ORYZA CERAMIDE

1. ORYZA CERAMIDE All Natural - 100% Rice Derived Ceramide Excellent Skin Rejuvenator オリザ油化株式会社

2. Rice (Oryza Sativa) • Rice (Oryza sativa) - widely grown and consumed grain in the Southeast Asia - rich in unique bioactive compounds - glycosphingolipids were extracted from rice bran & rice germ • Rice Ceramide: - brown extract from rice bran or rice germ - loaded with glycosphingolipids - similar to animal glycosphingolipids - plays an important roles in maintaining barrier function of skin

3. H CO H CO 3 3 HO C CHCOOC H HO C CHCOOH 30 49 H H R R 1 1 HO HO CH CH CH CH CH CH CH CH 3 3 3 3 3 3 3 3 R R CH O CH O 2 3 2 3 R R 3 3 CH OH OH 2 O O CH CH CH CH CH (CH ) CH CH ( CH ) CH 2 2 2 2 8 3 NH OH HO C R OH Fat ty acid O HO OH P O OH OH O OH O O HO HO OH P O O P OH HO OH HO OH HO O P O OH P OH HO O O O OH O P OH CH (CH ) CH OH NH CH CH CH COOH 3 2 26 2 2 2 2 2 CH 3 H C 3 CH(CH CH CH CH) CH CH CH CH (CHCH CH CH ) CH 2 2 2 2 2 2 2 2 2 2 2 2 H C 3 CH 3 CH CH 3 3 Rice (Oryza Sativa Linne) Physiological Compounds 2.γ- oryzanol 3.Ferulic Acid 1. Glycosphingolipid 4.　Tocotrienols 5.　Tocopherol 6.　Inositol 7. Phytosterol 8. Phytic Acid 9. Octacosanol 10. γ-amino butyric acid 11.　Squalane

4. Req. for Ceramide Content 1mg Ceramide 1kg Brown rice 0.0001% Rice Bran / Rice Germ 100g 0.001% Oryza 34mg 3.0% Ceramide-P Sources of Ceramide Grains Brown rice Bran Germ

5. ス フ ィ ン ゴ イ ド 塩 基 X C H - ( C H ) - C H = C H - C H C H - C H O 3 2 n 2 O H N H C R 脂 肪 酸 O X Name - H - H P O 2 4 + - H P O - O - C H - C H - N ( C H ) 2 2 2 3 2 - H P O - O - C H - C H - N H 2 2 2 2 - G l c o r - G a l N e u N A c α 2 → 3 G a l β 1 → 4 G l c β 1 - Variety of Sphingolipids Sphingoid base Fatty Acids Ceramide Sphingosine-1-phosphate Sphingolipid Sphingomyelin Cerebroside Glycosphingolipid Ganglioside

6. Structure of Skin & Biosynthesis of Ceramide Main component Stratum corneum Stratum granulosum Main component Langerhans cells Stratum spinosum Merkel’s cells Stratum basale Melanocytes

7. Structures of Various Ceramides

8. Stratum Corneum - Lipid Composition Ceramide Cerebroside Other Constituents 26% 55% Cholesterol ester 10% Free Fatty Acids 20%

9. Healthy skin with increasing age, n=34 Non-lesion skin, n=35 Atopic dermatitis skin Lesion skin, n=32 Reduction of ceramide (%) Based on µg/mg stratum corneum Ceramide/stratum corneum (µg/mg) Total Lipids Ceramide 1 Ceramide 2 Ceramide 3 Ceramide 6 Age Total Ceramide Ceramide 4&5 Changes in skin ceramide content with increasing age Reduction of ceramide content in patients with atopic dermatitis

10. Glycosphingolipids – HPLC spectra 96% Oryza Glycolipids (Lot No. 316) Time (minutes) Column: TSK GEL Si-60 + Develosil Si-30 Carrier: (C:M:W = 99:11:1), Flow: 3ml / min Qty of sample: 10mg/50ml Detector: ELSD (tube temp.:40°C, gas flow: 1.6L/min)

11. Whitening Effect of Oryza Ceramide • B16 melanoma cells – melanin hyperproducing cells • Oryza Ceramide  inhibited the production of melanoma cells  prevent hyperpigmentation / whitening effect • Whitening effect was compared with kojic acid, ellagic acid, arbutin  Rice Ceramide > Arbutin > Ellagic acid > Ascorbic Acid  Rice Ceramide < Kojic Acid

12. The Effect of Oryza Ceramide on B16 Melanoma Cells: Inhibition of melanin production Oryza Ceramide 5. Oryza Ceramide

13. The effect of Rice Glycoceramide vs. Kojic Acid On Fibroblasts Growth Control Kojic Acid Rice Glycoceramide Amount of Cell (104 cells/well) Control

14. Effect of Glycosphingolipids on Normal Human Dermal Fibroblasts (in vitro) [Comparison] Concentration ○100μg/ml ●300μg/ml Growth rate of dermal fibroblast cells (%) 1. Control 2. Rice derived 3. Konjack derived 4. Corn derived 5. Wheat derived Sample

15. Moisturizing Effect Of Ceramide (in vitro) [Comparison] Moisturising Effect (%) • Oryza Cramide • Konjack Ceramide • Wheat Ceramide • Control

16. Cosmeceutical Effect of Oryza Ceramide [Human Clinical Trials] • Collaboration with Dermatology Division, Medical Department, Osaka University & Medical Science Research Centre • Double blind trial: Subjects: 33 (6 Male, 27 Female, average aged 25.1±7.8) [skin texture: dry to dry xerotic skin] Dosage: Ceramide 40mg/day, Placebo 0mg/day Duration: 6 weeks Area of examination: whole body, left lower eye area, back area of neck, left upper arm inner area Method: Dermatological examination measurement of skin moisture, sebum & acidity level Microscopic illustration of skin surface

17. Effect of Oryza Ceramide (oral) on Dermatological Examination Face General • mean (non-parametric value) • Symptomatic indications: 0 ( no symptom); 1 (mild); 2 (moderate); 3 (severe) • Significant difference from the value of baseline, Wilcoxon test: *p<0.05, **p<0.01

18. The effect of Oryza Ceramide (oral) on skin moisture, sebum & pH Moisture Skin pH

19. Effect of Oryza Ceramide (oral): Visioscan Measurements Kurtosis : correction K, SE sm: suppleness SE r: roughness, SE sc: scaling, SE w: wrinkling Significant difference from that of baseline, Wilcoxon test: *p<0.05

20. Effect of Oryza Ceramide : Clinical Trial Dry scaly skin (+) Moisture (%) Moisturised skin (+) Bef. T’ment 3- week 6-week Bef. T’ment 3- week 6-week Changes of Moisture content on lower left eye area Changes of SE sc value at the back of neck area

21. Effect of Oryza Ceramide : Clinical Trial After Before 3D Microscopic Illustration of skin surface before and after treatment of Oryza Ceramide (oral) on the lower left eye area of a 23-yr old female

22. Latest Whitening Effect Of Rice Glycosphingolipid • Collaborated with University of Hokkaido • Objective: To evaluate the effect of rice derived glycosphingolipid on melanin pigment of melanocytes • Results: Rice Glycosphingolipid & its metabolites a. Inhibition of tyrosinase activity b. Inhibition of melanin pigment

23. Structure of Rice Glycosphingolipid In 2004, Oryza Oil & Fat Chemical Co., Ltd. collaborated with Hokkaido University to conduct research on “The Application of Rice Glycosphingolipids in Functional Food” グルコース Β-結合 Conjugate of -Glucose

24. ml ml ml ml ml ml ml ml Latest Whitening Effect Of Rice Glycosphingolipid Melanin Production Tyrosinase activity Oryza Ceramide  Tyrosinase activity Oryza Ceramide  melanin pigment production Oryza Ceramide  excellent skin rejuvenator with whitening effect

25. Latest Whitening Effect of Metabolites of Rice Glycosphingolipid Tyrosinase activity Melanin Production • Metabolites of Oryza Ceramide: • Tyrosinase activity • Melanin Production of melanocytes • Skin whitening effect

26. The Inhibition of Mast Cells Degranulation in RBL-2H3 mice among different sources of glycosphingolipids on Concentration of sample: 1g/ml Mean±SE N=6

27. The effect of Rice Glycosphingolipids on Skin Itchiness (Induction by Compound 48/80) No of Itchiness (% of control) [0.15%] group [0.3%] group [0.5%] group Experimental Method: 5-week old ICR mice divided into 4 groups were given free access to sample of rice glycosphingolipids. Compound 48/80 was applied to back of neck of mice 3-days later to induce itchiness. Mice were video 30 minutes later to view the reaction from compound 48/80 induction. Frequency of scratch were observed and counted.

28. Stability Heat Stability of Oryza Ceramide pH Stability of Oryza Ceramide Glycosphingolipid (%) Glycosphingolipid (%) Time (min)

29. Functional Applications • Whitening effect • Moisturizing effect • Skin rejuvenating effect • Anti-ageing effect • Anti-inflammatory effect • Anti-radical effect • Effective barrier function 100% Rice Derived – Highly suitable for oral & topical usage

30. ORYZA CERAMIDE