Lipout. Burns stored fat and achieves centimetric reduction BODY SCULPTURE

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April 14

Burns stored fat and achieves centimetric reduction

» The modern lifestyle Stress Sedentism Unbalanced diet EXCESS FAT + CELLULITE

» How to remove excess fat and cellulite? The fatty tissue is responsible for: 1. Absorption and accumulation of fat = body energy reservoir 2. Regulation of body metabolism and homeostasis 3. Body surface modeling It is also a source of cellulite, a condition affecting more than 85% of women. is an active that activates fat-burning cells in adipose tissue to reduce excess fat and thus refine the silhouette

» Types of adipocytes Adipose tissue is predominantly formed by adipocytes. Recent studies have demonstrated that there are 3 types of adipocytes: 1. White Single fat droplet Few mitochondria Fat-storing 2. Brown Multiple fat droplets Many mitochondria Fat-burning (thermogenesis) 3a. Active beige Brown-like Fat-burning (thermogenesis) 3b. Inactive beige White-like Fat-storing Recently discovered!

» Thermogenesis: a fat-burning process UCP1, the essential element for thermogenesis: Marker gene of beige and brown adipocytes. Indispensable for lipid burning. Thermogenesis consists in: 1. Stimulus that triggers the degradation of triglycerides and the intracellular release of fatty acids. 2. Activation of UCP1 by free fatty acids. 3. Stimulation of mitochondrial fatty acid oxidation. 4. Activation of thermogenesis = fat-burning. Cellular thermogenesis and respiration processes in beige and/or brown adipocytes

» Mechanism of action of acts on beige and white adipocytes: 1. Induces the expression of UCP1 in adipocytes to activate their browning (active state). 2. Increases β-oxidation of fatty acids thanks to the activation of thermogenesis, literally burning the accumulated fat. activates browning and thermogenesis processes for body fat reducing and cellulite erasing effects

», slimming and anti-cellulite marine active is obtained from an extract of Tisochrysis lutea: Unicellular microalgae (Haptophyta). Standardized in xanthophylls. Rich in polyunsaturated fatty acids or PUFA. Cultivated in closed photobioreactors with a low environmental impact. Renewable sources of active elements.

» In vitro efficacy: protocol To prove that activates beige adipocytes, browning of white adipoctyes and thermogenesis, we evaluated: 1. Expression of key genes in the browning and thermogenesis process. 2. Expression of thermogenic proteins, to confirm the previous step. 3. Biogenesis of new mitochondria. 4. Quantification of β-oxidation, to confirm the functionality of the induced changes. The study simulated both possible scenarios: 1. Cultures of preadipocytes from human subcutaneous adipose tissue, to evaluate the differentiation of beige preadipocytes to mature and active beige adipocytes. 2. Culture of human subcutaneous mature a priori white adipocytes to observe their conversion into active beige adipocytes.

Variation of mrna relative quantity (%) Variation of mrna relative quantity (%)» In vitro efficacy: expression of key genes in thermogenesis process Evaluation of the expression of key genes, at the highest concentration (0.56%) vs. control, increases the expression of UCP1, key gen on browning and thermogenesis. UCP1 mrna in preadipocytes UCP1 mrna in mature adipocytes 2500 2000 1500 1000 500 0 2094% 592% 170% 0.14 0.28 0.56 concentration (%) 600 500 400 300 200 100 0 537% 237% 91% 0.14 0.28 0.56 concentration (%) activates the genetic program of the inactive beige adipocytes in the a priori white and beige adipocytes

WITHOUT TREATMENT» In vitro efficacy: expression of key proteins in thermogenesis process Evaluation of the expression of key proteins increases UCP1 expression, by 50% in preadipocytes and by 378% in mature adipocytes. PREADIPOCYTES MATURE ADIPOCYTES Microscope images of UCP1 expression in preadipocyte cultures (left) and mature adipocytes (right), without treatment (upper) and treated with (lower) (UCP1 is clearly visible as yellowish-red spots)

WITHOUT TREATMENT» In vitro efficacy: biogenesis of new mitochondria Evaluation of new mitochondria biogenesis enhances VDAC protein expression, by 22% in preadipocytes and by 139% in mature adipocytes significant increase in the number of active mitochondria in the cells. PREADIPOCYTES MATURE ADIPOCYTES Microscope images of active mitochondria in the subcutaneous preadipocyte cultures (left) and mature adipocytes (right) without treatment (upper) and treated with (lower). The active mitochondria are stained with a specific marker, MitoTracker Red. The blue spots visible in the upper panel are cell nuclei. The active mitochondria are visible in the lower panel as bright yellow spots.

Variation in CCPM/µg protein (%) Variation in CCPM/µg protein (%)» In vitro efficacy: increase in fatty acid oxidation Evaluation of β-oxidation, at the highest concentration (0.56%) vs. control, increases β-oxidation by 118% in preadipocytes and by 173% in mature adipocytes. 160 140 120 100 80 60 40 20 0 β-oxidation in preadipocytes 150% 118% 62% 0.14 0.28 0.56 concentration (%) β-oxidation in mature adipocytes 200 150 100 50 0 173% 114% 40% 0.14 0.28 0.56 concentration (%) converts lipid-storing adipocytes into lipid-burning beige adipocytes

» In vivo efficacy: protocol Slimming and anti-cellulite efficacy of was evaluated: 61 volunteers, men and women, between 18 and 60 years of age, all with a body mass index (BMI) above 23 for women and above 25 for men.. Creamy gel with 3% was compared with the same formulation without an active ingredient (placebo). 21 women and 10 men applied the placebo and 20 women and 10 men applied the formulation with twice a day for 2 months. Women applied the product to their abdomen, hips and thighs in order to evaluate fat and cellulite reduction; men applied the product to their abdomen only in order to evaluate fat reduction.

Centimeters Centímeters» In vivo efficacy: reduction of fatty layer Measurements in centimeters reduces up to 5.7 cm thighs perimeter and up to 5.5 cm hips perimeter. 0,0 Average thigh perimeter D28 0,0 Average hips perimeter D28-0,5-0,5-1,0-0.9-1,0-0.8-1,5-2,0-2,5-3,0 1.0* -1.9 1.4* -1.3-2.7-1,5-2,0-2,5-3,0 0.7-1.5 1.3* -1.2-2.5 Placebo (*) statistically significant

Centimeters Centimeters» In vivo efficacy: reduction of fatty layer Measurements in centimeters reduces up to 5.0 cm abdomen perimeter in women and up to 10.4 cm in men. Average abdomen perimeter (women) D28 0,0 Average abdomen perimeter (men) D28 0,0-0,5-1,0-1,5-2,0-2,5-3,0 1.4* -0.6-2.0 2.2* -0.7-2.9-0,5-1,0-1,5-2,0-2,5-3,0 1.2-0.6-1.8 1.7-0.9-2.6 Placebo (*) statistically significant

» In vivo efficacy: reduction of fatty layer Standardized photos of the treated body areas D0 D0 D0 D28 D28 D28

» In vivo efficacy: reduction of fatty layer Thickness of the subcutaneous fat layer Ultrasonography images of the subcutaneous adipose tissue of the left hip of one of the volunteers D0 D28 significantly reduces body contours in both men and women by decreasing the thickness of subcutaneous adipose tissue

» In vivo efficacy: reduction of cellulite Reduction in the length of the dermis-hypodermis junction significantly reduced the length of the dermis-hypodermis junction by 21.7% compared with the placebo. significantly diminishes the length of dermis-hypodermis junction, visibly reducing cellulite

Percentage variation (%) Percentage variation (%)» In vivo efficacy: reduction of cellulite Skin roughness (female buttocks area) Sa: arithmetic average of peaks height and troughs depth present on skin surface. Sz: average of the 5 largest peaks and the 5 deepest troughs in the skin area. 0 Roughness (Sa) D28 0 Roughness (Sz) D28-5 -5-10 -10-15 -20-25 -30 5.7-18.8% 10.7* -13.1% -24.5% -23.8% -15-20 -25-30 -35 1.9-25.4% -27.2% 13.9* -18.0% -31.8% Placebo (*) statistically significant

» In vivo efficacy: reduction of cellulite Skin roughness (female buttocks area) D0 Images of skin roughness obtained using PRIMOS on D0, D28 and, using a color scale (red = the highest regions or peaks; blue = the lowest regions or troughs, green = the regions with a neutral level) D28 significantly smoothes out skin roughness and improves skin appearance

R7 variation (%) R7 variation (%)» In vivo: improvements in the skin s biomechanical properties Cutaneous elasticity (R7) increased the elasticity of the skin, both in the upper and deeper layers of the skin for all the body areas evaluated. 30 25 20 15 10 5 0-5 12.6* Elasticity in upper layers 24.0% 13.0% Elasticity in deeper layers 24.5% 18.9% 24.8* 10.9* 19.3* 10 8.0% 5.2% 5 0.4% 0 D28-0.8% D28 30 25 20 15 Placebo (*) statistically significant

F4 variation (%) F4 variation (%)» In vivo: improvements in the skin s biomechanical properties Cutaneous firmness (F4) 50 40 30 20 10 increased the firmness of the skin, both in the upper and deeper layers of the skin for all the body areas evaluated. 0 Firmness in upper layers 16.1* 31.7% 25.5* 15.5% D28 Placebo 17,7% 43,2% (*) statistically significant 30 25 20 15 10 5 0-5 -10 11.7* Firmness in deeper layers 5.6% 17.3% D28 28.9* -4.8% 24.1% improves the properties of the skin, blurring the appearance of cellulite

Temperature variation (ºC)» In vivo efficacy: activation of thermogenesis in subcutaneous adipose tissue Skin temperature Skin temperature 1,4 1,2 1,0 0,8 0,6 0,4 0.9* 0.3 1.2 1.3* 1.1 Placebo 0,2 0,0-0,2-0,4 Women -0.2 Men (*) statistically significant significantly increases the temperature of the skin, since it converts fat storing adipose tissue into thermogenic adipose tissue that actively burns fat

» In vivo efficacy: activation of thermogenesis in subcutaneous adipose tissue Skin temperature (thermographic images ) D0 D0 D0 35ºC 26ºC

» In vivo: subjective evaluation of the product s efficacy Subjective evaluation of vs. placebo Purchasing intention * Thigh perimeter reduction 100% 80% 60% 40% 20% 0% * Elasticity improvement Placebo Global evaluation Firmness improvement (*) statistically significant * Cellulite reduction

» Conclusions induces the browning of a priori white and inactive beige adipocytes, and stimulates thermogenesis, thus: Burns stored fat. Reshapes silhouette. Reduces body volume. Erases cellulite. Improves skin elasticity and firmness. activates the processes necessary to stimulate fat-burning, both in men and women, with slimming and anti-cellulite effects

» Cosmetic applications Slimming and body-shaping products. Prevention and treatment of cellulite. Complementary ingredient in body care products.

» Technical information INCI name: Caprylic / Capric Triglyceride, Plankton Extract, Tocopherol. Transparent liquid. Greenish brown color. Soluble in oil. Recommended dose: 1 3%. China approved.

PROVITAL. S.A. Pol. Ind. Can Salvatella Gorgs Lladó, 200 08210 Barberà del Vallès Barcelona (España) Tel. (+34) 93 719 23 50 info@provitalgroup.com www.provitalgroup.com