TAP CHI HOA HOC T. 49(6) 765-768 THANG 12NAM201I
CAC TECPENOIT VA PHENOLIC GLUCOSIT TU CAY K H 6 SAM MEM {BRUCEA MOLLIS WALL, ex KURZ)
Mai Hung Thanh Tiing', Bui I luu Tai', Ho Vict Diic', Young Ho Kim", Nguyen Manh Cirlmg' Vien Hda hoc cdc hpp chat thien nhien. Viin Khoa hoc vd Cong nghe Viit Nam
'Khoa Duoc Bql hoc Qudc gia Clmngnam. Daejeon. Hdn Quoc Din Toa sogn 31-10-2011
The isolation of two terpenoids, named la,6p-dihydroxy-4(H)-eudesmenc (I), niloticine (2), and two phenolic glucosides, dimethyl crenatin (3), and Iconuriside A (4) liom the stem and root o! Brucea mollis Wall, ex Ktirz, were reported. Their sUllctures were elucidated on the basis of spectral data, including ID, 2D NMR and MS.
l.DATVANDE
0 nudc ta, chi Brucea co hai loai Slu dau cut chupt (Brucea javanica) va Kho sam mem {Brucea mollis) dupe su dung trong y hpc dan gian 6i tr| sot ret, dau bung, u nhpt...[l]. Cac kit qua nghien cuu cho thay mpt so loai trong chi Brucea co kha nang khang ung thu rat manh, trong do c6 Brucea mollis.
Cac djch chiet tir cay Brucea mollis Co hoat tinh irc che manh cac dong ung thu phoi nguoi (A549), ung thu gan nguoi (Hep-G2) va ung thu vu (MCF-7) [2, 31-
Ddng bio ngirdi H'Mdng d Hoi Binh (Viet Nam) da su dung ciy B. mollis niu lay nudc tri chiy, ghe Id... Hp gpi nd li "Sua tdng la" hay "cay thudc trang". Tir cic ciy thudc chi Brucea, cic nha nghien ciru da tich ra duac nhieu hop chat cd hogt tinh thupc cic Idp chit khic nhau nhu quassinoit, ancaloit, tritecpenoit vi flavonoit. Cho dSn nay loii Brucea mollis cdn chua dupe nghien ciru nhieu ve hoa hpc vi hoat tinh sinh hpc.
CJ bii bio trudc [4], chiing tdi da cdng bd hai isoprenoit. Bii bio niy, trinh biy ket qui phan lip, xac djnh ciu true ciia hai tecpenoit vi hai phenolic glucoside tir than va re ciia ciy.
2. NGUYEN LIEU v A PHLTONG P H A P NGHIEN c u t ;
2.1. Nguyen lieu
Ciy Khd sam mem {Brucea mollis Wall, ex Kurz) dupe thu hii vio thing 3/2009 tgi Hda Binh.
Ten khoa hpc cay do TS. Trin Th6 Bich (Vien Sinh thii va Tai nguyen sinh vat) xac djnh [3]. Bd phgn thu li la, than vi rl cay. Miu ti6u ban ciia cay Khd sam mSm {Brucea molUs Wall, ex Kurz) dupe luu
tru tgi Vipn Sinh thii vi Tii nguyen sinh vat, ky hieu miu Ii VK 2211 (HN). Miu cay dupe thu vg, logi bd phin hdng, phai khd vi xay nhd thinh bdt.
2.2. Thiet bi va hoa chat
Phd khoi ion hda biii dipn tir ESI-MS dugc ghi tren may Agilent 6310 Ion Trap (Vien Hda hpc cic hap chat thien nhien), pho khoi phan giii cao HR- ESl-MS duac do tren may Varian MS (Vien Hda hpc). Phd cdng hudng tir hat nhin duac ghi tren may Bruker Avance 500 MHz (Vipn Hda hpc) vdi TMS li chit chuin ndi.
Sic ky Idp mdng TLC dupe thuc hien tren bin mdng silica gel Merck 60 F254, sac ky cpt vdi chat hap phu li silica gel pha thudng (Merck logi 40-63
^im) hogc pha dio (ODS, YMC (30-50 (im) va sic ky cpt Diaion HP-20.
2.3. Chiet tach va xac dinh cau true
Qui trinh chiet than re cay vi phan dogn bang cac dung mdi w-hexan, CH2CI2, EtOAc dugc md t i d bai bio trudc. Can CH2CI2 sau khi tich phan dogn bing cpt sac ky silica gel pha thudng thu dugc 9 phin dogn 11-19 [4].
Phin dogn 13 tiep tuc dugc tich tren cot silica gel pha thudng vdi hp dung mdi giii hip n- hexan:EtOAc 6/1, thu dugc 8 phan dogn F1-F8.
Phan dogn F6 (300 mg) dugc chgy tich chit tren cgt silica gel pha thudng vdi hp dung mdi nia giai diclometaniEtOAc 30/1 thu dugc chat 1 sach (22 mg). Phan doan F8 (l,lg) dugc tich chat tren cdt YMC vdi hp dung mdi rua giii MeOHmudc 1/1,5 - 2/1 - 4/1, thu dugc chit 2 sgch (100 mg).
Djch nudc cdn lai (D) dugc tach phin doan bang cpt Diaion HP-20 va lin lugt giii hip bing cac he
TCHH, T.49(6), 2011
dung moi MeOH:nudc 1/3 - l/I - 3/1 - 1/0, thu dupe bon phan doan D1-D4. Phan doan Dl (9 g) dirpc tach phan doan tren cpt silica gel pha thuong v6i h?
dung moi giai hSp diclometan:MeOH:nuac 4/1/0,1 thu dupe 12 phan do^n GI-G12. Phan doan Gil (500 mg) dupe chay sSc ky tren cpt silia gel pha thuong voi he dung moi rua giai axeton:diclometan:nudrc 3,5/1/0,1 Ihu dupe chat 3 sach (105 rag).
Opp hai phan doan G8-I-G9 (340 mg), chay tach chat tren cpt silia gel pha thuang vai hp dung moi rua giai a\cton:dicloinctan:nu(>c 6/1/0,1 thu dirpc chat 4 sach (10 mg)
ip,6a-dihydroxy-4(14)-cudcsmcnc (1):
C15H26O2 (M-238), tinh the hinh kim, mSu vang, tan trong CHCI,, Rr = 0,25 (it-hexan:EtOAc 2/1).
Pho 'H-NMR (500 MHz, CDCl,,), 8 (ppm): 5,01 (i, H,-I4), 4,74 (JT, Hb-14), 3,71 (/, J=10,0 Hz, H-6), 3,41 (rfrf,J= ll,5;4,5Hz, H-l), 2,32 ((/{/</, 7 = 13,5;
5,0; 2,5 Hz, H,,-3), 2,07 (dt, .1 = 13,5; 5,5 Hz, I li,-3), 2.24 (ra, H-l I), 1,90 (IH, m. H.-9), 1,84 (m, H.-2), 1,74 (d,J= 10,0 Hz, H-5), 1,53 (2H, m, Hr2, H.-8), 0,95 (d, J = 7,0 Hz, H-12), 0,87 (d, J = 7,0 Hz, H-
13), 0,70 (s. H-15), viing cac tin hieu bi che lap:
1,14-1,32 (3H, m. H-7, Hi,-8, »t-9).
Pho "C-NMR (125 MHz, CDClj), 6 (ppm);
146,2 (s, C-4), 107,8 ((, C-14), 79,0 (rf, C-l), 67,0 (d, C-6), 55,9 (rf, C-5), 49,3 (/, C-7), 41,7 (s, C-10), 36,3 (/, C-9), 35,1 (/, C-3), 31,9 ((, C-2), 26 (rf, C-11), 21,1 (j, C-13), 18,2 (/, C-8), 16,2 (q, C-12), 11,6 (q, C-15).
ESI-MS; in/z: 239,2 [M+ H]*.
Niloticine (2): C3oH4g03 (M=456), bpt mau vang, tan trong CHCI3. Rr = 0,22 (MeOH.nuoc 5/1).
Ph6 ' H - N M R (500 MHz, CDCI3), 5 (ppm): 5,32 (m, H-7), 3,57 (m, H-23), 2,76 (dt,J= 14,5; 5,5 Hz, H.-l), 2,66 (rf, J = 8,5 Hz, H-24), 2,27 (.t,J= 3,5 Hz, H-9), 2,23 (r, J = 3,5 Hz, Hb-1), 2,10 (/, y = 3,5 Hz, H-6), 2,05 (m, H.-I6), 2,02 (m, H.-2), 1,82 (m, H.- 12), 1,72 (d,J= 8,5 Hz, H-5), 1,66 (ra, H,-22), 1,65 (m, H-20), 1,64 (m, H-l 1), 1,58 (m, H-17), 1,50 (m, H-15), 1,49 (m, Hb-2), 1,44 (m, Hb-22), 1,42 (m, H r
Nguyen Mgnh Cuang vg congsu 12), 1,33 (s, H-26), 1,32 (s, H-27), 1,23 (m, Hb-16), 1,12 (.5, H-29), 1,05 is, H-28), 1,03 (s, H-30), 1,01 (s, 11-19), 0,96 (rf, J = 6,0 Hz, H-21), 0,82 (s, H-18).
Pho "C-NMR (125 MHz, CDCI3), 6 (ppm):
216,9 (.V, C-3), 145,7 (s, C-8), 1 18,0 (rf, C-7), 69,3 (/, C-23), 68,5 (rf, C-24), 60,3 (i, C-25), 53,3 (rf, C-17), 52.4 (rf, C-5), 51,2 (s, C-14), 48,5 (rf, C-9), 47,9 (s, C-4), 43,6 (s, C-13), 40,7 (/, C-22), 38,6 (/, C-2), 35,0 (V. C-10), 34,9 (/, C-l), 34,0 (/, C-15), 33,6 ((, C-12), 33,6 (rf, C-20), 28,8 (t, C-16), 27,4 (q, C-30), 24,9 (q, C-27), 24,6 (q, C-28), 24,4 (/, C-6), 21,8 («, C-l8), 21,6 (?, C-29), 19,9 (q, C-2), 19,8 (q, C-26), I8,3(/,C-11), 12,8 (f/, C-l9).
Pho IIR-ESl-MS (+) 457,36821 {[M+H]'), gia trj tinh toan ly thuyet M = 456,3603, C3oH4,03.
Dimetyl crenatin (3): CiiH;209( M = 346), dang bpt trang, tan trong MeOH.
Pho 'H-NMR (400 MHz, CD3OD), 6 (ppm):
6,69 (.!, H-4, H-6), 4,84 (rf, 7 = 8,0 Hz, H-P), 4,54 (j, 11-7), 3,84 (s, 1-OMe, 3-OMc), 3,76 (dd. J ' 10,1;
2,0 Hz , H.-6'), 3,66 (rfrf, J= 10,1; 4,3 Hz, Hb-6'), 3,47 (dt. J= 6,3; 2,3 Hz, H-2'), 3,41 (dt,J= 6,0; 1,75 Hz, H-3'), 3,40 (dt, J= 6,6; 1,75 Hz, H-4'), 3,19 (m, H-5').
Pho ''C-NMR (100 MHz, CD3OD), 5 (ppm):
154,2 (s, C-l, C-3), 139,7 (s, C-5), 135,2 (s, C-2), 105,6 (rf, C-4, C-6), 105,4 (rf, C-l'). 78,3 (rf, C-5'), 77,8 (rf, C-3'), 75,7 (rf, C-2'), 71,3 (rf, C-4'), 65,1 ((, C-7), 62,5 (/, C-6'), 57,0 (q, 1-OMe, 3-OMe).
Leonurisidc A (4): C„H,oO,(M =332), dang bpt trang, kho tan trong dung moi hiru co.
Ph6 'H-NMR (500 MHz, DMSO-ds), 5 (ppm):
6,05 (s, H-4, H-6), 4,63 (rf, J= 7,5 Hz, H-l'). 3,67(j, 1-OMe, 3-OMe), 3,58 (IH, rfrf, 7 = 10; 2,5 Hz, H.-6'), 3,41 (IH, Hb-6'), 3,16 (m, H-2'), 3,16 (m, H-3'), 3,12 (ra, H-4'), 3,00 (m, H-5').
Pho "C-NMR (125 MHz, DMSO-d,), 6 (ppm):
153,9 (s, C-5), 153,2 (rf, C-l, C-3), 127,5 (s, C-2), 103,5 (rf, C-l'), 93,8 (s, C-4. C-6), 77,0 (rf, C-5'), 76.5 (rf, C-3'), 74,2 (rf, C-2'), 70,0 (rf, C-4'), 61,0 (/, C-6'), 56,1 (q, 1-OMe, 3-OMe).
Hmh 1: Cju trUc cac hpp chat 1, 2, 3, 4
TCHH,T.49(6),20n 3. KET QUA VA T H A O LUAN
Hgp c h a t l dugc tich ra tir phan dogn CH2CI2 cua thin vi re ciy, cd dang tinh th6 hinh kim, miu vang, tan trong CHCI3. DD' kipn phd '^C NMR, DEPT cho biet phan tir hgp chit 1 cd 15 nguyen tir cacbon, bao gom: hai cacbon bgc IV, nam nhdm metin, nim nhdm metylen vi ba nhdm metyl. Du kien phd '^C-NMR cho thay cd mgt cacbon bic IV tgi 5 146,2 (C-4) va mgt cacbon metylen tai 6 107,8 (C-14) Cling vdi hai tin hipu singlet ciia hai proton d 5 5,01 (Ha-14) va 4,74 (Hb-14) tren pho ' H - N M R , dieu niy chirng td su cd mat ciia mdt nhdm cxo- metylen. Hai tin hieu doublet ciia hai nhdm metyl tgi 80,95 (H-12) vi 0,87 (H-13) du doin s\r cd m^t ciJa mpt nhdm iso-propyl trong phan tir 1 Ben cgnh dd, cac tin hieu metin proton d 6 3,41 {dd, J= 11,5; 4,5 Hz, H-l), 5 l , 7 4 ( t / , / = 10 Hz, H-5) vi 5 3,71 {t,J = 10 Hz, H-6) cho thay chung cd cau hinh axial. Cic nhdm metin vi metylen cdn lai nim trong khoing 5 1,1-2,4. Ket hgp dQ- kicn phd ESI-MS, ID-NMR vi d6i chieu vdi tii lipu tham khao, hgp chit 1 dugc xac djnh li ip,6a-dihydroxy-4(14)-eudesmene [5].
Cdc tecpenoit vd phenolic glucosit...
Hgp chat 2 dugc tich ra tur phan dogn CHjCb vdi hgp chat 1, d dang bgt miu ving, tan trong CHCI3. Pho HR-ESl-MS (+) cua hgp chit 2 cho peak ion gii phan tir d 457,36821 ([M+H]*), phii hgp vdi CTPT CjoH^gOj. Cac phd '^C-NMR vi DEPT cho biet phan tir cd mgt nhdm cacbonyl, tam nhdm metyl, tim nhdm metylen, biy nhdm metin vi siu nguyen tir cacbon bic IV. Dp bit bao hoi ciia phin tu niy li bay, trong khi dd phin tir cd bdn vdng, mgt lien ket ddi C-C tgi 5 118,0 {d, C-7) vi 5 145,7 (.s. C-8) vi mpt nhdm cacbonyl keton tgi S 216,9 {s, C-3). Ben canh dd, tren phd '^C-NMR xuat hien tin liicu cua cic cacbon lien ket vdi di td oxy nam d viang trudng thap tgi 6 68,5 {d, C-24), 60,3 (s, C-25).
Tir cic phan tich tren, vdng cdn lgi trong phan tu la d dgng epoxy tgo thanh tir lien ket giii'a C24-0-C-25.
Phan tich cic dir kien phd 1D-NMR vi ddi chieu vdi cac tai lipu tham khio [6, 7], hgp chat 2 dugc xac dinh la niloticine. Cau triic vi di! kien phd cua 2 dugc khing dinh bang du' kien phd hai chieu HMBC (hinh 2). Niloticine da dugc tich ra tir ciy Phellodendron chinense [6] vi Turraea nilotica [8].
Niloticine li chit trung gian trong qui trinh sinh tdng hgp cic quassinoit [7],
H J S O S ^ ' " ^ 5 OH H
Leonuriside A (4) Hinh 2: Tuang tic HMBC chinh ciia cic hgp chit 2,3 va 4
Hgp chit 3 cd dang bgt tring, tan trong MeOH va dugc tach ra tir phan dogn nudc cua than vi rl cay. Dii kien phd ' H , ' ^ C - N M R cho thiy hgp chit 3 cd ciu triic gdm mgt dudng glucoside vi mgt aglycon li vdng benzen the.
Phd ' H vi " C - N M R cho thiy cac tin hieu cua mpt nhdm thd dudng D-glucopyranosyl tgi 5 105,4;
78,3; 77,8; 75,7; 71,3; 62,5, trong do cacbon anome d 5 105,4 {d, C-l') va proton anome d 5 4,84 {d, H-
I') vdi hing sd tich J = 8,0 Hz, vi viy nhdm th6 dudng cd cau hinh p.
Phan aglycon ciia hgp chit 3 cd ciu triic tuong d6i die biet, dd la mgt vdng benzen the 1,2,3,5 cd tinh ddi xirng. Phd ' H-NMR xuat hien tin hieu singlet ciia hai proton nhan tham tuang duang nhau tgi 5 6,69 (2H, s, H-4 va H-6), dng vdi hai cacbon
tuang ducmg tgi 5 105,6 (C-4 va C-6). Ben cgnh dd, trong pho ' H , ' ^ C - N M R cdn cd tin hieu ciia hai nhdm methoxy tuong duang tgi 5 3,84 (6H, s) va 5 57,0 gin vao hai cacbon tuong ducmg C-1 va C-3 (5 154,2) vi mpt nhdm th6 hydroxymetylen tgi 5 4,54 (2H, s, H-7) lien k6t vdi C-5 (5 139,7). Goc dudng D-glucopyranosyl dugc gin vio vdng benzen tgi C-2 X\\k hipn qua tuong tic giiJa H-l' (5 4,84) vi C-2 (5
135,2) trong phd HMBC. Vj tri cic nhdm chuc trong hgp chit 3 dugc gin chinh xic bing phd HMBC. Tii cic phan tich tren va so sinh vdi tu lieu, hgp chit 3 dugc xic djnh la dimethyl crenatin [9].
Hgp chit 4 tach ra tir ciing phan doan nudc vdi 3.
Cic dCr lieu phd ' H va '^C-NMR ciia hgp chit 4 tuong tu nhu ciia hgp chit 3. Di6m khac biet li trong phan tir ciia hgp chit 4 khdng cd nhdm chuc
TCHH,T.49(6),201I
hydroxymetylen gin vio cacbon C-5, mi thay vio dd li nhdm OH. Chinh dieu nay da lim cho dg chuyen dich hda hgc ciia cacbon C-5 chuyen djch ve phia trudng thip tgi 5 153,9. Ben cgnh dd, du- kipn phd ciia 4 cung cho thay sy xuat hipn cic tin hipu cua mpt nhdm the dudng P-0-D-glucopyranosyl gin tren cacbon C-2. hai proton tham tuong duang d vj tri meta tgi 5 6,05 (211. .v, H-4, H-6) vi hai nhdm metoxy tuang duang tai 6 56,1 doi xirng nhau gin tren hai cacbon tucmg duang C-l vi C-3 (5 153,2).
Tir cic phin tich tren va so sinh vdi tu lipu. hgp chat 4 dugc xic djnh li leonuriside A [10], Chat niy di dugc phan Igp tir loii Bioscorea nipponica Mak., mgt ciy thudc d Trung Quoc [ 10].
Ngoii ra. dua tren cic dir lipu pho HMBC, HSQC ciia hgp chit 4 dii lipu phd "C-NMR ciia cic cacbon C-l, C-4 va C-5 da dugc gin lai chi'nh xac so vdi tii lieu da dugc cdng bo trudc day [10].
4. KET LUAN
Tir than va re ciy Kho sim mem (Brucea mollis Wall. e\ Kurz) di phin Igp vi xac djnh dugc ciu true ciia bdn hgp chit li ip,6a-dihydroxy-4(14)- eudesmene (1), niloticine (2), dimetyl crenatin (3) vi leonuriside A (4). Ci bdn chit deu lin diu tien phan lip tir chi Brucea.
Ldi cam OTI: Cdng trinh ndy dugc hodn thdnh vdi su hd trg kinh phi tic nhiem vu Nghi dinh thu Vi^t - Hdn f201I-2G13) vd de tdi cap Viin Khoa hpc vd Cdng nghi Viet Nam.
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3. Mai Hiing Thanh Tiing, Trin Thu Hudng, Nguyln Mgnh Cudng, Tr^n Th4 Bich, Nguyen Quoc Binh, Tdc dung gdy ddc le bao ung thu cua Id cdy Kho sdm mem {Brucea mollis). Tap chf Dugc lieu, 16(6) (2011) (dang in).
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ex Kurz), Tgp chi Khoa hpc vi Cdng ngh? (2012) (da nhin ding).
5 Tr^n Ding Thgch. Phgm VSn Cudng, Tran HQu Gidp, Doin Thj Mai Huong, Nguyln Thj Minh HJlng, Marc Litaudon. Lfi VSn Hgc, Nguyln VSn Hung. Chiu Van Minh, Thdnh phdn hda hpc qud cdy lanh cong long nmgt - Tgp chi Khoa hpc vi Cdng ngh§, 48(4A), 58- 63(2010).
6. Yan Chen, Wang Ye, Hao Xiao-Jiang, Triterpenes from the fruits of Phellodendron chinense Schneid var glabriusculum. Chin. J. Nat. Med., 7(1), 31-33 (2009).
7. Zhengming Guo, Suryanarayana Vangapandu, Robert William Sindelar, Larry Anthony Walker and Robert David Sindelar, Biologically active quassinoids and their chemistry, potential leads for drug design, Frontiers in Medicinal Chemistry. 4, 285-308 (2009).
8 Dulcie A Mijlholland and David A H Taylor, Prololimonoids from Turraea niloltca.
Phytochemisny, 27(4), 1220-1221 (1988).
9 Tetsuo Ozawa and Yoshinori Takino. Carbon-13 Nuclear Magnetic Resonance Spectra of phenolic glycosides isolated from Chestnut Galls, Agric Biol.
Chem., 43(6), 1173-1177(1979).
10. Jiayong Zhang, Haifeng Chen, Naili Wang, Xinsheng Yao, Identification of water-soluble components, that are not saponins, from Dioscorea nipponica Mak., Asian Joumal of Traditional Medicines, 2(2). 70-74 (2007).
Lien hi: Nguyen Manh Cirdng
Vipn Hda hpc cac hgp chit thien nhien Vien Khoa hpc vi Cdng nghe Viet Nam 8 Hoing Quoc Viet, Ciu Giiy, Hi Npi Email: [email protected].
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