Statistical Data of Dementia.

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1 Screening of Prolyl Endopeptidase Inhibitors from Medicinal Plants. Lee Si-young, Lee Jung-han, Paik Young-Sook College of Environment and Applied Chemistry, Kyung Hee University.

2 Statistical Data of Dementia.

3 Statistical Data of Dementia.

4 Extraction procedure. Sample MeOH Extract(3L/8hr/3times). Filteration & Evaporation. CHCl 3 layer O layer 90% MeOH layer n-hexane layer Ethyl Acetate layer O layer layer O layer Each layer Enzyme Assay. Discovered PEP activation layer.

5 Solvent Concentration. Absorbance(380nm) U PEP cont MeOH solvent effect 2.0 1ppm 2.5ppm 1.8 4ppm ppm ppm 16.6ppm Time(Sec)

6 Enzyme Assay. A. 100mM Phosphate buffer (ph 7.0 / 320/ 37, 30min UV cuvette) 2mM Z-Gly-Pro-pNA 32 Sample U PEP 32(380nm) B. 100mM Phosphate buffer(ph 7.0 / 384) Sample 16

7 Enzyme Assay. Control 100mM Phosphate buffer (ph 7.0 / 336/ 37, 30min UV cuvette) 2mM Z-Gly-Pro-pNA U PEP 32(380nm) Positive Control 100mM Phosphate buffer (ph 7.0 / 320/ 37, 30min UV cuvette) 2mM Z-Gly-Pro-pNA 32 Z -Pro-Prolinal U PEP 32(380nm) Inhibition(%) = A 380 of Control - (A B) A 380 of Control 100

8 Positive Control Positive control Absorbance ppb Z-Pro-Prolinal W avelength

9 0.1 U PEP(pH 7.0 / 37):10 min Interval nm 380nm 1.5 tim e in te rva l 1 0 m in /3 h r Absorbance W avelength

10 PEP control Absorbance(380nm) U PEP Control 10min/380nm Min

11 Absorbance O layer 4ppm CHCl 3 EA PEP Absorbance O layer 4ppm MeOH Hexane EA PEP 0.1

12 Absorbacne CHCl 3 layer 4ppm O EA PEP Absorbance O layer 4ppm EA Hexane MeOH PEP 0.1 second

13 Absorbacne 0.6 O layer 4ppm PEP 0.5 MeOH Hexane EA Absorbance 0.35 PEP Hexane layer 4ppm 0.30 MeOH EA 5 O

14 Absorbance EA layer 4ppm MeOH Hexane O PEP Absorbance 0.30 PEP Hexane layer 4ppm 5 MeOH EA 0 O

15 Absorbance 1.0 4ppm PEP CHCL 3 4ppm 0.8 EA 4ppm 0.6 Absorbance u pep cont MeOH layer 4ppm EA Hexane O PEP

16 Absorbacne O layer 4ppm MeOH Hexane EA PEP Absorbacne O layer 4ppm EA MeOH Hexane

17 Absorbance 0.6 PEP EA layer 4ppm 0.5 O Hexane MeOH 0.3 Absorbacne Hexane layer 4ppm MeOH O EA PEP 0.1

18 Absorbance PEP O layr 4ppm EA MeOH Hexane Absorbacne Hexane Layer 4ppm MeOH EA O PEP 5 0

19 Absorbance 0.7 PEP Hexane layer 4ppm 0.6 MeOH O 0.5 EA 0.3 Absorbance 0.35 PEP Hexane layer 4ppm 0.30 MeOH EA 5 O

20 Absorbacne(380nm) O Layer 4ppm EA MeOH Hexane Ginkgo leaves extracts 4ppm Time(Sec)

21 Percent inhibition of Compound 1.

22 PEP percent Inhibition.

23 Isolation procedure. n-hexane layer Silica gel Column Chromatography. Fr 1 Fr 2 Fr 3 Fr 4 Fr 5 Fr 6 Sephadex LH-20 C.C. Fr 5-a Fr 5-b Fr 5-c Prep TLC. Compound1

24 Compound 1 from Ginkgo biloba leaves Absorbance(380nm) ppm 3ppm 4ppm Compound 1 from Ginkgo L 0.1 Rf = 0.5 Time(Sec)

25 UV / IR Data Compound 1 in MeOH/ 1.5 ppm A max 0.79/301nm Absorbacne Wavelength

26 NMR Data ( 1 H, 13 C)

27 NMR Data (DEPT, COSY)

28 NMR Data (HMQC, HMBC)

29 EI + / HR-mass.

30 Determination of Chemical Structure. HO COOH Anacardic acid.

한약재품질표준화연구사업단 강활 ( 羌活 ) Osterici seu Notopterygii Radix et Rhizoma 생약연구과

한약재품질표준화연구사업단 강활 ( 羌活 ) Osterici seu Notopterygii Radix et Rhizoma 생약연구과 - 2 - - 3 - KP 11 Osterici seu Notopterygii Radix et Rhizoma Ostericum koreanum Maximowicz, Notopterygium incisum Ting, Notopterygium