DESC:FIELD OF THE INVENTION

The present invention relates to a synergistic antimicrobial composition involving anti microbial actives/ agents. More specifically, the present invention relates to a synergistic anti microbial composition including selective combination of cinnamic acid derivatives and any one compound selected from the group consisting of naphthoquinone derivatives and terpenophenol derivatives. The advanced antimicrobial composition is directed to provide synergistic inhibition of fungal and bacterial growth including those which are responsible for a number of skin infections. Advantageously, the present advancement also relates to a cosmetic, dermopharmaceutical and or pharmaceutical composition having such special antimicrobial activity especially including anti dandruff effect, stimulates hair growth and providing skin care benefits.

Background art
The human skin is the outer covering of the body and interfaces with the environment, playing an important role in immunity, protecting the body against pathogens and excessive water loss. Varieties of microorganisms colonize the skin surface, depending on region of the skin as moist, dry and sebaceous. It is well known that Corynebacteria , Staphylococci , ß-Proteobacteria, Flavobacteriales, Candida, Pityrosporum, Microsporum, Trichophyton(T.) Epidermophyton are some of the important microorganisms causing various skin infections.
Skin infections that have inflammation as one of the major mechanism/root cause are also well known to be seborrheic dermatitis, eczema, bacterial, viral, fungal infections, contact dermatitis and acne. Barber itch jock itch. athletes foot, ringworm are some of the different types of well known and common fungal infection .
Among various types of skin infections, dandruff is a very common scalp disorder, inflammatory in nature, affecting almost half of the population at the pre-pubertal age and of any gender and ethnicity. The presence of dandruff accompany telogen effluvium and exacerbate androgenetic alopecia. It has been observed that about 100-300 numbers of hairs were shed in dandruff sufferers instead of 50-100 in normal subjects. .
Older literature cites the fungus Malassezia furfur (previously known as Pityrosporum ovale) as the cause of dandruff. While this species does occur naturally on the skin surface of both healthy people and those with dandruff, in 2007 it was discovered that the responsible agent is a scalp specific fungus, Malassezia globosa, that metabolizes triglycerides present in sebum by the expression of lipase, resulting in a lipid byproduct oleic acid (OA). During dandruff, the levels of Malassezia increase by 1.5 to 2 times its normal level. Penetration by OA of the top layer of the epidermis, the stratum corneum, results in an inflammatory response in susceptible persons, which disturbs homeostasis and results in erratic cleavage of stratum corneum cells of the skin epidermis.
There are seven species of Malassezia - M. globosa, M. resticta, M. obtuse, M. sloofiae, M. sympodialis, M. furfur and M. pachydermatis, which are found to be associated with dandruff formation. The oily secretion of skin, metabolic by-products of skin micro-organisms and individual susceptibility are usually associated with dandruff.
The pathogenesis of dandruff involves hyper proliferation of keratinocytes, resulting in deregulation of keratinization. The corneocytes clump together, manifesting as large flakes of skin. Thus dead skin cells are shed in large, oily clumps, which appear as white or greyish patches on the scalp, skin and clothes. For people with dandruff, skin cells may mature and be shed in 2–7 days, as opposed to around a month in people without dandruff. The spectrum of dandruff is difficult to define because it blurs with seborrhoeic dermatitis and some other scaly conditions. The inflammation and extension of scaling outside the scalp exclude the diagnosis of dandruff from seborrhoeic dermatitis. Both in dandruff and seborrhoeic dermatitis, the population of Malassezia is not uniform throughout the skin surface and inside the stratumcorneum
The non-microbial cause for dandruff includes excessive exposure to sunlight, which is known to cause desquamation of the scalp. Minimal irritation of scalp due to over shampooing, frequent combing, use of certain cosmetic products, dusts and dirt also, to some extent, cause dandruff.
A number of natural anti dandruff agents are known such as coal tar (anti proliferative effect), neem (anti fungal effect), black pepper (dandruff fighting minerals zinc and selenium).
Normally the response of dandruff to treatment is swift, but transient. Although conventional antifungal drugs are available (Keratolytic agents, griseofulvin, selenium sulfide, azoles, allylamines and morpholine derivatives, Gentian violet 2.5%, Nystatin, Clatrimazole and amphotericin etc.,) an increasing resistance to these conventional compounds results in treatment failure. Use of herbal drugs for oral and tropical applications is also know. However, the skin infections caused by bacteria, virus, fungi still remains a challenge
In some cases, commercial antimicrobial compounds cannot provide effective control of microorganisms, even at high use concentrations, due to weak activity against certain types of microorganisms,. US Pat. No. 6,197,805 discloses a combination of 3-iodo-2-propynyl-butylcarbamate(IPBC) and 2-(methoxycarbonylamino) benzimidazole.

In this context some naturally occurring compounds have been explored for effective antimicrobial function .

Ethyl-p-methoxycinnamate (EPMC)
Ethyl-p-methoxycinnamate (EPMC) is a cinnamic acid derivative. EPMC has been isolated from Curcuma zedoaria and Hedychium spicalum and has been shown to inhibit a wide range of fungi.
US 20080113047A1 discloses, compositions containing extract of Hedychium spicalum are useful for treating Tinea infections by topical application. Extracts from rhizomes of Hedychium spicalum are particularly useful. The authors describes isolation and anti-dermatophytic activity of formulations containing EPMC.

Juglone
Juglone is a naphthoquinone, an aromatic organic compound that is commonly found in nature, major source being walnut tree, the most widespread naphthoquinones are juglone, lawsone, plumbagin, and lapachol. It has a distinct phenyl group and is not the only phenolic compound produced by the walnut tree; other phenolic compounds include flavonoids, terpenoids, other naphthoquinones and examples are Gallic acid, Caffeic acid, myricetin, and quercetin. Juglone is also the compound responsible for yellow pigmentation in the walnut tree.
Juglone has antifungal properties similar to some commercially available antifungal agents used to treat infections, such as athlete’s foot and ringworm. One study showed that juglone inhibits three key enzymes from Helicobacter pylori, a bacterium that can cause gastritis, peptic ulceration, and gastric cancer in humans. This bacterium affects approximately 50% of the world’s population and antibiotic resistance is a rising problem. Juglone could become a new antibacterial agent to treat infections from this bacterium

Bakuchiol
Bakuchiol is a meroterpene (a chemical compound having a partial terpenoid structure) in the class terpenophenol. It is found in Psoraleacorylifolia and in Otholobiumpubescens. It has been reported to have anticancer activity in pre-clinical models, possibly due to its structural similarity with resveratrol. Bakuchiol is an phytoestrogen, antioxidant, inhibitor of protein tyrosine phosphatase 1B (PTP1B). It also acts as an inhibitor of mitochondrial lipid peroxidation and inducible nitric oxide synthase (iNOS; NOS II) expression. Bakuchiol is also a DNA polymerase inhibitor which shows antimicrobial, anti-inflammation, anti-tumor, immunosuppressive, and hepatoprotective effects, as well as hypoglycemic and cytotoxic activity. It was recently found that bakuchiol is active against influenza virus with an IC50 of 0.2 µM.

US 20060251749Al describes isolation and purification of Bakuchiol and also its antifungal activity, particularly against the type of fungi that cause athlete's foot. Interestingly, part of its mechanism of action against fungi is via generation of ROS, which stands for reactive oxygen species, a type of free radical that, depending on the circumstances, can have a positive effect on health. The authors have clearly mentioned that Bakuchiol is not effective against Pityrosporum ovale a microorganism causing dandruff. Bakuchiol may also have a suppressive effect on melanin-producing cells in skin, which may make it a beneficial ingredient to lighten dark spots or improve uneven skin tone due to sun damage.

Thus , there is still a need for a broad spectrum antimicrobial composition involving synergistically special combinations of several antimicrobial actives for controlling various strains of microorganisms causing skin infections and simultaneously imparting skin benefits.

Objects of the invention:

It is thus the primary object of the present invention is to provide an anti-microbial compositions which is synergistic combinations of antimicrobial actives/ agents in an effective amounts which is safe and compatible to the skin and scalp.

Another object of the present invention is to provide a antimicrobial composition, which synergistically inhibit fungal growth.

Another object of the present invention is to provide for a antimicrobial composition having effective anti-microbial activity such as anti-bacterial etc.

Another object of the present invention is to provide a antimicrobial composition, which on one hand would provide synergistic anti dandruff action and on the other hand acts as a hair care benefiting agent such as prevention of hair fall, favouring to hair growth, Damage protection etc.

Another object of the present invention is to provide a antimicrobial composition, which on one hand would provide synergistic anti dandruff action and on the other hand acts as a skin care benefiting agent such as melanin inhibition, UV protection etc.

Another object of the present invention is to provide an anti-microbial composition in an effective amounts are safe and compatible to the skin and scalp in a cosmetically / dermopharmaceutically acceptable vehicle with or without other skin or hair care benefiting agents.

Another object of the present invention is to provide a anti-microbial compositions in an effective amounts in a safe and acceptable oral dosage form in a pharmaceutically acceptable vehicle with or without other anti microbial agents.

Another object of the present invention is to provide a anti-microbial compositions in an effective amounts which is beneficial for hair & skin can be left on the skin or scalp for considerable period of time using a suitable vehicles such as creams, lotions, oils, gels etc.

Another objective of the present invention is to provide for an improved cosmetic hair care composition which on one hand would be efficacious in controlling the growth of microorganism in the scalp and on the other hand stimulate hair growth to overcome the limitations and disadvantages observed in the existing anti dandruff and hair growth agents with better and wider consumers acceptance.

Yet another object of the present invention is to provide for skin and hair care composition comprising said synergistically special combinations of antimicrobial actives/ agents which can be obtained either synthetically or semi-synthetically.

Another object of the present invention is to provide cosmetic anti microbial composition which would be cost effective, and safe in having less or no side effects.

Summary of invention:
Thus according to the basic aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical composition comprising broad spectrum synergistic anti microbial/anti-fungal composition comprising atleast a compound selected from the cinnamic acid derivatives of Formula-1, with any one compound selected from the group consisting of Naphthoquinone derivatives of Formula-2, or terpeneophenol derivatives of Formula -3;
FORMULA-1:

where in R1 , R2 = C1-C14 alkyl/ C1-C14 alkene/ C1-C14 alkyne;
FORMULA -2:

Such that
When R1 = OH, R2 = H OR C1-C6 alkyl or C1 – C6 Alkene and R3 = H
When R1 = C1-C4 alkyl, R2 = H and R3 = OH
When R1and R2 = H, R3 = OH
FORMULA -3:

Where in R1 = C12 – C15 alkyl or C12 – C15 alkene with 2 to 4 Carbon carbon double bond or C12 – C15 alkyne with 2 to 3 carbon carbon triple bond.
In accordance with another preferred aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical composition comprising of Ethyl-p-methoxy cinnamate of Formula-4 with any one compound selected from the group of compounds of Formula -5 and Formula -6, wherein the compounds of Formula 4 , 5 and 6 are as represented hereunder:
FORMULA -4: C12H14O3. Ethyl 4-Methoxycinnamate; Ethyl p-methoxycinnamate; (E)-Ethyl 3-(4-methoxyphenyl)acrylate; Ethyl 3-(4-methoxyphenyl)acrylate; Ethyl trans-4-methoxycinnamate.

FORMULA-5: C10H6O3. Juglone, 5-Hydroxy-1, 4-naphthalenedione, 5-Hydroxy-1,4-naphthoquinone, 5-Hydroxy-p-naphthoquinone, 5-hydroxynaphthoquinone , Regianin, , Nucin, NCI 2323, Oil Red BS

FORMULA -6: C18H24O Bakuchiol, 4-[(1E,3S)-3-ethenyl-3,7-dimethylocta-1,6-dienyl] phenol

In accordance with another aspect of the present invention there is provided a cosmetic and/or dermopharmaceuticalcomposition comprising a synergistically coacting anti microbial /anti-fungal combination of Ethyl-p-methoxycinnamate of Formula-4 with any one compound selected from Juglone of Formula-5 and Bakuchiol of Formula-6.

Yet another aspect of the present invention relates to a cosmetic and/or dermopharmaceutical composition wherein said compound selected from the cinnamic acid derivatives of Formula-1: said any one compound selected from the group consisting of Naphthoquinone derivatives of Formula-2, and terpeneophenol derivatives of Formula -3 is in the range of 9.9:0.1 to 0.1:9.9 preferably 9:1 to 1:9 .

In accordance with another aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical antimicrobial composition, having synergistic anti dandruff action and related hair care benefits.
In accordance with another preferred aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical composition comprising a suitable vehicle including aqueous, anhydrous, emulsions, creams, lotions, Surfactants, oils etc., wherein in the vehicle is in range from 5 to 99.9%, preferably from 20 to 85%, more preferably from between 1 to 99% by weight.

In accordance with another aspect of the present invention there is provided a cosmetic and / or dermopharmaceutical composition comprising hair and/or skin care benefiting agents and cosmetically/ pharmaceutically acceptable vehicles.
In accordance with another aspect of the present invention there is provided an anti-microbial compositions comprising a leave on product including oil or a wash off product including surfactant.

In accordance with another aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical composition, comprising hair benefit agents or mixture of hair benefiting agents including Hair fall reducing agents, Hair growth promoting agents, anti-aging, sebum controlling/ reducing agents, plasticizers, elastomers, pigments, antioxidants, chelating agents, and perfumes, as well as organic/ inorganic sunscreens and including sunscreens as UV diffusing/ protection agents and optionally, adjunct minor components /ingredients including coloring agents, opacifiers, pH adjusting agents and perfumes etc in an amounts in the range from 0.01 % up to 50 % by weight, preferably 0.1 to 20% by weight, more preferably 0.2 to 10% by weight of the composition.

In accordance with another aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical composition, comprising skin benefit agents or mixture of skin benefiting agents including anti-aging, wrinkle-reducing, skin whitening and sebum controlling/ reducing agents plasticizers, elastomers, pigments, antioxidants, chelating agents, and perfumes, as well as organic/ inorganic sunscreens and including sunscreens as UV diffusing/ protection agents and optionally, adjunct minor components /ingredients including coloring agents, opacifiers, skin lightening agents, pH adjusting agents and perfumes etc in an amounts in the range from 0.01 % up to 50 % by weight, preferably 0.1 to 20% by weight, more preferably 0.2 to 10% by weight of the composition.

In accordance with yet another preferred aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical composition comprising said broad spectrum synergistic anti microbial/anti-fungal composition in amounts of 0.01 wt % to 99.9 wt. %, preferably 0.1 wt % to 50wt % and more preferably 0.5 wt % to 5 wt % ;and
a cosmetically acceptable vehicle with or without other hair and skin benefiting agents.

According to yet another preferred aspect of the present invention there is provided a cosmetic and/or dermopharmaceutical composition for topical use comprising a leave-on or a wash-off product adapted for topical delivery in the form of hair oil, hair gel, hair lotion, hair spray, shampoo, creams, ointments, emulsions, gels, lotions, face wash, oils, sticks, sprays, soaps, packs, wraps, woven or nonwoven wipes, powders, films or patches etc., as a vehicle for topical application of the said synergistically acting anti microbial composition.
According to yet another preferred aspect of the present invention there is provided a pharmaceutical composition for oral or topical use comprising a product adapted for topical delivery in the form of tablets, capsules, syrup, liniment, hair oil, hair gel, hair lotion, hair spray, shampoo, creams, ointments, emulsions, gels, lotions, face wash, oils, sticks, sprays, soaps, packs, wraps, woven or nonwoven wipes, powders, films or patches etc., as a vehicle for oral or topical application of the said synergistically acting anti microbial composition.

Detailed Description of the Invention
As discussed hereinbefore the present invention provides for a synergistic anti microbial composition comprising a synergistically special combination of atleast one cinnamic acid derivative and any one compound selected from the group consisting of naphthoquinone derivatives and terpenophenol derivatives. Preferably said composition comprises a synergistic combination including EPMC:Juglone and EPMC:Bakuchiol as the anti microbial active/agent combinations preferably as effective anti dandruff combination and particularly provides for cosmetic anti microbial compositions comprising the same in effective amounts present in combination with or without other skin or hair care benefiting agents. More particularly, the present invention relates to cost-effective cosmetic or dermopharmaceutical compositions for topical applications that are especially effective towards management of infections affecting skin or scalp wherein said anti microbial active ingredient or pure compounds are obtained either synthetically/semi-synthetically.
The said composition on one hand would provide synergistic anti dandruff action and on the other hand acts as a hair & /or skin care benefiting agent such as prevention of hair fall, favouring hair growth, damage protection, melanin inhibition, UV protection etc. which on one hand would provide synergistic anti dandruff action and on the other hand acts as a hair & /or skin care benefiting agent such as prevention of hair fall, favouring to hair growth, damage protection, melanin inhibition, UV protection etc.

The synergistic anti microbial composition of the present invention is used in an effective amounts in a safe and acceptable oral or topical dosage form in a pharmaceutically acceptable vehicle with or without other anti microbial agents.

Antimicrobial actives selectively involved in the present advancement are discussed hereunder:
Cinnamic acid derivatives are generally recognized for their antimicrobial activities. Ethyl-p-methoxy cinnamate (EPMC) is a cinnamic acid derivative which has been isolated from Curcuma zedoaria and has been shown to inhibit a wide range of fungi. EPMC inhibits the growth of Trichophyton rubrum, Aspergillus niger, Saccharomyces cerevisiae and Epidermophyton floccosum at a concentration less than 10 µg/ml; A. fumigatus, Penicillium purpurogenum, Trignoposis variabilis, Microsporum gypseum, Sclerotium rolifsii, Geotricular candiade, Fusarium oxysporum and Helminthosporium oryzale at a concentration less than 25 µg/ml; and Candida krusei and T. mentagrophytes At a concentration less than 50 µg/ml. The spores of T. rubrum lose viability or ability to germinate when exposed to its ethanolic solution (30 µg / ml) for 2 hours (SK Gupta et al. Lloydia 39 (4), 218-222. Jul-Aug 1976).
The specific mode of action of EPMC is based in part on cellular energy depletion, rather than nonspecific membrane-disrupting effects of other phenolic antimicrobial agents. EPMC has also been reported to have larvicidal, antineoplastic, anti-inflammatory, insect repellent properties and acts as chemopreventive agent against mice’s fibrosarcoma.

Juglone is a naphthoquinone, an aromatic organic compound with a distinct phenyl group and is commonly found in nature, major source being walnut tree. The most wide spread naphthoquinones are juglone, lawsone, plumbagin, and lapachol. Juglone is the compound responsible for yellow pigmentation in the walnut tree. Juglone has antifungal properties similar to some commercially available antifungal agents used to treat infections, such as athlete’s foot and ringworm. One study showed that juglone inhibits three key enzymes from Helicobacter pylori, a bacterium that can cause gastritis, peptic ulceration, and gastric cancer in humans. Recent data suggest juglone could be a promising chemo-preventive agent for human intestinal neoplasia and thus anti-tumour properties have been reported.

Bakuchiol is a meroterpene (a chemical compound having a partial terpenoid structure) in the class terpenophenol. It is found in Psoralea corylifolia and in Otholobium pubescens. Bakuchiol is a phytoestrogen, having antioxidant, antimicrobial and cytotoxic activity. Bakuchiol isolated from P. corylifolia has shown activity against numerous gram-positive and Gram-negative oral pathogens. It was able to inhibit the growth of Streptococcus mutans under a range of sucrose concentrations, pH values and in the presence of organic acids in a temperature-dependent manner and also inhibited the growth of cells adhered to a glass surface. It has antifungal activity, particularly against the type of fungi that cause athlete's foot. Its mechanism of action against fungi is via generation of reactive oxygen species (ROS) , which have a positive effect on health. Bakuchiol may also have a suppressive effect on melanin-producing cells in skin, which may make it a beneficial ingredient to lighten dark spots or improve uneven skin tone due to sun damage.

Advantageously, the actives of Formula 4, Formula 5 and Formula 6 can be sourced synthetically or semi synthetically which were surprisingly and selectively found to be unexpectedly and specially efficacious in terms of the desired mechanism for inhibiting the dandruff causing organism that is additionally endowed with favorable hair growth., skin lightening and sunscreen properties.
Minimum inhibitory concentration (MIC) values of the three actives against M. furfur were studieed and studies revealed that just because the two compounds have inhibitory activity against M.furfur, the same may not lead to any synergistic inhibitory activity by combining the compounds . Thus while it is evident that Bakuchiol is having very high MIC against M.furfur > 500µg/ml, however, surprisingly its combination with EPMC revealed excellent synergistic inhibition. Under the present advancement the synergy is further demonstrated by interpreting the FIC (Fractional inhibitory concentration) of the three actives in different combination.
A wide variety of cosmetic preparations are suitable, in particular skin care products like creams, ointments, pastes, lotions etc., for the use according to the invention of the said composition . Examples of other preparations which are suitable according to the invention and which may be mentioned are skin care, face wash. Accordingly, these are always preparations which are applied to the skin for a shorter or longer time depending on the actual purpose for which they are used. Addition of the compounds according to the invention then affects simultaneous skin lightening. However, it is also possible to produce cosmetic or medicinal preparations which are used primarily or exclusively for the purpose of eliminating/mitigating other skin pigmentation disorders incorporating these actives.
The cosmetic compositions prepared according to the invention are supplied as shampoos, body wash etc. These can be clear liquids, opaque liquids (with pearly lustre effect), in cream form, gel-like or else in powder form or in tablet form, and as aerosols.
The ingredients typically employed in such a composition are surfactants, emulsifiers, emollients, silicones, thickeners, antioxidants, sunscreen agents chelating agents, perfumes, opacifiers, colors, other skin lightening agents, herbal extracts, / compounds, pH adjusting agents and water to qs. The detergent raw materials on which these shampoos are based can be anionic, cationic, nonionic and amphoteric in nature and also be present in combinations of these substances.
The composition may contain usually employed vehicle such as may be aqueous, anhydrous or an emulsion. Preferably, the compositions are aqueous or an emulsion, especially water-in-oil or oil-in-water emulsion, preferentially oil in water emulsion. Water when present will be in amounts which may range from 5 to 99%, preferably from 20 to 85%, optimally between 40 and 80% by weight.
According to a still further aspect of the invention the cosmetic/ dermopharma-ceutical composition may contain various other skin benefit agents such as plasticizers, elastomers, calamine, pigments, antioxidants, chelating agents, and perfumes, as well as organic/ inorganic sunscreens and including such sunscreens as UV diffusing/ protection agents.
A still further aspect of the invention the cosmetic composition of the present invention may optionally contain other adjunct minor components /ingredients such as including coloring agents, opacifiers, skin lightening agents, pH adjusting agents and perfumes. Amounts of these other adjunct minor components may range anywhere from 0.0001 % up to 20 % by weight of the composition.

The composition may further include suitable skin benefiting agents comprising anti-aging, wrinkle-reducing, skin whitening and sebum controlling/ reducing agents. Examples of these include alpha-hydroxy acids and esters, beta-hydroxy acids and esters, polyhydroxy acids and esters, kojic acid and esters, ferulic acid and ferulate derivatives, vanillic acid and esters, dioic acids (such as sebacic and azeleic acids) and esters, retinol, retinal, retinyl esters, hydroquinone, t-butyl hydroquinone, mulberry extract, licorice extract, and glycosides of benzyl protocatechuate derivatives other than the derivatives discussed herein above.
Suitable cosmetic carriers are well known to one skilled in the art. The cosmetic bases may be any bases which are ordinarily used for skin benefit agents and are not thus critical. Specific preparations of the cosmetics to which the skin benefit agents of the invention is applicable include creams, ointments, emulsions, lotions, washes, masks, packs, oils, sprays / aerosols and wipes. Cream bases are, for example, beeswax, cetyl alcohol, stearic acid, glycerine, propylene glycol, propylene glycol monostearate, polyoxyethylene cetyl ether and the like. Lotion bases include, for example, oleyl alcohol, ethanol, propylene glycol, glycerine, lauryl ether, sorbitan monolaurate and the like.
The cosmetically acceptable vehicle may act as a diluent, dispersant or carrier for the skin beneficial ingredient of the composition of the invention, so as to facilitate their distribution when the composition is applied to the skin.
Besides water, relatively volatile solvents may also serve as carriers within compositions of the present invention. Most preferred are monohydric C1-C3 alkanols. These include ethyl alcohol, methyl alcohol and isopropyl alcohol. The amount of monohydric alkanol may ranges from 1 to 70 %, preferably from 10 to 50 %, and optimally between 15 to 40 % by weight.
Emollient material also serves as cosmetically acceptable carriers. These may be in the form of silicone oils and synthetic esters. The amount of the emollient material ranges anywhere from 0.1 to 50 %, preferably between 0.5 and 30 % by weight.
Examples of anionic detergent substances of this type which may be mentioned are: C10 -C20 -alkyl- and alkylenecarboxylates, alkyl ether carboxylates, fatty alcohol sulfates, fatty alcohol ether sulfates, alkylolamide sulfates and sulfonates, fatty acid alkylolamide polyglycol ether sulfates, alkanesulfonates and hydroxyalkanesulfonates, olefinsulfonates, acyl esters of isethionates, alpha-sulfo fatty acid esters, alkylbenzenesulfonates, alkylphenol glycol ether sulfonates, sulfosuccinates, sulfosuccinic monoesters and diesters, fatty alcohol ether phosphates, protein-fatty acid condensation products, alkyl monoglyceride sulfates and sulfonates, alkyl glyceride ether sulfonates, fatty acid methyltaurides, fatty acid sarcosinates, and sulforicinoleates. These compounds and their mixtures are used in the form of their salts which are soluble in water or dispersible in water, for example the sodium, potassium, magnesium, ammonium, mono-, di- and triethanolammonium and analogous alkylammonium salts.
Examples of suitable cationic surfactants are quaternary ammonium salts such as di (C10 -C24 -alkyl) dimethylammonium chloride or bromide, preferably di (C12-C18 -alkyl) -dimethylammonium chloride or bromide; C10-C24 alkyldimethyl- ethylammonium chloride or bromide; C10-C24 alkyltrimethyl-ammonium chloride or bromide, preferably cetyltrimethylammonium chloride or bromide and C10 -C24 -alkyltrimethylammonium chloride or bromide and other commonly available materials.
Examples of suitable nonionic surfactants which can be used as detergent substances are: fatty alcohol ethoxylates (alkylpolyethylene glycols); alkylphenol polyethylene glycols; alkyl mercaptan polyethylene glycols; fatty amine ethoxylates (alkylaminopolyethylene glycols); fatty acid ethoxylates (acylpolyethylene glycols); polypropylene glycol ethoxylates; alkanol amides, fatty acid alkylolamides (fatty acid amide polyethylene glycols); sucrose esters; sorbitol esters and polyglycol ether.
For the purposes of the invention, the term amphoteric surfactant is understood to refer to the compounds chosen, alone or as a mixture, from alkyl amphocarboxyglycinates, alkyl amphocarboxypropionates, alkyl ampho-diacetates, alkyl amphodipropionates, alkyl amphoglycinates, alkyl ampho-propionates, alkyl iminopropionates, alkyl iminodipropionates, alkyl ampho-propylsulphonates, alkylbetaines, alkylamidopropylbetaines, alkylsultaines and alkylamidopropylhydroxysultaines.
Thickners: Thickeners intended to give a sufficient consistency and improved cosmetic qualities. Among the known thickeners, carbomers (acrylic acid homopolymers crosslinked with an allylic ether of pentaerythritol, of sucrose or of propylene), PEG esters, laurylpyrrolidone, acrylic copolymers and cationic polymers of natural or synthetic origin, including certain polyquaternium compounds give very satisfactory results in the application considered, in particular bearing in mind that viscosities of greater than 1000 mPa.s, preferably between 1000 mPa.s and 30,000 mPa.s, could be obtained with these specific products ("Carbopol from BF Goodrich, "Atlas G-1821" from ICI, "Surfadone 300" from ISP, "Acrysol" from Rohm & Haas, "Jaguar.162" and "Rhodicare" from Rhone-Poulenc, "Ucare Polymer Jr. 400" from Amerchol, "Mackernium" from Jan Dekker/MacIntyre and "Gafquat N" from ISP). These specific thickeners and mixtures there of (not restricted) make it possible to obtain acceptable viscosities when they are used at concentrations of between 0 and 5% by weight relative to the whole composition, preferably between 0.2 and 3% by weight.
For the preparation of the cosmetic preparations the active substance is dissolved under stirring at a temperature in the range between 20 and 90oC, preferably at room temperature, in the detergent substance used. Subsequently, the further additives are added.
In the event of alcohol containing cosmetic preparations the said synergistic anti microbial composition is dissolved in the alcohol at a temperature in the range between 20 and 40oC, preferably at room temperature. Subsequently, the further additives are added. In the event of hair rinses and oil-in-water emulsions the said synergistically special anti microbial composition is dissolved in the fatty phase, which means together with the oil and the emulgator, at a temperature in the range between 70 and 90oC, preferably at 75o C. Subsequently, hot water is added and the emulsion is stirred and cooled.
The shampoos are produced in a manner known per se by mixing the individual components and-where necessary further processing appropriate for the particular type of preparation. Some of these wide varieties of possible formulations are described by way of example in the exemplary embodiments.
The skin lightening preparations according to the invention can also be supplied in the form of aqueous and aqueous alcoholic skin lotions also those in gel form, and in aerosol form as body spray, as well as in the form of skin care and skin dressing creams and gels. Ethanol and isopropanol are preferably used as alcohols.
The skin lightening active substance is incorporated in the preparations according to the invention in amounts which are normally between about 0.1 and about 15%. Within this range, the concentrations of the specific preparations depend on the purpose for which they are used. Certain formulations such as, for example, concentrates which must be diluted before they are used have considerably higher concentrations.
For preparations which remain on the skin, such as, for example, creams, lotions etc., lower concentrations of the said combination is to be used, for example of about 0.001 to about 15%, preferably 0.001 to 5%. They are expediently used in higher concentrations if the cosmetic preparations act, where appropriate after dilution, for only a short time on the hair and scalp, such as, for example, face wash, shampoos or hair rinses. In these cases, for example, preferable concentrations are of about 0.001 to about 15%, more preferably about 0.001 to about 10%.
The advantages and utility of the present invention is further illustrated by way of the following non-limiting examples. The stated amounts are based on weight.
Examples
Example 1: Synergistic anti dandruff activity of EPMC with Juglone and Bakuchiol
The antifungal susceptibility testing (MIC) of M. furfur strains was performed by broth microdilution CLSI M27-A3 using Sabouraud Dextrose Broth with 1% Tween 80 (1, 2). MIC was determined by double dilution method, for the neat compound it was tested from 1000 µg/ml to 1.95 µg/ml and for the combination samples concentration starts from 500 µg/ml to 0.97 µg/ml. [a) Clinical and Laboratory Standards Institute. Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts. 3rd ed. Wayne, PA: CLSI; 2008. M27-A3. b) Iatta R, Immediato D, Montagna MT, Otranto D, Cafarchia C. In vitro activity of two amphotericin B formulations against Malassezia furfur strains recovered from patients with bloodstream infections. Med Mycol. 2015 Apr;53(3):269-74.]
Table -1 provides the MIC values for the three test compounds, all the experiment were performed in triplicate.
TABLE-1 MIC (M. furfur) of pure compounds
S.No Sample details MIC Value in µg/ml
Test-1 Test-2 Test-3
1 EPMC 125 125 125
2 Juglone 31.25 31.25 31.25
3 Bakuchiol >500 >500 >500

Table -2 provides MIC values of the combinations, all the combinations were tested in triplicates, for each test MIC of the combination (column named as MIC for each Test) and the MIC values of the antimicrobials when tested in combination with other (column named as A+B for each Test) was given.
TABLE -2 Synergy study data
S.No Sample Detail Test-1 Test-2 Test-3
MIC A+B MIC A+B MIC A+B
1 EPMC : Juglone 9:1 7.81 7.03+0.78 3.90 3.51+0.39 7.81 7.03+0.78
2 EPMC : Juglone 7:3 7.81 5.46+2.34 7.81 5.46+2.34 15.62 10.9+4.68
3 EPMC : Juglone 1:9 15.62 1.56+14.06 31.25 3.12+28.12 31.25 3.12+28.12
4 EPMC : Bakuchiol 9: 1 15.62 14.06+1.56 7.81 7.03+0.78 15.62 14.06+1.56
5 EPMC : Bakuchiol 7:3 62.50 43.75+18.75 31.25 21.8+9.37 62.50 43.75+18.75
6 EPMC : Bakuchiol 1:9 125.00 12.5+112.5 125.00 12.5+112.5 125.00 12.5+112.5
Fractional inhibitory concentration (FIC):
Calculated for each Antimicrobial combination according to the formulas
FIC of Drug A (FIC A) = MIC drug A when tested in combination with drug B
MIC of Drug A alone
FIC of Drug B (FIC B) = MIC drug B when tested in combination with drug A
MIC of Drug B alone
FIC = FIC A + FIC B
Interpretation of the results was based on the given below criteria (Planta Med.2011;77:1168-82 ; Aro et al. BMC Complementary and Alternative Medicine ,2016,16: 385),
1. Synergy = 0.5
2. Additive effect > 0.5 and = 1.0
3. Indifference > 1 and = 4.0
4. Antagonism > 4.0

Table – 3 provides the Fractional inhibitory concentration values for each combination tried.

TABLE -3 FIC calculation
S.No Sample Detail Fractional inhibitory concentration ( Test-1 Data)
A (EPMC) B (Juglone/Bakuchiol) FIC-A FIC-B FIC Result
1 EPMC : Juglone 9:1 7.03 0.78 0.05 0.02 0.07 Synergistic
2 EPMC : Juglone 7:3 5.46 2.34 0.04 0.07 0.11 Synergistic
3 EPMC : Juglone 1:9 1.56 14.06 0.01 0.4 0.41 Synergistic
4 EPMC : Bakuchiol 9: 1 14.06 1.56 0.11 0 0.11 Synergistic
5 EPMC : Bakuchiol 7:3 43.75 18.75 0.35 0 0.35 Synergistic
6 EPMC : Bakuchiol 1:9 12.5 112.5 0.1 0.2 0.3 Synergistic

The combination of Bakuchiol having MIC value of > 500µg/ml, with EPMC having MIC value of 125µg/ml gives FIC index of 0.1-0.4. It is evident from the above that Bakuchiol is having very high MIC against M. furfur > 500µg/ml, however, in the combination (S. No. 5-8) it showed excellent synergistic inhibition,
Example -2 Synergistic anti dandruff activity of EPMC with Bakuchiol in surfactant base.
Table -4 EPMC : Bakuchiol Synergy Study (With surfactant base)
S.No Sample details MIC µg/ml A+B A FIC-A B FIC-B FIC
1 Compound (A+B) 1:9 (1.5% in surfactant) 125 12.5+112.5 12.5 0.1 112.5 0.1125 0.2125
2 Compound(A+B) 3:7 (1.5% in surfactant) 125 37.5+87.5 37.5 0.3 87.5 0.0875 0.3875
3 EPMC (C) (1.5% in surfactant) 125 µg/ml
4 Bakuchiol (D) (1.5% in surfactant) 1000 µg/ml
5 Surfactant > 58.3 mg/ml
Note: A denotes EPMC pure compound, B denotes Bakuchiol pure compound, C denotes EPMC with surfactant base and D denotes Bakuchiol with surfactant base. Required amount of A+B mixture prepared as per the ratio, using this mixture, 1.5% dispersion in surfactant prepared and tested MIC values.

Table - 4 provides MIC values of the select combinations, in a surfactant base along with FIC index, it is evident that the combinations were able to produce synergistic activity in the surfactant base as well.
Example -3: Preparation of shampoo with Anti dandruff actives:
Table-5. AD Shampoo preparation
Sl. no. Ingredient %W/W
1 Sodium Laureth Sulpahte (SLES) 59.542
2 Cocomonoethanalomide (CMEA) 0.500
3 Cetostearyl alcohol 0.150
4 Cocoamido propyl betain (CAPB) 1.500
5 Guar Gum 0.100
6 Sodium Citrate 0.030
7 Sodium benzoate 0.120
8 Silicon fluid 0.500
9 Fragrance 0.500
10 Colouring agents 0.058
11 Alkylpolyglucoside 1.000
12 Ethyl p-methoxy Cinnamate (EPMC) 4.500
13 Bakuchiol 0.500
14 Nacl (25%) 5.500
15 DM water 25.500
Total 100.000

STEP WISE PREPARATION:

Sl. no. Table -6. STEP-1 (PHASE-I)
Ingredient %W/W
1 SLES 10.000
2 CMEA 0.500
3 Cetostearyl alcohol 0.150
4 DM water 29.000
Mix the ingredients and heat to 75 °C

Sl. no. Table-7. STEP-2 (PHASE - II)
Ingredient %W/W
1 SLES 49.542
2 CAPB 1.500
3 Guar gum 0.100
4 Sodium Citrate 0.030
5 Sodium benzoate 0.120
6 Silicon fluid 0.500
Cool the Phase-I to room temperature and add the Phase -II ingredients one at a time with stirring

Sl. no. Table-8. STEP-3. PHASE - III
Ingredient %W/W
1 Alkylpolyglucoside (Vehicle) 1.000
2 EPMC 4.500
3 Bakuchiol 0.500
Mix Phase III ingredients @ 50°C with high speed stirring until uniform and add to Phase I+II


Sl. no. Table-9. STEP-4. PHASE - IV
Ingredient %W/W
1 Fragrance 0.500
2 Colour 0.014
3 Nacl(25%) 5.500
4 DM water 31.500
Add Phase IV ingredients one by one with stirring and mix until uniform.

Shampoo with 0.5% climbazole is prepared by keeping 0.5g climbazole as Phase-III ingredient.
Shampoo without actives and with vehicle is prepared by keeping vehicle as Phase-III ingredient.
Shampoo without actives (Base) is prepared by following Step-1, Step-II and Step-IV. With out Phase-III ingredients.
Example -4: Anti dandruff activity of EPMC & Bakuchiol combination in shampoo system
MIC of four samples prepared by Example-3 above were studied. The results are as follows:
Table -10 EPMC : Bakuchiol Antidandruff Study (In shampoo base)
S.No Sample details MIC µg/ml
1 Shampoo base with out vehicle and actives >10,000
2 Shampoo base with vehicle >10,000
3 EPMC:Bakuchiol shampoo (7.8 :1) 5000
4 Shampoo with 0.5 % Climbazole >10,000
Thus it is evident that, the synergistic combination is delivering antidandruff benefit in a shampoo formulation as well.
Example-5: Solubility of the combination in various vehicles.
EPMC : Bakuchiol (9:1) & EPMC : Bakuchiol (1:9) were prepared and dissolved in various vehicles @ 20% level. These mixtures can be incorporated into an oil base. Oil prepared in these way can be used as a leave on anti microbial product. The combination is freely soluble in PEG-7, Diisopropyl adipate, C12-C15 alkyl benzoate, Isopropyl palmitate, Isopropyl myristate, Caprylic capric triglyceride, Crodamol TM PTIS (Pentaerythrityl tetraisostearate).
Thus it is evident that, the synergistic combination can be utilised as a leave-on antimicrobial product as well.
It is thus possible by way of the present invention to provide for a synergistic antimicrobial composition comprising a synergistically special combination of cinnamic acid derivatives and any one one compound selected from the group of naphthoquinone derivatives and terpenophenol derivatives in effective amounts having surprisingly special inhibiting of fungal /microbial infections for effective safe and wide scale applications and uses .
,CLAIMS:We Claim :
1. A cosmetic and/or dermopharmaceutical composition comprising broad spectrum synergistic anti microbial/anti-fungal composition comprising atleast a compound selected from the cinnamic acid derivatives of Formula-1, with any one compound selected from the group of Naphthoquinone derivatives of Formula-2, and terpeneophenol derivatives of Formula -3.
FORMULA-1:

where in R1 , R2 = C1-C14 alkyl/ C1-C14 alkene/ C1-C14 alkyne
FORMULA -2:

Such that
When R1 = OH, R2 = H OR C1-C6 alkyl or C1 – C6 Alkene and R3 = H
When R1 = C1-C4 alkyl, R2 = H and R3 = OH
When R1and R2= H, R3 = OH
FORMULA -3:

Where in R1 = C12 – C15 alkyl or C12 – C15 alkene with 2 to 4 Carbon carbon double bond or C12 – C15 alkyne with 2 to 3 carbon carbon triple bond.

2. A cosmetic and/or dermopharmaceutical composition as claimed in claim1 comprising Ethyl-p-methoxycinnamate of Formula-4 with any one compound selected from the group of compounds of Formula -5 and of Formula -6, wherein the compounds of Formula 4 , 5 and 6 are as represented hereunder
FORMULA -4: C12H14O3. Ethyl 4-Methoxycinnamate; Ethyl p-methoxycinnamate; (E)-Ethyl 3-(4-methoxyphenyl)acrylate; Ethyl 3-(4-methoxyphenyl)acrylate; Ethyl trans-4-methoxycinnamate.

FORMULA-5: C10H6O3 Juglone, 5-Hydroxy-1, 4-naphthalenedione, 5-Hydroxy-1,4-naphthoquinone, 5-Hydroxy-p-naphthoquinone, 5-hydroxynaphthoquinone , Regianin, , Nucin, NCI 2323, Oil Red BS

FORMULA -6: C18H24O Bakuchiol, 4-[(1E,3S)-3-ethenyl-3,7-dimethylocta-1,6-dienyl] phenol

3. A cosmetic and/or dermopharmaceutical composition as claimed in anyone of claims 1 to 2 comprising a synergistically co acting anti microbial /anti-fungal combination of Ethyl-p-methoxycinnamate of Formula-4 with any one compound selected from Juglone of Formula-5 and Bakuchiol of Formula-6.

4. A cosmetic and/or dermopharmaceutical composition as claimed in anyone of claims 1 to 3 wherein said compound selected from the cinnamic acid derivatives of Formula-1: said any one compound selected from the group consisting of Naphthoquinone derivatives of Formula-2, and terpeneophenol derivatives of Formula -3 is in the range of 9.9: 0.1 to 0.1 to 9.9 preferably 9 :1 to 1:9

5. A cosmetic and/or dermopharmaceutical composition, as claimed in anyone of claims 1- 4, having synergistic anti dandruff action and related hair care benefits.

6. A cosmetic and/or dermopharmaceutical composition as claimed in anyone of claims 1 to 5 comprising a suitable vehicles including aqueous, anhydrous, emulsions, creams, lotions, Surfactants, oils etc., wherein in the vehicle is in range from 5 to 99.9%, preferably from 20 to 85%, more preferably from between 1 to 99% by weight..
7. A cosmetic and/or dermopharmaceutical composition as claimed in anyone of claims 1-6 comprising hair and/or skin care benefiting agents and a cosmetically/pharmaceutically acceptable vehicles.
8. A cosmetic and/or dermopharmaceutical composition as claimed in anyone of claims 1 to 7 comprising hair benefit agents or mixture of hair benefiting agents including Hair fall reducing agents, Hair growth promoting agents, anti-aging, sebum controlling/ reducing agents, plasticizers, elastomers, pigments, antioxidants, chelating agents, and perfumes, as well as organic/ inorganic sunscreens and including sunscreens as UV diffusing/ protection agents and optionally, adjunct minor components /ingredients including coloring agents, opacifiers, pH adjusting agents and perfumes etc in an amounts in the range from 0.01 % up to 50 % by weight, preferably 0.1 to 20% by weight, more preferably 0.2 to 10% by weight of the composition.

9. A cosmetic and/or dermopharmaceutical composition as claimed in anyone of claims 1 to 7 comprising skin benefit agents or mixture of skin benefiting agents including anti-aging, wrinkle-reducing, skin whitening and sebum controlling/ reducing agents plasticizers, elastomers, pigments, antioxidants, chelating agents, and perfumes, as well as organic/ inorganic sunscreens and including sunscreens as UV diffusing/ protection agents and optionally, adjunct minor components /ingredients including coloring agents, opacifiers, skin lightening agents, pH adjusting agents and perfumes etc in an amounts in the range from 0.01 % up to 50 % by weight, preferably 0.1 to 20% by weight, more preferably 0.2 to 10% by weight of the composition.

10. A cosmetic and/or dermopharmaceutical composition, as claimed in anyone of claims 1-9 comprising a leave on product including oil or a surfactant based wash off product .

11. A cosmetic and/or dermopharmaceutical composition as claimed in anyone of claims 1 to 10 comprising said broad spectrum synergistic anti microbial/anti-fungal composition in amounts of 0.01 wt % to 99.9 wt. %, preferably 0.1 wt % to 50wt % and more preferably 0.5 wt % to 5 wt % ; and a cosmetically acceptable vehicle with or without other hair and skin benefiting agents.

Dated this the 7th day of April, 2017 Anjan Sen
Of Anjan Sen and Associates
(Applicants Agent)