TECHNICAL FIELD
[001] The subject matter described herein in general relates to the field of hair growth compositions, and in particular relates to interleukin (TL)-4 and transforming growth factor (TGF) - P inhibitor compositions for promoting hair growth.
BACKGROUND OF INVENTION
[002] Hair follicles are external appendages of the skin. Normal hair growth and prevention from hair loss are of health and aesthetic importance. Hair loss management and hair growth improvement offer great market potential worldwide. Currently, finasteride and minoxidil are the only two FDA approved drugs for hair growth, one for oral consumption and the other for topical application, respectively. However, due to number of side effects there is a growing quest for drugs or actives of natural origin, which are in general considered safe. [003] Recent literature suggests the use of actives having plant origin for use in promoting hair growth. For example, naringenin and hesperetin (Madaan et al., 2017. American J. Dermatol. Venerol. 6(3): 51-57), retinol (Yoo et al., 2007. Bio Pharm Bull. 30:21-26), alga (Ishige sinicola) (Kang et al., 2013, Mar Drugs. 11:1173-1799), Asiasari radix (Rho et al., J. Dermatol. Sci. 2005. 38(2): 89-97), Panax ginseng (Matsuda, et al., 2003. Phytotherapy Res. 17(7): 89-97; Park et al., 2011. J. Ethnopharmacology. 138(2): 340-344), and Eclipta alba (Roy et al., 2008. Arch. Dermatol. Res. 300(7): 357-364; Datta et al., 2009, J. Ethnopharmacology. 124(3): 450-456) are shown as useful hair growth inducers.
[004] Flavonoids are a group of secondary metabolites in plants and fungus. Some flavonoids are known to promote hair growth. For example, Baicalin enhances the proliferation of dermal papilla cells (DPC) and hair growth in mice (Shin et al., 2015, Naunyn Schmiedebergs Arch. Pharmacol. 388: 583-586). Epigallocatechin-3-gallate (EGCG) stimulates DPC proliferation (Kwon et al., 2007, Phytomedicine. 14: 551-555) and protects against dihydrotestosterone that has role in scalp balding (Shin et al., 2016, Ann. Dermatol. 28: 327-334). Quercetin promotes hair growth in vivo in mouse (Wikramanayake et al., 2012, Cell Stress Chaperones. 17: 267-274). Apigenin is an antioxidant with anti-inflammatory and

anti-tumor properties. It inhibits TGF-pi, which is an inhibitor of hair growth.
Apigenin promotes proliferation of human dermal papilla cells and epidermal
keratinocytes stimulate elongation of hair follicles in vitro (Hu et al., 2009, Arch
Dermatol. Res. 301(5): 381-385).
[005] Although numerous compositions, derived from plant sources, have been
described in the past, there still is a continuing interest in providing novel hair
growth compositions which demonstrate efficacy in improving hair volume and
stimulating hair growth.
SUMMARY OF THE INVENTION
[006] In an aspect of the present invention, there is provided a composition
comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein
naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a
range of 4-9: 1.5-2.5:0.75-1.5.
[007] In another aspect of the present invention, there is provided a process for
preparing a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10;
and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said
composition is in a range of 4 -9: 1.5 -2.5: 0.75 -1.5; said process comprising: i)
obtaining naringenin; ii) obtaining genistein; iii) obtaining coenzyme Q10; iv)
obtaining biotin; and v) contacting naringenin, genistein, coenzyme Q10, and biotin
to obtain the composition.
[008] These and other features, aspects, and advantages of the present subject
matter will be better understood with reference to the following description and
appended claims. This summary is provided to introduce a selection of concepts in
a simplified form. This summary is not intended to identify key features or
essential features of the claimed subject matter, nor is it intended to be used to limit
the scope of the claimed subject matter.
BRIEF DESCRIPTION OF THE DRAWINGS
[009] The detailed description is described with reference to the accompanying
figures. The same numbers are used throughout the drawings to reference like
features and components.

[0010] Figure 1 depicts the effect of combination of actives (naringenin, genistein,
and coenzyme Q10) with and without biotin, at a defined w/w ratio, on
transforming growth factor (TGF) - P2 gene expression, in accordance with an
embodiment of the present subject matter.
[0011] Figure 2 depicts the effect of composition comprising naringenin, genistein,
biotin, and coenzyme Q10 at varying w/w ratios on interleukin (IL) - 4 gene
expressions, in accordance with an embodiment of the present subject matter.
[0012] Figure 3 depicts gene expression analysis often genes on Sequencing Bead
Array (SB A), in accordance with an embodiment of the present subject matter.
[0013] Figure 4 depicts gene expression analysis of nine genes on SAB array, in
accordance with an embodiment of the present subject matter.
[0014] Figure 5 depicts the effect of composition of the present disclosure on
Bcl2/Bax gene ratio, in accordance with an embodiment of the present subject
matter.
[0015] Figure 6 depicts an in vitro hair growth assay, in accordance with an
embodiment of the present subject matter.
DETAILED DESCRIPTION OF THE INVENTION
[0016] Those skilled in the art will be aware that the present disclosure is subject to
variations and modifications other than those specifically described. It is to be
understood that the present disclosure includes all such variations and
modifications. The disclosure also includes all such steps, features, compositions,
and compounds referred to or indicated in this specification, individually or
collectively, and all combinations of any or more of such steps or features.
Definitions
[0017] For convenience, before further description of the present disclosure,
certain terms employed in the specification, and examples are delineated here.
These definitions should be read in the light of the remainder of the disclosure and
understood as by a person of skill in the art. The terms used herein have the
meanings recognized and known to those of skill in the art, however, for
convenience and completeness, particular terms and their meanings are set forth
below.

[0018] The articles "a", "an" and "the" are used to refer to one or to more than one
(i.e., to at least one) of the grammatical object of the article.
[0019] The terms "comprise" and "comprising" are used in the inclusive, open
sense, meaning that additional elements may be included. It is not intended to be
construed as "consists of only".
[0020] Throughout this specification, unless the context requires otherwise the
word "comprise", and variations such as "comprises" and "comprising", will be
understood to imply the inclusion of a stated element or step or group of element or
steps but not the exclusion of any other element or step or group of element or
steps.
[0021] The term "including" is used to mean "including but not limited to".
"Including" and "including but not limited to" are used interchangeably.
[0022] Ratios, concentrations, amounts, and other numerical data may be presented
herein in a range format. It is to be understood that such range format is used
merely for convenience and brevity and should be interpreted flexibly to include
not only the numerical values explicitly recited as the limits of the range, but also
to include all the individual numerical values or sub-ranges encompassed within
that range as if each numerical value and sub-range is explicitly recited.
[0023] The term "at least one" is used to mean one or more and thus includes
individual components as well as mixtures/combinations.
[0024] For the purposes of the present disclosure, the term "transparent" refers to
allowing light to pass through so that objects behind can be distinctly seen.
[0025] For the purposes of the present disclosure, the terms "thickener",
"humectant", "emulsifier", "hair oil", "neutralizer", "soothing agent",
"preservative", "revitalizing agent", "conditioning agent", "moisturizing agent",
"antidandruff agent", "hair cell stimulator", "diluent", "silicone oil", "antioxidant",
refer to the generally known components used in the art. The specific components
of the above-described categories have been mentioned in the present disclosure
wherever applicable. Wherever not mentioned, it can be construed that any
component falling under the categories can be used as part of the composition.

[0026] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the disclosure, the preferred methods, and materials are now described. All publications mentioned herein are incorporated herein by reference. [0027] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally-equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein. [0028] Apoptosis is a programmed cell death mechanism involved in elimination of cells that have performed their biological function during development and tissue homeostasis. It is an energy intensive process, initiated by a variety of stimuli, including withdrawal of growth factors, loss of cell adhesion, stimulation of pro-apoptotic receptors and DNA damage (Afford S and Randhawa S, 2000, Mol. Pathol. 53:55-63).
[0029] Hair Follicles cycle through three successive stages termed, anagen, catagen and telogen. Anagen, the growing stage of scalp hair follicle lasts about 2-7 years followed by catagen, the regression phase and telogen, the resting phase. At catagen stage, hair follicle detaches from hair bulb (dermal papilla) and this stage lasts for 2-4 weeks. Telogen is the resting phase and at this stage the detached hair (during catagen) falls off and the hair follicle unit rests for about 3 months. In general, on a healthy scalp, about 85-90% of hair follicles will be in anagen stage, while 10-15% remain in telogen stage. Generally, hair loss can be due to several factors including, internal factors like body physiology, hormonal changes, alopecia, androgenetic alopecia, diseases and stress, and external factors like lifestyle, excessive abrasion of scalp and pollution, to name a few. While, some low hair loss is common and part of a healthy life, excessive hair loss is not healthy. Promoting hair growth and minimizing hair loss are of paramount importance both for health and economic reasons.

[0030] Coordinated action of various biological pathways and molecules orchestrate the hair follicle cycle (Alonso and Fuchs, 2005, J. Cell Sci. 2006 119: 391-393; Rishikaysh et al., 2014, Int. J. Mol. Sci. 15: 1647-1670), and dysregulation of these pathways leads to abnormalities. Prominent pathways involved in hair biology include WNT, hedgehog, notch and bone morphogenetic protein (BMP) signaling. Proper positioning and placing of hair follicles is mediated by ectodysplasin A receptor (EDAR)-BMP signaling and their transcriptional interactions (Mou et al., 2006. Proc. Natl. Acad. Sci. 103: 9075-9080). Vascular endothelial growth factor (VEGF)-mediated angiogenesis controls hair follicle size and growth (Yano et al., J. Clin. Invest. 2001, 107(4): 409-417). Sonic hedgehog signaling (Callahan et al., 2004, Genes Dev. 18(22): 2724-2729; Mill et al., 2003, Genes Dev. 17: 282-294; St-Jacques et al., 1998, Curr. Biol. 8(19): 1058-1068) is essential for anagen initiation. Few other molecules required for maintenance of anagen include SGK3 and MSX2 (Alonso et al., 2005, J. Cell Biol. 170(4): 559-570; Ma et al., 2003, Development. 130: 379-389). Some growth factors essential for anagen include insulin-like growth factor (IGF), epidermal growth factor (EGF), fibroblast growth factor (FGF) and platelet derived growth factor (PDGF) (Sten and Pauss, 2001. Physiol. Rev. 81: 449-494). BMP signaling is essential in hair follicle differentiation (Botchkarev et al., 1999, Nat. Cell Biol. 1: 158-164; Kulessa et al., 2000, EMBO J. 19: 6664-6674), hair follicle morphogenesis, postnatal regeneration, modulating hair follicle cycling, hair matrix precursor cell proliferation and differentiation (Botchkare and Kishimoto, 2003. J. Investig. Dermatol. Symp. Proc. 8:46-55). Noggin is an inhibitor of BMPs and expressed by the mesenchyme. It is involved in hair follicle morphogenesis in embryos and promotes hair growth postnatally (Jamora et al., 2003. Nature. 422: 317-322; Kobielak et al., 2003. J. Cell Biol. 163: 609-623). Enhanced BMP signal activation or targeted inactivation of Noggin (BMP antagonist) results in significantly retarded hair follicle induction and progressive baldness (Botchkarev et al., 1999. Nature Cell Biol. 1: 158-164). Signaling by WNT-beta catenin proteins (Gat et al., 1998, Cell. 95: 605-614; Huelsken et al., 2001, Cell. 105: 533-545; Lo Celso et al., 2004, Development. 131: 1787-1799; Lowry et al., 2005, Genes Dev.

19: 1596-1611; Van Mater et al., 2003, Genes Dev. 17: 1219-1224; Zhang et al.,2009, Dev. Cell. 17: 49-61); Rabbani et al., 2011, Cell. 145: 941-955) and WNT proteins including WNT3a, WNT5a, WNTlOa, WNTlOb, frizzled7, disheveled 2, GSK3P, P-catenin and LEF1 are known to be associated in hair follicle development (Kishimoto et al., 2000, Genes Dev. 14: 1181-1185). The PDGFRA gene encodes platelet-derived growth factor receptor alpha (PDGFRA), which belongs to the family of receptor tyrosine kinases. The signaling pathway stimulated by PDGFRA controls many important cellular processes such as cell growth and division and cell survival. During hair follicle induction, to counteract the BMP inhibition, epithelial PDGF activates mesenchymal Platelet derived growth factor receptor alpha (PDGFRa). This, in combination with Sonic hedgehog (SFIH) signaling activates Noggin secretion by mesenchymal (dermal) cells, which in turn inhibits BMP signaling to promote induction, organogenesis and cellular differentiation (Gao et al., 2008, Genes Dev. 22: 2111-2124). [0031] Transition from anagen to catagen stage is mediated by growth factors like FGF5 and EGF, neurotrophins like BDNF and TFG beta family proteins like TFGP and BMPRIa (Andl et al., 2004, Development. 131: 2257-2268; Foitzik et al., 2000, FASEB J. 14: 752-760; Hansen et al., 1997, Am. J. Pathol. 150: 1959-1975; Hebert et al., 1994, Cell. 78: 1017-1025; Schmidt-Ullrich and Paus, 2005, Bioessays. 27: 247-261). During quiescence (catagen and telogn), hair follicle stem cells reside in hair bulge (Cotsarelis et al., 1990, Cell. 61: 1329-1337) and actively proliferate during transition from catagen to anagen stage. Several genes currently used as biomarkers of stem cell proliferation and differentiation are also essential in hair biology. CK19 is expressed during hair follicle budding in fetal and adult hair follicles (Schirren et al., 1997. Am. J. Dermatopathol. 19(4): 335-40). Recently it has been shown that the stem cells contribute to balding. The bald scalp although retains hair follicle stem cells, lacks CD200-rich and CD34-rich hair follicle progenitor cells (Garza et al., 2011, J. Clin. Invest. 121(2): 613-22). [0032] The telogen to anagen transition is facilitated by dermal papilla and hair follicle cell cross talk and interplay between WNT pathway, and BMP inhibitory factors (Kulessa et al., 2000. EMBO J. 19: 6664-6674; Greco et al., 2009. Cell

Stem Cell. 4: 155-169; Rabbani et al., 2011, Cell. 145: 941-955; Botchkarev et al., 2001. FASEB J. 15: 2205-2214; Zhang et al., 2006, Stem Cells. 24: 2826-2839; Enshell-Sejiffers et al., 2010, Dev. Cell. 18: 633-642). The hair follicle biology is complex interplay between various pathways and many of these molecules can be used as biomarkers in hair growth, and hair care product development. The effect of some of the most important genes on hair growth and their function in hair follicle cycle is herewith provided below in Table 1.

[0033] Conventional methods to stimulate hair growth include administration of drugs such as minoxidil which are associated with several side effects. Certain other conventional methods involve the use of soy protein isolates or powdered protein isolates that are known to have adverse effects on hair growth. Dietary supplements, vitamins, and plant source extracts for healthy growth of hair and nails are ineffective once the end user stops consumption of these supplements. Therefore, there still exists a need to develop novel compositions that overcome the drawbacks associated with prior art. Accordingly, objective of the present disclosure is to develop a cost-effective hair growth composition derived from plant sources that can effectively promote hair volume and growth. Accordingly, the present disclosure provides for a composition comprising naringenin, genistein, biotin, and coenzyme Q10, which when used in defined w/w ratios is effective in inhibiting IL-4 production and TGF-P2 expression, and consequently lower the induction of apoptosis in follicular keratinocytes that responsible for hair and nail growth.
[0034] Naringenin is a flavanone found mainly in variety of herbs and fruits, predominantly in grape fruit. It is known to have anti-inflammatory and analgesic effects. In addition, it has been shown to induce cell proliferation, protect against oxidative damage, induce vascular endothelial growth factor in hair follicle dermal papilla cells, and human keratinocytes (Kumar and Pandey, 2013, The Scientific World Journal. Article No. 162750).
[0035] Genistein is a phytoestrogen belonging to the subgroup of isoflavones. It is commonly found in several plants like, lupin, favabeans, soyabeans, kudzu, psoralea, coffee, and two species of shrubs, Flemingia vestita and Fleminga

macrophylla. Genistein is a potent inhibitor of protein-tyrosine kinase and topoisomerase-II and used as antineoplastic and antitumor active. [0036] Ubiquinone-50, also referred as coenzyme Q10 is a well-known cosmetic agent especially in skin care mainly due to its antioxidant potential. In cultured hair follicle keratinocytes, coenzyme Q10 stimulates hair keratin genes that are down-regulated during anti-aging process of hair follicles. In placebo-controlled clinical study in human volunteers of over 40 years, shampoo and tonic enriched with CoQIO enhanced the hair keratin gene expression (Giesen et al., 2009, Int. J. Cosmetic Sci. 31(2): 154-155) showing its potential as a hair growth active. [0037] Biotin, also termed as vitamin B7, is a water-soluble vitamin. It has multiple roles in fat, carbohydrate metabolism, cell growth and protein synthesis in body. Of late, several formulations either cosmetic or food include vitamin B7 with a focus on hair and nail health. Although research on direct effect of biotin on hair growth is limited, several reports of clinical success do exist, which show improvement in hair and nail health upon biotin supplementation in individuals with hair and nail pathology, like uncombable hair and brittle nail (Patel et al., 2017, Skin Appendage Disorder. Review. 3(3): 166-169).
[0038] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5. In another embodiment of the present disclosure, the naringenin to genistein to coenzyme Q10 weight ratio in said composition is in the range of 7-9:1.75-2.25-0.9-1.1. In yet another embodiment of the present disclosure, the naringenin to genistein to coenzyme Q10 weight ratio in said composition is 8:2:1.
[0039] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5: 0.01-1. In another embodiment of the present disclosure, naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in the range 7-9:1.75-2.25:0.9-1.1:0.01-1. In yet another embodiment of the present disclosure, naringenin to

genistein to coenzyme Q10 to biotin weight ratio in said composition is 8:2:1:1. In an alternate embodiment of the present disclosure, naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is 8:2:1:0.01. [0040] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5: 0.01-1.
[0041] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2%) with respect to the composition, and biotin has a weight percentage in a range of 0.0007%>-0.2%> with respect to the composition. In another embodiment of the present disclosure, naringenin has a weight percentage in the range of 0.6-0.9%) with respect to the composition, genistein has a weight percentage in the range of 0.1-0.3% with respect to the composition, coenzyme Q10 has a weight percentage in the range of 0.05-0.1%) with respect to the composition, and biotin has a weight percentage in the range of 0.001-0.1%) with respect to the composition. In yet another embodiment of the present disclosure, naringenin has a weight percentage of 0.8%> with respect to the composition, genistein has a weight percentage of 0.2% with respect to the composition, coenzyme Q10 has a weight percentage of 0.1% with respect to the composition, and biotin has a weight percentage of 0.001% with respect to the composition. In an alternate embodiment of the present disclosure, naringenin has a weight percentage of 0.8% with respect to the composition, genistein has a weight percentage of 0.2% with respect to the composition, coenzyme Q10 has a weight percentage of 0.1% with respect to the composition, and biotin has a weight percentage of 0.1% with respect to the composition.
[0042] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a

range of 4-9: 1.5-2.5: 0.75-1.5, and wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2%) with respect to the composition, and biotin has a weight percentage in a range of 0.0007%>-0.2%> with respect to the composition.
[0043] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5: 0.01-1; and wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2%) with respect to the composition, and biotin has a weight percentage in a range of 0.0007%>-0.2%> with respect to the composition.
[0044] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 8:2:1, wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2%) with respect to the composition, and biotin has a weight percentage in a range of 0.0007%>-0.2%> with respect to the composition.
[0045] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 8:2:1, and wherein naringenin has a weight percentage of 0.8% with respect to the composition, genistein has a weight percentage of 0.2% with respect to the composition, coenzyme Q10 has a weight percentage of 0.1% with respect to

the composition, and biotin has a weight percentage of 0.1% with respect to the composition.
[0046] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the composition further comprises at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, silicone oil, antioxidant, and combinations thereof. In an embodiment, the thickener is a carbomer; the humectant is glycerine; the emulsifier is at least one of polysorbate -20, microcrystalline wax, ceteareth-20, cetearyl alcohol, and combinations thereof; the hair oil is coconut oil; the silicone oil is polydimethyl siloxane polymer; the antioxidant is at least one of naringenin, genistein, coenzyme Q10, and combinations thereof; the neutralizer is triethanolamine; the soothing agent is allantoin; the preservative is phenoxyethanol, methylparaben, ethylparaben, n-butylparaben, isobutylparaben, and combinations thereof; the antidandruff agent is climbazole; the conditioning agent is PEG - 12 demethicone; the revitalizing agent is magnesium aspartate, zinc gluconate, copper gluconate; the hair cell stimulator is caffeine, biotin, and combinations thereof; the moisturizing agent is D-panthenol; and the diluent is water.
[0047] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5; and wherein the composition further comprises at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof.
[0048] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5:0.01-1, and wherein the composition further

comprises at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof.
[0049] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is 8:2:1, wherein the composition further comprises at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof. [0050] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5:0.01-1, and wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is 8:2:1, and wherein the composition further comprises at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof. [0051] In an embodiment of the present disclosure, there is provided a composition as described herein, wherein the composition further comprises at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof, and wherein the thickener has a weight percentage in a range of 0.1-0.2% with respect to the composition, the humectant has a weight percentage in a range of 0.5-2% with respect to the composition, the emulsifier has a weight percentage in a range of 4-8% with respect to the composition, the hair oil has a weight percentage in a range of 8-12% with respect to the composition, the silicone oil has a weight percentage in a range of 4-6% with respect to the composition, the

neutralizer has a weight percentage in a range of 0.05-0.2% with respect to the composition, the soothing agent has a weight percentage in a range of 0.05-0.2% with respect to the composition, the preservative has a weight percentage in a range of 0.3-1%) with respect to the composition, the revitalizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the conditioning agent has a weight percentage in a range of 3-10% with respect to the composition, the antidandruff agent has a weight percentage in a range of 0.3-1% with respect to the composition, the hair cell stimulator has a weight percentage in a range of 0.5-2% with respect to the composition, the moisturizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, and the diluent has a weight percentage in a range of 65-75%> with respect to the composition. In another embodiment of the present disclosure, thickener has a weight percentage in a range of 0.14-0.16%) with respect to the composition, the humectant has a weight percentage in a range of 0.75-1.25%) with respect to the composition, the emulsifier has a weight percentage in a range of 5-7% with respect to the composition, the hair oil has a weight percentage in a range of 9-11% with respect to the composition, the silicone oil has a weight percentage in a range of 4-6%o with respect to the composition; the antioxidants have a weight percentage of 1.0-1.2% with respect to the composition; the neutralizer has a weight percentage in a range of 0.75-0.2%) with respect to the composition, the soothing agent has a weight percentage in a range of 0.75-0.2%) with respect to the composition, the preservative has a weight percentage in a range of 0.4-0.6%) with respect to the composition, the revitalizing agent has a weight percentage in a range of 0.05-0.15%) with respect to the composition, the conditioning agent has a weight percentage in a range of 4-6% with respect to the composition, the anti dandruff agent has a weight percentage in a range of 0.4-0.6%) with respect to the composition, the hair cell stimulator has a weight percentage in a range of 0.5-1.5%) with respect to the composition, the moisturizing agent has a weight percentage in a range of 0.09-0.15%) with respect to the composition, and the diluent has a weight percentage in a range of 68-72%> with respect to the composition.

[0052] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; d) biotin; and e) at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is 4-9: 1.5-2.5: 0.75-1.5; and wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2% with respect to the composition, and biotin has a weight percentage in a range of 0.0007%-0.2% with respect to the composition, and wherein the thickener has a weight percentage in a range of 0.1-0.2% with respect to the composition, the humectant has a weight percentage in a range of 0.5-2% with respect to the composition, the emulsifier has a weight percentage in a range of 4-8% with respect to the composition, the hair oil has a weight percentage in a range of 8-12% with respect to the composition, the silicone oil has a weight percentage in a range of 4-6%) with respect to the composition, the neutralizer has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the soothing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the preservative has a weight percentage in a range of 0.3-1% with respect to the composition, the revitalizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the conditioning agent has a weight percentage in a range of 3-10% with respect to the composition, the antidandruff agent has a weight percentage in a range of 0.3-1% with respect to the composition, the hair cell stimulator has a weight percentage in a range of 0.5-2%> with respect to the composition, the moisturizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, and the diluent has a weight percentage in a range of 65-75%> with respect to the composition.
[0053] In an embodiment of the present disclosure, there is provided a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; d) biotin; and e) at least

one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 0.4-9: 1.5-2.5: 0.75-1.5:0.01-1, and wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2%) with respect to the composition, and biotin has a weight percentage in a range of 0.0007%>-0.2%> with respect to the composition, and wherein the thickener has a weight percentage in a range of 0.1-0.2% with respect to the composition, the humectant has a weight percentage in a range of 0.5-2% with respect to the composition, the emulsifier has a weight percentage in a range of 4-8% with respect to the composition, the hair oil has a weight percentage in a range of 8-12% with respect to the composition, the silicone oil has a weight percentage in a range of 4-6%) with respect to the composition, the neutralizer has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the soothing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the preservative has a weight percentage in a range of 0.3-1% with respect to the composition, the revitalizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the conditioning agent has a weight percentage in a range of 3-10% with respect to the composition, the antidandruff agent has a weight percentage in a range of 0.3-1% with respect to the composition, the hair cell stimulator has a weight percentage in a range of 0.5-2% with respect to the composition, the moisturizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, and the diluent has a weight percentage in a range of 65-75%> with respect to the composition.
[0054] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5, said process comprising: i)

obtaining naringenin; ii) obtaining genistein; iii) obtaining coenzyme Q10; iv) obtaining biotin; and v) contacting naringenin, genistein, coenzyme Q10, and biotin to obtain the composition.
[0055] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5: 0.01-1; said process comprising: i) obtaining naringenin; ii) obtaining genistein; iii) obtaining coenzyme Q10; iv) obtaining biotin; and v) contacting naringenin, genistein, coenzyme Q10, and biotin to obtain the composition.
[0056] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin; wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5: 0.01-1, and wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2% with respect to the composition, and biotin has a weight percentage in a range of 0.0007%-0.2% with respect to the composition; said process comprising: i) obtaining naringenin; ii) obtaining genistein; iii) obtaining coenzyme Q10; iv) obtaining biotin; and v) contacting naringenin, genistein, coenzyme Q10, and biotin to obtain the composition.
[0057] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; d) biotin; and e) at least one carrier, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5; said process comprising: i) obtaining naringenin; ii) obtaining genistein; iii) obtaining coenzyme Q10; iv) obtaining biotin; v) obtaining at least one carrier; and vi) contacting naringenin, genistein, coenzyme Q10, biotin; and the at least one carrier to obtain the composition.

[0058] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; d) biotin; and e) at least one carrier, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5: 0.01-1; said process comprising: i) obtaining naringenin; ii) obtaining genistein; iii) obtaining coenzyme Q10; iv) obtaining biotin; v) obtaining at least one carrier; and vi) contacting naringenin, genistein, coenzyme Q10, biotin; and the at least one carrier to obtain the composition.
[0059] In an embodiment of the present disclosure, there is provided a process for preparing a composition comprising: a) naringenin; b) genistein; c) coenzyme Q10; d) biotin; and e) at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, the moisturizing agent, antidandruff agent, hair cell stimulator, diluent, and combinations thereof, wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2%) with respect to the composition, and biotin has a weight percentage in a range of 0.0007%>-0.2%> with respect to the composition, and wherein the thickener has a weight percentage in a range of 0.1-0.2%) with respect to the composition, the humectant has a weight percentage in a range of 0.5-2%> with respect to the composition, the emulsifier has a weight percentage in a range of 4-8%> with respect to the composition, the hair oil has a weight percentage in a range of 8-12% with respect to the composition, the silicone oil has a weight percentage in a range of 4-6%o with respect to the composition, the neutralizer has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the soothing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the preservative has a weight percentage in a range of 0.3-1% with respect to the composition, the revitalizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the conditioning agent has a weight percentage in a range of 3-10% with respect to the composition, the antidandruff agent has a

weight percentage in a range of 0.3-1% with respect to the composition, the hair
cell stimulator has a weight percentage in a range of 0.5-2% with respect to the
composition, the moisturizing agent has a weight percentage in a range of 0.05-
0.2%) with respect to the composition, and the diluent has a weight percentage in a
range of 65-75%> with respect to the composition; said process comprising: i)
obtaining naringenin; ii) obtaining genistein; iii) obtaining coenzyme Q10; iv)
obtaining biotin; v) obtaining at least one carrier; and vi) contacting naringenin,
genistein, coenzyme Q10, biotin; and the at least one carrier to obtain the
composition.
[0060] In an embodiment of the present disclosure, there is provided a composition
as descried herein, wherein the composition inhibits transforming growth factor
(TGF)-P expression and interleukin (IL)-4 expression.
[0061] In an embodiment of the present disclosure, there is provided a composition
as descried herein, wherein the composition leads to hair growth.
[0062] In an embodiment of the present disclosure, there is provided a composition
as descried herein, wherein the composition is in a form of serum.
[0063] In an embodiment of the present disclosure, there is provided a composition
as descried herein, wherein the composition is in a form of oil.
[0064] In an embodiment of the present disclosure, there is provided a composition
as descried herein, wherein the composition is in a form of cream.
[0065] Although the subject matter has been described with reference to specific
embodiments, this description is not meant to be construed in a limiting sense.
Various modifications of the disclosed embodiments, as well as alternate
embodiments of the subject matter, will become apparent to persons skilled in the
art upon reference to the description of the subject matter. It is therefore
contemplated that such modifications can be made without departing from the spirit
or scope of the present subject matter as defined.
EXAMPLES
[0066] The disclosure will now be illustrated with working examples, which is
intended to illustrate the working of disclosure and not intended to take
restrictively to imply any limitations on the scope of the present disclosure. Unless

defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply. [0067] The working examples described herein clearly depict the effect of the composition of the present disclosure on transforming growth factor (TGF) -P, and interleukin (IL) - 4 gene expressions. The present disclosure is directed to a composition comprising naringenin, genistein, biotin, and coenzyme Q10, which when used in defined w/w ratio's is effective in downregulating TGF-P, and IL-4 production, and consequently lower the induction of apoptosis in follicular keratinocytes that responsible for hair and nail growth. MATERIALS AND METHODS
[0068] The ingredients naringenin, genistein, and coenzyme Q10 were procured from Biogen Idee Biotech, Pvt. Ltd. India. Biotin was procured from Sigma. Example 1: RNA extraction and Real-time PCR
[0069] For this purpose, HaCaT cells were seeded onto a 12-well plate at a density of 1 x 105 cells/ well and incubated at 37°C with 5% C02, for about 24 hours (till 70-80% confluency is reached). The cells were then treated with actives in at individual concentration as well as in combinations of different w/w ratios. The cells were simultaneously treated with different combinations of both the actives and incubated at 37°C, with 5% CO2 for 24 hours. Post incubation, the media from the wells were discarded, and the cells were washed with 1ml of lx phosphate buffer saline (PBS) followed by addition of 1 ml of trizol reagent (Ambion, Life Technologies, USA) in each well. The plate was incubated at -80°C overnight and RNA isolation was performed next day in the same trizol reagent. The steps followed for the RNA isolation were as per the manufacturer's protocol (Ambion, Life Technologies, USA). The resulting RNA was quantified using TECAN, Infinite M200 Pro, USA, a multimode microplate reader. cDNA was synthesized

from the RNA, using High Capacity cDNA reverse transcription kit 200 reactions (Applied Biosystems, USA). The expression levels of target gene, TGF - p, and IL-4 was determined by real time PCR using ABI7500 Real Time PCR system from Applied Biosystems USA. GAPDH was used as endogenous control for the present study. The sequences of the primers are herewith provided below in Table
The PCR program was as follows: (a) preheating for 2 minutes at 50°C and heating for 10 minutes at 95°C (b) 40 cycles of denaturation, annealing and extension at 95°C for 15 sec and 60°C for 1 minute (c) melting curve analysis at 95°C for 15 seconds, 60°C for 1 minute, 95 °C for 30 seconds and 60 °C for 15 sec. The sxpression levels of TGF - p, and IL-4 genes were normalized to GAPDH gene sxpression in each sample.
Example 2: Sequencing Bead Array (SBA) analysis
[0070] Following the informed consent, human hair follicles were plucked from the scalp of healthy donors using a standardized protocol by medical doctors at MSCR Pvt. Ltd., Bangalore. Hair follicles were collected and maintained in William's E medium (Invitrogen, CA, USA) until use. Thirty hair follicles were treated with a combination of naringenin, genistein and coenzyme Q10 in the ratio (8:2:1) at a concentration of 20ug/ml and 0.1% of biotin. Control group included follicles treated with vehicle control (DMSO). Follicles were treated for 72 hours and collected in a microcentrifuge tube. Further, the hair follicles were washed with lxPBS and stored in RNA later solution (Thermo Scientific, USA. Cat. No.

AM7020). Total RNA was isolated from treated human hair follicles according to the protocol supplied by the manufacturer (RNeasy mini kit, Qiagen, Cat. No. 74124). The first strand of cDNA was reverse-transcribed from the total RNA (350ng/reaction) using the RT2 first strand kit (Qiagen). PCR was performed in 20ul reactions containing SYBR green master mix (2x RT2 SYBR Green Mastermix, Cat. No. 330523, Qiagen). Briefly, cDNA was added to 25ul reaction mixture containing 2x RT2 SYBR Green Mastermix, random primers and 10X buffer as per manufacturer's protocol. The PCR was performed on custom SAB Array plates (Qiagen) at heating conditions as follows: 10 min at 95°C, 40 cycles of 15sec at 95°C and 60sec at 600C, followed by melting curve analysis. GAPDH was used as housekeeping gene. Data was analyzed for comparison of fold change gene expression between control group and that in treatment group. Example 3: In vitro hair growth assay
[0071] Hair follicles from the scalp of healthy donors were plucked using a standardized protocol after taking their consent by medical doctors. Hair follicles were collected and maintained in William's E medium (Invitrogen, CA, USA) till use. Following a wash in lxPBS (Sigma), the hair follicles were trimmed to about lcm from the bulb using scissors without harming the root. Only those hair follicles with an intact bulb and root sheath (RS) were selected for this assay. Twenty hair follicles were treated with 1:50 diluted hair tonic. Hair tonic was prepared in-house comprising of combination of ingredients including naringenin, genistein and coenzyme Q10 in the ratio (8:2:1) at a concentration of 20ug/ml and 0.1% of biotin. Similarly, twenty hair follicles were used for treatment with market product (at 1:50 dilution), Williams E medium including supplements (glutamine 2mM, hydrocortisone 10 ng/ml, insulin 10 ug/ml) and Williams E medium without supplements. Follicles were cultured in 24-well plates in a standard cell culture incubator. Single hair follicles were maintained in a free-floating and isolated state in each well. On day zero, the length of each hair follicles was measured under the microscope (Leica DMI 4000B) using Leica Q Win Image processing and analysis software V.3.5.0. Medium change was performed on every alternate day and the length of hair follicles were measured subsequently. The day the hair follicles were

inoculated was considered as day zero and hair length was measured for each hair follicle. On day 7, the final measurement was performed and the same was subtracted from the corresponding measurement on day zero. The difference in length (in micrometer) for each hair follicle was calculated. The average increase of all hair follicles in a treatment was taken as hair growth and the standard errors were calculated. Example 4: Formulation
[0072] The formulation as provided below was prepared in which all the constituents, were combined in different fractions, as provided below in Table 3.

Example 5: Process of preparation of the formulation
[0073] The components required for preparing the composition as described herein were all commercially procured. The formulations Al as disclosed in Example 4, was prepared by mixing all the constituents namely naringenin, genistein, coenzyme Q10, biotin, and obtaining carriers comprising carbomer, glycerine, polysorbate 20, coconut oil, microcrystalline wax, polydimethyl siloxane polymer, ceteareth-20, cetearyl alcohol, triethanolamine, allantoin, phenoxyethanol, methylparaben, ethyl paraben, n-butylparaben, isobutyl paraben, magnesium aspartate, zinc gluconate, copper gluconate, PEG-12 dimethicone, climbazole, caffeine, D-panthenol, and water, at a temperature range of 25 - 27 °C to obtain the composition.
RESULTS AND DISCUSSION
Evaluating the effect of the composition on TGF-p and IL-4 gene expression [0074] In order to evaluate the effect of the biotin in the composition on TGF-P gene expression, two different combinations of actives (wherein one combination contained biotin, and the other combination did not contain biotin) at defined w/w ratio was exposed to HaCaat cells. The results of this study are presented in Figure 1. DMSO was used as a control. 'N', 'G', 'C, and 'B' as used in the Figures 1 and 2 correspond to naringenin, genistein, coenzyme Q10, and biotin. The active combination of naringenin, genistein and coenzyme Q10 at 8:2:1 ratio shows about 65% inhibition of TGF-P gene expression in comparison to the control. However, the combination including naringenin, genistein, coenzyme Q10 at 8:2:1 ratio with biotin at 0.001% shows around 70% inhibition of TGF-P gene expression. This data suggests that the apoptosis induction by the combination comprising biotin is lower than that by the other combination including only three actives, naringenin, genistein and coenzyme Q10 at 8:2:1.
[0075] Based on the results observed in Figure 1, experiments were conducted at varying w/w ratios and their effect on percentage reduction in IL-4 was observed. The results are herewith presented below in Table 4, and Figure 2. Table 4

[0076] As can be seen in Table 4, and Figure 2, for the various w/w ratios of naringenin, genistein, coenzyme Q10, and biotin tested, it can be observed naringenin, genistein and coenzyme Q10 at 8:2:1 ratio with biotin at 0.001% shows lower IL-4 gene expression (signifying increased inhibition) compared to that in the other combination of 3:3:3:0.5 ratio with 0.0005% of biotin. This suggests that the combination of 8:2:1 ratio with biotin at 0.001% has the least potential to induce apoptosis, and consequently offer great benefits for hair growth applications.
Gene expression profile analysis
[0077] Gene expression profile analysis showed that 19 genes required for hair growth are upregulated in HaCaaT cells treated with the composition of the present disclosure (8:2:1 ratio of naringenin, genistein and coenzyme Q10 with 0.1% biotin), as can be observed in Figure 3, and Figure 4. Gene expression ranged from 1.96-fold in the WNT10 gene to 72-fold in the HGF gene. Expression levels of nine genes namely, MK167, WNT3A, SCF, SUR1, ALP1, IGF1, EDAR, PDGRA and HGF was very high ranging from 12.3-fold to 72-fold. Other ten genes namely, WNT10B, CD200, CK19, NOGGIN, APCDD1, CD271, NANOG, WNT10A, KIR6.1 and SUR2B showed moderately high expression levels. The expression of each of these genes was normalized to the control (GADPH) The role of these genes in hair follicle biology is mentioned in the table 1. These genes are involved in WNT pathway, EDAR pathway, PDGF pathway, cell proliferation, stem cell proliferation and maintenance, and vasodilation. In addition, the expression of Bcl2 gene, which is an apoptosis inhibitor is upregulated and the expression of Bax gene, an apoptosis promoter is downregulated. Hence, in effect, the ratio of Bcl2/Bax is 5.1, which is much higher than that observed in the control treatment,

as can be observed in Figure 5. The term "CDQB" as mentioned in Figure 5 and Figure 6 correspond to the composition of the present disclosure. The sequences of primers are herewith provided below in Table 3.
In vitro hair growth assay
[0078] Figure 6 corroborates the results observed in gene expression analysis. Enhanced hair growth was observed in hair follicles treated with 1:50 dilution of ITC hair tonic formulation (composition of the present disclosure comprising 8:2:1 ratio of naringenin, genistein and coenzyme Q10 with 0.1% biotin). The hair growth was higher than that observed in market product. When compared to the negative control wherein the hair follicles were treated with Williams E medium without supplements, the hair growth was significantly higher. The hair growth in treatment with hair tonic was almost similar to that in positive control wherein the hair follicles were treated with Williams E medium including supplements. [0079] Overall, the data suggests that the composition of the present disclosure, i.e., naringenin, genistein, coenzyme Q10 and biotin at w/w ratio of 8:2:1:0.01, is most effective in lowering and TGF - P, and IL-4 expression, thereby lowering the potential to induce apoptosis in keratinocytes, and consequently promote hair growth. Further, the composition of the present disclosure is also effective in upregulating genes involved in hair follicle cycle, that are responsible for promoting hair growth. Advantages of the present disclosure:
[0080] The present disclosure discloses a composition comprising a) naringenin; b) genistein; c) coenzyme Q10; and d) biotin, wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5. The composition of the present disclosure can be effectively deployed in the

cosmetic industry as it reduces transforming growth factor (TGF) - P, and interleukin (IL)-4 gene expression; and consequently, lower the induction of apoptosis in follicular keratinocytes, thereby promoting hair and nail growth. The composition has unique potential benefit for anti-hair loss or hair growth applications at defined w/w ratios. Further, the compositions of the present disclosure utilize non-synthetic components; and are therefore safe for application, with minimal side effects.

I/We Claim:
1. A composition comprising:
(a) naringenin;
(b) genistein;
(c) coenzyme Q10; and
(d) biotin,
wherein naringenin to genistein to coenzyme Q10 weight ratio in said composition is in a range of 4-9: 1.5-2.5: 0.75-1.5.
2. The composition as claimed in claim 1, wherein naringenin to genistein to coenzyme Q10 to biotin weight ratio in said composition is in a range of 4-9: 1.5-2.5:0.75-1.5: 0.01-1.
3. The composition as claimed in claim 1, wherein naringenin to genistein to coenzyme Q weight ratio in said composition is 8:2:1.
4. The composition as claimed in claim 1, wherein naringenin has a weight percentage in a range of 0.5%-1.0% with respect to the composition, genistein has a weight percentage in a range of 0.1%-0.4% with respect to the composition, coenzyme Q10 has weight percentage in the range of 0.05-0.2% with respect to the composition, and biotin has a weight percentage in a range of 0.0007%-0.2% with respect to the composition.
5. The composition as claimed in claim 4, wherein naringenin has a weight percentage of 0.8% with respect to the composition, genistein has a weight percentage of 0.2% with respect to the composition, coenzyme Q10 has a weight percentage of 0.1% with respect to the composition, and biotin has a weight percentage of 0.001% with respect to the composition.
6. The composition as claimed in any one of the claims 1-5, wherein the composition further comprises at least one carrier selected from a group consisting of thickener, humectant, emulsifier, hair oil, silicone oil, neutralizer, soothing agent, preservative, revitalizing agent, conditioning agent, antidandruff agent, hair cell stimulator, moisturizing agent, diluent, and combinations thereof.

7. The composition as claimed in claim 6, wherein the thickener has a weight percentage in a range of 0.1-0.2% with respect to the composition, the humectant has a weight percentage in a range of 0.5-2% with respect to the composition, the emulsifier has a weight percentage in a range of 4-8% with respect to the composition, the hair oil has a weight percentage in a range of 8-12% with respect to the composition, the silicone oil has a weight percentage in a range of 4-6% with respect to the composition, the neutralizer has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the soothing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the preservative has a weight percentage in a range of 0.3-1% with respect to the composition, the revitalizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, the conditioning agent has a weight percentage in a range of 3-10% with respect to the composition, the antidandruff agent has a weight percentage in a range of 0.3-1% with respect to the composition, the hair cell stimulator has a weight percentage in a range of 0.5-2%) with respect to the composition, the moisturizing agent has a weight percentage in a range of 0.05-0.2%) with respect to the composition, and the diluent has a weight percentage in a range of 65-75%> with respect to the composition.
8. A method to prepare the composition as claimed in claim 1, said process comprising:

(a) obtaining naringenin;
(b) obtaining geni stein;
(c) obtaining coenzyme Q10;
(d) obtaining biotin; and
(e) contacting naringenin, genistein, coenzyme Q10, and biotin to obtain the composition.
9. A method to prepare the composition as claimed in claim 6, said method
comprises:
(a) obtaining naringenin;

(b) obtaining geni stein;
(c) obtaining coenzyme Q10;
(d) obtaining biotin;
(e) obtaining the at least one carrier; and
(f) contacting naringenin, genistein, coenzyme Q10, biotin, and the at least one carrier to obtain the composition.
10. The composition as claimed in any one of the claims 1-7, wherein the composition inhibits TGF-P expression and IL-4 expression.