FIELD OF THE INVENTION
The present invention relates to anti- hair loss formulation. More particularly the present invention relates to an anti hair loss formulation for topical application.
BACKGROUND & PRIOR ART OF THE INVENTION
Hair loss is a condition in which the hair falls at an abnormal rate eventually leading to partial or complete loss of hair. The scalp has at an average 100,000 strands of hair. A healthy scalp sheds hair continuously and new hair replaces them. However when the scalp is not able to produce new hair to replace the shed hair, baldness develops. The medical term for hair loss is "alopecia".
There are different types of hair loss based on the type and cause. The most common type is androgenic alopecia also know as male pattern baldness (when it occurs in men) or female pattern baldness (when it occurs in women). This is a permanent type of hair loss which is incurable and is caused by heredity and genetics. Other types are usually temporary and may be a symptom of serious illness, skin infection, stress, use of certain drugs, thyroid diseases etc. The hair loss halts once the underlying cause has been identified and treated.
GB2471504 relates to an aqueous or alcoholic solution comprising of caffeine to be used in healing skin cuts and cracks. Caffeine is known to have a property of healing and/or repair and/or regeneration and/or rejuvenation of damaged skin and/or body tissue, and/or muscles and/or organs of the body and/or other part of the body including hairs within the ears using caffeine and/or any other central nervous system stimulant and/or any other stimulant by application directly to the target area. It is further mentioned in the particular patented document that caffeine along with minoxil is known to improve the condition of sensory hair inside the ear.
JP4193821 relates to the use of caffeine preferably with minoxidil for the treatment of dandruff. This composition is also known to act as a hair nourishing cosmetic. In the patented document it is mentioned that when minoxidil is used with caffeine in a particular percentage, it is known to give synergistic effect.

Cell proliferation plays an important role in hair growth. Minoxidil, one of the most commonly used active molecules in hair growth products is shown to have proliferative effect.Mechanism of action of minoxidil involves dilation of blood vessels and enhanced potassium channel activity, resulting in improved blood circulation and nutrient supply to hair follicles. Minoxidil has a lot of drawbacks that have been reported with time. These may include skin rash, irritated eyes, redness and tenderness of the treated area, and also allergic reaction. Allergic reaction is generally more severe than the other side effects and can include things like rash, hives, chest pain, swelling, fainting, dizziness, rapid heartbeat, weight gain, and others.
Hence there is a need for an improved formulation that provides effective treatment against hair fall that is capable ofovercoming the drawbacks of minoxidil.
OBJECTIVE OF THE INVENTION
The primary objective of the present invention is to overcome the drawbacks of the prior art.
Another objective of the present invention is to provide a composition that is effective against hair fall and can enhance hair growth.
SUMMARY OF THE INVENTION
The present invention relates to an anti hair loss composition effective against hair loss comprising
a) Caffeine
b) Kopexil and
c) cosmetically acceptable excipients
wherein said caffeine and kopexil being present in the range of 1: 5 to 1:20, preferably, 1:9 to 1:16.

BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 shows the graphical representation of Cell proliferation assay (WST-1 assay) with different concentrations of caffeine and Kopexil separately and in combination.
Figure 2 shows the graphical representation of In- Vitro Hair Growth Assay with different concentrations of caffeine and Kopexil separately and in combination.
Figure 3Measurement of ln-Vitro Hair growth with different concentrations of caffeine and Kopexil separately and in combination.
Figure 4Measurement of ln-Vitro Hair growth with different concentrations of caffeine and Kopexil separately and in combination and comparison of control verses tonic (caffeine and kopexil at 1:15 ratio along with other nutrients).
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a composition for treatment of hair fall by topical application. The composition actively comprise of caffeine and kopexil in a specific ratio. Caffeine is a naturally-occurring compound found in several plants and has a beneficial effect on hair follicles which in turn promotes healthy hair. Caffeine has the ability to interact with hair follicles and helps to guide a follicles' behavior which regulates the hair growth. As a result, caffeine is helpful in restoring hair growth or in other words helps prevent abnormal hair loss. There are several ways to expose your hair follicles to caffeine and potentially stimulate hair growth. Caffeine consumed via food and beverages enters the bloodstream, and eventually reaches the hair follicles. Also hair follicles can absorb caffeine directly via topical application of caffeine-enriched hair products. This is particularly beneficial as it allows the hair follicles to expose to a high dose of caffeine, without causing the side effects that can occur due to high-dose caffeine ingestion. As a result hair product containing caffeine representsa cure for hair loss.
Kopexil is a minoxidil derivative and increases the volume of hair in the growth stage by working on the deep structure of the roots. It rejuvenates the hair roots so that healthy hair growth can persist, kopexil corrects the problem of hair roots that causes blood vessels to

compress and the life span of the hair follicle to shorten, kopexil helps in softening of the collagen.
The composition of the present invention comprising caffeine and kopexil in a specific ratio is tested on hair growth in two mediums namely Williams E medium and DMEM medium. The compositionin a specific ratio shows enhanced hair growth over the control in both the medium.
The actives (caffeine and kopexil) of the present invention are present in the ratio range of 1: 5 to 1:20, preferably, 1:9 to 1:16.
The said actives in the specified range are present in the composition along with other nutrients in the combined activity of the actives and nutrients through a number of mechanisms e.g. vasodilation (improving circulation & nutrient supply), counteracting the DHT inhibition, increased cell proliferation, helping in perifollicular fibrosis at physiological condition, thereby resulting in improved hair growth.
The composition of the preset invention is formulated into anti hair loss shampoo, anti hair loss tonic and anti hair loss cream for enhancing hair growth and in turn curing alopecia. Such formulations use caffeine and kopexil as active agents along with some cosmetically acceptable excipients.
Cosmetically acceptable excipients can be selected from cleansing agent, foam boosters, aesthetic improving agents, thickeners, preservatives, conditioning agents, humectants, additives, fragrance color, pH adjusters, viscosity adjuster, chelating agents, diluents, solubilizers, emulsifier.
Cleansers are used in shampoo formulation to clean the scalp of oil, dirt etc. so that the active agent can effectively act on the hair follicle and prevent hair fall.
Preferable cleansers used in the shampoo formulation are Anionic such as but not limiting to Sodium Laureth Ether Sulphate, Sodium Lauryl Sulphate, Ammonium Laureth Ether Sulphate, Ammonium Lauryl Sulphate. Said cleansers present in the formulation ranges between 6 to 22 % cone w/w.

Preferable foam boosters such as but not limiting to Lauryl sulfobetaine, Myristylsulfobetaine, Palmitylsulfobetaine, Cocamidopropylbetaine, Polyethylene glycols of different molecular weights.Preferred amount of foam booster present in the formulation ranges between 0.01 to 10% cone w/w.
Preferable aesthetic improving agents such as but not limiting to Ethylene Glycol Distearate, Ethylene Glycol monostearate, Mica,Titanium Dioxide.Preferred amount of aesthetic improving agents present in the formulation ranges between 0.01 to 5% cone w/w.
Preferable thickeners include but are not limiting to Polyacrylic acids & derivatives (Carbopols), emulsifiers, thickening waxes, xanthan gum.Preferred amount of thickeners present in the formulation ranges between 0.1 to 4% cone w/w.
Preferable preservatives include but are not limiting to DMDM Hydantoin, Methylchloroisothiazolinone, Methylisothiazolinone, Phenoxyethanol, Methylparaben, Ethylparaben, Butylparaben, Propylparaben, Isobutylparaben.Preservatives present in the formulation ranges between 0.01 to 1% cone w/w.
These preferably include but are not limiting to Silicones, silicone derivatives, Guar Hydroxypropyltrimonium Chloride, Polyquaterniums. Preferred amount of conditioning agent present in the formulation ranges between 0.01 to 20% cone w/w.
Preferably humectants include but are not limited to Glycerine, propylene glycol, 1,2,6-hexanetriol, triethyelene glycol, butylene glycol, hexanediol, Pentanediol. Humectants present in the formulation ranges between 0.1 to 10% cone w/w.
PreferredColors are but not limiting to oil colors, waters soluble colors, pigments. Preferred amount of color present in the formulation ranges between 0.001 to 2% cone w/w
Chelating Agents such as but not limiting to Sodium trihydroxy EDTA, Disodium EDTA, Tetra sodium EDTA, Disodium Dihydrogen EDTAare preferable.Preferred amount of chelating agents in the formulation ranges between 0.05 to 2% cone w/w
Preferred amount of additives is 0.001 to 1 % cone w/w.

Preferred amount of fragrance used is 0.01 to 3% cone w/w.
Preferred pH adjustment agents such as but limiting to sodium hydroxide, triethanolamine, aminomethylpropanol, citric acid, orthophosphoric acid.
Preferred viscosity adjusters are(but not limiting to) sodium or ammonium xylene or cumenesulphonate, sodium chloride, ammonium chloride.
Actives such as but not limiting to Magensium Aspartate & Zinc Gluconate, Copper Gluconate are preferred. Amount of active preferred is 0.01 to 2% cone w/w.
The invention is now described with the help of the following examples which are provided solely to illustrate the present invention and are not intended to limit the scope of the invention.
Example 1:
Cell proliferation assay (WST-1 assay)
Cells go into senescence and apoptosis when grown in medium without growth factor. However when certain ingredients are added to these cells the senescence is reversed and cell proliferation improved. Wstl Assay measures the metabolic activity of cells in culture and indicates the growth promoting effect of tested ingredient in comparison to controls. Reduction of Wst-1 (tetrazolium salt) to colored formazan compounds by succinate-tetrazoliumreductase in viable cells provides an accurate method to measure cell viability and proliferation.
Method:
HaCaT cells were plated at 1x104 cells/well and 0.75 xl04 cells/well, respectively on 96 well plates. After 24h of culture with medium containing growth supplements at 37°C the culture medium was removed from the wells and the treatments with ingredients/actives at various concentrations were carried out. Negative controls included wells treated with appropriate culture medium without growth supplements. The vehicle control included medium without growth supplements with solvent. The positive controls were the wells treated with cell culture medium with growth supplements. Cells were treated in quadruplicates for each treatment. Wst-1

reagent was added at 10% of the total volume of culture medium on 3d, 4d and 5d intervals. Three hours after addition of Wst-1 reagent the absorbance was measured at 460nm on spectrophotometer (Varioscan Flash, Thermo Scientific).

Actives are compared against control, NC, Combination of Caffeine + Kopexil at 1+15 μm is 140.577% of the NC value with P value 0.01336.
Results: As illustrated in Figure 1.

• Caffeine at l0μm shows enhanced proliferation effect while kopexil shows no proliferation effect at 15 μm.
• On testing the combinations of caffeine and kopexil, it was found that the combination of kopexil and caffeine at 15and lμm respectively shows enhanced proliferation effect.
• While the combination of kopexil and caffeine at 15and l0μm respectively does not show proliferation effect.
• Thus the caffeine which alone at 10 μm produced proliferative effect failed to show similar effect with 15 |im kopexil, when kopexil alone does not show any effect at all. On the other hand when 1 μm of caffeine is used with the same 15 μm of kopexil, an unexpected increase is seen over caffeine 10 μm plus kopexil 15 μm.
Example 2
In- Vitro Hair Growth Assay:
In- vitro Hair Culture and Maintenance:
The hair samples for in- vitro hair growth assay are inoculated in 24/48- well plates with 500ml of reconstituted Williams E medium supplemented with Insulin, hydrocortisone, L-Glutamine and antibiotics.. Each hair sample is maintained free- floating and isolated in each well. The medium is changed every alternate day. In case of treatments, the actives to be tested are added in the medium at desired concentrations and fresh treatments given on alternate days .
Measurement of In-Vitro Hair growth:

The day the hair samples are inoculated is considered as day 0 and each hair samples' length is measured using the scale bar in the software for the microscope. The picture of each sample is also taken each day. The scale bar can be merged on to the picture. Each day after the inoculation at an interval of 24 hrs, each hair sample is measured and picture taken.
Calculation of In- Vitro Hair Growth:
To calculate the growth of hair sample, each day's length is subtracted from Day 0 length for respective hair sample (eg D1-D0, D2- DO... D10-D0 etc). In case of treatments each set has 10 hair follicles (for statistical significance), average of them is calculated and taken as the growth (increase in length). SDs and p values are calculated.


*Control- Williams E medium with hair growth supplements.

• When tested on in- vitro hair growth assay caffeine shows improved hair growth while
kopexil shows no effect on in- vitro hair growth.
• On testing the combination of kopexil and caffeine at 15+10μm and 15+1 μm respectively, it
was observed that kopexil and caffeine at 15+1 urn showed significant increase in in-vitro hair
growth.
• While the combination at 15+ l0μm of kopexil and caffeine respectively did not show
significant change over control.
• Anti- hair loss tonic formulation containing kopexil and caffeine, when used alone in place of
media, maintained the hair in anagen phase and showed significant hair growth over day 0.



• When tested on in- vitro hair growth assay Tonic shows improved hair growth compared to placebo-base formulation.
Example 3: Method of Preparation:
Caffeine and Kopexil are incorporated in the cosmetic formulation in the last stage post completion of the product formulation.
(1) For Leave-On Single-Phase formulations such as but not Limiting to Hair Tonic
(i) Take the diluent alcohol in a suitable container and go on adding all the ingredients one by one ensuring complete dissolution of each ingredient.
(ii) Post-addition of all ingredients, caffeine and Kopexil are added to the formulation to get the desired Hair Tonic formulation.
(2) For Leave-on Emulsion-based products such as but not limiting to Leave-on Hair Cream
or Hair Leave-On
(i) Take all water-phase ingredients in one container and heat till 75°C to 80°C.
(ii) Take all the oil-phase ingredients in another container and heat till 75°C to 80°C.
(iii)When both the oil and water-phases reach the desired temperature, add the oil phase to the water phase and homogenize for 30 to 60 min. Stop heating the formulation and add all other ingredients in the formulation, once temperature reaches 45°C, giving enough time for each ingredient to get either dissolved or dispersed in the product.

(iv)Post-addition of all ingredients, caffeine and Kopexil are added to the formulation to get the desired Hair Leave-on Cream or Hair leave-On formulation.
(3) For Rinse-off formulations such as but not limiting to Shampoos
(i) Thickening agents are dispersed in a suitable solvent from amongst the ingredients used in the product separately in a sidepot.
(ii) Anioinic Surfactant is diluted in water to suitable concentration and then the thickening agent dispersed in the sidepot is introduced in the surfactant solution. All other ingredients are then added one by one giving enough time for each ingredient to get dissolved or dispersed in the shampoo.
(iii) Post addition of all ingredients, caffeine and Kopexil are then added to the formulation to get the desired Hair Shampoo formulation.

WE CLAIM:
1. An anti hair loss composition effective against hair loss comprising
d) Caffeine
e) Kopexil and
f) cosmetically acceptable excipients
said caffeine and kopexil being present in the range of 1: 5 to 1:20, preferably, 1:9 to 1:16.
2. The anti hair loss composition as claimed in claim 1, wherein cosmetically acceptable excipients is selected from cleansing agent, foam boosters, aesthetic improving agents, thickeners, preservatives, conditioning agents, humectants, additives, fragrance color, pH adjusters, viscosity adjuster, chelating agents, diluents, solubilizers, emulsifier.
3. The anti hair loss composition as claimed in claim2, wherein cleansing agent is selected from Sodium Laureth Ether Sulphate, Sodium Lauryl Sulphate, Ammonium Laureth Ether Sulphate, Ammonium Lauryl Sulphate.
4. The anti hair loss composition as claimed in claim 2, wherein foam boosteris selected from Lauryl sulfobetaine, Myristylsulfobetaine, Palmitylsulfobetaine, Cocamidopropylbetaine, Polyethylene glycols of different molecular weights.
5. The anti hair loss composition as claimed in claim 2, wherein aesthetic improvement agent is selected from Ethylene Glycol Distearate, Ethylene Glycol monostearate, Mica, Titanium Dioxide.
6. Th eanti hair loss composition as claimed in claim 2, wherein thickener is selected from Polyacrylic acids & derivatives (Carbopols), emulsifiers, thickening waxes , xanthan gum.

7. The anti hair loss composition as claimed in claim 2, wherein preservative is selected from DMDM Hydantoin, Methylchloroisothiazolinone, Methylisothiazolinone, Phenoxyethanol, Methylparaben, Ethylparaben, Butylparaben, Propylparaben, Isobutylparaben.
8. The anti hair loss composition as claimed in claim 2, wherein conditioning agent is selected from Silicones, silicone derivatives, Guar Hydroxypropyltrimonium Chloride, Polyquaterniums.
9. The anti hair loss composition as claimed in claim 2, wherein humectants is selected from Glycerine, propylene glycol, 1,2,6-hexanetriol, triethyelene glycol, butylene glycol, hexanediol, Pentanediol.

10. The anti hair loss composition as claimed in claim 2, wherein additives are selected from vegetable oils and /or other plant extracts.
11. The anti hair loss composition as claimed in claim 2, wherein color is selected from oil colours, waters soluble colors, pigments.
12. The anti hair loss composition as claimed in claim 2, wherein pH adjuster is selected fromfrom sodium hydroxide, triethanolamine, aminomethylpropanol, citric acid, orthophosphoric acid.
13. The anti hair loss composition as claimed in claim 2, wherein viscosity adjuster is selected from sodium or ammonium xylene or cumenesulphonate, sodium chloride, ammonium chloride.
14. The anti hair loss composition as claimed in claim 2, wherein chelating agent is selected from Sodium trihydroxy EDTA, Disodium EDTA, Tetra sodium EDTA, Disodium Dihydrogen EDTA.
15. The anti hair loss composition as claimed in claim 2, wherein the diluent is water or ethyl alcohol.
16. The anti hair loss composition as claimed in claim 2, wherein solubilizersis selected from PEG-7-Glyceryl Cocoate, PEG 40 Hydrogenated Castor Oil, Polysorbates.

17. The anti hair loss composition as claimed in claim 2, wherein emulsifiers is selected from Polysorbates, Ethoxylated fatty alcohols, Cetyl alcohol, Cetostearyl alcohol.
18. The anti hair loss composition as claimed in claim 2, wherein the thickener is present in the ranges between 0.1 to 4% cone w/w.
19. The anti hair loss composition as claimed in claim 2, wherein the preservative is present in the ranges between 0.01 to 1% cone w/w.
20. The anti hair loss composition as claimed in claim 2, wherein the chelating agent is present in the ranges between 0.05 to 2% cone w/w.
21. The anti hair loss composition as claimed in claim 2, wherein the fragrance is present in the ranges between 0.01 to 3% cone w/w.
22. The anti hair loss composition as claimed in claim 2, wherein the humectants is present in the ranges between 0.1 to 10% cone w/w.
23. The anti hair loss composition as claimed in claim 2, wherein the emulsifier is present in the ranges between 0.01 to 10% cone w/w.
24. The anti hair loss composition as claimed in claim 2, wherein the conditioning agent is present in the ranges between 0.01 to 20% cone w/w.
25. The anti hair loss composition as claimed in claim 2, wherein the additives is present in the ranges between 0.001 to 1% cone w/w.
26. The anti hair loss composition as claimed in claim 2, wherein the diluent is present in the ranges between 1 to 100% cone w/w.