NNAGRAM. 64, rue Anatole France LEVALLOIS-PERRET - FRANCE TEL.: ++33.(0) FAX.: ++33.(0) TENSENNA VEGETAL TENSOR

Transcription

1 Σ NNAGRAM 64, rue Anatole France LEVALLOIS-PERRET - FRANCE TEL.: ++33.(0) FAX.: ++33.(0) TENSENNA VEGETAL TENSOR WHEAT PROTEINS COMPLEX

2 1. INTRODUCTION TENSENNA is a complex of vegetal proteins extracted from wheat having a tensor effect together with moisturising, softening, and anti-ageing properties.

3 2. DESCRIPTION OF TENSENNA The preparation of TENSENNA calls upon advanced exclusive technology. It is obtained by aqueous extraction of two soluble proteins from wheat gluten. These proteins are albumin and globulin. ELECTROPHORESIS CHARACTERISATION The electrophoresis enables a separation and determination of proteins, and thus allows to study their molecular weight distribution. The results show that TENSENNA is composed of proteins with a molecular weight comprised between and daltons. The majority of these proteins have a molecular weight close to daltons which is corresponding to the albumin. These results thus confirm that TENSENNA is composed of non-hydrolysed albumin and globulin. ISO-ELECTROFOCALISATION A first iso-electrofocalisation is realised between ph 3 and ph 9 and shows the presence of iso-electric points between ph 3 and ph 6. A second iso-electrofocalisation is realised between ph 3.5 and ph 5.9 and shows a major band at ph 5.1, and the presence of iso-electric points between ph 4 and ph 5.9.

4 PHYSICOCHEMICAL CHARACTERISTICS Appearance: Colour: Odour: limpid liquid yellow characteristic, very slight ph (direct): Dry extract: 4 % Gardner index: Proteins content: 3 % ± 10 % Amino-nitrogen: 0.15 % Starch: Absence Preservation: 0.5 % Phenonip 0.1 % DMDM Hydantoin Total germs: Yeast and Mould: Pathogens: < 10 CFU /ml < 10 CFU /ml Absence

5 3. DERMO-COSMETIC PROPERTIES OF TENSENNA MOISTURISING With their high molecular weight, albumin and globulin have impressive water - retention capacities and are thus able to ensure an interesting moisturising and protective film-forming activity when applied on skin. TENSOR The tensor effect of TENSENNA has been proved by an in vivo study described in chapter 4. The analysis of cutaneous prints under electronic microscope show a significant decrease in depth of crow s foot wrinkles. ANTI-AGEING TENSENNA is also proved to limit and prevent skin ageing and inflammation thanks to anti-lipase and anti-radicals properties. - ANTI-LIPASE The TENSENNA is proved to inhibit the lipase, an enzyme directly involved in the breakdown of the triglycerides into free fatty acids and diglycerides. These free fatty acids are known to induce a lipid and protein (collagen, keratin...) peroxidation, inducing a damage to the epidermis. The use of TENSENNA on the skin reduces the action of this enzyme and thus limits and prevents the skin ageing process. We have performed an anti-lipase test with TENSENNA as shown in chapter 5.

6 - ANTI-RADICALS The skin functions as a barrier to daily insults from the atmosphere. Thus, superimposed on the chronological ageing process are the effects of a long term exposure to the environment, the most damaging factors being the sun s UV and IR radiations, pollution, and smoke. Free radicals are continuously generated by the cells during exposure of the skin to this environment, potentially creating damage to the epidermis as well as to the dermis (oxidative damage including lipid peroxidation, and reduction of the skin defences). This leads to premature ageing (wrinkling and sagging of the skin) especially in sun-exposed areas, and increased reactivity. There is evidence that protection from oxidative damage caused by daily environment exposure can lead to long term benefits for the skin by slowing down the ageing process. Several investigators have published experimental data showing the protective effect of topically applied free radical scavengers (compounds capable of inhibiting free radical reactions) in reducing acute oxidative damage to the skin. Long-term beneficial effects are a highly significant protection against thinning and a much slower loss of elasticity. The process of wrinkle formation is slowed down and some repair of the damages is allowed to take place. We have performed a test specifically showing the anti-radicals activity of the TENSENNA, as described in chapter 6.

7 4. CLINICAL EFFICIENCY OF TENSENNA We have realised a first study, conducted at Saint Louis Hospital, Service of Dermatology, Paris. A clinical test was realised on 17 individuals versus a placebo. The results are very good: 94% of the individuals estimate to feel the tensor effect of TENSENNA. 33 % of the individuals have spontaneously declared to feel the softening and soothing effect of TENSENNA. A second study was carried out in the Anatomo-Pathologic Service of the Pitié- Salpétrière Hospital in Paris to objectivate the tensor effect of the ENNAGRAM s Vegetal Tensor (TENSENNA). They have analysed cutaneous prints of crow s foot wrinkles skin sections under electronic microscope (x 65). Results show a decrease in depth after application of the Vegetal Tensor (TENSENNA) (see following page).

8 Crow s foot wrinkles before treatment with TENSENNA Crow s foot wrinkles after treatment with TENSENNA

9 9 5. INHIBITION OF THE LIPASE BY TENSENNA DESCRIPTION OF THE TEST We use a dosage kit for the triglycerides, which contains a lipase. We use some plasma, which contains triglycerides, as a substrate for the enzyme. When the lipase is added to the substrate, enzymatic reactions lead to the formation of a coloured compound derived from glycerol. The concentration of this compound can be detected using spectrophotometry, and is proportional to the activity of the enzyme. We can thus compare the action of the lipase, with or without the presence of an inhibitor. METHOD We compare the results obtained in both reaction mediums (with or without TENSENNA) by measuring the absorbance. TEST Plasma + Lipase + TENSENNA CONTROL Plasma + Lipase DETERMINATION OF THE RESULTS The percentage of inhibition is calculated as follows: A = Absorbance % Inhibition = A control - (A test - A blank test) x 100 A control

10 10 RESULTS The tests are realised on four batches of TENSENNA. We obtain the following result: % INHIBITION = 50 % CONCLUSION TENSENNA inhibits the lipase activity. It reduces 50 % of the lipid hydrolysis induced by the lipase.

11 11 6. EVALUATION OF THE ANTI-RADICALS EFFECT OF TENSENNA DESCRIPTION OF THE TEST The determination of the anti-radicals effect is based on inhibition or decrease of cytochrome C reduction velocity by adding a test molecule into the reactive medium. Superoxide anion (O 2 - ) is produced by action of xanthine oxydase on xanthine. This reaction conduces, in absence of molecules able to catch it, to the reduction of cytochrome C. Reduced cytochrome C formation is followed on a spectrophotometer at 550 nm, in presence or in absence (control) of anti-radicals molecules. The reaction can be represented as follow:

12 12 METHOD Enzymatic kinetics are performed and followed on a spectrophotometer Philips PU 8740 UV /visible with automatic carriage of samples. Enzyme control curve : In a phosphate buffer ph 7.8 at 25 C, we add successively xanthine, cytochrome C, and then xanthine oxydase. After homogenisation, kinetic is followed at 550 nm during a minimum period of 3 minutes. Initial velocity of the reaction is noted: Vo Xanthine oxydase concentration is determined so as to obtain: DO (550 nm) = ± /min Inhibition curves : In a phosphate buffer ph 7.8 at 25 C, we add successively xanthine, cytochrome C, and then TENSENNA. Xanthine oxydase is added. After homogenisation, kinetic is followed at 550 nm during a minimum of 3 minutes. Initial velocities of the reactions are noted: Vi

13 13 DETERMINATION OF THE RESULTS The spectrophotometer gives results expressed as DO /min. The percentage of inhibition is determined in comparison with the enzyme control curve. % Inhibition = 100 x (1- Vi) Vo RESULTS TENSENNA is tested in the reactive medium. We obtain the following result: Inhibition of cytochrome C reduction (% compared to control enzyme curve) TENSENNA 48 % CONCLUSIONS In the enzymatic system studied here, the reduction of cytochrome C is generated by production of superoxide anions in the reactive medium. Results show that TENSENNA can inhibit the reduction of cytochrome C by catching superoxide anions (free radicals) present in the reactive medium. As a consequence, topical applications of TENSENNA will limit and even stop the degradation of connective tissue macromolecules due to radicals attacks.

14 14 7. COSMETIC USE OF TENSENNA IDENTIFICATION INCI (EU): Triticum Vulgare Gluten INCI Name (US): Triticum Vulgare (Wheat) Gluten CAS No.: Einecs No.: SAFETY The product is safe from both the toxicological & pharmacological points of view. It is proved non-irritant for both eye and skin at 100% concentration. SUGGESTED DOSAGE Tensor effect: % 50 % in aqueous solutions (up to 80 %) 30 % in gels Softening effect: % Moisturising activity: % Protective activity: 1-5 % COMPATIBILITY TENSENNA is not compatible with propylene glycol, alcohol, glycerol, and vegetal extracts. It is moderately compatible with detergents. DIRECTIONS Add the TENSENNA in a solution or emulsion at a temperature below 40 C.

15 In the case of clear solutions, it is preferable to salt and buffer at first the lotion at ph 8 in order to be as far as possible from the iso-electric points. 15 STORAGE Keep the product in the unopened container, at cool temperature (10 20 C), protected from light, heat and moisture. After opening or for long term storage, keep at 4 C.