Synergetic compositions for women health

Abstract

The present invention relates to an oral extract composition comprising Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) extract composition, and its process of preparation. Also relates to the oral extract composition formulation comprising with synergistic combinations of the Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) in suitable doses and at least one nutraceutically and / or pharmaceutically accepted excipient for, improvement, and maintenance of women health more preferably in improving of hot flashes, night sweats, poor sleep, anxiety, sexual dysfunction, mood changes, Genitourinary symptoms in overall improving the quality of life, serum estradiol, progesterone, follicle stimulating hormone (FSH), and cortisol during the period of menopause (perimenopause and menopause). The oral extract composition formulation comprising with synergistic combinations of the Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) in suitable doses and at least one nutraceutically and / or pharmaceutically accepted excipient is used for improving Brain-derived neurotrophic factor (BDNF) levels in women during the period of menopause. The present invention, further related to the extraction process of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.), also provides the process for the preparation of formulation composition. This invention further relates to the extraction process from Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) either as individual herbs or co-extraction of the two herbs or mixing the extracts at various stages of the individual extracts.

Description

[0001] “SYNERGETIC COMPOSITIONS FOR WOMEN HEALTH”

[0002] HELD OF THE INVENTION

[0003]

[0001] The present invention relates to the oral nutritional extract composition comprising combinations of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.). The composition comprising Shatavari (Asparagus racemosus Willd.') and Fenugreek (Trigonella foenum-graecum L.) is used for women's health, more preferably during menopause. The above-mentioned combination surprisingly exhibited a synergistic effect and outperformed the individual constituents by over 50% w / w in measurable benefits like increased Brain Derived Neurotrophins. The invention further relates to the extraction process from Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.), either as individual herbs or co-extraction of the two herbs or mixing the extracts at various stages of the individual extracts. The oral nutritional extract composition of Shatavari (Asparagus racemosus Willd) and Fenugreek (Trigonella foenum-graecum L.) also relates to the formulation composition and the process for the preparation of the formulation composition with suitable oral doses to the subject in need thereof.

[0004] BACKGROUND OF THE INVENTION

[0005]

[0002] Menopause is a normal process in woman above 45 years of age. Menopause is the permanent cessation of menstruation due to loss of ovarian follicular function. Clinically, menopause is diagnosed after 12 months of amenorrhea, so the time of the final menses is determined retrospectively. A common initial indicator is the onset of menstrual irregularity. The biology underlying the transition to menopause includes central neuroendocrine changes as well as changes within the ovary, the most striking of which is a profound decline in follicle numbers.

[0006]

[0003] Perimenopause is period of changing ovarian function that precedes the final menses. During perimenopause, levels of estrogen, a key female hormone, start to decrease. During this period, menstrual cycles may get lengthen or shorten. During menopause, the intricate interplay of changing hormonal levels, specifically estrogen and progesterone, renders women more vulnerable to a spectrum of symptoms including hot flashes, sleep problems, vaginal dryness, urogenital infections, osteoporosis, early onset of coronary heart disease, anxiety, depression, diabetes, and cognitive difficulties etc.

[0004] Over recent years, many nutritional candidates have proved to be an effective remedy to handle such body changes. Such nutritional candidates include but not limited to Asparagus racemosus Willd (Shatavari), Trigonella foenum graecum (Methi), Bacopa monnieri (Brahmi), Berberis aristata (Indian barberry), Butea monosperma (Flame-of-the-forest), Oroxylum indicum (Indian trumpet tree), Aegle marmelos ((Wood apple), Cyperus rotundus (Java Grass), Sphaeranthus indicus (Gorakhmundi) etc.

[0007]

[0005] The main treatment for decreasing the menopause symptoms is hormone replacement therapy (HRT), which replaces hormones that are at low levels. Doses and routes of hormone replacement therapy vary by indication while complications of hormone replacement therapy depend on the dosage regimen used. Thus, it is of prime importance to use the right dose for improving the menopause symptoms such as hot flushes, brain fog, joint pains, mood swings and vaginal dryness.

[0008]

[0006] Shatavari or Asparagus racemosus contains compounds (phytoestrogens - steroidal saponins) that have structural similarity to estrogen and are known for their estrogen-like effects, which play a crucial role in alleviating the discomfort associated with menopause. Thus, this herb is one of the major health tonics and most popular Ayurvedic herbs to treat ailments of women during menopause period. In Ayurveda, Shatavari is known to predominantly balance the Pitta dosha followed by Vata dosha. Pitta dosha is the biological energy responsible for digestion and balancing or regulating all metabolic and hormonal activities in the human body. The traditional use of Shatavari highlights the benefits in managing women’s hormonal disorders and therefore making it a prime candidate for further research.

[0009]

[0007] Fenugreek or Trigonella foenum- graecum is another herb that contains phytoestrogen compounds that could modulate hormones without resulting in any harmful side effects. Fenugreek seeds also contain compounds like protodioscin, diosgenin, which have estrogen-like properties which help in regulating the hormonal balance.

[0010]

[0008] Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) both also possess anti-inflammatory properties attributed to the saponins and phytonutrients present therein. Asparagus provides a wide variety of antioxidant nutrients, including vitamin C, beta-carotene, vitamin E, and the minerals zinc, manganese, and selenium. Fenugreek helps in preventing oxidative stress.

[0011]

[0009] United States Publication No. 2007 / 0122494 Al discloses pharmaceutical or medicinal preparation comprising a mixture of herbs including Glycine max, Asparagus racemosus, Boerhavia diffusa, Withania somnifera, Saraca indica, and Coleus forskohlii for the treatment of hormone dependent conditions and circulatory conditions in the human body.

[0012]

[0010] United States Publication No. 2023 / 0089704 Al discloses fenugreek extract composition containing at least one of diosgenin, gitogenin, sarsasapogenin, dioscin, protodioscin, pseudoprotodioscin, apigenin or a derivative thereof, luteolin or a derivative thereof, nicotinic acid or a derivative thereof, and kaempferol. This reference further discloses a process for preparing the composition using alcohol extraction method.

[0013] [Oi l] Indian Publication No. 202341014559 discloses and ethanolic extract of Asparagus racemosus Willd root and its use in treatment of Polycystic ovary syndrome (PCOS).

[0014]

[0012] United States Patent Publication No. 8,461,102 B2 discloses compositions, and kits for treatment or prevention of symptoms of hormonal variation associated with menopause, surgery, anti-estrogen drugs and / or androgen deprivation therapy, such as hot flashes, night sweats, and insomnia. The pharmaceutical composition of this reference includes an effective amount of a pharmacologically active agent for treating or preventing symptoms of hormonal variation, wherein the agent binds to the NMDA receptor and thereby modifies the activity of dopamine and / or serotonin and at least one receptor selected from a serotonin type 2 A (5-HT2A) receptor and a dopamine type 2 (D2) receptor.

[0015]

[0013] Chinese Publication No. 101896189 A discloses edible composition for enhanced immunity comprising theanine or a source of theanine and herb selected from Shankhpushpi, Shatavari, or a mixture thereof.

[0016]

[0014] International Publication No. WO 2023091390 Al discloses herbal composition of Asparagus racemosus Willd and Withania somnifera. The composition of the invention is used for the treating postmenopausal women.

[0017]

[0015] United States Patent Publication No. 9,855,308 B2 discloses nutraceutical composition comprising extract of Trigonella foenum- graecum, at least one bioavailable form of magnesium and vitamin E useful for amelioration of peri- and post-menopausal symptoms in women. This reference also discloses the process for producing a nutraceutical composition by fractionating the fenugreek seeds for targeted extraction and enrichment of desired phytonutrients for delivering beneficial effect on postmenopausal women. The reference further elaborates on preparation of Fenugreek Husk Extract.

[0018]

[0016] Indian Publication No. 202021046021 discloses herbal composition for the management of polycystic ovarian syndrome (PCOS) wherein the herbal composition comprises powder extracts of Caesalpinia bonduc, Cissampelospareira, Tinospora cordifolia, Asparagus racemosus Willd, Anethum sowa, Zingiber officinale, Piper nigrum, Piper longum, Holarrhenaantidysentrica and Carum bulbocastanum.

[0019]

[0017] International Publication No. WO 2017007833 Al discloses healthful supplements including at least four nutrients selected from an amino acid blend, a coconut extract, a vegetable glycerin extract, an alcohol blend, a cannabinoid blend, a hemp plant extract, a Boswellia serrata extract, a curcuma / turmeric blend, black cumin seed, an Artemisia ludoviciana extract, an Astragalus extract, a fenugreek extract, a mushroom extract blend.

[0020]

[0018] International Publication No. WO 2016098120 Al discloses single herb extract of fenugreek seeds and its oral dosage form. The composition of the invention is used for the treatment of PCOS (Poly Cystic Ovarian Syndrome).

[0021]

[0019] Indian Publication No. 202011038468 discloses herbal composition comprising an extract or powder of Asparagus racemosus Willd, Withania somnifera, Pueraria tuberosa, Glycyrrhiza glabra, Moringa oleifera, Cissus quadrangularis, Bacopa monnerei, Saracaindica, Symplocosracemosa, Sugar and Glycine max. The composition of the invention is prepared for the treatment pre and / or post-menopausal conditions in women.

[0022]

[0020] International Publication No. WO 2008125120 A2 discloses an extract obtainable from Trigonella foenum-graecum and compositions comprising this extract. Another aspect of the present invention relates to the use of the extract of Trigonella foenum- graecum for or in the preparation of disinfactants or cleansing products.

[0023]

[0021] United States Patent Publication No. 8,754,205 B2 discloses an extract obtained by a commercially viable process for the extraction of furostanolic saponins from fenugreek seeds.

[0024]

[0022] United States Patent Publication No. 8,337,915 B2 discloses fenugreek seed extract composition for lowering blood cholesterol in a non-diabetic patient. This reference further provides a method for preparing such compositions. The compositions of this invention are advantageously used in methods for treating a non-diabetic individual having a disorder associated with high cholesterol.

[0025]

[0023] United States Publication No. 2006 / 0147561 Al discloses herbal nutraceutical formulation for diabetics, and its process for the preparation of the formulation. The formulation of this reference comprises the base product of a microwave roasted seed powder mixture from selected genera of Glycine, Cicer, Phaseolus, Cyamompsis, Mucuna, Hordeum, Amaranthus and Fagopyrum, fortified with herbs / medicinal plants selected from the genera of Gymnema, Momordica, Syzgium, Pterocarpus, Trigonella, Cinnamomum, Withania, Coccinia, Pueraria, Asparagus, Boerhaavia and Aegle.

[0026]

[0024] Chinese Patent Publication No. 101559157 B discloses method for fractional extraction of asparagus saponin and asparagus amylose using asparagus slag.

[0027]

[0025] Indian Publication No. IN 202111041937 discloses nutraceutical herbal supplement to support feminine health wherein the supplement comprises Foeniculum vulgare, Prunus and Asparagus racemosus Willd.

[0028]

[0026] International Publication No. WO 2006107745 A2 discloses herbal composition for the treatment of menopause wherein the composition is a mixture of herbs, an extract of a mixture of herbs or a mixture of herbal extracts. The herbal composition of this reference comprises a mixture of Herba Scutellaria Barbata, Radix Sophora Subprostratae, Radix Anamarrhena, Semen Glycine Sojae, Radix Glycyrrhiza, Rhizoma Rhei, Fructus Tritici Levis, Radix Astragali, Radix Rehmania, Fructus Ligustri Lucidi, Semen Zyziphi Spinozae, Plumula Nelumbif|is, Poria Cocos, Rhizoma Alismatis, Cortex Moutan Radicis, Fructus Corni, Radix Achyranthis, Concha Ostrea, Radix Aspargi, Radix Pueraria, Radix Atractylodis Macrocephala and Herba Epimedia or an extract of said mixture.

[0029]

[0027] In light of the available state of arts, not limiting to the above stated prior arts, there exists a need to develop the more potent composition, which is more synergetic and efficacious to benefit the women health, preferably during menopause.

[0030]

[0028] The present invention surprisingly provides the improved efficacy with synergetic effect due to the combined effect of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonellafoenum-graecum L.), resulting in a more effective improvement for hormonal balancing during menopause.

[0031]

[0029] Also, the present invention surprisingly provides an improved efficacy with synergetic effect due to the presence of preferred percentages of active constituents of Shatavari Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.), resulting in a more effective formula for improving hormonal balance during menopause.

[0032]

[0030] Thus, the required dosage to derive the full benefit of the combination is significantly lower than the individual extracts of the two herbs.

[0033]

[0031] The present invention also provides a series of flexible extraction processes to extract out the active constituents from Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonellafoenum-graecum L.

[0034]

[0032] The present invention further provides the formulation composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum) with suitable excipients and the process for the preparation thereof.

[0035] FIGURES OF THE INVENTION

[0036] Figure 1 : Serum Metabolic Profile

[0037] Figure 2 & 3: Effect of supplementation on Body and Tail temperature in ovariectomized rats

[0038] Figure 4: Neuropeptides that mediate the physiology of hot flashes

[0039] Figure 5: Effect on Serum BDNF levels

[0040] Figure 6: Effect on Serum Dopamine levels

[0041] Figure 7 : Effect on Serum Cortisol levels

[0042] Figure 8: Effect on Oxidative stress and antioxidant system

[0043] Figure 9: Effect on serum hormones

[0044] Figure 10: Effect on BDNF in ovariectomized rats

[0045] SUMMARY OF THE INVENTION

[0046]

[0033] The present invention provides a nutritional extract composition comprising synergistic combinations of the Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonellafoenum-graecum L.).

[0047]

[0034] Also, the present invention encompasses the extract composition comprising of synergistic combinations of the Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonellafoenum-graecum L.) and at least one nutraceutically and / or pharmaceutically accepted excipient for, improvement, and maintenance of women health more preferably in improving of hot flashes, night sweats, poor sleep, anxiety, sexual dysfunction, mood changes, genitourinary symptoms in overall improving the quality of life during the period of menopause. Also, the composition further encompasses the improvement of serum estradiol, progesterone, follicle stimulating hormone (FSH), cortisol, and Brain-derived neurotrophic factor (BDNF) levels in women during the period of menopause.

[0048]

[0035] The present invention further discloses the nutritional extract composition comprising the selective and standardized active constituents of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonellafoenum-graecum L.) as a whole composition and with the suitable ratios and / or ranges and with at least one nutraceutically accepted excipient to form diverse formats of formulation doses for, improvement, and maintenance of women health more preferably during menopause.

[0049]

[0036] The present invention also provides a nutritional composition comprising combinations of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) which may be formulated in different forms like orally administrable solid, semisolid, liquid forms, selected from, but not limited to dosages such as, powders, granules, pellets, beadlets, caplets, tablets, capsules, soft gel capsules, solution, emulsions, suspensions, oil suspensions, dispersions, dispersible powders, gummies, and similar dosage forms.

[0050]

[0037] The present invention also provides a process for extracting Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) in a single solvent process and the formulation dosages process

[0051] DETAILED DESCRIPTION OF THE INVENTION

[0052]

[0038] Menopause is a natural decline in reproductive hormones when a woman reaches the age between 45 to 51 years, and it marks the natural end of fertility. Menopause is signaled by 12 months since the last menstruation. Menopause is started with perimenopause period and is that period during which normal ovulation cycles gradually lead to cessation of menses. As the ovulatory cycles get lengthened and become more irregular, the level of estrogen may initially increase, but will eventually drop with the onset of menopause.

[0053]

[0039] Around the time of menopause and perimenopause, females mostly experience physical symptoms such as hot flashes, poor sleep, night sweats, vaginal dryness, and a reduced sex drive. It can also lead to anxiety, changes in mood, and genitourinary symptoms or sexual dysfunction. Menopause is also marked by ovarian failure and estrogen deprivation, wherein estrogen deprivation affects the various organs in the body such as central nervous system and urogenital system. The decrease in estrogen in the body results vasomotor symptoms, sleep disruption, depression and anxiety, weight gain, increased fat and waist circumference, decreased sexual desire, vaginal dryness, vulvar itching and burning, urinary tract infection and bone, skin, hair changes including decreased skin thickness, decreased skin elasticity, decreased skin hydration, increased skin wrinkling, hair loss and osteoporosis.

[0054]

[0040] Menopause is a natural process, and treatments majorly focus on symptomatic relief. Till date, various nutritional remedies were identified including Asparagus racemosus Willd (Shatavari), Trigonella foenum graecum (Methi), Bacopa monnieri (Brahmi), Berberis aristata (Indian barberry), Butea monosperma (Flame-of-the-forest), Oroxylum indicum (Indian trumpet tree), Aegle mammelose ((Wood apple), Cyperus rotundus (Java Grass), Sphaeranthus indicus (Gorakhmundi) etc.

[0055]

[0041] Amongst these available remedies, Asparagus racemosus Willd (Shatavari), Trigonella foenum graecum (Methi / fenugreek) are more viable options for women health during menopause because these are well established women health ingredients, widely used in food products and are found to be safe for women.

[0056]

[0042] Asparagus racemosus Willd is also known as Shatavari and is a species of Asparagus native from Africa through southern Asia, including the Indian subcontinent, to northern Australia. It belongs to Asparagaceae family and is rich in nutrients such as steroidal saponins which is known as Shatavarins and thus is also known as female reproductive tonic. Shatavari helps to manage physical and emotional stress. It also possesses antioxidant and anti-inflammatory activity and plays an important role in hormone balancing by acting as phytoestrogen that can bind to estrogen receptors. Asparagus racemosus also reduces symptoms of menopause such as hot flashes, night sweats insomnia and vaginal dryness.

[0043] Asparagus racemosus Willd is known as potent immunomodulatory plant which helps in increasing Thl (IL-2, IFN-g) and Th2 (IL-4) cytokines, increasing cellular and humoral immune response and decreasing pro-inflammatory cytokines (IL-6 and TNF). Asparagus racemosus Willd is also established as potent antioxidant that increases glutathione, superoxide dismutase, and catalase levels in the brain and decreases nitric oxide, malondialdehyde, and cytokine synthesis in the brain.

[0057]

[0044] Fenugreek is one of the oldest medicinal plants cited in Ayurveda and Chinese medicine and is used for lactation stimulant, and laxatives, postmenopausal symptoms. Fenugreek is scientifically known as Trigonella foenum graecum and is established as an antioxidant, anti- inflammatory, antidiabetic, anti-obesity, anti-hyper-lipidemic activity in women’s health.

[0058]

[0045] Trigonella foenum graecum belongs to Fabaceae family possessing phyto- estrogenic properties.

[0059]

[0046] Trigonella foenum graecum has various health benefits including increased breast milk production, decreased cholesterol levels in blood, decreased high blood sugar and hemoglobin Ale levels and ameliorates menopausal discomforts related to vasomotor, urogenetic, psychological issues.

[0060]

[0047] Fenugreek is also known to possess phytoestrogenic activities that increases expression of the ER genes and induced proliferation of MCF-7, decreases vasomotor symptoms (reduced hot flash symptoms) in menopausal women (through antiinflammatory effect), decreases metabolic and inflammatory alterations in ovariectomized rat (management of menopause), decreases ge-related symptoms of androgen decrease, increases testosterone levels and improves sexual function in healthy aging males (Testofen), decreases the symptoms of PCOS (Furocyst®) and decreases postmenopausal symptoms and helps to establish hormonal balance (FenuSMART™).

[0061]

[0048] Fenugreek is also established to provide protection against metabolic conditions by increasing insulin-stimulated glucose transport, increasing phosphorylation of insulin receptor IR- and IRS-1, decreasing suppressor of cytokine signaling-3 and normalizing the glucose uptake, increasing glucose uptake through upregulation of GLUT-2 & SREBP1C, decreasing hyperglycemia and diabetic condition by promoting the adipocyte differentiation (Diosgenin), decreasing diabetic neuropathic conditions, decreasing blood glucose levels, decreasing inflammation-stimulated IRS-1 phosphorylation, increasing insulin sensitivity, decreasing ROS-induced inflammation, decreasing NFkB, JNK 1 / 2, ERK 1 / 2) & P38MAPK, decreasing MDA in the serum and levels in kidney to reduce the LPO and oxidative stress in diabetic rats, increasing protection of kidney by restoring the uneven thickening of glomerulus caused by diabetes, restoring the biochemical changes such as cholesterol, triglycerides, urea, uric acid, creatinine, and ALT to normal levels.

[0062]

[0049] The present invention provides the synergetic composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.). The said composition is used for benefitting the stress-mediated reproductive disorders in women. The said synergetic composition is used for hormonal balancing and for treating hot flashes, night sweats, poor sleep, anxiety, sexual dysfunction, mood changes, genitourinary symptoms, and overall improving the quality of life. Also, this synergetic composition further provides the improvement of Brain-derived neurotrophic factor (BDNF) levels in women during menopause. BDNF levels are reliable markers that reflect the overall neuroplasticity, energy homeostatic and is known to act by influencing eating pattern, physical activity and improved neuronal plasticity and thus offer protection against various metabolic issues that increase during menopause. In postmenopausal women, elevated BDNF has been shown to correlate with better associative and verbal memory (REF) and improves neuronal survival.

[0063]

[0050] The present invention further provides the composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) as a whole and / or in suitable ratios and / or ranges. It additionally provides the active constituents selected from Protodioscin, optionally Trigonelline, Flavone glycosides as vitexin and vicenin-1, extracted from Fenugreek, and Shatavarin IV, extracted from Shatavari, and saponins as trigoneosides and glycosides collectively and / or individually from Fenugreek, and Shatavari.

[0064]

[0051] The present invention additionally provides that the Asparagus racemosus Willd but not limited to Asparagus densiflorus, Asparagus Setaceus, Asparagus Plumosus and Asparagus Sprengeri with total Shatavarins and Shatavarin IV as the markers and adjusting their levels to the desired ranges disclosed in this invention. Furthermore, the present invention also provides the Fenugreek (Trigonella foenum-graecum L.) but not limited to Trigonella emodi Benth, Trigonella gracilis Benth, Trigonella balansae Boiss. & Reut, Trigonella glabra Thunb, Trigonella obcordata Benth, Trigonella occulta Delile ex Ser., with the protodioscin as the markers and adjusting their levels to the desired ranges disclosed in this invention.

[0065]

[0052] The active constituent structures from the Shatavari (Asparagus racemosus Willd) and Fenugreek (Trigonella foenum-graecum L.) are depicted below:

[0053] The inventors of the invention conducted rigorous experimentation to arrive at a selective range of constituents such as Protodioscin, optionally Trigonelline, Flavone glycosides as vitexin and vicenin-1, extracted from Fenugreek, and Shatavarin IV, extracted from Shatavari, and the saponins collectively from Fenugreek, and Shatavari.

[0066]

[0054] In one embodiment, the present invention further provides that the fenugreek extract is enriched and / or standardized to 1 to 50% of the composition, which further comprises protodioscin in at least 3%, Flavone glycosides as vitexin, and vicenin / vicenin-1 in at least 0.1%, saponin as a trigoneoside IVa, and glycosides in at least 10% of the extract composition.

[0067]

[0055] In one more embodiment, the present invention further provides that the shatavari extract is enriched and / or standardized to 1 to 50% of the composition, which further comprises shatavari as shatavarin IV in at least 3% and saponin in at least 10% of the extract composition. The saponin measured in the composition is considered as a collective and / or individually from the fenugreek and Shatavari extracts composition.

[0068]

[0056] The present invention provides the extraction process for Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) in a single solvent wherein both the extract is processed separately and / or combinedly, but more preferably in combination of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.).

[0069]

[0057] The present invention further provides the oral extract composition is enriched and / or standardized to 1 to 100% of the total extract composition, which further comprises protodioscin in at least 3 %, Flavone glycosides as vitexin, and vicenin / vicenin-1 in at least 0.1%, saponin as a trigoneoside IVa, and glycosides in at least 10% shatavari as shatavarin IV in at least 3% and saponin from shatavari is in at least 10% of the extract composition of the extract composition.

[0070]

[0058] In one of the aspects of the present invention is to provide the oral extract composition is enriched and / or standardized to 1 to 90% of the total extract composition, which further comprises protodioscin from about 3 to about 30%, Flavone glycosides as vitexin, and vicenin / vicenin-1 from about 0.1 to about 10%, saponin as a trigoneoside IVa, and glycosides and other saponins from about 10 to about 64%, and shatavari as shatavarin IV from about 3 to about 30 % of the extract composition.

[0071]

[0059] In one of the aspects of the present invention is to provide the oral extract composition is enriched and / or standardized to 1 to 80% of the total extract composition, which further comprises protodioscin from about 3 to about 20%, Flavone glycosides as vitexin, and vicenin / vicenin-1 from about 0.1 to about 8%, saponin as a trigoneoside IVa, and glycosides and other saponins from about 10 to about 44%, and shatavari as shatavarin IV from about 3 to about 20% of the extract composition of the extract composition.

[0072]

[0060] In one more of the aspects of the present invention is to provide the oral extract composition is enriched and / or standardized to 1 to 70% of the total extract composition, which further comprises protodioscin from about 3 to about 15%, Flavone glycosides as vitexin, and vicenin / vicenin-1 from about 0.1 to about 6%, saponin as a trigoneoside IVa, and glycosides and other saponins from about 10 to about 40%, and shatavari as shatavarin IV from about 3 to about 15% of the extract composition of the extract composition.

[0073]

[0061] The present invention provides the extraction process for Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) combinedly in a ratio of 50:50 to70:30 % w / w of Shatavari and Fenugreek, which comprises the following steps: a) mixing the pulverized shatavari root and fenugreek seeds; b) adding aqueous alcoholic mixture; c) combining with 50% alcoholic extract; d) separating the extract into three fractions, namely water, 38 to 40% of alcoholic fraction and 90 to 95% alcoholic fraction, by using resin column chromatography; e) discarding the water and, 38 to 40% of alcoholic fraction; f) concentrating the 90 to 95% alcoholic fraction and converting it into the dry powder.

[0074]

[0062] The present invention also provides the extraction process for Shatavari and Fenugreek, and mixing both raw materials at the macella stage, followed by the process mentioned above. The details are provided in the examples.

[0063] The present invention additionally provides the extraction process for Shatavari and Fenugreek, separately, in line with the process mentioned above, and blended in the last stage to provide the composition. The details are provided in the examples.

[0075]

[0064] The present invention also provides the d formulation composition ShatavariAsparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.). More preferably, a Shatavari and Fenugreek f is formulated using excipients selected from the group of, but not limited to, solvents, anticaking agents, bioavailability enhancers, solubilizers, emulsifiers, binders, fillers, optionally pH modifiers, and coating agents, and / or the combination thereof.

[0076]

[0065] In one embodiment of the invention, the composition described herein can be available in orally administrable solid, semisolid, and liquid forms, selected from, but not limited to, dosages such as, powders, granules, pellets, beadlets, caplets, tablets, capsules, soft gel capsules, solution, emulsions, suspensions, oil suspensions, gummies, dispersions and the like.

[0077]

[0066] The compositions, as described herein, are comprised of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) extract and at least one excipient, such as pharmaceutically and / or nutraceutically accepted excipients, which is formulated more preferably as granules, powder, and / or beadlet.

[0078]

[0067] In one aspect of the invention, the present invention also comprises a process for the preparation of the oral composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) in the form of powder, wherein the process comprises: a) adding a suitable solvent to shatavari & fenugreek extract in a clean vessel under stirring to obtain uniform dispersion; b) adding suitable carrier in step (a) under stirring to obtain uniform dispersion; c) spray drying the solution obtained from step (b); and d) adding an anti-caking agent to the spray-dried powder obtained in step (c).

[0079]

[0068] In one aspect of the invention, the solvent used for the preparation and / or extraction of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) extract composition is selected from the group consisting of isopropyl alcohol, acetone, methanol, alcohol, ethyl acetate, ethanol, methylene dichloride, water, and their combination thereof.

[0069] In another embodiment of the invention, the carrier employed in preparation of composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) is selected from the group consisting of hydroxy propyl methyl cellulose and other cellulose derivatives, Starch, modified starch, Chitosan, Gums i.e. Gum Arabic, Xanthan Gum etc., polyvinyl alcohol, polyvinylpyrrolidone, dextran, pectin, PLGA: Poly (lactic-co-glycolic acid), Polyethylene glycol, maltodextrin, and their combination thereof.

[0080]

[0070] In another embodiment, the anticaking agent used in preparation of composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) composition is selected from the group consisting of silicon dioxide, dicalcium phosphate, magnesium stearate, stearic acid, mannitol, and their combination thereof.

[0081]

[0071] In another aspect of the present invention, the process for the preparation of the composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) in the form of beadlet, wherein the process comprises - a) adding solvent in a clean vessel along with shatavari and fenugreek extract, bioenhancer, solubilizer, and emulsifier under stirring to obtain uniform dispersion; b) adding binder and solvent in another vessel; c) adding filler and anticaking agent to form a uniform powder blend in separate vessel; d) adding solutions prepared in step (a) in the dry blend of step (c) to obtain the wet mass; e) adding solution prepared in step (b) in wet mass obtained in step (d); f) passing the wet mass through extruder followed by spheronizer; g) drying and sifting the granules obtain in step (f). h) preparing the coating solution using coating agent and solvent; i) loading the granules obtained in step (g) in fluidized bed coater to coat with solution obtained in step (h); and j) drying the granules obtained in step (i).

[0082]

[0072] In another embodiment, the bioavailability enhancer employed in preparation of compositionof Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) is selected from the group consisting of d- limonene, mono & diglycerides, and phospholipids such as lecithin / phosphatidyl choline, botanical extracts, Vegetable Oils, and their combination thereof.

[0083]

[0073] In one aspect of the invention, the solubilizer used in the preparation ofcompositionShatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) is selected from the group consisting of MCT oil, polysorbate, propylene glycol alginate, sugar alcohols, sugar esters, phospholipid, Vitamin E TPGS (d- a-Tocopheryl polyethylene glycol 1000 succinate), beta cyclodextrin, and their combination thereof.

[0084]

[0074] In another aspect of the invention, the emulsifier used in preparation of composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) composition is selected from the group consisting of steric acid, phosphatidylcholine, polysorbate, sugar alcohols, glycerol & derivatives, and their combination thereof.

[0085]

[0075] In another aspect of the invention, the binder used in the preparation of composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) is selected from the group consisting of hydroxy propyl methyl cellulose and other cellulose derivatives, pregelatinized starch, and their combination thereof.

[0086]

[0076] In another embodiment, the present invention also provides the process of preparation of the composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) in the form of granules / beadlets, wherein the process comprises - a) adding solvent in clean vessel along with shatavari and fenugreek extract, carrier, filler and optionally pH modifier under stirring and / or homogenizing to obtain uniform dispersion; b) passing the dispersion obtained in step (a) through dyno mill to reduce the size and to form uniform dispersion; c) passing the dispersion obtained in step (b) through spray congealer to form the granules; d) drying and sifting the granules obtain in step (c). e) preparing the coating solution using coating agent and solvent; f) loading the granules obtained in step (d) in fluidized bed coater to coat solution obtained in step (e); and g) drying the granules obtained in step (f).

[0087]

[0077] In one aspect of the invention, the filler employed in preparation of the composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) is selected from the group consisting of microcrystalline Cellulose, hydroxypropyl methyl cellulose, and other cellulose derivatives, Maize Starch, Lactose, Maltose, and their combination thereof.

[0088]

[0078] In another aspect of the invention, optionally the pH modifier used in the preparation of composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) is selected from the group consisting of sodium bicarbonate, citric acid, trisodium citrate, lactic acid, L-arginine, calcium carbonate, magnesium carbonate, and their combination thereof.

[0089]

[0079] In another embodiment, the coating agent used in the preparation of composition of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) is selected from the group consisting of stearic acid, glyceryl distearate, shellac, zein, acrylate polymer, cellulose polymer, and their combination thereof.

[0090]

[0080] The present invention is explained in detail by referring to examples, which are not to be construed as limitative.

[0091] Materials, Experimental Method, and Analysis

[0092] A. Animals

[0093]

[0081] This study involved to examine the Estrogenic activity, Vasomotor hot flashes and Stress and anxiety caused by menopause. Ovariectomized rats are used for study design.

[0094] B. Experimental Condition

[0095] TFG - Trigonella foenum graecum AR - Asparagus racemosus Willd NC: normal control;

[0096] OVX: ovariectomized rats;

[0097] OVX+TA1: ovariectomized rats + TFG (30 mg / kg bw) + AR (30 mg / kg bw) combination 1; OVX+TA2: ovariectomized rats + TFG (30 mg / kg bw) + AR (15 mg / kg bw) combination 2; 0VX+TA3: ovariectomized rats + TFG (15 mg / kg bw) + AR (30 mg / kg bw) combination 3; OVX+TA4: ovariectomized rats + TFG (40 mg / kg bw) + AR (30 mg / kg bw) combination 4; OVX+TA5: ovariectomized rats + TFG (30 mg / kg bw) + AR (40 mg / kg bw) combination 5; OVX+IAT1: ovariectomized rats + Trigonella + Asparagus IAT1 (30 mg / kg bw); OVX+IAT2: ovariectomized rats + Trigonella + Asparagus IAT2 (60 mg / kg bw).

[0098] * IAT30 and IAT60: Integrated extracts ofTRG and ARS (Shatavari and Fenugreek composition) at 30 mg and 60 mg / KG body weight, respectively.

[0099] C. Study Details and Their Results

[0100]

[0082] Menopauses causes decreased estrogen level which leads to various lipid metabolic disorders and metabolic syndromes including cardiovascular diseases (CVD) and Type 2 diabetes (T2D). The study results are as discussed below: a) Serum Metabolic Profile

[0101]

[0083] Menopause in ovariectomized rats used in the study design caused decreased estrogen level and dyslipidemia resulting in increased triglycerides and cholesterol level. Triglycerides and cholesterol level were later observed after administering the various formulations as discussed in experimental conditions. There was a significant reduction in both Triglycerides and Cholesterol levels in supplementation with a combination of Trigonella and Asparagus extracts at the raw level, as provided in the examples, to exert maximum effects seen with the integrated actives technology (IAT) combination of Trigonella and Asparagus extracts. The results are as shown below in Figure 1.

[0102]

[0084] The depicted Whisker plots represent the median and min-max values. ANOVA and Tukey’s post-hoc test was used for comparing the results among different treatment groups. Different small letters (a-f) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. b) Effect of supplementation on Body and Tail temperature in ovariectomized rats

[0103]

[0085] In rats, the tail is the primary heat exchange organ due to its extensive vascularization, lack of fur, and high surface area to volume ratio. Cutaneous vasodilation is the hallmark of hot flashes wherein cutaneous vasodilation of the tail can be monitored by measuring tail skin temperature. Dysregulation of hormonal and neurotransmitter system due to lack of estrogen lead to pulses of increase in body temperature.

[0104]

[0086] Body and Tail temperature was measured twice a day during supplementation period, and it was observed that there was a small but consistent increase in body and tail skin temperature in OVX rat which was moderately reduced by combined extracts supplementation with maximum reduction in case of I AT extracts. Results are as shown in Figure 2 and Figure 3.

[0105]

[0087] The Effects of different combinations of Trigonella foenum graecum and Asparagus racemosus Willd on body and tail temperature in ovariectomized (OVX) rats. The error lines point out the standard error of the mean. Data are presented as the mean and standard deviation, a-f: Means in the same line without a common superscript differ significantly (p<0.05, ANOVA and Tukey post hoc test). c) Neuropeptides that mediate physiology of hot flashes

[0106]

[0088] The proposed negative and positive feedback mechanism exerted by ovarian steroids on the regulation of kisspeptin neurons. In rodents, Kissi neurons are in both AVPV and ARC and stimulate (C) GNRH neurons. In the ARC, Kissi neurons project directly to GNRH cell bodies and their terminals in the median eminence. Moreover, estradiol (E2) inhibits (K) ARC Kissi neurons consistent with negative feedback control of GNRH neurons. In the AVPV, E2 stimulates (C) Kissi neurons, facilitating positive feedback regulatory control and the preovulatory GNRH / LH surge. AVPV Kissi neurons only project directly to GNRH cell bodies. Effect of neuropeptides are shown in Figure 4. Significant effects were observed for supplementation with simple combination of Trigonella and Asparagus extracts with maximum effects seen with the integrated actives technology (IAT) combination of Trigonella and Asparagus extracts.

[0107]

[0089] The densitometric analysis of the relative intensity according to the control group of the western blot bands was performed with [3-actin normalization to ensure equal protein loading. Blots were repeated at least three times (n=3) and a representative blot is shown (D). The error lines point out the standard error of the mean. ANOVA and Tukey’ s post hoc tests were used to compare the results among different treatment groups. Different small letters (a-h) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. d) Effect on Serum BDNF levels

[0090] Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survival and growth, serves as a neurotransmitter modulator, and participates in neuronal plasticity. Menopause is associated with significantly declined serum BDNF levels in the women. There was a significant increase in expression of BDNF in brain tissues in animals supplemented with combination of Trigonella and Asparagus extracts with maximum increase in case of IAT extracts. Results are as shown in Figure 5.

[0108]

[0091] The densitometric analysis of the relative intensity according to the control group of the western blot bands was performed with [3-actin normalization to ensure equal protein loading. Blots were repeated at least three times (n=3) and a representative blot is shown (D). The error lines point out the standard error of the mean. ANOVA and Tukey’s post hoc tests were used to compare the results among different treatment groups. Different small letters (a-h) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. e) Effect on Serum Dopamine levels

[0109]

[0092] Dopamine is known as “happy hormones” which regulates mood, emotion and sleep. Low estrogen levels result in reduced dopamine which is responsible for stress, anxiety and lack of sleep during menopause. Supplementation of experimental conditions significantly restored serum dopamine levels with maximum effect in case of IAT extracts. Results are as shown in Figure 6.

[0110]

[0093] The depicted Whisker plots represent the median and min-max values. ANOVA and Tukey’s post-hoc test was used for comparing the results among different treatment groups. Different small letters (a-g) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. f) Effect on Serum Cortisol levels

[0111]

[0094] Cortisol (stress hormone) regulates response to stress wherein prolonged increase in cortisol levels may lead to stress, anxiety & some chronic diseases. Cortisol is also responsible for multiple symptoms observed during menopause including stress, anxiety, lack of sleep and hot flashes. Ovariectomy in rats increased cortisol levels and there is a significant decrease in serum cortisol levels during supplementation particularly with IAT extract with maximum response at 60 mg / KG body weight dose Results are as shown in Figure 7.

[0112]

[0095] The depicted Whisker plots represent the median and min-max values. ANOVA and Tukey’s post-hoc test was used for comparing the results among different treatment groups. Different small letters (a-g) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. g) Effect on Oxidative stress and antioxidant system

[0113]

[0096] Menopause causes estrogen deficiency resulting in Oxidative stress which, in turn, is responsible for metabolic derangements, chronic inflammation, hot flashes, stress and anxiety.

[0114]

[0097] Free oxygen radical’s breakdown phospholipids of the cell wall that lead to accumulation of malondialdehyde (MDA). Serum MDA is the biomarker of Oxidative Stress and responsible for many disease conditions. Oxidative stress was increased in OVX rat and supplementation significantly reduced MDA levels with highest reduction in case of IAT extracts. Results are as shown in Figure 8.

[0115]

[0098] Menopause causes estrogen deficiency resulting in decreased estrogen level and decreased antioxidant which, in turn, is responsible for development of a variety of symptoms and pathologies that characterize menopause.

[0116]

[0099] Antioxidants like Super oxide dismutase (SOD), Catalase (Cat) Glutathione peroxidase (GPx) act to prevent lipid peroxidation. There was a significant restoration of both SOD, GPx and Cat serum levels with supplementation in ovariectomized rat with maximum effect in case of IAT extracts. Results are as shown in Figure 8.

[0117]

[0100] The Effects of different combinations of Trigonella foenum graecum and Asparagus racemosus Willd on MDA (A), SOD (B), CAT (C) and GSH-Px (D) in ovariectomized (OVX) rats. The depicted Whisker plots represent the median and min-max values. ANOVA and Tukey’s post-hoc test were used for comparing the results among different treatment groups. Different small letters (a-g) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. h) Effect on serum hormones

[0118]

[0101] Menopaue is characterized by increase in Follicle-stimulating hormone (FSH) and Euteinizing hormone (EH) levels, and decrease in 17[3-estradiol (estrogen) and progesterone levels. These hormone levels were significantly improved with maximum effect in case of I AT extracts.

[0119]

[0102] The Effects of different combinations of Trigonella foenum graecum and Asparagus racemosus Willd on FSH (A), LH (B), 17B-Estradiol (C), Progesterone (D), in ovariectomized (OVX) rats. The depicted Whisker plots represent the median and min- max values. ANOVA and Tukey’s post-hoc test were used for comparing the results among different treatment groups. Different small letters (a-g) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. i) Effect on BDNF in ovariectomized rats

[0120]

[0103] Brain-derived neurotrophic factor (BDNF) plays an important role in neuronal survival and growth, serves as a neurotransmitter modulator, neuronal plasticity and may modulate stress and anxiety during menopausal condition. Menopause is associated with significantly declined serum BDNF levels in the women. Both ARS and TRG have shown to exert neuroprotective effects by enhancing BDNF expression possibly due to their estrogenic activity. Decreased levels of BDNF observed after menopause were restored by hormone therapy indicative of the modulatory effect of estrogen on BDNF expression.

[0121]

[0104] We have observed that BDNF expression in brain is reduced in rats by ovariectomy which was significantly restored by IAT extract as compared to individual components (ARS & TRG)

[0122]

[0105] The densitometric analysis of the relative intensity according to the control group of the western blot bands was performed with [3-actin normalization to ensure equal protein loading. Blots were repeated at least three times (n=3) and a representative blot is shown. The error lines point out the standard error of the mean. ANOVA and Tukey’s post hoc tests were used to compare the results among different treatment groups. Different small letters (a-h) above the bars indicate statistical differences between groups. There is no difference between groups with the same letter, whereas there is a difference between groups with entirely different letters. j) Summary of effect of supplementation in ovariectomized rats

[0123]

[0106] Statistical method used is Tukey’s test (The number indicates degree of significant difference between experimental groups).

[0124]

[0107] Combination of Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum- graecum L.) extract was supplemented as a blend (Comb) prepared from the provided processes was supplemented to ovariectomized rats for 15 days. Animals were sacrificed at the end of the study, and different parameters were measured. A combination of blends was used at different ratios of Shatavari and Fenugreek, whereas IAT (Shatavari and Fenugreek composition) was dosed at 30 and 60 mg KG body weight.

[0125]

[0108] Parameters for Hot flashes: Strongly positive outcome for combination as well as IAT at higher concentration with significant down regulation of tail temperature and expression of neuronal markers of hot flashes (KP, NKB, GnRH, c-FOS and TRPV1).

[0126]

[0109] Parameters for stress and anxiety: Strongly positive outcome for IATS at both doses as compared to blends of extracts with significant down regulation of serum Dopamine levels and expression of BDNF levels in brain tissue that play a significant role in management of stress and anxiety.

[0127]

[0110] Parameters for Oxidative stress: Strongly positive outcome for Integrated actives IAT (composition of present invention: Shatavari (Asparagus racemosus Willd.) and Fenugreek (Trigonella foenum-graecum L.) extracts as compared to combination (blends) with significant reduction in serum levels of MDA while increased SOD Cat and GSH-Px levels.

[0128]

[0111] Parameters for hormonal balance: Strongly positive outcome for IAT extracts as compared to combination (blends) with significant reduction in serum FSH and LH levels while increased 17[3-estradiol (estrogen) and progesterone levels.

[0129]

[0112] Parameters for metabolic profile: Strongly positive outcome for IATs as compared to blends significant down regulation of serum cholesterol and triglyceride levels.

[0130]

[0113] Biochemical parameters indicate that the extracts in individual and combination doses are safe.

[0131]

[0114] IAT extracts at dose of 60 followed by 30 mg KG body weight demonstrated maximum restoration of normalcy as compared to ovariectomised condition while IAT extracts of Trigonella and Asparagus at a dose of 30 mg / KG BW as well as IAT at 60 mg / KG BW is significantly more efficacious as compared to blends of extracts at all the measured parameters.

[0132]

[0115] The present invention is further described in details with following examples, but is not restricted to them.

[0133] Example 1:

[0116] Asparagus root powder and fenugreek seed powder were mixed in the ratio of 6:4 (total weight 100 g) and extracted with 50% EtOH (500 mL) at 60-65°C for 3h. The extraction process was repeated with 50% EtOH (2 x 500 mL). The combined extracts were evaporated under reduced pressure to get 40-50% TDS. The concentrated solution was loaded on column, packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol.

[0117] Ethanol fraction was concentrated to obtained brown coloured solid (3.1 g). HPLC method with two markers (Protodioscin: 18 %; Shatavarin IV: 22 %). Further, hydroxypropyl methylcellulose (69.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.4% and shatavarin IV 6.6% in the powdered extract.

[0134] Example 2

[0135]

[0118] Asparagus root powder and fenugreek seed powder was mixed in the ratio of 7:3 (total weight 100 g) and extracted with 50% EtOH (500 mL) at 60-65°C for 3h. The extraction process was repeated with 50% EtOH (2 x 500 mL). The combined extracts were evaporated under reduced pressure to get 40-50% TDS. The concentrated solution was loaded on column, packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtained brown coloured solid (3.1 g). HPLC method with two markers (Protodioscin: 12.3 %; Shatavarin IV: 26.8 %). Further, hydroxypropyl methylcellulose (53.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.6% and shatavarin IV 12.3% in the powdered extract.

[0136] Example 3

[0137]

[0119] Asparagus root powder and fenugreek seed powder was mixed in the ratio of 7:3 (total weight 100 g) and extracted with 50% EtOH (500 mL) at 60-65°C for 3h. The extraction process was repeated with 50% EtOH (2 x 500 mL). The combined extracts were evaporated under reduced pressure to get 40-50% TDS. The concentrated solution was loaded on column, packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtained brown coloured solid (3.1 g). HPLC method with two markers (Protodioscin: 6.7%; Shatavarin IV: 14.2%). Further, hydroxypropyl methylcellulose (19.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.36% and shatavarin IV 11.36% in the powdered extract.

[0138] Example 4:

[0120] Asparagus root powder and fenugreek seed powder were mixed in the ratio of 6:4 (total weight 100 g) and extracted with 50% EtOH (500 mL) at 60-65°C for 3h. The extraction process was repeated with 50% EtOH (2 x 500 mL). The combined extracts were evaporated under reduced pressure to get 40-50% TDS. The concentrated solution was loaded on column, packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtained brown coloured solid (3.8 g). HPLC method with two markers (Protodioscin: 8.3%; Shatavarin IV: 10.6%). Further, hydroxypropyl methylcellulose (31.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.6% and shatavarin IV 7.2% in the powdered extract.

[0139] Example 5:

[0140]

[0121] Asparagus root powder and fenugreek seed powder were mixed in the ratio of 1:1 (total weight 100 g) and extracted with 50% EtOH (500 mL) at 60-65°C for 3h. The extraction process was repeated with 50% EtOH (2 x 500 mL). The combined extracts were evaporated under reduced pressure to get 40-50% TDS. The concentrated solution was loaded on column, packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtained brown coloured solid (4.2 g). HPLC method with two markers (Protodioscin: 14.8%; Shatavarin IV: 24.2%). Further, hydroxypropyl methylcellulose (60.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.7% and shatavarin IV 9.4% in the powdered extract.

[0141] Example 6:

[0142]

[0122] Asparagus root powder and fenugreek seed powder were mixed in the ratio of 1:1 (total weight 100 g) and extracted with 70% EtOH (500 mL) at 60-65°C for 3h. The extraction process was repeated with 70% EtOH (2 x 500 mL). The combined extracts were evaporated under reduced pressure to get 40-50% TDS. The concentrated solution was loaded on column, packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtained brown coloured solid (3.8 g). HPLC method with two markers (Protodioscin: 15.6%; Shatavarin IV: 14.3%). Further, hydroxypropyl methylcellulose (62.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.7% and shatavarin IV 5.3% in the powdered extract.

[0143] Example 7 :

[0123] Asparagus root powder and fenugreek seed powder was mixed in the ratio of 7:3 (total weight 100 g) and extracted with 30% EtOH (500 mL) at 60-65°C for 3h. The extraction process was repeated with 30% EtOH (2 x 500 mL). The combined extracts were evaporated under reduced pressure to get 40-50% TDS. The concentrated solution was loaded on column, packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtained brown colour solid (4.1 g). HPLC method with two markers (Protodioscin: 9.2%; Shatavarin IV: 18.9%). Further, hydroxypropyl methylcellulose (36.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.8% and shatavarin IV 12.1% in the powdered extract.

[0144] Example 8 (Micelle process)

[0145]

[0124] Asparagus roots and fenugreek seeds (100 gm each) were extracted with 50% ethanol (500ml) at 60-65 °C for 3h separately. The extraction process was repeated for both the raw materials with 50% EtOH (2 x 500 mL) to get asparagus miscella and fenugreek miscella. The individual miscella was evaporated under reduced pressure to get 40-50% TDS. Asparagus miscella and fenugreek miscella were blended at this stage in the required ratio. This combined miscella was loaded on a column, which was packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtain a coloured solid (3.1 g). HPLC method with two markers (Protodioscin: 8.7%; Shatavarin IV: 28.2%). Further, hydroxypropyl methylcellulose (34.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 5.6% and shatavarin IV 18.4% in the powdered extract.

[0146] Example 9 (Micelle process)

[0147]

[0125] Asparagus roots and fenugreek seeds (100 gm each) were extracted with 50% ethanol (500ml) at 60-65 °C for 3h separately. The extraction process was repeated for both the raw materials with 50% EtOH (2 x 500 mL) to get asparagus miscella and fenugreek miscella. The individual miscella was evaporated under reduced pressure to get 40-50% TDS. Asparagus miscella and fenugreek miscella were blended at this stage in the required ratio. This combined miscella was loaded on a column, which was packed with polystyrene divinylbenzene resin. The column was eluted with water followed by ethanol. Ethanol fraction was concentrated to obtain a coloured solid (3.1 g). HPLC method with two markers (Protodioscin: 28.6%; Shatavarin IV: 12.9%). Further, hydroxypropyl methylcellulose (44.5% w / w) and colloidal silicon (0.5% w / w) were added in the extract so to get protodioscin 15.8% and shatavarin IV 7.1% in the powdered extract.

[0148] Example 10 (Blend process)

[0149]

[0126] Asparagus roots and fenugreek seeds (100 gm each) are extracted with 50% ethanol (500ml) at 60-65 °C for 3h separately. The extraction process was repeated for both the raw materials with 50% EtOH (2 x 500 mL) to get asparagus miscella and fenugreek miscella. The individual miscella was evaporated under reduced pressure to get 40-50% TDS. Asparagus miscella and fenugreek miscella were loaded on different two columns separately which were packed with polystyrene divinylbenzene resin. The columns were eluted with water followed by ethanol. Hydroxypropyl methylcellulose was added to ethanol fractions. These liquids were dried separately and obtained light brown to brown powders. These dried powders were blended in required ratios to get protodioscin 5.5% and shatavarin IV 7.5% in the blended powder.

[0150] Example 11 : Shatavari and Fenugreek formulation Composition (Beadlet Preparation)

[0151]

[0127] Phosphatidylcholine, and Fenugreek & Asparagus extract, were weighed and added to the ethanol under stirring condition to prepare a uniform dispersion. Hydroxypropyl methylcellulose, was weighed and added to the purified water in separate vessel under stirring condition to prepare solution. The obtained Hydroxypropyl methylcellulose solution was then added into Fenugreek & Asparagus extract dispersion to form a uniform dispersion. Microcrystalline cellulose and Colloidal silicon dioxide were weighed accurately and loaded in RMG container and dry mixed at appropriate speed to form a uniform blend. The obtained Fenugreek & Asparagus extract was added gradually to the dry blend in RMG to obtain wet mass / wet granule. The wet mass was then passed through an extruder to obtain extrudes. Extrudes were loaded on a pitch spheronizer plate to obtain spherical beadlets / pellets. Spherical beadlets were loaded in a tray dryer for drying. Dried beadlets were sifted to obtain desired particle size distribution

[0152] Example 12: Shatavari and Fenugreek formulation Composition (Powder Preparation)

[0153] Procedure:

[0128] Hydroxypropyl methylcellulose was weighed accurately and added to the Fenugreek

[0154] & Asparagus extract dispersion obtained from the extraction process under stirring conditions to prepare a uniform dispersion. The required quantity of purified water was added to the obtained dispersion. The dispersion was spray dried to obtain free -flowing powder. The anticaking agent was added to the spray-dried powder. Example 13: Shatavari and Fenugreek formulation Composition (Gummies)

[0155] Procedure:

[0156]

[0129] All the ingredients, as given in above table, were weighed. 19 gm of water was taken in the mixing bowl and starch was mixed thoroughly until smooth and lump free starch mixture was formed. Isomalt (16.48g), maltitol syrup, and stevia were added in the starch mixture. The mixture was stirred well until all ingredients were fully dissolved and then transferred the slurry from the bowl into a heating pan. The slurry was heated until it reaches 60+2° Brix.

[0157]

[0130] Pectin gel was prepared in separate beaker by mixing pectin and 10g isomalt with continuous stirring with the remaining water (21.6 ml) as hot water (75-80°C) until a smooth, lump-free gel was formed.

[0158]

[0131] The formed pectin gel was then added to the pan once the mixture was reached to 60+2° Brix and stirred thoroughly to avoid any lumps. The cooking and stirring were continued until the mixture were reached to 55+2° Brix. (Visual consistency was stringy / elastic drop).

[0159]

[0132] Blend of fenavari, sunflower oil and soy lecithin was prepared and added to the formed mixture with continued cooking until the mixture was reached to 60+2° Brix (approx. 95°C). On reaching 60+2° Brix (approx. 95°C)., heat was turned off to form the gummy mixture.

[0160]

[0133] Flavours and acids (citric and malic acid) were added to 1 ml water and then mixed well with the gummy mixture. The final mixture was poured into greased silicone moulds [Silicone moulds were greased with food-grade mineral oil] and moulds were rested for 24 hours at room temperature in open condition; followed by demoulding of the gummies and proceeded to packaging.

Claims

Claims1. An oral extract composition comprising synergistic combinations of the Shatavari (Asparagus racemosus Willd.). to Asparagus species, comprising but not limited to Asparagus densiflorus, Asparagus Setaceus, Asparagus Plumosus, and Asparagus Sprengeri, and Fenugreek (Trigonella foenum-graecum L.), to fenugreek species, comprising but not limited to Trigonella emodi Benth, Trigonella gracilis Benth, Trigonella balansae Boiss. & Reut, Trigonella glabra Thunb, Trigonella obcordata Benth, Trigonella occulta Delile ex Ser.

2. An oral extract composition of the Shatavari and Fenugreek as claimed in claim 1 , comprising protodioscin from about 3 to about 30%; Flavone glycosides as vitexin, and vicenin / vicenin- 1 from about 0.1 to about 10%; saponin as a trigoneoside IVa, and glycosides and other saponins from about 10 to about 64%; and shatavari as shata varin IV from about 3 to about 30% of the extract composition.

3. The oral extract composition as claimed in claim 1, comprises protodioscin from about 5 to about 15%; Flavone glycosides as vitexin, and vicenin / vicenin- 1 from about 0.1 to about 3%; saponin as a trigoneoside IVa, and glycosides and other saponins from about 30 to about 64%; and shatavari as shatavarin IV from about 5 to about 15% of the extract composition.

4. An oral extract composition comprising synergistic combinations of the Shatavari and Fenugreek and / or separately for the improvement, and maintenance of women health.

5. The oral extract composition as claimed in claim 4 is used for improving the hot flashes, night sweats, poor sleep, anxiety, sexual dysfunction, mood changes, genitourinary symptoms, and the quality of life, serum estradiol, progesterone, follicle-stimulating hormone (FSH), and cortisol during the period from perimenopause to menopause.

6. The oral extract composition as claimed in claim 4 is used for improving Brain-derived neurotrophic factor (BDNF) levels in women from perimenopause to menopause.

7. The extraction process claimed in claim 1, for Shatavari and Fenugreek comprises: a) mixing the pulverized shatavari root and fenugreek seeds; b) adding an aqueous alcoholic mixture; c) combining with 50% alcoholic extract; d) optionally, asparagus roots and fenugreek seeds were extracted with 50% ethanol separately to form micellar solutions and mixed in a suitable ratio;e) separating the extract into three fractions, namely water, 38 to 40% of alcoholic fraction, and 90 to 95% alcoholic fraction, by using resin column chromatography; f) discarding the water and 38 to 40% of the alcoholic fraction; g) concentrating the 90 to 95% alcoholic fraction and converting it into the dry powder.

8. The extraction process for Shatavari and Fenugreek, as claimed in claim 7, is extracted separately and blended in a suitable ratio to form the extract composition.

9. The composition claimed in claiml, is in the form of granules, powder, and / or beadlet comprise Shatavari and Fenugreek extract and at least one excipient such as pharmaceutically and / or nutraceutically accepted excipients.

10. A powder composition comprising Shatavari and Fenugreek extract is prepared by the process comprising- a) adding suitable solvent and shatavari & fenugreek extract in a clean vessel under stirring to obtain uniform dispersion; b) adding a suitable carrier in step (a) under stirring to obtain a uniform dispersion; c) spray drying the solution obtained from step (b); and d) adding an anti-caking agent to the spray-dried powder obtained in step (c).

11. A beadlet composition comprising Shatavari, and Fenugreek extract is prepared by the process comprising- a) adding solvent in a clean vessel along with shatavari and fenugreek extract, bioenhancer, solubilizer, and emulsifier under stirring to obtain uniform dispersion; b) adding binder and solvent in another vessel; c) adding filler and anticaking agent to form a uniform powder blend in separate vessel; d) adding solutions prepared in step (a) in the dry blend of step (c) to obtain the wet mass; e) adding the solution prepared in step (b) to the wet mass obtained in step (d); f) passing the wet mass through the extruder followed by the spheronizer; g) drying and sifting the granules obtained in step (f). h) preparing the coating solution using coating agent and solvent; i) loading the granules obtained in step (g) in fluidized bed coater to coat with solution obtained in step (h); and j) drying the granules obtained in step (i).

12. A beadlet composition as claimed in claim 9 comprises a bioavailability enhancer (bioenhancer) selected from the group consisting of d-limonene, mono & di- glycerides, and phospholipids such as lecithin / phosphatidyl choline, botanical extracts, Vegetable Oils and their combination thereof.

13. A beadlet composition as claimed in claim 9 comprises a solubilizer selected from the group consisting of MCT oil, polysorbate, propylene glycol alginate, sugar alcohols, sugar esters, phospholipid, Vitamin E TPGS (d-a-Tocopheryl polyethylene glycol 1000 succinate), beta cyclodextrin, and their combination thereof.

14. A beadlet composition as claimed in claim 9 comprises an emulsifier selected from the group consisting of steric acid, phosphatidylcholine, polysorbate, sugar alcohols, glycerol & derivatives, and their combination thereof.

15. A beadlet composition as claimed in claim 9 comprises a binder selected from the group consisting of hydroxy propyl methyl cellulose and other cellulose derivatives, pregelatinized starch, and their combination thereof.

16. A granule composition comprising Shatavari, and Fenugreek extract is prepared by the process comprising- a) adding solvent in a clean vessel along with shatavari and fenugreek extract, carrier, filler, and optionally pH modifier under stirring and / or homogenizing to obtain uniform dispersion; b) passing the dispersion obtained in step (a) through a dyno mill to reduce the size and to form a uniform dispersion; c) passing the dispersion obtained in step (b) through a spray congealer to form the granules; d) drying and sifting the granules obtain in step (c). e) preparing the coating solution using the coating agent and solvent; f) loading the granules obtained in step (d) in fluidized bed coater to coat solution obtained in step (e); and g) drying the granules obtained in step (f).

17. A granule composition as claimed in claim 14 comprises optionally a pH modifier selected from the group consisting of sodium bicarbonate, citric acid, trisodium citrate, lactic acid, L- arginine, calcium carbonate, magnesium carbonate, and their combination thereof.

18. A granule composition as claimed in claim 14 comprises a coating agent selected from the group consisting of stearic acid, glyceryl distearate, shellac, zein, acrylate polymer, cellulose polymer, and their combination thereof.

19. The compositions as claimed in claims 8, 9 and 14 comprises solvent selected from the group consisting of isopropyl alcohol, acetone, methanol, alcohol, ethyl acetate, ethanol, methylene dichloride, water and their combination thereof.

20. The compositions as claimed in claims 8 and 14 comprises carrier selected from the group consisting of hydroxy propyl methyl cellulose and other cellulose derivatives, Starch, modified starch, Chitosan, Gums i.e. Gum Arabic, Xanthan Gum etc., polyvinyl alcohol, polyvinylpyrrolidone, dextran, pectin, PLGA: Poly (lactic-co-glycolic acid), Polyethylene glycol, maltodextrin and their combination thereof.

21. The compositions as claimed in claims 8 and 9 comprise an anti-caking agent selected from the group consisting of silicon dioxide, dicalcium phosphate, magnesium stearate, stearic acid, mannitol, and their combination thereof.

22. The compositions as claimed in claims 9 and 14, comprise filler selected from the group consisting of microcrystalline Cellulose, hydroxypropyl methyl cellulose, and other cellulose derivatives, Maize Starch, Lactose, Maltose, and their combination thereof.

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