Sepicap MP

SEPICAP MP – a vehicle for amino acids and panthenol – represents a new multifunctional, protective active substance for effective action against the kind of hair damage that results from everyday stress.

SEPICAP MP is based on a completely novel way of combining certain components: it is produced by conjugating a fatty acid chain (cocoyl) with the amino acids which are essential for the capillary fiber (glutamic acid, aspartic acid, alanine and glycine), and subsequently combining this conjugate with provitamin B5 (Panthenol) conjugated with water-soluble silicone.

2– SIMPLE IDEA, PROVEN EFFICACY

The idea is simple: SEPICAP MP has been designed to help the hair resist the kind of stress factors routinely encountered in everyday life such as exposure to UVA irradiation (e.g. sunlight), chemical treatments (e.g. perms, dying, bleaching, etc.), regular heat styling, brushing and pollution.

THE CONSEQUENCES OF STRESS?
THE MULTIFUNCTIONAL, THERMO-ACTIVATED PROTECTION OF SEPICAP® MP

HOW TO PROTECT HAIR AGAINST THE EFFECTS OF STRESS ? Multifunctional and thermo-activated, SEPICAP MP protects hair against the consequences of stress. Biovector for both amino acids and panthenol, SEPICAP MP : • Reduces oxidative stress by inhibiting the generation of peroxides, • Protects the root by reversing the inhibition of cell division and by restoring normal protein metabolism • Protects the shaft by preserving the keratin's integrity • Prevents flaking of scales. Since it is thermo-activated , it is most effective while blow-drying and heat styling your hair.

SEPICAP MP restructures and protects stressed hair. Proven efficacy on stressed and damaged hair, demonstrated both with ex vivo and in vitro tests.

3 – TEST NO. 1: THOROUGH PROTECTION OF THE HAIR AGAINST OXIDATIVE STRESS

Stress causes the production of free radicals by introducing the formation of lipid peroxides at both the surface of the hair shaft and within the follicle itself . These peroxides are particularly pernicious because they in turn induce a chain reaction of free radical formation (a Fenton reaction) which leads to widespread cell damage, protein degradation, and ultimately serious damage to the hair shaft.

• Damage to the root and the hair.

Therefore, to achieve thorough protection of all your hair, a capillary product must be able to reduce the levels of lipid peroxides formed on the surface of the shaft (i.e. it should be able to scavenge free radicals).

3.1 – The protective capacity of SEPICAP MP with respect to peroxides formed at the surface of stressed hair

Principle (ex vivo experiment)

The protective capacity of SEPICAP MP was estimated by measuring the levels of peroxide present at the surface of hair samples exposed to stress:

• Irradiation with UVA (25 J/cm 2 ).

Protective activity was measured after the topical application of a concentration of 1% of the commercially available product (diluted in water). Both preventive (i.e. application before irradiation) and curative (i.e. application after irradiation) activities were measured.

Lipid peroxide levels were measured using dichlorofluorescein which only fluoresces when it is in its oxidized form. Quantities of lipid peroxide measured at the surface of the hair are expressed in FU (arbitrary fluorescence units) per milligram of hair.

Procedure

This ex vivo study involved taking ahir samples from normal, healthy volunteers. Lipid was removed from the hair using 70° ethanol after which it was all thoroughly rinsed. The samples were treated as follows:

• 10 samples were neither irradiated nor treated with SEPICAP MP

• 10 samples were irradiated but not treated at all (control)

• 10 samples were treated with 1% SEPICAP MP and then irradiated (preventive activity)

• 10 samples were irradiated and then treated with 1% SEPICAP MP (curative activity)

Peroxide levels were assayed 20 hours after irradiation. All the hair samples were rinsed weighed and incubated with dichlorofluorescein. Fluorescence was measured on a Fluoroskan II machine and the readings were normalized to the weight of the hair samples analyzed.

Protective capacity with respect to the formation of lipid peroxides at the surface of UVA irradiation hair.

FU: Fluorescence units / mg of hair

NT: Non treated

Pc: Commercially available product

Irradiation of the hair samples induced a 100% increase in the levels of peroxides detected (i.e. +318 FU):

• Preventive treatment of the hair with 1% SEPICAP MP (pc) reduced peroxide levels by a factor of 43% compared with untreated irradiated samples ( +181 FU induced compared with untreated hair;

• Curative treatment of the hair with 1% SEPICAP MP (pc) reduced peroxide levels by a factor of 71% compared with untreated irradiated samples (+93 FU induced compared with untreated hair).

• SEPICAP MP effectively protects against peroxides generated at the surface of stressed hair.

• Its protective function is more marked when it is used curatively (71%) but nevertheless, its efficacy is still impressive when it is used preventively (43%).

3.2 – Free radical – scavenging activity of SEPICAP MP compared with that of reference species

Its free radical-scavenging activity is the property that underlies the protective capacity of SEPICAP MP with respect to peroxides generated at the surface of the hair.

The potency of its scavenging activity has been compared to that of the standard species used as benchmarks in the field, e.g. Vitamin C (the most common reference compound, and Panthenol (often included in hair care products).

Principle (in vitro experiment)

This in vitro test is designed to measure free radical scavenging activity based on its quenching of superoxide anion.

A substance's activity is estimated on the basis of the decrease in the rate of reduction of cytochrome C induced by the presence of the test compound in the reaction mixture.

Superoxide anion generated by a xanthine oxidase/xanthine couple, will reduce cytochrome C as long as there is not any species present which is capable of sequestering and detoxifying it. The rate of reduction of cytochrome C can be followed spectrophotometrically at 550 nm. In each experiment, the activity of the putative free radical scavenger was estimated by comparing reactions in its presence (experimental) and in its absence (control).

For each species tested, the decrease in the rate of generation of the colored product (which is proportional to the amount of superoxide anion being generated) in the experimental reaction compared with the rate in the control is expressed as a percentage.

For any substance being tested, the percentage inhibition of the rate of generation of the colored product therefore corresponds to the percentage inhibition of superoxide anion (= free radical scavenging activity).

Protection against free radicals

Free radical scavenging efficacy (%)

am : active matter

• The free radical scavenging activity of SEPICAP MP is exactly the same as that of vitamin C and superior to that of panthenol which is commonly used in products designed to protect the hair.

4 – TEST NO.2: DEEP HAIR PROTECTION

• Stimulates cell division

• Restores normal protein metabolism

Following oxidative stress, free radicals are produced and attack the cells in the hair follicles. This results in both a reduced rate of division and impaired protein metabolic function. As a consequence, the condition of the hair may be compromised.

SEPICAP MP exerts a two-fold action deep in the root of the hair:

It reverses the stress-induced reduction in the rate of cell division.	It restores normal protein metabolic function (necessary for cellular defense and repair mechanisms)

An experimental model for studying the hair root

The two-fold protective action of SEPICAP MP was investigated in an experimental model based on heat –stressed keratinocytes (undifferentiated cells of the hair follicle).

• In both parts of the experiment, the PREVENTIVE activity of SEPICAP MP was compared with that of unconjugated PANTHENOL alone.

4.1 – Stimulation of cell division (preventive activity)

Principle (in vitro experiment)

The effect of the test compounds on the rate of cell division was measured by fluorimetric assay of the amount of DNA produced in a culture of normal, human keratineocytes exposed to thermal stress (a temperature of 50°C maintained for a period of 20 minutes).

The amount of DNA measured corresponds to the rate of cell division in the culture.

The cells were grown to the 60% confluence and then co-cultured with the test substances for a period of 24 hours. The amount of DNA present in each well was measured in order to gauge the effect of the presence of the test substances on the rate of cell division.

Stimulation of the rate of division of stressed keratinocytes

am: active matter

NT: non-treated

Exposing cells to thermal stress results in a reduced rate of cell division (-20%). It is easy to imagine how, if such a thermal attack is relatively regular (as in the case with heat-styling and blow drying), permanent damage to the hair could result.

• Preventive treatment with SEPICAP MP reverses the stress-induced reduction in the rate of cell division (+53%).

• In this respect, it is more active then unconjugated Panthenol alone (a reversal of only +17%).

4.2 – Protective action with respect to the protein metabolic function of heat-stressed hair root cells (preventative activity)

The protective activity of SEPICAP MP with respect to protein metabolic function was investigated using the same heat-stressed keratinocyte model.

Principle (in vitro experiment)

The activity of SEPICAP MP with respect to protecting protein metabolic function was estimated by colorimetric assay of interacellular protein content (expressed in micrograms per milliliter) after 24 hours of contact with the various test substances.

Again, the cells were grown to 60% confluence and then co-cultured with the test substances for a period of 24 hours.

: active matter

NT : non-active

Subjecting the cells to thermal stress induced a reduction in protein content (-28%).

• Preventative treatment with SEPICAP MP reversed the stress-induced reduction in the rate of protein metabolism (+87%).

• In this respect, it is more active then unconjugated Panthenol (only +17%) which is a recognized reference material for this activity.

This activity is all-important because cells thus protected will be able to react to stressful attacks more rapidly and will therefore be able to defend and repair themselves more effectively (by stepping up their rate of protein synthesis).

• In times of stress, SEPICAP MP acts deep at the hair room to maintain normal rates of cell division and to enhance the celss' capacity to defend themselves by stimulating protein sythesis. Thereby, it helps keep hair in good condition.

• Its activity is superior to that of unconjugated Panthenol alone.

5 – TEST NO.3: PROTECTION OF THE HAIR SHAFT AGAINST KERATIN BREAKDOWN

SEPICAP MP preserves proteins that are normally broken down in response to oxidative stress (either due to UVA irradiation or thermal shock);

Its protective activity due to its capacity to prevent the breakdown of hair proteins (most importantly keratin) was assessed using a 1% concentration of the commercially available product on UVA or heat-stressed hair samples; protein breakdown was followed by assaying intrinsic tryptophan fluorescence.

• Stress-induced protein degradation is associated with a reduction in the natural, inherent fluorescence intensity due to tryptophan.

Procedure (ex vivo experiment)

The hair samples were impregnated with a 1% solution of SEPICAP MP in water for 10 minutes and then air-dried for 30 minutes. Control samples were treated in parallel with water alone.

Hair samples were subjected to UVA irradiation in a VILBER LOURMAT apparatus (at an intensity of 25 joules / cm2).

Other samples were subjected to heat stress by placing them in a jet of air at a temperature of 90°C for one hour, followed by a 15-minute relaxation phase at room temperature.

Measurement of hair protein fluorescence
The intensity of the fluorescence due to tryptophan (i.e. protein) was measured using a CD60 DESAGA photodensitometer.

The amount of fluorescence emitted from a pre-fixed area of hair (350 mm2) was measured in a longitudinal direction (equivalent to moving from the root out towards the end). The fluorescence reading was expressed in arbitrary fluorescence units normalized to the weight of the hair sample.

Protection of keratin structure in UVA-stressed hair

NT : non treated

Results expressed in FU/g = intrinsic fluorescence due to tryptophan per gram of hair

Non treated : the protein in the irradiated hair tresses was damaged as reflected by a decrease in the fluorescence reading (-193 FU). In the hair tresses which has been pre-treated with 1% SEPCAP MP, protein damage was reduced by a factor of 60% (only – 73 FU).

• When hair is subjected to UVA irradiation, preventive application of 1% SEPICAP MP significantly protects its proteins from breakdown.

Protection of keratin structure in heat-stressed hair

NT : non treated

Results expressed in FU/g = intrinsic fluorescence due to tryptophan per gram of hair

Non treated: the protein in the heat shocked hair samples was damaged as reflected by a decrease in fluorescence reading (-163 FU). In hair samples which had been pre-treated with 1% SEPICAP MP, protein damage was reduced by a factor of 35% (-107 FU).

• When hair is subjected to thermal stress, preventive application of a concentration of 1% SEPICAP MP significantly protects its proteins from breakdown.

• 1% of SEPICAP MP efficiently prevents UVA or heat-stress induced protein breakdown. Preventive application protects the natural structure of the hair shaft.

6 – TEST No.4: PROTECTION AGAINST STRESS-INDUCED FLAKING

The effects of stress (especially the generation of free radicals which attack both the root and the shaft) compound to cause flaking of the surface of the capillary fiber. This results in dull, brittle and badly damaged hair.

In the light of SEPICAP MP's multifunctional protective activity at the level of both root and shaft, we developed a way of measuring its ability to inhibit the kind of flaking that is induced by thermal stress.

Principle (ex vivo experiment)

The capacity of a concentration of 3% SEPICAP MP (the commercially available product) to inhibit flaking was investigated using hair damaged by exposure to thermal stress. The product's protective activity was estimated by examining and comparing treated and untreated hair samples by scanning electron microscopy.

Procedure

• Normal volunteers came to the laboratory to give hair tresses

• 5 hairs from each volunteer were soaked for 10 minutes in either water alone (placebo) or in a 3% solution of SEPICAP MP in water. All samples were air-dried

• Tresses were subjected to standardized heat shock (a temperature of 90°C for a period of 1 hour)

• The effect of the product on the appearance of the surface of the hair fibers was then assessed by examining the samples by scanning electron microscopy and recording micrographs

Protective activity in terms of the inhibition of flaking (scaling)

In every case, thermal stress induced flaking at the surface of the hair. Preventive application of 3% SEPICAP MP (the commercially available product) markedly inhibited the extent of detachment of the flakes: SEPICAP MP had a clear SMOOTHING and COVERING EFFECT.

• SEPICAP MP has preventive activity with respect to stress-induced flaking. The hair's natural smoothness and shine is therefore restored.

7 – TEST NO.5: THERMO-ACTIVATED EFFECT

The efficacy of SEPICAP MP is enhanced when the hair is actually under stress.

Principle (in vitro experiment)

The capacity of SEPICAP MP to stimulate cellular protein metabolism was measured by comparing heat-stressed and unstressed cultures of keratinocytes.

• The cells were grown to 60% confluence and then co-cultured for 24 hours with the active substances.

• The degree to which protein metabolism was being protected was assessed by colorimetric assay of the protein content of the cell culture.

• Exposure to a high temperature was carried out in the presence of the active substances (the interest here being the product's preventive activity).

Stimulation of protein metabolism in stressed and unstressed cells

SEPICAP MP: “thermo-activated” effect

Increased level (%) of cellular protein	+80% on heat-stressed cells

am: active matter

SEPICAP MP is associated with increased protein content in cells subjected to heat stress.

• Heat stress enhances the efficacy of SEPICAP MP.

This “thermoactive” effect is easy to understand because an unstressed cell has no reason to increase its protein content. Such a response is only likely to enhance cellular survival rates in the event of some external attack, like heat shock.

• SEPICAP MP is all the more effective when the hair is actually being stressed, especially if the stress is thermal.

• Therefore, the product's efficacy is optimized during heat-styling.

MULTIFUNCTIONAL, THERMOACTIVE PROTECTION WITH SEPICAP MP

8 – SAFETY

patch test

When SEPICAP MP at a concentration of 5% (with respect to the commercially available product) was applied in an occlusive patch test (FINN Chambers) to volunteer's skin, it proved to be NON IRRITANT;

• DERMASCAN Report no. 99954

RBCA Test

At a concentration of 2% (the concentration of the stock solution), SEPICAP MP caused neither hemolysis nor denaturation: therefore it is considered to be NON IRRITANT;

• Seppictox Report RBCA754

HET CAM test

SEPICAP MP was evaluated according to the protocol stipulated in the JORF of 26/12/96. at a concentration of 0.2% (dilution 1/10 in comparison to in vivo method), SEPICAP MP is NON IRRITANT;

• Seppictox HET-CAM.1945.

9 – REGULATORY DATA

INCI name: Sodium Cocoyl Amino Acids and Potassium Dimethicone PEG-7 Panthenyl Phosphate;

EINECS: 2690872 / 2905232 / 2913505 / 2904789

Potassium Dimethicone PEG-7 panthenyl Phosphate is not concerned (polymer)

CAS: 68187-32-6 / 90170-86-8 / 90387-74-9 / 90170-45-9 /

10 – ANYLYTICAL DATA

EP: European Pharmocopoeia

• The only specifications which are guaranteed are those which are given in the Certificate of Analysis supplied with each delivery of the product.