{ARTOCARPUS HETEROPHYLLUS LAMK.) VIET NAM THANH HOC CAY MfT DAI

TAP CHI HOA HQC T. 49(4) 451-454 THANG 8 N A M 2011

THANH

P H A N H O A

HOC

G 6

CAY MfT DAI

{ARTOCARPUS HETEROPHYLLUS LAMK.) COA VIET NAM

P H A N

I - CAC Hgp

C H A T

FLAVONOIT

Nguyen Chi Bao', Vo Thi Phirong', Pham Thi Minh Phu-ong', Nguyen Thi Hoang Anh^

Tran Van Sung^

Khoa Hda hgc, Dgi hgc Suphgm - Dgi hgc Hue

^Vi^n Hda hgc - Vi^n Khoa hgc vd Cdng ngh$ Vi4t Nam Den Toa soan 25-11-2010

Abstract

From the ethyl acetate extract of Artocarpus heterophyllus's wood 4 compounds were isolated. Their structures were elucidated as norartocarpetin, isoartocarpesin, cycloartocarpesin and cyanomaclurin by analysis of MS, NMR spectra and comparison with the published data.

1. M6 D A U 2. THI/C NGHIEM Cay Mit dai co tfn khoa hgc la Artocarpus 2. L Thiet bi va hoa chat heterophyllus, thuge hg Dau tam (Moraeeae) la mgt

eay kha quen thugc vdi nhan dan ta. Ngoai gia tri Phd egng hudng tir hgt nhan NMR dugc ghi trfn lam thuc pham cua qua Mit thi gan nhu tat ca cac bg may Bruker Avanee 500 MHz vdi TMS lam chat ndi phan cua cay duge sir dung trong y hgc dan gian chuan cho ' H va tin hifu dung moi lam chuan cho Vift Nam va mgt so nudc. Vi du d nudc ta go va la '^C-NMR. Phd khoi ESI-MS duge do trfn may Mit dugc dung lam thuoc an than, ehira huyft ap cao Agilent LC-MSD-Trap SL. Sic ky ban mong (TLC) hay chira nhiing trudng hgp co quap bang each uong dugc thue hifn trfn ban mong silica gel Merck 60 nude mai gd Mit len mieng da nhap cd thfm it nudc F254 trang san, thudc hifn la vanilin trong axit [1]. Cac nha khoa hgc Nhgt ban va Indonesia da phat sunfuric dgc. Sac ky cdt six dung silieal gel cd hgt hien ra hogt tfnh dgc vdi cac ddng tf bao ung thu ac 0,063 - 0,200 mm.

tfnh B16 (B16 melanoma cells) ciia djch chiet

metanol go Mft thu hai tgi Indonesia va tir djch chift 2.2. Chiet mau thvc vat va phan lap cac chat nay hg da phan lap dugc 9 flavonoit, trong do co 8

hgp chat isoprenyl flavonoit. Nhom tac gia nay da Mau dugc thu hai tgi Ha Ngi vao thang 3 nam nghien ciiru moi quan hf cau true - hogt tfnh dgc vdi 2010. Go than cay Mft (A. heterophyllus Lamk.) sau cae dong tf bao B16 ciia II hgp chit flavonoid va khi thu hai ve dugc riia sgch, bao nho va phoi kho isoprenyl flavonoit, kft qua chi ra rang cac nhom the trong bdng ram rdi say trong tii sly d 40°C sau dd isoprenyl lam tang rd rft hogt tfnh cua cac hgp chat xay thanh bgt. 6850 g mau bgt go dugc ngam chiet 3 flavonoid, ngoai ra so nhom the isoprenyl va vj trf lin trong hon hgp MeOH:H20 (85:15) d nhift do kft noi Clia ehiing trong moi phan tu eiing la yfu to phong. Sau khi cit logi dung moi dudi ap suit giam, anh hudng dfn hogt tfnh [2]. Trong bai bao trudc djch nudc cdn lgi dugc chiSt lin lugt vdi cac dung day chiing toi da cong bd vifc phan lap va xac djnh moi cd do phan cue tang din n-hexan, etyl axetat, n- clu triie cua 2 isoprenyl flavonoit va 1 tritecpen tir butanol. Cat logi dung moi dudi ap suit giam thu vo than cay Mft [3]. Bai bao nay cdng hd kft qua dugc cac can chiet tuong ung, trong do can chift khao sat djch chift etyl axetat gd Mft thu hai tai Ha etyl axetat cd khoi lugng la 250 g.

ngi. Tir djch chift nay 4 hgp chit da dugc phan Igp, Liy 50 g can ghift etyl axetat phan tach bing sic clu triie ciia chiing dugc xac djnh la mgt flavonoit: ky cgt trfn chat hap phu la silica gel, hf dung moi norartocarpetin (1), mdt isoprenyl flavonoit: rua giai la n-hexan:EtOAc (85:15 0:100, v/v) va isoartocarpesin (2) cimg vdi cycloartocarpesin (3) va EtOAc:MeOH (100:0 50:50, v/v), thu dugc 11 cyanomaclurin (4).. phSn dogn, ky hifu la AHGEI-AHGEII.

451

TCHH, T.49(4), 2011

Phan dogn AHGE7 (1,884 g) thu dugc khi nia giai vdi hf dung moi n-hexan:EtOAc = 50:50. Trong qua trinh eo quay dudi dung mdi, phan dogn nav xuat hifn chit rin mau vang nhgt. Lgc, rira va ket tinh lgi chit rin dd bing EtOAe thu dugc 256,9 mg chit 3 dudi dgng bgt mau vang nhgt, Rf = 0,46 (CH2Cl2:MeOH = 95:5). ESI-MS: 353 [M+H]' ' H - NMR (CD3OD, 500MHz): 5H = 7,77 (IH, d, 8,8Hz,

H-6'); 7,17 (IH, s, H-3); 6,67 (111, d, lOHz, H-l");

6,48 (IH, dd, 2,4 & 8,8Hz, H-5'); 6,44 (IH, d, 2,4Hz, H-3'); 6,42 (IH, s, H-8); 5,71 (IH, d, lOHz, H-2"); 1,47 I6H,s,3"-(CH3)2].

Phan dogn AHGE8 sau khi co quay duoi bdt dung moi thay co ket tiia chat rin mau vang nhgt;

Igc, rira nhieu lan va tien hanh ket tinh Igi trong EtOAc thu dugc 154,9 mg chit 2, Rf = 0,29 (CH2Cl2:MeOH = 95:5). 'H-NMR (CD3OD, 500 MHz): 5H = 7,74 (IH, d, 8,8Hz, H-6'); 7,14 (IH, s, H-3); 6,69 (IH, dd, 16,2 & 7,IHz, H-2"); 6,55 (IH, dd, 16,2 & l,OHz, H-l"); 6,46 (IH, s, H-8); 6,45 (IH, dd, 8,8 & 2,3Hz, H-5'); 6,42 (IH, d, 2,3Hz, H- 3'); 2,42 (IH, m, H-3"); 1,10 [6H, d, 6,7Hz, 3"- (CH3)2].

Phan dogn AHGE9 sau khi tifn hanh sac ky cgt silica gel trong hf CH2Cl2:MeOH = 90:10 thu dugc 3 phan dogn (AHGE9.1-AHGE9.3).

Phan dogn AHGE9.2 kft tinh lgi trong hon hgp CH2Cl2:MeOH thay cd kft tua xuat hifn. Lgc liy phan ket tua nay va tien hanh sac ky cot Sephadex LH-20 nhifu lan vdi dung mdi MeOH thu dugc 50,8mg chat 1 d dgng ran, mau vang nhgt, Rf = 0,32 (CH2Cl2:MeOH = 93:7). ' H - N M R (CD3OD, 500 MHz): 5H = 7,77 (IH, d, 8,8 Hz, H-6'); 7,15 (IH, s, H-3); 6,44 (IH, d, l,8Hz, H-8); 6,43 (IH, d,

Nguyen Hodng Anh vd cgng sy 2,3Hz, H-3'); 6,47 (IH, dd, 2,3 & 8,8Hz, H-5');

6,20 (lH,d, 1,8 Hz, H-6).

s i c ky cgt Sephadex LH-20 phan dogn AHGE9.3, MeOH thu dugc 306,6 mg chit 4 d dgng bgt khong mau, Rf= 0,29 (CH2Cl2:MeOH = 90:10).

'H-NMR (CD3OD, 500 MHz): 5H = 7,12 (IH, d, 8,3Hz, H-11); 6,38 (IH, dd, 8,3 & 2,4Hz, H-10);

6,22 (IH, d, 2,4 Hz, H-8); 5,93 (IH, d, 2,IHz, H-2);

5,78 (IH, d, 2,1 Hz, H-4); 5,27 (IH, dd, 3,0 & 2,0 Hz, H-I3); 5,03 (IH, dd, 3,0 & 2,0 Hz, H-15); 4,22 (IH, t, 3,0 Hz, H-l4). ^C-NMR (CD3OD, 125MHz):

5c = 160,50 (s, C-3); 160,28 (s, C-1); 159,68 (s, C- 9); 155,72 (s, C-7); 155,69 (s, C-5); 132,96 (d, C- 11); 113,94 (s, C-6); 109,68 (d, C-10); 103,65 (d, C- 8); 100,37 (s, C-10); 96,32 (d, C-2); 95,34 (d, C-4);

72,89 (d, C-13); 66,77 (d, C-15); 64,14 (d, C-14).

3. KET QUA VA T H A O LUAN

Pho ' H-NMR cua chat 1 cho thiy phan tii co 6 proton thom, gom cd 3 tin hifu cua vdng thom bi thf 3 lin d 5H 7,77 (d, J = 8,8 Hz); 6,47 (dd, I = 2,3 va 8,8 Hz); 6,21 (d, J = 2,3 Hz); mgt tfn hifu singlet d 5H 7,15 va 2 tfn hifu doublet cd tuong tac meta d 5H 6,43 (d, J = 2,3) va 6,42 (d, J = 2,3). Pho '^C-NMR cho tfn hifu ciia 15 nguyfn tii cacbon, gdm co 1 nhdm cacbonyl d 5c 184,36; 6 cacbon bgc 4 co lifn ket vdi oxy (159,45 165,80) ciing vdi 6 nhdm CH va 2 cacbon bgc 4 cgng hudng trong vimg thom tir 94,84 130,98 ppm. Cac sd lifu pho ciia chat 1 dgc trung cho hgp chat flavon va chiing hoan toan dong nhat vdi sd lifu ciia norartocarpetin da dugc phan lap trudc day tir re Artocarpus heterophyllus [4].

^ " ^ ^ ^ ^ OH

5! 4f OH O

1: Norartocarpetin

3: Cycloartocarpesin

HO. 9

OH O 2: Isoartocarpesin

"-0-^5 4: Cyanomaclurin

Chit 2 cho pic ion phan bi tgi m/z = 353 [M H]' Pho ' H va '^C-NMR cua chit 2 rat tuong tir nhu cua trong phd khdi ESI-MS. Tir so lifu phd khdi va phd 1, nhung trong phan tiir cd chira them mgt nhdm A^- NMR cd thf dua ra cdng thiic phan tu la CzoH.gOe. isopentenyl thf hifn qua cac tfn hifu d 8H 1,11 [6H,

452

TCHH, T.49(4), 2011

d, J = 6,8 Hz, (CH3)2]; 2,44 (m, -CH<, H-3"); 6,55 (dd, 1,0; 16,2 Hz, H-l"); 6,69 (dd, 7,1; 16,2 Hz, H- 2"). Vj trf noi kft cua nhdm A'-isopentenyl dugc xac djnh la d C-6 eua vong A thf hifn qua vifc mat tin hifu H-6 va H-8 xuat hifn dudi dgng mgt tfn hifu singlet d 5H= 6,46 trong phd ' H-NMR va thay vi tin hifu doublet cua C-6 d 9^ ,83 ppm trong chat 1, thi pho '^C-NMR cua chat 2 cho tfn hifu nay dudi dgng singlet d 110,17 ppm. Pho '^C-NMR cho tfn hifu cua 20 nguyfn tir cacbon, trong dd ngoai cac tin hifu cua khung flavon thi nhom A'-isopentenyl dugc the hifn qua 3 tin hifu CH d 5c = 117,4 (C-1"); 142,53 (C-2"); 34,30 (C-3") va 2 nhdm metyl d 5c = 23,21 [C-4" va C-5"]. Cau true ciia chat 2 duge xac djnh la isoartocarpesin nhd so sanh vdi so lifu pho trong tai lifu [5]. Chat nay da dugc phan lap trudc day tir go A. heterophyllus. Mgt s5 hogt tfnh cua

isoartocarpesin da dugc cong bo nhu hogt tfnh iic chf Tyrosinase a nam (mushroom Tyrosinase inhibitory activity) vdi gia trj IC50 < 50 ^M, hifu qua hon axit kojic, mdt chat da bift tir lau la cd hogt tinh ire chf Tyrosinase.

Thdnh phdn hda hgc gd...

Chit 3 cho pie ion phan tir d m/z = 353 [M+H]*

trong pho khdi ESI-MS. Cong thiie phan tii dugc rut ra tir pie ion phan ttr va so lifu pho NMR la C2oH,606. Pho ' H va '^C-NMR cho thiy chat 3 cung CO khung flavonoid nhung trong phan tii eo mat nhom 2,2-dimetylehromen thf hifn qua eae tfn hieu dgc trung eiia 2 proton vinyl d 5H 6,67 (d, J = 10 Hz, H-l"); 5,71 (d, J = 10 Hz, H-2") va 2 nhom metyl singlet d 5H 1,47 (6H, s). Cac tin hifu eon Igi dae trung cho khung flavonoit, bao gom 15 tin hieu carbon vdi mgt nhom cacbonyl; tin hifu singlet d 5H 7,17 CO the gan cho H-3 ciia khung flavonoid; cac tfn hifu cua 2 vong thom: mgt vong the 3 lan [5H 7,77 (IH, d, 8,8Hz); 6,48 (IH, dd, 2,4 & 8,8 Hz); 6,44 (IH, d, 2,4 Hz)] va mgt vong thS 5 lln d 5H (6,42, IH, s). Cau true cua chat 3 dxioc xac djnh la cycloartocarpesin khi so sanh vdi so lifu trong tai lifu [5], chat nay da dugc phan lap trudc day tir go A. heterophyllus.

So lifu pho '•'C-NMR cua cac chat 1 3 va norartocarpetin, isoartocarpesin, cycloartocarpesin theo tai lifu dugc dua trong bang 1.

Bdng 1: So lifu pho C-NMR cua cac chat 1, 2, 3 va norartocarpetin, isoartocarpesin, cycloartocarpesin

C 2 3 4 5 6 7 8 9 10 1' 2' 3' 4' 5'

&

1"

2"

3"

3 "-Me

1 (CD3OD) 163,06 s 109,09 d 184,36 s 160,31 s 99,83 d

165,80 s 94,84 d 163,31s 105,17 s 110,74 d 159,45 s 104,17 d 164,16 s 108,31 d 130,98 d

Norarto-earpetin (CD3OD + axeton-de)

[4]

163,1 109,1 184,2 160,2 99,8

165,6 94,8

163,3 105,2 110,7 159,4 104,2 163,9 106,4 131,0

2 163,74 s 108,25 d 184,46 s 163,74 s 110,17s 163,24 s 94,22 d

157,10 s 104,97 s 110,74 s 160,24 s 104,14 d 163,20s 109,03 d 130,89 d 117,40 d 142,53 d 34,30 d 23,21 q

Isoarto-earpesin (CD3OD)

[5]

164,0 108,4 184,7 163,5 110,3 163,5 94,4

157,3 105,1 110,9 160,5 104,3 162,99 109,2 131,0 117,6 142,7 34,5 23,4

3 (CD3OD) 160,39 s 108,33 d 184,44 s 164,31 s 106,39 s 163,34 s 95,91 d

156,95 s 105,84 s 110,63 s 160,74 s 104,22 d 158,51 s 109,17 d 131,06 d 116,09 d 129,50 d 79,15 s 28,52 q

Cycloarto- carpesin (DMS0)[5]

158,5 s 106,8 d 182,1 s 162,0 s 104,5 s 162,0 s 94,8 d

155,5 s 104,5 s 108,4 s 159,0 s 103,2 d 156,4 s 108,1 d 129,8 d 114,6 d 128,8 d 77,9 s 27,8 q

453

TCHH, T.49(4), 2011

Chit 4 cho pic ion phan tir d m/z = 289 [M+H]*

trong pho khoi ESI-MS. Cong thirc phan tir ciia chat 4 dugc rut ra tir so lifu pho khoi va cgng hudng tir hgt nhan la CisH^Oe. Ph6 ' H-NMR cho thay cac proton cua hai vong thom vdi hf spin ABX: 5H = 7,12 (IH, d, 8,3 Hz); 6,38 (IH, dd, 2,4 & 8,3 Hz);

6,22 (IH, d, 2,4 Hz) va hf spin AB: 5,,= 5,93 (IH, d, 2,2Hz); 5,78 (IH, d, 2,2 Hz). Ngoai ra pho 'H-NMR eon cho lin hifu proton cua 3 nhom metin gin vdi oxy d 5H = 5,27; 5,03 va 4,22. Pho '-^C-NMR chi ra sg CO mat ciia 15 nguyfn tii cacbon trong phan tir, trong do CO 12 cacbon cgng hudng trong viing thom tir 95.34 den 160,5 ppm vdi 5 cacbon bgc bon c6 gin vdi oxy d viing trudng thap. Bfn cgnh do, sg co mgt eua 3 nhom metin eo gan vdi oxy d 5c = 64,14;

66,77 va 72,89 ciing dugc thf hifn d trong phd. Cac so lieu pho phan tich trfn day dac trung cho flavonoit ngung ty (condensed flavonoid) hiem g^p trong thien nhien. So lifu pho ciia chit 4 hoan toan phil hgp vdi so lifu pho ciia chat cyanomaclurin, chat nay da duge phan lap trude day tir A.

heterophyllus [5]. Cac hgp chat tuong tg nhu cyanomaclurin cung duge tim thiy trong loai Peltophorum afrieanum, hg Leguminosae [6]. Hing so tuong tae J ^ H = 3,0 Hz va JMJS = 3,0 Hz chirng to vong C va D CO cau hinh cis [6]. So lifu phd ' H

Nguyen Hodng Anh vd cgng sy va '^C-NMR Clia chat 4 dugc dua trong phin th\rc nghifm.

Ldi cam on: Nhom tdc gid xin ehdn thdnh cdm an Bg Khoa hgc vd Cong ngh$ dd tdi trg cho nghien cuu ndy thong qua De tdi Nghf dinh thu v&i Y, 2008

2010.

T A I L I $ U THAM K H A O

1. VO van Chi. Tir dien cay thu6c Vift Nam. Nxb. Y hpc (1999).

2. Enos Tangke Arung, Keisuke Yoshikawa, Kuniyoshi Shimizu, Ryuichiro Kondo. Fitoterapia, 81, 120 -123 (2010).

3. V6 Thj Phuang, Nguyen Chf Bao, Nguyen Thj Hoang Anh, Trin Van Sung. Thanh phin hoa hpc vo than cSy Mft dai (Artocarpus heterophyllus Lamk.) cua Vift Nam. Tap chf Hoa hpc 2010 (da nhan dang).

4. Chun-Nan Lin, Chai-Ming Lu, Pao-Lin Huang.

Flavonoids from Artocarpus heterophyllus.

Phytochemistry, 39, 1447 1451 (1995).

5. Zong-Ping Zheng, Ka-Wing Cheng, James Tsz-Kin To, Haitao Li, Mingfti Wang. Mol. Nutr. Food Res., 52, 1530- 1538(2008).

6. Magrieta Bam, Daneel Ferreira, Edward V. Brandt.

Phytochemisny, 27, 3704 - 3705 (1988).

Lien he: Nguyen Thj Hoang Anh Vifn Hoa hge,

Vifn Khoa hge va Cong nghf Vift Nam 18 Hoang Quoc Vift, Clu Giiy Ha Ngi Email: hoanganhvhh@gmail.eom.

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