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1 RYZA IL & FAT CHEMICAL C., LTD. SAKURA EXTRACT Ver.1.4MK

2 Ver. 1.4MK 1)

3 Ver. 1.4MK Sakura 1-caffeoyl---D-glucopyranoside quercetin 3---D-glucopyranoside 1) :, 4, (1995). 2

4 Ver. 1.4MK 3

5 Ver. 1.4MK 1 AGEsAdvanced Glycation End Products AGEs RCH H 2 NR RCNR () RCCHNHR Pentosidine Fructoselysine. CML 1. AGEs 4

6 Ver. 1.4MK AGEs 2 AGEs 2. AGEs AGEs AGEs 3 3. AGEs 5

7 Ver. 1.4MK AGEs Dyer 2) AGEs AGEs AGEs AGEs CML Pentsidine CML Pentsidine 4. AGEs AGEs AGEs AGEs CML AGEs 2) Dyer D.G. et al. Accumulation of maillard reaction products in skin collagen in diabetes and aging. J. Clin. Invest., 91, (1993). 6

8 Ver. 1.4MK 1-caffeoyl-- -D-glucopyranosidequercetin 3-- -D-glucopyranoside 5 H H H H H H H H H H H 5. 7

9 Ver. 1.4MK AGEs ,4) 100 µg/ml AGEs 10 µg/ml AGEs 1 6 AGEs 2 IC50 1. AGEs AGEs % IC50 3µg/mL µg/ml -14.6± ± ± ±0.7** 42.6±3.2** > ± ±0.1** 19.5±0.3** 25.0±0.3** 30.0±0.4** > ±0.1** -8.6±0.1** -8.9±0.1** -3.7±0.1** 11.6±0.1** > ± ±0.4* -7.8± ± ±0.4** > ±0.2** -2.0± ±0.1** 45.0±0.5** 80.3±0.7** ±0.1* 27.6±0.5** 49.8±0.7** 74.2±1.1** 100.8±0.6** ± ± ±0.3** 50.8±0.4** 91.7±1.7** ± ±0.3** 43.7±0.7** 74.6±0.7** 103.9±3.6** ± ± ±1.7** 67.7±1.6** ± ±0.3** 19.3±0.5** 0.4± ±0.1** ± ± ± ±0.7* 91.8±27.3* ± ± ± ±2.2* 52.7±4.1** ±0.5* 30.4±2.8** 61.5±7.5** 87.5±7.2** 98.2±7.3** ± ±0.1* 38.9±1.4** 96.1±4.3** 32 * 3 Dunnett *: p < 0.05, **: p <

10 Ver. 1.4MK AGEs % ** ** ** ** ** ** µg/ml 3 **p < AGEs 3) Lee E.H. et al. Inhibitory effect of the compounds isolated from Rhus verniciflua on aldose reductase and advanced glycation endproducts. Biol. Pharm. Bull. 31, (2008). 4) Nakamura K. et al. Acid-stable fluorescent advanced glycation end products: Vesperlysines A, B, and C are formed as crosslinked products in the Maillard reaction between lysine or proteins with glucose. Biochem. Biophys. Res. Commun. 232, (1997). 9

11 Ver. 1.4MK AGEs 5,6) 5 AGEs AGEs AGEs Control Non 10 µg/ml AGEs 1 10 µg/ml AGEs AGEs AGEs AGEs AGEs 5) Kueper T. et al., Vimentin is the specific target in skin glycation. J. Biol. Chem. 282, (2007). 6) Cantero A.V. et al., Methylglyoxal induces advanced glycation end product (AGEs) formation and dysfunctionof PDGF receptor-β: implications for diabetic atherosclerosis. FASEB J. 21, (2007). µ µ 7. AGEs 10

12 Ver. 1.4MK 7) (100, 1000 µg/ml) 89 7) Kueper T. et al. Vimentin is the specific target in skin glycation. J. Biol. Chem. 282, (2007). 11

13 Ver. 1.4MK Normal () Control () 100 g/ml () 1000 g/ml () Nomal Control Normal Control Control 100 g/ml 1000 g/ml

14 Ver. 1.4MK AGEs AGEs 1 carboxymethyl lysine (CML)-collagen 8) AGEs % 1µg/mL ± ±4.2* 26.2±0.5* 37.6± ±2.7* 17.2± ± ± ± ± ± ± ± ± ±1.5* 39.0±1.1* 121.5±5.4** -18.9± ± ± ± ± ±4.4* 104.8±34* * 5 CML-collage Dunnett *: p < 0.05, **: p < ) Alikhani Z. et al. Advanced glycation end products enhance expression of pro-apoptotic genes and stimulate fibroblast apoptosis through cytoplasmic and mitochondrial pathways. J. Biol. Chem. 280, (2005). 13

15 Ver. 1.4MK % µg/ml 5 **p < 0.01*p <

16 Ver. 1.4MK B16 B16 3 B16 C 3. B (µg/ml 100± ± ± ± ± ± ± ± ± ± ± ±0.5 * % of Control 15

17 Ver. 1.4MK B16 % µg/ml 11. ± n = 5**: p<

18 Ver. 1.4MK -P P -P 5 3 -P mg/ 30 ml 4. -P 3 -P ph67 ph % 2% 5% 10% 20% 17

19 Ver. 1.4MK -PpH ph -P 0.5%pH P 0.5%pH % 0.2% 18

20 Ver. 1.4MK TA100 TA98 Ames 1%EpiSkin MTT IL-1 ECVAM Risk Phrase 38 1%HCE 1% CLIPA 1%

21 Ver. 1.4MK very good -P 5.0g/100g 43.4g/100g 4.5g/100g 0.0g/100g 1 1.5g/100g 0.0g/100g 2.5g/100g 0.0g/100g 86.5g/100g 56.4g/100g 2 378kcal/100g 138kcal/100g 3 0.0g/100g 0.0g/100g 50mg/100g 0.4mg/100g 0.1g/100g 0.0g/100g (+++) 3 4; 9; ; 2; 3; 2; 2; 2;

22 Ver. 1.4MK -P 5 kg 20 kg -PC 5 kg -LC 5 kg -P-PC -LC 21

23 Ver. 1.4MK -P / //// /// * / //// /// * -PC INCI -LC INCI Dextrin (and) Prunus Lannesiana Flower Extract (and) Ascorbic Acid (and) Malic Acid Water (and) Butylene Glycol (and) Prunus Lannesiana Flower Extract (and) Ascorbic Acid (and) Malic Acid BG 22

24 Ver. 1.4MK Prunus lannesiana 2.0% 0.05% 2.0 % HPLC 0.05 % HPLC 10.0 % 1 g1052 (1) 20 ppm (2) 1 ppm /g /g BGLB 69 % 25 % 3 % 3 % 100 % 23

25 Ver. 1.4MK Prunus lannesiana (1) 20 ppm (2) 1 ppm /g /g BGLB 56.0 % 43.4 % 0.2 % 0.2 % 0.2 % % 24

26 Ver. 1.4MK Prunus lannesiana 2.0% 0.05% 2.0 % HPLC 0.05 % HPLC 10.0 % 1 g1052 (1) 20 ppm 2 (2) 1 ppm /g /g BGLB 69 % 25 % 3 % 3 % 100 % 25

27 Ver. 1.4MK Prunus lannesiana 1,3-BG 30 µl 3.5 ml 0.2 ml 0.4 ml (1) 10 ppm 2 (2) 1 ppm /g /g BGLB % BG % 1.00 % 0.12 % 0.12 % % 26

28 Ver. 1.4MK TEL(0586) () FAX(0586) URL/ F TEL (03) FAX (03) * * * * p.26 LC 20 ppm10 ppm p

29 Ver. 1.4MK 28

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