Skin lightening becomes clearer! Undecylenoyl phenylalanine: a pure, unique lightening molecule (SEPPIC PATENT)

Transcription

1 SEPIWHITE TM MSH Skin lightening becomes clearer! Undecylenoyl phenylalanine: a pure, unique lightening molecule (SEPPIC PATENT) Novel mode of action: alpha-msh antagonist Lightening activity proven on cultured melanocytes, on reconstructed human epidermis and on healthy volunteers More effective than the tested reference molecules (hydroquinone, arbutine, kojic acid, magnesium ascorbyl phosphate) Excellent tolerance at the usage level (2%) Easy to use in all types of formulation (stable and colourless finish formula) 1

2 CONTENTS 1- Structure of SEPIWHITE MSH Page 4 2- SEPIWHITE MSH: a novel mode of action Page 5 3- Action of SEPIWHITE MSH on the pigment cascade induced by alpha-msh Page Affinity for MC1R alpha-msh receptors Page Inhibition of adenylate cyclase Page Reduction of intracellular camp content Page Inhibition of protein kinase A Page Effect on tyrosinase activity Page Lightening action of SEPIWHITE MSH on cultured melanocytes Page Spontaneous inhibition of melanogenesis in cultured melanocytes Page Inhibition of melanogenesis melanocytes cultivated with alpha-msh Page Inhibition of melanogenesis in melanocytes irradiated with UVB Page Lightening action of SEPIWHITE MSH on reconstructed pigmented human epidermis Page Method Page Results on the Inhibition of the melanin content on the pigmentent reconstructed epidermis Page Results on the measured lightening effect on the pigmented reconstructed epidermis Page Lightening action of SEPIWHITE MSH on volunteers : tan removal test Page Lightening efficacy of SEPIWHITE MSH on asian panel Page Tolerance Page Regulatory data Page Cosmetic applications Page 30 2

3 Skin pigmentation is an international preoccupation. Natural or photo-induced ageing, hormonal disorders (contraceptives, pregnancy, menopause, etc.), repeated exposure to the sun and irritation or inflammation reactions lead to the appearance of skin pigment problems. The complexion is not uniform and highly unattractive marks appear on the skin. Many cosmetic products claiming a lightening action in fact have very mediocre effectiveness and non-negligible toxicity, and are difficult to formulate. SEPPIC has developed a major lightening research program, focusing its attention on the effects of a precursor molecule and major factor in the regulation of skin pigmentation, melanotropin (or alpha-msh). Melanotropin controls tyrosinase activity, melanin (eumalanin and phaeomelanin) synthesis and melanosome transfer. Pigmentation control Developing a concept based on innovation, effectiveness and tolerance, SEPPIC has recently developed a new lightening active ingredient, SEPIWHITE MSH. A new molecule, this active ingredient has a complete and innovative mode of action. An alpha-msh antagonist, SEPIWHITE MSH is our solution for combating pigment problems. Maintaining skin integrity, it acts upstream of melanogenesis on all the steps in the pigment cascade induced by melanotropin. Its depigmenting effectiveness has been proven in vitro and in vivo on healthy volunteers. 3

4 1 STRUCTURE OF SEPIWHITE MSH Our development strategy Alpha-MSH (or melanotropin) is one of the key molecules responsible for melanin synthesis. It acts in the pigment cycle to stimulate melanogenesis via a type 1 membrane receptor: MC1R (the importance of the MC1R receptor in the control of melanogenesis has been extensively described in the literature). Studies conducted on human skin have shown that subcutaneous injection of α-msh induces hyperpigmentation. Moreover, α-msh activity is increased in hyperpigmented areas or in areas exposed to UV radiation. Subjects with pale phototypes who show little tanning under the effect of UV radiation have few MC1R receptors in their melanocytes. These MC1R receptors, activated by α-msh and its analogues, are inhibited by natural antagonists present in the skin, the AGRPs (agouti-related protein). These proteins have a phenylalanine in a key position which plays an essential role in binding to MC1R receptors. They thus inhibit melanogenesis. αmsh + - MC1R Agouti-related protein AGRP (Phenylalanine in key position) Melanocyte Melanin Structure of SEPIWHITE MSH In view of these findings about the involvement of melanotropin in the regulation of melanogenesis, SEPPIC research has developed SEPIWHITE MSH, an undecylenoylphenylalanine. With its lipoaminoacid form, a common molecular structure in the skin, SEPIWHITE MSH has remarkable skin bioaffinity. It is thus well tolerated at the suggested utilization concentrations. SEPIWHITE MSH is an easily-formulable white powder, and has a fully-defined structure. End products containing SEPIWHITE MSH are stable, and are white or colourless. 4

5 Sepiwhite MSH structure 2 SEPIWHITE MSH: A NOVEL MODE OF ACTION How is melanotropin involved in the regulation of melanogenesis? By binding to its type 1 receptors, α-msh (melanotropin) stimulates the α subunit of the stimulatory G protein. The latter activates adenylate cyclase, increasing the intracellular camp content. The camp activates protein kinase A, which then phosphorylates tyrosinase. The phosphorylated tyrosinase becomes active and stimulates melanogenesis. Melanotropin action 5

6 SEPIWHITE MSH, an alpha-msh antagonist SEPIWHITE MSH acts as a melanotropin inhibitor and thus competes with this key molecule in upstream control of melanogenesis. All the steps leading to melanin synthesis are then inhibited by SEPIWHITE MSH:. affinity for the MC1R melanotropin receptor,. inhibition of adenylate cyclase,. reduction of intracellular camp content,. inhibition of protein kinase A,. inhibition of melanogenesis. SEPIWHITE MSH an original mode of action! α-msh antagonist The lightening activity of SEPIWHITE MSH has been proven not only in vitro (mode of action) but also on reconstructed pigmented human epidermis and in vivo on healthy volunteers. 6

7 3. ACTION OF SEPIWHITE MSH ON THE PIGMENT CASCADE INDUCED BY ALPHA-MSH => SEPPIC BioLightening expert report CA0003RF-GB The effects of SEPIWHITE MSH were compared with various reference lightening agents: hydroquinone, kojic acid, arbutine and magnesium ascorbyl phosphate (VCPMG). The effects of undecylenic acid and phenylalanine (which are the starting materials of SEPIWHITE MSH) have also been characterized Affinity for MC1R α-msh receptors Method (binding test) Type 1 α-msh receptors were isolated from B16/F1 murine melanocyte cell membranes. The type 1 α-msh receptors (MC1R) were incubated with a radioactive ligand (high affinity for the receptor) in the presence or absence of the products. Control cultures were incubated in the absence of the product and in the presence of the radioactive ligand. Incubation with: MC1R receptors SEPIWHITE MSH or phenylalanine or undecylenic acid or arbutine or kojic acid or VCPMG or control (no active matter) Radioactive ligand* % affinity of the products for the α-msh receptor compared with control (according to the displacement of bound ligand* from the receptor) A product with the capacity to bind to MCR1 receptors displaces the radioactive ligand from its binding sites on the receptors and, as a result, reduces the quantity of radioactivity. 7

8 Results SEPIWHITE MSH is the only product capable of binding to alpha-msh receptors, in comparison with the tested reference products (arbutine, kojic acid, VCPMG). As neither undecylenic acid nor phenylalanine is capable of binding to the receptor, the effectiveness of SEPIWHITE MSH is a consequence of its lipoamine form Inhibition of adenylate cyclase Method (biochemical test) The camp produced by the action of adenylate cyclase on ATP was measured by a radioimmunological assay. ATP Adenylate cyclase Incubation with: camp % adenylate cyclase activity compared with control SEPIWHITE MSH or phenylalanine or undecylenic acid or arbutine or kojic acid or VCPMG or control (without active matter) 8

9 A product with the capacity to inhibit adenylate cyclase reduces the quantity of radioactivity. Results SEPIWHITE MSH is the best inhibitor of adenylate cyclase, in comparison with the tested reference products. Undecylenic acid, but not phenylalanine, also has an inhibitory effect, which demonstrates the specific involvement of the lipoamine form in this inhibition Inhibition of intracellular camp content Method (cell culture) Melanocyte cultures were incubated in the presence of the products. Cultures were incubated without product. The intracellular camp content was measured immunochemically. 9

10 B16/F1 melanocytes Incubation with: SEPIWHITE MSH or hydroquinone or arbutine or kojic acid or VCPMG or control (no active matter) Measurement of intracellular camp content (immunochemistry) Results SEPIWHITE MSH reduces (-34%) the intracellular camp content. Its action is considerably greater than that of the reference agents tested (arbutine, kojic acid and VCPMG). This result confirms the inhibition of adenylate cyclase by SEPIWHITE MSH in cultured melanocytes. 10

11 3.4 - Inhibition of protein kinase A Method (biochemical test) Protein kinase A (PKA) phosphorylates proteins, including tyrosinase. Like most enzymes tyrosinase must be phosphorylated to be active. The activated enzyme was incubated in the presence of the products, radioactive ATP and histone (= protein substrate). Control cultures were incubated in the absence of the product and in the presence of radioactive ATP and histone. Histone + ATP* PKA Incubation with: Histone-P* % PKA activity compared with control (radioactivité) SEPIWHITE MSH or phenylalanine or undecylenic acid or arbutine or kojic acid or VCPMGor control (no active matter) A product capable of inhibiting protein kinase A reduces the quantity of labelled phosphorylated histone. Results 11

12 Of the products tested, SEPIWHITE MSH is the only PKA inhibitor (total inhibition, 100%). This inhibition is due to its lipoamine form (no inhibition by undecylenic acid or phenylalanine alone) Effect of SEPIWHITE MSH on tyrosinase activity Method (biochemical test) Tyrosinase was incubated in the presence of the products and in the presence of its substrate, L-tyrosine. Control cultures were incubated in the absence of the product and in the presence of tyrosinase and L-tyrosine. The quantity of DOPA-QUINONE formed was measured by spectrophotometry at 490 nm. Tyrosinase L Tyrosine Incubation with: L-DOPA Tyrosinase DOPA-QUINONE (spectrophotometry) % tyrosinase activity compared with control SEPIWHITE MSH or phenylalanine or undecylenic acid or arbutine or kojic acid or VCPMG or hydroquinone or control (no active matter) Results 12

13 SEPIWHITE MSH is a very effective inhibitor of tyrosinase activity. Its effectiveness is comparable to that of the reference products. Undecylenic acid and phenylalanine have significantly lower anti-tyrosinase activity than SEPIWHITE MSH, demonstrating the benefits of the lipoamine form for this activity. Given its chemical structure, undecylenoyl-phenylalanine, and the fact that phenylalanine is the precursor of L-tyrosine, the natural substrate of tyrosinase, SEPIWHITE MSH may act as a false substrate for the tyrosinase. An alpha-msh antagonist, SEPIWHITE MSH has a novel mode of action compared with the reference lightening agents commonly used. Acting as a melanotropin inhibitor, it acts on all the steps in the pigment cascade. α-msh + MC1R SEPIWHITE MSH - Competition with α-msh on the MC1Rreceptors ProtGαS Adenylate Cyclase Inhibition of adenylate cyclase ATP camp Protein Kinase A Reduction of the intracellular camp content Inhibition of PKA Inactive tyrosinase Active tyrosinase-p Inhibition of tyrosinase MELANOGENESIS The inhibition of melanogenesis (melanin synthesis) has been confirmed in cell culture and in reconstructed pigmented human epidermis. 13

14 4. LIGHTENING ACTION OF SEPIWHITE MSH ON CULTURED MELANOCYTES (IN VITRO) The lightening activity of SEPIWHITE in comparison to the reference active ingredients used customarily (arbutine, kojic acid, VCPMG, hydroquinone) has been proven on B16 melanocytes cultured under three types of conditions: 1. spontaneous lightening action on melanocytes => inibition of basal melanogenesis by SEPIWHITE MSH 2. lightening action on melanocytes cultivated with alpha-msh => inhibition by SEPIWHITE MSH of the melanogenesis induced by alpha MSH => proof of its antagonistic effect towards alpha-msh 3. lightening action on melanocytes following UVB irradiation => inhibition by SEPIWHITE MSH of the melanogenesis induced by UVB In these three types of test, the lightening activity was determined by measurement of the melanin content. 4.1 Inhibition of basal melanogenesis within melanocytes cultivated alone : Time scale: The effectiveness of the active ingredients under test was assessed after three days of incubation with melanocytes. D0 D1 D2 D3 Days Incubation of the products ASSESSMENT OF THE EFFECTS 14

15 Results SEPIWITHE MSH Skin lightening effect on melanocytes SEPIWHITE MSH inhibits very effectively basal melanogenesis (-66%) compared with the control group (untreated). Its action is considerably greater than that of arbutine, kojic acid or VCPMG. The melanin reduction effect of hydroquinone is due essentially to cytotoxic activity (the morphology of the melanocytes appears substantially altered under the microscope). In contrast, SEPIWHITE MSH maintains the morphological integrity of the melanocytes Inhibition of melanogenesis induced by alpha-msh within melanocytes : Time scale: The effectiveness of the active ingredients under test was assessed after three days of incubation with melanocytes cultured in the presence of alpha-msh. D0 D1 D2 D3 Days Incubation of the products in the presence of 10 nm α-msh ASSESSMENT OF THE EFFECTS 15

16 Results for melanin (mg/ml) : 0,04 Melanin (mg/ml) 0,03 0,02 0,01 * * * 0 Control αmsh 10 nm SEPIWHITE 40 ppm HQ 5 ppm KA 40ppm + αmsh 10nM AR 40 ppm VCPMG 40ppm * results significantly different (p<0.05) from the alpha-msh group) HQ: hydroquinone KA: kojic acid AR: arbutine - Alpha-MSH stimulates melanogenesis. This stimulation increased the melanin content by 113% compared with that of the untreated control group. This result validates the melanin-stimulating effect of melanotropin. - SEPIWHITE MSH completely inhibited the melanin content induced by the presence of alpha-msh. - The melanin content induced by the presence of alpha-msh was inhibited by hydroquinone (100%) and by arbutine (48%). Kojic acid and VCPMG did not have a significant inhibitory effect on melanotropin-induced melanogenesis. SEPIWHITE MSH is thus a very effective inhibitor of the melanogenesis induced by melanotropin (reduction of the melanin content). This result confirms its alpha-msh antagonist effect on cell cultures. 16

17 4.3 - Inhibition of melanogenesis induced by UVB within melanocytes : Time scale: The effectiveness of the active ingredients under test was assessed after three days of incubation with melanocytes cultured in the presence of UVB radiation. D0 D1 D2 D3 D4 Days UVB irradiation (4x) ASSESSMENT 20 mj/cm 2 OF THE EFFECTS Incubation of the products a. Results for melanin (mg/ml) 0,1 0,09 Melanin (mg/ml) 0,08 0,07 0,06 0,05 0,04 Control UVB SEPIWHITE 4x20 mj/cm 2 30ppm * TOXIC HQ 5ppm * KA 30ppm * AR 30ppm * VPMG 30ppm UVB (4x20 mj/cm 2 ) * results significantly different (p<0.05) from the UVB group HQ: hydroquinone KA: kojic acid AR: arbutine UVB radiation increased the melanin content by 150% compared with the nonirradiated control group, which confirms its melanin-stimulating effect. SEPIWHITE MSH was a very effective inhibitor of the melanogenesis induced by UVB radiation (-66%). Its action was greater than that of kojic acid (-55%), VCPMG (-53%) or hydroquinone (toxic in the presence of UVB radiation). 17

18 SEPIWHITE MSH has thus a highly-significant inhibitory effect on both basal melanogenesis and melanogenesis stimulated by melanotropin or UVB radiation. SEPIWITHE MSH Skin lightening effect on melanocytes Melanocytes cultivated alone Melanocytes + α MSH Melanocytes + UVB SEPIWHITE MSH inhibits melanogenesis Without Sepiwhite With Sepiwhite 5. LIGHTENING ACTION OF SEPIWHITE MSH ON RECONSTRUCTED PIGMENTED HUMAN EPIDERMIS 5.1 Method The lightening activity of SEPIWHITE MSH formulated at 2% has been confirmed on pigmented reconstructed epidermis (3D model of organized human skin) after 7 days of topical application. Its activity was compared with those of arbutine and kojic acid, also formulated at 2%, and that of hydroquinone formulated at 0.1% (because of its cytotoxicity). 18

19 Histological section of the epidermis (Skinetic phototype 4) The lightening activity was assessed according to two parameters : 1. The melatin content was measured by spectrophotometry at 500 nm. 2. The colour of the reconstructed human epidermis (phototype IV) was assessed using a colorimeter. The parameters L*, a*, b* were measured: L*: luminosity index. a*: colour spectrum from green to red. b*: colour spectrum from blue to yellow. SEPIWHITE MSH Skin lightening effect on reconstructed epidermis 19

20 5.2 Results on the inhibition of the melanin content on the pigmented reconstructed epidermis SEPIWHITE MSH effectively reduced the melanin content in reconstructed pigmented epidermis (-24%). Its action was similar to that of arbutine (-24%) and kojic acid (-33%) and greater than that of hydroquinone (-9%). The melaninreducing effect of SEPIWHITE MSH has thus been confirmed on a 3D reconstructed epidermis model. 20

21 5.3. Results on the measured lightening effect on the pigmented reconstructed epidermis The lightening activity was measured by determination of the change in the colour of the epidermis. SEPIWHITE MSH was the only agent that had a measurable lightening effect at the surface of the skin from 7 days of application ( L, a, b). Its activity was considerably greater than that of arbutine, kojic acid or hydroquinone. The excellent effectiveness of SEPIWHITE MSH is very likely to be related to its lipoamine form, its amphiphilic character giving it early effectiveness in the reconstructed skin model in comparison with the other reference molecules. 21

22 6. LIGHTENING ACTION OF SEPIWHITE MSH ON VOLUNTEERS: TAN REMOVAL TEST => DERMSCAN expert report no (formula 6895 and placebo 6894) The effectiveness of SEPIWHITE MSH formulated at 2% was tested on 14 volunteers in comparison with a placebo. The skin pigmentation of the subjects was stimulated previously by UV irradiation (solar spectrum). After acquisition of a tan, SEPIWHITE MSH formulated at 2% was applied for 42 days. The return to normal pigmentation on the area treated with Sepiwhite, in comparison with a placebo area and an untreated area, was assessed by colorimetric measurement of the skin: measurement of the parameters L, a, b and ITA, where ITA is the individual typology angle (integration of L and b). A lightening agent must increase this parameter. ITA = [Arc tan ((L-50)/b)] x 180/Π Time scale: The backs of caucasian volunteers were irradiated at 0.8 MED (minimal erythema dose) on the first day (D0) then 48 and 72 hours later (D2 and D4). When the tan appeared (D7), the products were applied every day for six weeks on the irradiated areas (until D49). The skin colour, and in particular the return to normal pigmentation, were assessed on D49. D0 D2 D4 D7 D49 Days UV irradiations TOPICAL APPLICATION ASSESSMENT OF 0.8 MED OF THE PRODUCTS THE EFFECT 2x per day 22

23 Skin colour change results on D49 : % variation compared with the untreated area Placebo SEPIWHITE MSH 2% -4 L a b ITA Colorimetric parameters (D49) * *significative results (p<0,07) compared to non treated area SEPIWHITE MSH showed excellent lightening effectiveness on healthy volunteers compared to the placebo : It improves the return to normal pigmentation characterized by a reduction of b (in 86% of volunteers) and an increase of ITA (in 57% of volunteers). The placebo did not have any significant effect. 23

24 7. LIGHTENING EFFECT OF SEPIWHITE MSH TESTED ON ASIAN VOLUNTEERS => DERMSCAN expert report no (formula 6911) The objective of this study is the lightening evaluation of SEPIWHITE MSH formulated at 2% (formula 6911) on 30 Asian volunteers (test performed in Bangkok). Formula 6911 (described in the cosmetics applications) was applied to the face twice a day for 3 months. The lightening effectiveness of SEPIWHITE MSH was assessed before treatment (D0), after treatment for two months (D56) and after treatment for three months (D84) according to five parameters: 1. Observation of melanin particles at the skin surface => samples of stratum corneum cells taken using D-Squames; 2. Measurement of the luminance, parameter L* in chromametry; 3. Measurement of the skin colour (melanin index) using a Mexameter; 4. Dermatological examination of hyperpigmented marks; 5. Overall assessment of formula 6911 by the panelists (in terms of effectiveness, comfort and tolerance). 24

25 7.1 Decrease of melanic pigments at the skin surface By the second month of treatment, there were fewer melanic pigments located in the corneocytes (surface cells of the stratum corneum). The skin is visibly clearer. 7.2 Even skin tone effect Determination of the luminosity (parameter L*) by chromametry: By the second month of treatment, SEPIWHITE MSH significantly increases even skin tone (increase of L* parameter). The ITA (Individual Typological Index) also increases significantly from the second month of treatment. The tone is sublimated, more radiant. 25

26 7.3 Lightening effect Measurement of skin colour reduction (melanin index) using a Mexameter: By the second month of treatment, the skin was significantly lightened (decrease of the melanin index). The pigmented areas fade away, the tone becomes more uniform. 7.4 Lightening effect on pigmented areas : Clinical and visual examination done by a dermatologist : By the second month of treatment, the hyperpigmented areas are visibly and significantly lightened. 26

27 7.5 Overall assessment of formula 6911 by the panelists after three months of treatment 93% of the volunteers appreciated formula 6911 for its effectiveness and comfort. Moreover, tolerance was excellent on all the persons tested. 7.6 Summary of the lightening effectiveness of SEPIWHITE MSH formulated at 2% and tested on Asian volunteers Glowing and uniform, the tone is visibly rejuvenated. 27

28 SEPIWHITE MSH, a new generation of lightening molecule! Through a novel and demonstrated mode of action, SEPIWHITE MSH lights up the complexion while limiting pigment-related problems. An alpha-msh antagonist, SEPIWHITE MSH is a very effective inhibitor of melanogenesis The complexion is more uniform, age spots disappear. Prepare to be enlightened! 28

29 8 TOLERANCE The good tolerance of SEPIWHITE MSH at 2% has been assessed using cosmetics methods. Skin patch test 1. Tested on Caucasian volunteers : SEPIWHITE MSH was assessed in an occlusive patch test (FIHN CHAMBERS) on 20 volunteers. It is considered non-irritant for 48- hours exposure at a concentration of 2% in water (ph = 6.3). DERMSCAN expert report no LCE Tested on Japanese volunteers : SEPIWHITE MSH was assessed in an occlusive patch test (FIHN CHAMBERS) on 20 volunteers. It is considered non-irritant for 48- hours exposure at a concentration of 2% in parafin oil (ph = 6.3). IEC expert report no. J LCA 03019B Diluted in water and tested in occlusive patch test, SEPIWHITE MSH may eventually cause some skin irritations on Japanese volunteers. Conditions of use on Asian volunteers: After three months of treatment with two applications per day on the face, formula 6911 containing 2% of SEPIWHITE MSH was very well tolerated by all the panellists (in vivo study performed in Bangkok). => DERMSCAN expert report no (formula 6911) Sensitization Under the experimental conditions of this protocol by MARZULLI and MAIBACH, SEPIWHITE MSH did not cause any primary or cumulative irritation reaction, nor any skin sensitization. SEPIWHITE MSH is not sensitizing. => ASTER expert report no. PC2796LCA02009 Inside elbow iterative test Tested on 20 subjects with normal or sensitive skin for 15 days, 2% SEPIWHITE MSH in water was well tolerated. => PERITESCO expert report no. PO1713D LCE02018a RBCA (Red Blood Cell Assay) Tested at 1%, SEPIWHITE MSH is considered non-irritant. It caused no haemolysis or denaturation. SEPPICtox expert report RBCA0918a Ames test According to the Ames test, SEPIWHITE MSH is considered non-mutagenic. => SAFEPHARM expert report no. SPL

30 The advised maximum level of SEPIWHITE MSH is 2% in cosmetics applications. The MSDS data must be consulted before the use of SEPIWHITE MSH. 9 REGULATORY DATA INCI name: Undecylenoyl phenylalanine CAS No.: ELINCS No.: underway registration (SEPIWHITE MSH is a new molecule subject to notification) Japan: underway registration for quasi-drug products. 10 COSMETIC APPLICATIONS => SEPPIC GALENIC expert report LCO03024-GB SEPIWHITE MSH can be used easily in gel, gel-cream, simple and multiple emulsion, lotion, tonic and hygiene product formulas. Completely stable, its use is particularly recommended at 2% in any type of formula where lightening activity is sought. Complexion uniformity anti-ageing care Complexion enhencing care Age spot removal Skin Lightening make-up removers Skin Lightening cleansers Skin Lightening powder SEPIWHITE MSH is fully compatible in formulation with other lightening active ingredients such as the AHAs or kojic acid (formulas with ph < 5), arbutine, hydroquinone or VCPMG (formulas with ph > 7). SEPIWHITE MSH is also fully compatible in formulation with sun screens. Formulation studies have proven the excellent stability of SEPIWHITE MSH formulated at 2%. No change of colour or odour has been observed in the end formulas over time or according to temperature. 30

31 SEPIWHITE MSH can be formulated very easily in clear, colourless lotions, whether they are aqueous, hydroalcoholic or hydroglycolic. Advice on using SEPIWHITE MSH in formulation: Introduction into the aqueous or oil phase at T 60 C ph < 5 => incorporation into all types of formula 5 < ph 6.2 => addition of ethanol advised ph 6.2 => incorporation into any type of formula 31

32 REFRESHING ILLUMINATING WATER 6912A Hydroalcoholic formula: A SEPIWHITE MSH (Undecylenoyl phenylalanine - SEPPIC) 2.00% Water QS % Tromethamine 0.80% B Glycerine Ethanol Fragrance SEPICIDE HB (Phenoxyethanol/Methylparaben/Ethylparaben/Propylparaben/Butylparab en - SEPPIC) SEPICIDE CI (imidazolidinyl urea - SEPPIC) 5.00% 10.00% 0.10% 0.30% 0.20% Procedure Melt the SEPIWHITE MSH in hot water, neutralize, and cool this phase A under slow stirring. When cold, add phase B. Comments SEPIWHITE MSH SEPICIDE HB/CI New lightening active ingredient acting through a novel mode of action. An alpha-msh (melanotropin) antagonist, it reduces the synthesis of melanin pigments effectively while maintaining skin integrity. Preservative system. Characteristics Appearance Clear ph 7 Viscosity < 50 mpa.s Stability Stable at RT/40 C/50 C 32

33 SMOOTH ALCOHOL-FREE ILLUMINATING WATER 6912 Alcohol-free formula: SEPIWHITE MSH (Undecylenoyl phenylalanine - SEPPIC) 2.00% A Water QS % Tromethamine 0.80% B Glycerine Fragrance SEPICIDE HB (Phenoxyethanol/Methylparaben/Ethylparaben/Propylparaben/Butylparab en - SEPPIC) SEPICIDE CI (imidazolidinyl urea - SEPPIC) 5.00% 0.10% 0.30% 0.20% Procedure Melt the SEPIWHITE MSH in hot water, neutralize, and cool this phase A under slow stirring. When cold, add phase B. Comments SEPIWHITE MSH SEPICIDE HB/CI New lightening active ingredient acting through a novel mode of action. An alpha-msh (melanotropin) antagonist, it reduces the synthesis of melanin pigments effectively while maintaining skin integrity. Preservative system. Characteristics Appearance Clear ph 7 Viscosity < 50 mpa.s Stability Stable at RT/40 C/50 C 33

34 LIGHT INFUSED COMPLEXION EXPERT SKIN CARE 6911 Formula (in vivo tested on asian volunteers during 3 months) A MONTANOV 202 (Arachidyl alcohol and behenyl alcohol and 2.00% arachidylglucoside SEPPIC) MONTANOV 68 (Cetearyl alcohol and cetearyl glucoside - SEPPIC) 2.00% Isononyl isononanoate 8.00% LANOL P (Glycol palmitate - SEPPIC) 2.00% B Water QS 100% C D Procedure SEPIWHITE MSH (Undecylenoylphenylalanine - SEPPIC) Tromethamine Magnesium aluminium silicate SIMULGEL EG (sodium acrylate/acryloyldimethyltaurate copolymer and isohexadecane and Polysorbate 80 - SEPPIC) Cyclomethicone 2.00% 0.65% 1.00% 2.00% 4.00% 0.30% SEPICIDE HB (Phenoxyethanol/Methylparaben/Ethylparaben/Propylparaben/Butylparaben - SEPPIC) SEPICIDE CI (imidazolidinyl urea - SEPPIC) 0.20% Fragrance DL-α-Tocopherol 0.30% 0.05% Melt A at C in a double-walled tank to prevent any overheating. Heat B to C. Introduce all the melted phase A into the water, then start the homogenizer (rotor/stator turbine). Add phase C. After a few minutes, stop the homogenizer and cool gradually with moderate stirring to prevent any recrystallization of MONTANOV 202. Introduce the constituents of D at about 30 C. Comments SEPIWHITE MSH MONTANOV 202 MONTANOV 68 LANOL P New lightening active ingredient acting through a novel mode of action. An alpha-msh (melanotropin) antagonist, it reduces the synthesis of melanin pigments effectively while maintaining skin integrity. Glucolipid emulsifier of vegetable origin. Produces emulsions with a light, evanescent feel, which are very easy to apply and penetrate quickly. These emulsions leave the skin soft and non-oily. Glucolipid emulsifier of vegetable origin. In combination with MONTANOV 202, used to modulate emulsion texture and suppleness as required. Texture agent does not leave any oily feeling easy to rinse. Improves texture and does not soap up upon application. 34

35 SIMULGEL EG SEPICIDE HB/CI Thickening and emulsifying agent in very easy to use liquid form (no predispersion or neutralization). SIMULGEL EG gives gel-creams with a silky, unctuous texture which are easy to apply and rapidly absorbed by the skin. Preservative system. Characteristics Appearance white cream. ph approximately 6.3 Viscosity 35,000 mpa.s BROOKFIELD LVT spindle 4 6 rpm. Stability stable at RT/40/50 C/cycles Notes Fragrance: NINALIX (QUEST) 35

36 ANTI AGEING SKIN CARE EVEN COMPLEXION 6930 Formula containing Magnesium ascorbyl phosphate A MONTANOV 202 (Arachidyl alcohol and behenyl alcohol and 2.00% arachidylglucoside SEPPIC) SIMULSOL 165 (Glyceryl stearate and PEG100 stearate - SEPPIC) 3.00% Diisopropyl adipate 10.00% Jojoba oil 2.00% LANOL P (Glycol palmitate - SEPPIC) 2.00% B Water QS 100% SEPIWHITE MSH (Undecylenoylphenylalanine - SEPPIC) Carbomer 2.00% 0.20% C SIMULGEL EG (sodium acrylate/acryloyldimethyltaurate copolymer and isohexadecane and Polysorbate 80 - SEPPIC) 2.00% D Magnesium ascorbyl phosphate Tromethamine 3.00% 1.25% E SEPICIDE HB 0.30% (Phenoxyethanol/Methylparaben/Ethylparaben/Propylparaben/Butylparaben - SEPPIC) SEPICIDE CI (imidazolidinyl urea - SEPPIC) 0.20% Fragrance DL-α-Tocopherol 0.30% 0.05% F Tromethamine QS ph=7.5 Procedure Melt A at C in a double-walled tank to prevent any overheating. Melt the SEPIWHITE and disperse the carbomer in hot water at C. Introduce all the melted phase A into phase B, then start the homogeniser (rotor/stator turbine). Add phase C. After three or four minutes, add phase D. After a few minutes, stop the homogeniser and cool gradually under moderate stirring. Introduce the constituents of E at about 30 C. Adjust the ph on completion of emulsification if necessary. Comments SEPIWHITE MSH MONTANOV 202 SIMULSOL 165 New lightening active ingredient acting through a novel mode of action. An alpha-msh (melanotropin) antagonist, it reduces the synthesis of melanin pigments effectively while maintaining skin integrity. Glucolipid emulsifier of vegetable origin. Produces emulsions with a light, evanescent feel, which are very easy to apply and penetrate quickly. These emulsions leave the skin soft and non-oily. Self-emulsifying base. Can be used in synergy with an emulsifier from the MONTANOV range to obtain unctuous, stable creams. 36

37 LANOL P SIMULGEL EG SEPICIDE HB/CI Texture agent does not leave any oily feeling easy to rinse. Improves texture and does not soap up upon application. Thickening and emulsifying agent in very easy to use liquid form (no predispersion or neutralization). SIMULGEL EG gives gel-creams with a silky, unctuous texture which are easy to apply and rapidly absorbed by the skin. Preservative system. Characteristics Appearance thick cream ph approximately 7.5 Viscosity 50,000 mpa.s BROOKFIELD LVT spindle 4 6 rpm. Stability stable at RT/40/50 C/cycles Notes Fragrance: MIMI X (QUEST) Carbomer: ULTREZ 10 (NOVEON) 37

38 SPRAYABLE LIGHTENING CARE 6924 Formula A MONTANE 60 (Sorbitan stearate - SEPPIC) 2.00% MONTANOX 60 (Polysorbate 60- SEPPIC) 3.00% Octyl palmitate 15.00% B Water QS 100% C SEPIWHITE MSH (Undecylenoylphenylalanine - SEPPIC) Tromethamine Magnesium aluminium silicate SIMULGEL EG (sodium acrylate/acryloyldimethyltaurate copolymer and isohexadecane and Polysorbate 80 - SEPPIC) 2.00% 0.80% 1.00% 0.50% E SEPICIDE HB 0.30% (Phenoxyethanol/Methylparaben/Ethylparaben/Propylparaben/Butylparaben - SEPPIC) SEPICIDE CI (imidazolidinyl urea - SEPPIC) 0.20% Fragrance DL-α-Tocopherol 0.30% 0.05% F Tromethamine QS ph=6.3 Procedure Melt A at 75 C in a double-walled tank to prevent any overheating. Melt the SEPIWHITE and disperse the magnesium aluminium silicate in hot water at 75 C. Introduce all the melted phase A into phase B, then start the homogeniser (rotor/stator turbine). Add phase C. After a few minutes, stop the homogeniser and cool gradually under moderate stirring. Introduce the constituents of E at about 30 C. Adjust the ph on completion of emulsification if necessary. Comments SEPIWHITE MSH New lightening active ingredient acting through a novel mode of action. An alpha-msh (melanotropin) antagonist, it reduces the synthesis of melanin pigments effectively while maintaining skin integrity. MONTANE/MONTANOX 60 Pair of emulsifiers giving fluid, stable emulsions. SIMULGEL EG SEPICIDE HB/CI Thickening and emulsifying agent in very easy to use liquid form (no predispersion or neutralization). SIMULGEL EG gives gel-creams with a silky, unctuous texture which are easy to apply and rapidly absorbed by the skin. Preservative system. Characteristics Appearance white fluid ph approximately 6.3 Viscosity 300 mpa.s BROOKFIELD LVT spindle 2 6 rpm. 38

39 Stability stable at RT/40/50 C/cycles Notes Fragrance: WHITE DEAD NETTLE RS (TECHNICO FLOR) Magnesium aluminium silicate: VEEGUM HS (NOVEON) 39

40 SUBLIM COMPLEXION PRESSED POWDER 6907A Formula A SEPIWHITE MSH (Undecylenoylphenylalanine - SEPPIC) 2.00% MICROPEARL M310 (Cross linked polymethylmethacrylate- SEPPIC) 5.00% Mica 50.00% Talc 32.50% Iron oxide yellow 0.50% B SEPICALM VG (Sodium palmitoyl proline and Nymphea alba flower extract - SEPPIC) 5.00% C Dimethicone 5.00% Procedure Weigh all the powders (phase A) and grind them dry in a knife cutting mill (split the grinding time into two equal parts and scrape the edges of the bowl between the two periods). Add phase B and grind for the same time as for phase A alone. Add phase C and repeat the same grinding operation as for phase B. The powder prepared in this way is pressed in pots using a KENWALL manual compactor with a pressure of 120 bar. Comments SEPIWHITE MSH SEPICALM VG New lightening active ingredient acting through a novel mode of action. An alpha-msh (melanotropin) antagonist, it reduces the synthesis of melanin pigments effectively while maintaining skin integrity. Its powder form is ideally suited to the formulation of cosmetic powders. Lipoamine soothing active ingredient, rich in water lily extract. Particularly suited to the requirements of sensitive or sun-damaged skin. It effectively relieves stressed skin and soothes irritation due to the use of certain active ingredients. MICROPEARL M310 Lipodispersible powder with excellent absorbent and matt-finishing properties. 40

41 Characteristics Appearance Stability pale yellow powder stable at RT/40 C Notes Talc: Talc 00C (LUZENAC) Mica: Mica1000 (SCIAMA) Iron oxide yellow: SICOVIT yellow 10 E172 (BASF) Dimethicone: DC cps (DOW CORNING) 41

42 FLASH COMPLEXION FLUID 6931 Formula A SEPIWHITE MSH (Undecylenoylphenylalanine - SEPPIC) 2.00 % Ethanol % MICROPEARL M310 (Methyl methacrylate crosspolymer - SEPPIC) 3.00 % B Water QSP 100 % Carbomer 0.25 % Xanthan gum 0.20 % C Isononyl isononanoate 5.00 % D SEPICIDE HB (Phenoxyethanol/Methylparaben/Ethylparaben 0.30 % /Propylparaben/Butylparaben - SEPPIC) SEPICIDE CI (Imidazolidinyl urea - SEPPIC) 0.20 % Fragrance 0.30 % DL α Tocopherol 0.05 % E Tromethamine QS ph=7.2 Procedure Solubilize SEPIWHITE MSH into Ethanol under mixing and add MICROPEARL M310. Disperse carbomer and xanthan gum in water and add C in this phase B, under mixing. Add phase D and adjust the ph at the end. Comments SEPIWHITE MSH MICROPEARL M310 SEPICIDE HB / CI A new lightening active ingredient acting through an original pathway. Alpha MSH antagonist ( Melanotropin ), it decreases effectively the melanic pigments by respecting the cutaneous integrity. Lipodispersible powder with excellent absorbent and matifying properties. A preservative system. Characteristics Appearance light gel ph Approximately 7.2 Viscosity 6,000 mpa.s BROOKFIELD LV S4-6rpm. Stability stable at room temperature, 40 and 50 C, freeze Thaw cycles Notes Fragrance : NINALIX X (QUEST) Xanthan gum : KELTROL T (KELCO) Carbomer : ULTREZ 10 (NOVEON) 42

43 Nota The analytical specifications warranted are only those mentioned on the certificate of analysis supplied with each delivery of the product. Except as set forth above, SEPPIC* makes no warranties, whether express, implied or statutory, as to the product which is the subject of this document. Without limiting the generality of the foregoing, SEPPIC* makes no warranty of merchantability of the product or of the fitness of the product for any particular purpose. Buyer assumes all risk and liability resulting from the use or sale of the product, whether singly or in combination with other goods. The information set forth herein is furnished free of charge and is based on technical data that SEPPIC* believes to be reliable. It is intended for use by persons having technical skill and at their own discretion and risk. Since conditions of use are outside SEPPIC*'s control, SEPPIC* makes no warranties, express or implied, and assumes no liability in connection with any use of this information. Nothing herein is to be taken as a license to operate under or a recommendation to infringe any patents. * SEPPIC being: and, depending on the country : SEPPIC S.A. SEPPIC UK Ltd SEPPIC ITALIA Srl 75, quai d Orsay 50 Salisbury Road Via Quarenghi Paris cedex 07 PO Box Hounslow Milano FRANCE TW4 6SH - ENGLAND ITALY Tel. : +33 (0) Tel. : Tel. : Fax : +33 (0) Fax : Fax : SEPPIC Inc. SEPPIC China 30, Two Bridges Road, suite 210 Room 510 Jin Tai Building Fairfield, New Jersey South Mao Ming Road USA Shanghai CHINA Tel. : Tel. : +86 (21) Fax : Fax : +86 (21) SEPPIC GmbH ABC Tower Köln Ettore - Bugatti - Str Köln-Porz Germany Tel. : +49 (0) Fax : +49 (0) Subsidiary of the AIR LIQUIDE group C/2144/GB/02/July