy)-3-methoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) showed restoration of normal follicles but the number of normal follicles increased on administration of nano-conjugates. NF=Normal follicle; DF=Developing follicle; GF= Graffian follicle.

DETAILED DESCRIPTION OF THE INVENTION
Throughout the disclosure the following terms, unless otherwise indicated, shall be understood to have the following meanings:
In the present specification, the term polycystic ovary syndrome (PCOS) is a diseased condition in reproductive women characterized by the development of multiple cysts in ovaries due to excess of androgens.
Luteinizing hormone (LH) is a reproductive hormone responsible for stimulation of the process of ovulation in the menstrual cycle in women.
Follicle-stimulating hormone (FSH) is a reproductive hormone that stimulates ovaries to produce eggs.
Gonadotropin-releasing hormone (GnRH) is a hormone released by hypothalamus of the brain responsible for stimulation of release of various reproductive hormones such as FSH, LH and Progesterone.
Hirsutism is a diseased condition in women characterized by excessive hair growth in a male-like pattern, such as on face, chest and back.
Acne is a skin condition that arises due to clogging of the skin pores by hair, bacteria, dirt, dead skin, etc.
Insulin resistance is a diseased condition wherein the muscles and liver do not respond well to insulin produced by the body.
Irregular menses is reffered to a condition wherein the menstrual bleeding occurs before or after the expected time according to normal cycle.
Obesity is referred to excessive fat in body, leading to body overweight.
Testosterone is a male reproductive hormone, responsible for the development of secondary sex characters in male.
Progesterone is a pregnancy maintaining hormone in female body.
Estrogen is a female reproductive hormone, responsible for the development of secondary sex characters in female.
Metformin is an antihyperglycemic drug administered to control blood glucose levels in order to control elevated glucose in diabetic condition.
Gliptins is a class of antidiabetic drugs intended to reduce elevated blood glucose levels.
Finasteride is an anti-androgenic drug that inhibits the biosynthesis of androgens by inhibiting 5?-reductase enzyme.
Spironolactone is an aldosterone receptor antagonist.
Curcumin is a yellow coloured phytoconstitutent found in turmeric.
Hyperglycaemia means abnormally elevated blood glucose levels.
Hyperlipidaemia means abnormally elevated blood triglyceride levels.
G2/M phase is an intermediate stage in the life of a cell characterized by rapid cell growth
Cell cycle is the complete lifecycle of a cell from cell division to the development of a daughter cell for next cell division.
AkT is a serine threonine kinase enzyme responsible for stimulation of cell growth and proliferation.
p38 MAPK is a p38 mitogen activated protein kinase enzyme responsible for many cellular processes including inflammation, cell differentiation and death.
Cancer is pathological condition characterized by uncontrolled proliferation of cells.
Nuclear Factor-kB inhibitor is transcription regulator that is responsible for a cell’s immunie response and survival.
Angiogenesis is the process of development of new blood vessels around a tumour to facilitate blood supply to the abnormally growing tumour cells.
Letrozole is a class of drugs that inhibits aromatase enzyme to manage estrogen dependent diseases such as breast cancer.
The compounds of formula (S) may contain asymmetric carbon atom and some of the compounds of this invention may contain one or more asymmetric centres and may thus give rise to stereoisomers and diastereomers. Although the compound of formula (S) is not depicted with any stereo centres, the present invention includes such stereoisomers and diastereomers, as well as the racemic and resolved, enantiomerically pure R and S stereoisomers, as well as other mixture of the R and S stereoisomers. The compound of formula (S) may exhibit geometrical isomerism including cis-trans and Z-E forms. So, the present invention comprises the individual geometrical isomers and stereoisomers.
Such isomers or diastereomers can be separated from their mixtures, by the application or adaptation of known methods, for example chromatographic techniques and recrystallization techniques, or they are separately prepared from the appropriate isomers of their intermediates. All forms are within the scope of the invention.
The present invention relates to synthesis of novel (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione of formula (S) or its pharmaceutically acceptable salts, solvates, isomers, enantiomers, any sort of formulation, and uses thereof;

(S)
wherein;
Unsubstituted/substituted biphenyl moiety is linked to hydroxyl group of (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione, with an ethereal linkage bridged with ethanone linker;
R1 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R2 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R3 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
X1 and X2, Y1 and Y2, Z1 and Z2 independently represent hydrogen or together form a chemical bond.
The compounds of general formula (S) comprises;
Table 1: Chemical structure and IUPAC name of compound Sa
Comp. No. Chemical Structure IUPAC name
Sa
(1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione

In another embodiment, the present invention discloses a process of making of novel (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione of formula (S) which comprises at least one of the following reactions;
i. Synthesis of 1-(1, 1’-biphenyl]-4-yl)-2-chloroethan-1-one (III) by condensation of biphenyl (I) with chloroacetyl chloride (II), in presence of anhydrous aluminium chloride in carbon disulfide (step i, Scheme 1)
ii. Synthesis of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) by condensation of (III) with (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (IV), in the presence of base anhydrous K2CO3 in solvent acetone (step ii, Scheme 1)
iii. Preparation of nano-conjugates of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) by solvent evaporation method, using dichloromethane as a common solvent for Sa and polymer, carboxymethyl cellulose (CMC)
In step (i), an accurate quantity of biphenyl (I) (1.54 g; 0.01 mol) was weighed and placed in a 250 mL triple neck round bottom flask, 30 mL of carbon disulfide (CS2) was added to the above flask and stirred continuously to form a clear solution. To the above reaction mixture finely powdered aluminium chloride (AlCl3; 1.33 g; 0.01 mol) was added with continuous stirring. Chloroacetyl chloride (II) (0.8 ml; 0.01 mol) was added to the above reaction mixture dropwise drop using dropping funnel. The reaction was continued for 6 h at room temperature with continuous stirring. TLC was used to monitor the reaction. After completion of the reaction, 5% hydrochloric acid (200 mL) was added to quench the reaction and the compound was filtered under vacuum and washed with water to remove traces of hydrochloric acid. The residue thus obtained was recrystallized using ethanol to yield yellow crystals of 1-(1, 1’-biphenyl) -4yl) -2-chloroethan-1-one (III).
In the step (ii), a mixture of 1-(1, 1’-biphenyl]-4-yl)-2-chloroethan-1-one (III) (2.3 g, 0.01 mol), (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (3.68 g, 0.01 mol), anhydrous potassium carbonate (K2CO3) (2.76 g, 0.02 mol) was taken in acetone (50 mL) in 250 mL round bottom flask and refluxed for 12 h. TLC was used to monitor the reaction. The reaction mixture was filtered while hot and the solvent was evaporated under reduced pressure to obtain the residue. The residue was mixed with crushed ice with continuous stirring. The precipitated product was filtered, washed with distilled water, dried and recrystallized using ethanol to yield pure compound (Sa).
In the step (iii), nano-conjugates of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) were prepared using the solvent evaporation method. Dichloromethane was selected as a common solvent for Sa as well as the polymer. Both the components were dissolved in the solvent in equal proportions and evaporated under reduced pressure to yield the nano-conjugates of Sa.
The pharmaceutical composition of compounds of general formula (S) or its pharmaceutically acceptable salts, their isomers, enantiomers, solvates along with pharmaceutically employed excipients are administered in an effective therapeutic amount to a subject in need either orally or parenterally.
The active compound of general formula (S) will be present in such pharmaceutical compositions in amounts sufficient to provide the desired dosage amount. The compounds of general formula (S), for oral administration, can be combined with a suitable solid or liquid carrier or diluent and other pharmaceutically acceptable excipients to form capsules, tablets, powders, syrups, solutions, suspensions and the like. For parenteral administration, the compounds can be combined with sterile aqueous or organic media to form injectable solutions.
In another embodiment, the present invention provides pharmaceutical composition comprising the compounds of general formula (S) or its pharmaceutically acceptable salts, their isomers, enantiomers, solvates along with pharmaceutically employed excipients which may be useful in the treatment of PCOS, ovarian cancer, breast cancer, or other forms of cancer. The composition can be formulated as tablets, capsules, powders, syrups, solutions, suspensions and such like or as parenteral injectable.
In another embodiment, the in vivo anti-PCOS activity of compound (Sa) was studied in Letrozole induced PCOS rats. Oral administration of compound (Sa) (nano-conjugates) exhibited significant reduction in ovarian cysts compared to control group.
In yet another embodiment, the histopathological view of PCOS ovaries was observed (Fig 1). In control rats, prominent cysts were observed during the 21 days’ treatment period, which could be attributed to PCOS. Letrozole treatment resulted in ovarian cysts. Compound Sa showed reduced bulk of ovarian cysts, which indicated a beneficial effect of the treatment of Sa, compared to letrozole treatment.

The following example, which includes preferred embodiments, will serve to illustrate the practice of this invention, it being understood that the particulars shown are by way of example and for purpose of illustrative discussion of preferred embodiments of the invention.
Example 1: 1-(1, 1’-Biphenyl]-4-yl)-2-chloroethan-1-one (III)
An accurately weighed quantity of biphenyl (I) (1.54 g; 0.01 mol) was placed in a 250 mL triple neck round bottom flask, 30 mL of carbon disulfide (CS2) was added to it and stirred continuously to form a clear solution. To the above reaction mixture, finely powdered aluminum chloride (AlCl3;1.33 g; 0.01 mol) was added with continuous stirring. Chloroacetyl chloride (II) (0.8 ml; 0.01 mol) was further added dropwise using dropping funnel. The reaction was continued for 6 h at room temperature with continuous stirring. The reaction was monitored using thin layer chromatography. After completion of the reaction, 5% hydrochloric acid (200 mL) was added to quench the reaction and the compound was filtered under vacuum and washed with water to remove traces of hydrochloric acid. The residue thus obtained was recrystallized using ethanol to yield yellow crystals of 1-(1, 1’-Biphenyl) -4yl)-2-chloroethan-1-one (III).

Example 2: (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa)
A mixture of 1-(1, 1’-biphenyl]-4-yl)-2-chloroethan-1-one (III) (2.3 g, 0.01 mol), (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (3.68 g, 0.01 mol), anhydrous potassium carbonate (K2CO3) (2.76 g, 0.02 mol) was taken in acetone (50 mL) in 250 mL round bottom flask and refluxed for 12 h. TLC was used to monitor the reaction. The reaction mixture was filtered while hot and the solvent was evaporated under reduced pressure to obtain the residue. The residue was mixed with crushed ice with continuous stirring. The precipitated product was filtered, washed with distilled water, dried and recrystallized using ethanol to yield pure compound (Sa).

Example 3: Nano-formulation of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa)
Accurately weighed quantities of Sa and selected polymer, Carboxymethyl cellulose were taken in equal proportions and dissolved in a common solvent, dichloromethane. The solution was further subjected to solvent evaporation under reduced pressure to yield nanoparticles of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa)

BIOLOGICAL EVALUATION
In vivo Anti-PCOS Activity:
The anti-PCOS activity of compound Sa was determined using Letrozole induced PCOS rat model. Letrozole was taken as reference standard. PCOS was induced in female wistar rats by 1mg/kg oral dose of letrozole to all rats for 21 days. Induction of PCOS was confirmed by increased body weight and vaginal smear examination. Histopathological examination showed the presence of ovarian cysts.
Histopathology of polycystic ovaries of rats:
The animals were sacrificed after the completion of studies and their ovaries were subjected to histopathological studies, which revealed that nano-conjugates of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) showed excellent anti-PCOS activity, as compared to compound Sa alone.

We claim;

1. (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) as anti-PCOS agent with general formula (S) or its pharmaceutically acceptable salts, solvates, isomers, enantiomers, any sort of formulation, and uses thereof;

(S)
wherein;
Unsubstituted/substituted biphenyl moiety is linked to hydroxyl group of (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione, with an ethereal linkage bridged with ethanone linker;
R1 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R2 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R3 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
X1 and X2, Y1 and Y2, Z1 and Z2 independently represent hydrogen or together form a chemical bond.
2. The compounds of general formula (Sa) according to claim 1 comprises;
(1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa).
3. The process for preparation of unsubstituted/substituted (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione of general formula (S) or its pharmaceutically acceptable salts, solvates, isomers, enantiomers thereof according to claim 1

(S)
wherein;
Unsubstituted/substituted biphenyl moiety is linked to hydroxyl group of (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione, with an ethereal linkage bridged with ethanone linker;
R1 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R2 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
R3 is independently selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH) or their salts; substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones;
X1 and X2, Y1 and Y2, Z1 and Z2 independently represent hydrogen or together form a chemical bond.
The compounds of general formula (S) according to claim 1 comprises; (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa);
Comprising at least one of the following reactions (step i, ii and iii);
i. Synthesis of 1-(1, 1’-biphenyl]-4-yl)-2-chloroethan-1-one (III) by condensation of biphenyl (I) with chloroacetyl chloride (II), in presence of anhydrous aluminium chloride in carbon disulfide (step i, Scheme 1)
ii. Synthesis of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) by condensation of (III) with (1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (IV), in the presence of base anhydrous K2CO3 in solvent acetone (step ii, Scheme 1)
iii. Preparation of nano-conjugates of (1E,6E)-1-(4-(2-(biphenyl-4-yl)-2-oxoethoxy)-3 methoxy phenyl)-7-(4-hydroxy-3-methoxyphenyl)hepta-1,6-diene-3,5-dione (Sa) by solvent evaporation method, using dichloromethane as a common solvent for Sa and polymer, carboxymethyl cellulose (CMC)
4. The process according to claim 3, wherein the substitution at R3 of compound having general formula (S) in step (ii) is selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH); substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones.
5. The process according to claim 3, wherein the substitution at R2 is selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g.,-CH3); (un)-substituted or substituted linear or branched aryl; (un)-substituted or substituted heteroaryl; (un)-substituted or substituted alkylaryl; (un)-substituted or substituted alkylheteroaryl; (un)-substituted or substituted –CN; (un)-substituted or substituted -SO3H; (un)-substituted or substituted amines; (un)-substituted or substituted nitro or nitrites or nitrates; (un)-substituted or substituted esters or acids and the like.
6. The process according to claim 3, wherein the substitution at R3 of compound having general formula (S) in step (ii) is selected from hydrogen; (un)-substituted or substituted linear or branched alkyl (e.g., -CH3, -C2H5, C3H7, C4H9, -CH2-CH2-Br, -CH2-CH2-CH2-Cl, -CH2-CH2-CH2-Br); (un)-substituted or substituted aryl or (un)-substituted or substituted alkylaryl (e.g., -CH2-C6H5, -CH2-p-Cl-C6H4); substituted or (un)-substituted, linear, branched hetero or cyclic alkyl, alkenyl, or alkynyl; substituted or (un)-substituted aryl or heteroaryl, halogen, substituted or (un)-substituted alkoxy; substituted or (un)-substituted hydroxyl; substituted or (un)-substituted cyano; substituted or (un)-substituted formyl; substituted or (un)-substituted acyl; substituted or (un)-substituted carboxylic acid (e.g., -CH2-COOH); substituted or (un)-substituted carboxylate; substituted or (un)-substituted primary or secondary or tertiary amide; substituted or (un)-substituted secondary or tertiary carbamate; substituted or (un)-substituted urea; substituted or (un)-substituted carbine; substituted or (un)-substituted ester (e.g., -COOC2H5, -CH2-COOC2H5); substituted or (un)-substituted thiol; substituted or (un)-substituted primary or secondary or tertiary amine; substituted or (un)-substituted thioether; substituted or (un)-substituted sulfinyl group; and substituted or (un)-substituted sulfonyl group; substituted or (un)-substituted ketones.
7. The process according to claim 3, wherein base in steps (ii) is selected from alkali or alkaline earth metal carbonates.
8. The process according to claim 3, wherein the solvent in step (ii) is selected from glacial acetic acid, lower alcohols, anhydrides, aldehydes, ketones, polar aprotic solvent such as acetonitrile THF, acetone, DMF and the like.
9. The process according to claim 3, wherein base in steps (ii) was performed between (Sa) and substituted/unsubstituted alkyl/aryl halide.
10. The process according to claim 3, wherein X1 and X2, Y1 and Y2, Z1 and Z2 independently represent hydrogen or together form a chemical bond.