e}8hs}a] 938 6s. 791~794(1994) Yakhak Hogji Vol. 38, No. 6 UG Hee SHHESO Bees ofo|edt Seb Ata Maas HAS UE DE A ea the eH T, ssa (Received July 26, 1994) Inhibition of Aromatic L-Amino Acid Decarboxylase (AADC) by Some Phenolic Compounds from Medicinal Plants Shi Yong Ryu’, Yong Nam Han and Byung Hoon Han Natural Products Research Institute, Seoul National University, Seoul 110-460 “Korea Research Institute of Chemical Technology, Taejeon 305-606, Korea Abstract Sixteen kinds of naturally occurring phenolic compounds including 5 stilbenes, 7 flavo- noids and 4 anthraquinones were examined in the inhibitory activity against rat liver AADC(aromatic L-amino acid decarboxylase) in vitro, using 5-hydroxytryptophan as a substrate. Three hydroxys- tilbenes , resveratrol 1, rhapontigenin 3 and piceatanol §, which were known to be monoamine oxidase A inhibitors, exhibited a significant inhil ion against AADC(C.=20, 8 and 5 uM, respe- ctively). By the comparison of the activity of each phenolic compound, it was suggested that the 3'4'-dihydroxypheny! group of stilbenes or flavones was the best pharmacophore for the AADC inhibitory activity. Keywords [] AADC, Aromatic amino acid decarboxylase, AADC inhibitor, Rapontigenin, Resveratrol, iceatanol, Stilbene, Flavone, MAO, Monoamine oxidase. WaSoho|eAPEEALRA(AADC: aromatic L- amino acid decarboxylase, EC 4.11.28) 91% DOPA $HRALE-A(DDC:DOPA decarboxylase}. = ¥e}4->j tyrosine, DOPA (dihydroxyphenylala- nine) 3! S-hydroxytryptophan “§ 3 aromatic L- amino acid $2 @"H418}4] tyramine, dopamine serotonin (5-hydroxytryptamine) 3 28h: & 4&4] SHA} epinephrine, norepinephrine, dopamine “| 2} catecholamine % serotonin3} Z SAAAUTEASS) B-dol lel WEAal FBS assets et. Aah SE ©) RANS-S Se]spi) AesHS en- zyme inhibitore& Ao 4+e| ato}eaaat She Fal Bo] VALS B09S2] YRS YL o} 2] MeOHFSES ARR sho} APY Aaa ARIS Ws} B Ast SABRheum undula- +e Sa BU EE 9) AS 791 tum, Polygonaceae), 4:%\(Perilla frutescens, Labia- tae), 9141 2(Artemisia capillaris, Compositae), 7+ (Chrisanthemum indicum, Compositae), &22(Po- Iygonum cuspidatum, Polygonaceae) -§°] 72833} BAUNSS AHL VSS UF UME” HE AAp % ARZHE KLANHEVS Be] staat B2MMHAGS ARE sho] activity-guided isola- tionahdel oe} Rh avsS Yeh Bde & FAG dat SHA BY BAS OEPHe resvera- trol 1, rhapontigenin 3° hydroxystilbene™) #}4} Be BVT ES AS TeYA oh eh BAS °}@ hydroxystilbene4] 3}4}243-2 4st] flavo- noid 8 anthraquinone‘+ -§ 7}% phenolic com- pound} 4)2hy AADC) a} K>4¢]) d}s}e} wa a}sta} Qe}. obSe} ol SHALES ole] awe Bag BH *13]2] monoamine oxidase-A(MAO- Ajo Habe]sS BAU apes Hea Wi 9baoa AALS: DEY VE 2} HESS] AADC*aS}S} MAO-AA ALPS AB HBAS asic. alee AADC(Aromatic L-amino acid decarboxylase) BS AsO] SB—Umezawa F2] willl ae} ¥12]2] liver homogenate # Ae]8}o] Aa 2S AME S AS 150~250 g2} SDA $d MAS BEA Be} UWS) Ae ASA} 001M phosphate buffered saline(PBS, pH 7.0)2.2 AAI F FR 1 g's 0.003M mercaptoethanol 2 mi& 7}3}0) 2¥-2b waring blender(Omni mixer 17105, Dupont instrument) F¥8}34¢}. °1 44] ACHAAL 700 g& 20EAE MIRAR HABEAS A ste} 18,000 ge ala} 20321 MAlBeystseh. ARS AS vjelat pellets PBS 5 m/e} Be}]y —15c freezero] FAMWstT £4] VLA} defrosta}e} Bade et. HAG 05 mi, 24 10m! Y VAY 05 m/s 2] Bee Yat 375 YEA] 154221 incubations} Xt F 4mM 5-hydroxytryptophan 0.5 mi 7}8} 2 AF}e} 90%2} incubation} ach. BEAL py- ridoxal phosphate(cofactor, Sigma Co.)$} iproniazid phosphate(MAO inhibitor, Sigma.Co)# 42} ¥-8-4 FA 2] “F=7} 0.1 mM, 10 mM} B+ gS ¥] 2] 0.1M tris-HCL buffer(pH 85)9)) Fo) Aaa & AST VSS OST STOMA SHAE BS HES SUIANZ FES] AUF 700 ge S¥2t UAlze] tT BSA 20 mS Ashe} ve] Ev} Amberite CG-50(H type) column(0.6%4.0 cm)el] 4-0} 91}. 40 mi 0]4t2] 44#+-S. Amberite resin& 3248] 4 AIG FAN acetic acid 3m 71514 SSAS BS alate} 277 nmel}7] F(A; absorbance) S 47] ataeh. A4yLsample) °] so] Wes Qeyaal | 2) SHS 7H AAT control), W741 Hal WSEdGl4] 7/4444 (5-hydroxytryptophan)-& +2 SA) 8 Tblank, zero time) ¥ 7] Al $2] buffer solutions YS JYRVE(sample control) & qa aleastsou} Ze 2 duplicate = AM] BAAS Mlsigder A2VHA% Inhibition) ® he Ale oe} Aleeh sich. ar (Aste Aen cate) AenesrBume 10? AASHS ASHES A] dimethylsulfo- xide(DMSO)l 1 SHAS sooo, WS SoA] DMSOSE7} 05% ARS Aah ee SS sic. CAS Ne doles] Baal $e EqE ao] Washes) 50% BAA SE Cu) & Abeebsic}. Hes Seo) eel- Veo] see 4S AR SHES 212} 24 Hresveratrol 1 and piceid 2), a) AX(chapontigenin 3, rhaponticin 4, chrysopha- nol 13, physcion 14, emodin 18 and emodin-glu- coside 16), 2}Mluteolin 8, apigenin 9, acacetin 11 and diosmetin 12) = 2}8Kquercetin 6, rutin 7 and kaempferol 10) S2=44] Bey? $2 Eelsl 3+ HE)S ©1214} Y-U(piceatanol 5A” HFSS Abeebach % Inhibition= weaa wy ae WF opel GEE A(AADC) A} RE ATG] BU RAMAAMAES ALS Ae} activity-gui- ded isolation Yo] Fe} Gat Sl SARTO Spe} eI 2S] BAARH I and 3S ERE SF] sti Ibeneal SHES #1Z84) Oo) SHES HARE FRPAE 71D UE 744} flavonoid B 4¥2) anthraquinone™) 3}4+3-S-<4 o}é}e} 242} 4) 43h u) AADC AVIS Z4}8+4] UskeHTable 1. o] & stilbeneal 3HHEG] alee $pet KAAMITS Ue 2>) A719 SH QA SAA we} BYES] ao] wolssich. & sti- Ibene#742] 3-hydroxyl7|-Rio glucose7} 233 piceid 2 4 rhaponticin 49] 2-4 Bde] 712} 241S|91.204, hydroxyl7]7} 47H 194%) piceatanol 5 3! 37H] hydroxyl7|s} 1712) methoxyA]87] 71a rhapontigenin 3¢ 242} hydroxyl7IRt 374 4] ‘We resveratrol 16]] 3] s}o] Fab WJo] Babs} ae}. Fs] Ag] 3541 92], BR] 347A All 2b hydroxyl717} 219H2 piceatanol 5&7 $4°@b AZ VoIFIch. Flavonoidal SHEE stilbene SHHESo] vate] topo e eet ays yehy B27} $8] ©] SARS] 5,703} BH] 34" $A)ol] J. Pharm. Soc. KoreaAa sd SHEE S| Wes} opel ah etal Mae 793 ‘Table 1—Enzyme inhibitory activity of some naturally occurring phenolic compounds against AADC(aromatic L- amino acid decarboxylase) and MAO-A(monoamine oxidase-A) in vitro Ry ( ee Ry Ry wot OR ou 8 @) ©) Ry wy on LU J on Ry ay % () @ Compound RI R2 RS. ICM)" = ‘AADC MAO-A 1 H -H 20 2 2 -Glu -H >1000 35 stilbene (a) 3.0 -H CH, 8 4 4 Glu CHS >1000 >1000 5-H -H 5 15 6-H —On 80 100 7 -H -OH 800 >1000 8 H -OH 80 100 flavonoid (6) 9 H -H >1000 10 1-H -H NT 10 un CH = -H >1000 >1000 eee 12 —CHs —OH 1000. 1000 B H —H >1000 >1000 4-H —OCH, >1000 >1000 anthraquinone (-) 1S —H -OH 200 50 16 Glu -0n 400 1000 —isoflavonoid (@"* 17 0.03, “ICiy Value of each compound was determined as a concentration(uM) that caused 50% inhi ponding enzyme activity in vitro. ** The ICy, value of orobol against AADC was measured by Umezawa et al 22} hydroxyl7|7} 9]@241 quercetin 63} luteolin 86] vin @Eat AR Meltste} = anthraqui- none” SHAEES AGS} 6CRI.8CR)Y $12] hy droxyl7|7} 1,3-diol type®= 219491 emodin 15 8 =Lwhedal onto) oF et Ags EAL WS HE SHARES 719) Qa eh] Zatateh. Se] WAS UeAHS SHAERG, 3, 5, 6, 8, 15 and 16)& 74} 2 7]2EAlab,c)2] Yehoate} a AES 20] WolFaL YE s}-yRt Agel] 272) Vol. 38, No. 6, 1994 hydroxyl7]7} 1,3-diol AYU we} 2)R=I°} 31 SL Bato) 2742} hydroxyl7]7} 22} 3.4 $A) e4 1, 2diolQ@AZ APN SAG, 6 and 8) 7 + tt BAS dehisteh. U4]7] Umezawa 2 + $9 VE MF LS He AADCAMAS Get We WAALS S Tel s}e] Bw shahied) ole] EPR B-YYLS-S Vp isoflavonoida] BHF) do 2S 7h AAU BAS MIE HS AR] 5,741 lol 42} hydroxyl7|7} 1,3-diol Belbd SAS: Hod: DBE = ATs} 312 BH] 34" 42)9)) hydroxyl7|7+ L2-diol JAZ AYN WE orobol 17¢]g12.%4, orobol 172] Bt} 3'eel] 2]RtS1 hydroxyl7|7t °) ‘WA genistein (,7,4,-trihydroxyisoflavone)®] 73-7 Bo] VASA ALTCo=200 uM) BRS + EHUSICH ols} Ze ES) FM APE oO] | Umezawa $0] #8} ¥}H = isoflavonoid”) 3 HES Nola BA he} EATS Sahe] peal fs} ze] flavonoid stilbene”) 8}9He-E°] AADCA ARTS Hebe] NAS We WS AR UG FF leh. Ua ARM] VI vps 0} stilbene flavonoid] S4HES= 2419) sol) Eas} MAO- A(monoamine oxidase-A)o] Hate] ja Az}S YehH st sl2c} 2} SHEE) MAO-A} SEES Table lol HAIL Us eh. elas of 3-H Bol AADCAA M2] Aol) Bo} Fahel uf she hie ds) Fea API Basa. , stilbeneal SHES] 44 Bats] 4°e ated t MS} hydroxyl7]XE ALR resveratrol 1] 2719) hydroxyl7|7} 442} 3'4°X Alel| 1.2-diol Gaz >| 243] rhapontigenin 3 ¥ piceatanol $Y} 74 9 ALS UEWZ Web. flavonoid SHAKES] 2 +1 apigenin 9 5 kaempferol 10°] quercetin 6 ‘3 luteolin BUC} FR RYS Mela glo] stil- bene SHES] AF5} oa yl9} Fas + BWV} ASNT USS YF ach BAP ALRSHHES AMEE 26 phenolic compound] ols} ze) 2 7s SoA ee AE PR 7} als] Bol) date] AE ChE PRAY Aol Be} AHASS UES Se Fa BoLEe Ab Alelmy ole} abe dalsk: ALP o]S AADC 3! MAO-AK29] XE7|41 S29 Atel Sele BYS & + eee} ARach HAS] Be 9) Ae UHI] AP]7} eo s}gt A stole} oleh Egy 2 wu 1) Han, Y. N, Cheong, M. S, Ryu, S. ¥., Kim, T. H. and Han, B, H.: Studies on the inhibitory compo- nents against aromatic L-amino acid decarboxylase. Abstracts of the 17th annual convention of Korean Society of Pharmacognosy. p.56. (1986). 2 Ryu, S. Y,, Han, Y..N. and Han, B. H.: Monoamine Oxidase-A inhibitors from medicinal plants. Arch Pharm. Res, U1, 230 (1988). 3) Lovenberg, W., Weisbach, H. and Udenfriend. S. ‘Aromatic L-Amino Acid Decarboxylase. J Biol Chem, 237, 89 (1962). 4) Umezawa, H., Tobe, H., Shibamoto, N., Nakamura, F,, Nakamura, K,, Matsuzaki, M. and Takeuchi, T.: Isolation of isoflavones inhibiting DOPA decarbo- xylase from Fungi and Streptomyces. J: Antibiotic, 18, 947 (1975). 5) Han, Y. N,, Noh, D. B. and Han, D. S.: Studies on the Monoamine oxidase inhibitors. from medicinal plants 1, Isolation of MAO-B inhibitors from Ch- rysanthemum indicum. Arch, Pharm. Res, 10, 132 (1987). 6) Bachelor, F. W., Loman, A. A. and Snowdon, L. R.: Synthesis of pinosylvin and related heartwood sti- Ibenes. Canadian J. Chem, 48, 554 (1970). 7) Drewes, S. E. and Fletcher, I. P.: Polyhydroxysti- Ibenes from heartwood of Schotia brachypetala. J. Chem. Soc. Perkin 1. 961 (1974). 8) Baker, D, Downing, D. F,, Floyd, A. J, Gilbert, B, Ollis, W. D. and Russel, R. C.: Synthesis of isof- Javones. V. Irigenin and tectorigenin. J. Chem. Soe C. Organic Chemistry, 1219 (1970). 9) Tobe, H., Naganawa, H,, Takita, T., Takeuchi, T. and Umezawa, H.: Structure of a new isoflavone from mmyces inhibiting DOPA decarbo- 5, 29, 623 (1976) J. Pharm, Soc. Korea