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TRANSCRIPT
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Nghi&r crnr bio ch6 vi6n nang cfmg Bio nh6n khang
dufirg huy6t va th* dQc tinh cip cria chd phim'
Nghi€n crru thhnh ptrln trOa hgc vi tdc {rng rtc ch6
t€ Ua'o ung thu cria ciy Bt dC tia(Uvaria grandiflora
Robxb. Ex Hornem - Annonaceae).
NghiOn crnr ti tQ, su phen b6 vh mirc itQ kh6ng thu6c
khdng sinh cta c6c chring Enterobacterirceae sinh
ESBL. CASBAPENEMASE tai BQnh viQn 103 giai
dsan 2010 - 2013.
Tdc dung hg glucose m6u cta cao l6ng sinh dia trOn
m6 hlnh chuQt nhit dfi th6o tluong thgc nghiQrn'
NghiOn crnr bio ctr6 tl6u ph6n nano artesunat.
Ph6t hiQn ilQt bi6n gen gdy bpnh cr Thalassemia bingky thuAt multiplex-PCR.
Nghi0n ctru ung dung kg thuit nested RT-PCR nong
chAn dodn nhi€m Rubella tru6c sinh.
Nghi0n cuu bidu hiQn dAu 6n insulin trong qu6 trinh
noOi "iy
bift h6a d bao giic ming tii nguoi thanh tii
bio tpy tlio.
Ung dgng c6ng ngh$ sinh hqc trong sinh t6ng hqp
mQt s5 flavonoid glycosides c6 hont tinh y duo. c hgc'
Nghi0n cuu dic di6rn hinh th6i mO u trOn chuQt thi6u
nul mi6n dich mang kh6i ung thu thanh quan nguli.
NghiOn crnr t6ng hqp mQt s5 o6n xu.it cta Quinazoline.
NghiOn cfru sin xuit ecotin miniproinsulin u d bioEscherichia coli tiit6 hgp.
Nghi€n cuu phuong ptr,6p phin tich acid benzoic, acid
sorbic vi vir fmg dpng khio sit thgc trErg st dUng
trong mQt s6 togi thgc phim trOn dia ben Hai Duong.
Nghi0n crnr biro ch6 gel ra mit krsitu chua Gentarnicin
0,3Vo vd Dexamethason 0,17o.
Nguy€nThi Kim Oanh
Ktuing Thi Hoa
DdnsThu Hiine
U Thi Bfch Hiin
Hd Thi Thu Vdn
NSuyen I'€ Vdn
NguydnVdn An
Trin QuinhTrangHodng Thanh Loan
Nguydn HqnhTh*yfr,nfuiin nQp
BniThi Minh Phuqng
DdnS Tiiin Trutug
Ii Hda Y€n
NguydnViiit HiiiuNsuydnThi Hd
NguydnVdnTilng
Nsrryin Htty Thwin
NguydnThi Binh
NguydnThi Crtc
NgO Thanh Httong
Mai ElwngTrd
Pham Thi H6ng,
HulnhThi NSpc Di6m
Trang
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NcHmN cuU TITANH rHAN n6^1, ngc vArAc DUNc {rc crn[ rr sAo rJNc rHrt cuA iav nr) nn rie
(UVARIA GRANDIFLOM RO)G. EX HORNEM. ANNONACEAE)
r6nr rAr
ThS. L/e Thi Bich niin*HrA+g ddn: PGS.TS. Nguy*n Thi Hodi*
Trong quri uinh tim tridu ui thrlc srl dgng cdy thu6c cua ddng bio pako, v6n Ki€u, tinh Qudng Tri, c6y Btr de da(uvaria grandiflora) dE duo. c nghiOn criu sing loc vi cho rh6y tac aung gay dgc ,6 ;;;"g;u 11BUT) n€n m6 hinh invitro' Blt b6o niy nhim nghi€n crlu thinh pran noa hgc vi ia. a;nt, noet tinr, ric chii rsi,tr cta ciic ph6n do4n vi c6chqp chir ph6n lQp tu cAy BD d€ tia.D6i t'gng nghi€n cr?': ph6n tr€n,nit tt6t ciy Bil d€ tfa thu h6i t4i tinh eunng Tri.
. , "oY*| phdp nghiGn crtu: chiilt xu6t uang cdc phuong phdp ngdm chi6t, chitit ldng - t6ng, rin - long. phan lfpbdng cdc phuong ph6p: sic kf c9t pha thudng, pnu aao, rli"-ti .Oi Sephadex LH-20. ttreo oot.qua uinh phin tgp bingsnc kf lop mdng. X6c 9it :fo trfc beng cac frructng pnap p.h6 cQng hudng tu hpt nh6n mQt chi6u (rH-NMR, ,3c-NM&
DEPT), hai chi0u G{MBC, HSQC, co5v, r.ioesvj ua;f6 kh6, phrn mir rtiQn tu (Esr - Ms). X6c trinh ho4t tinh riccn6 tgUT bnng md ttnh in vitro.
K6t qui: DE ph6n l0p a hqp ch6t tu cay Bir dd tfa li uvarilactam, 3-o-debenzoylzeylenon, znylenonvi pipoxid.Ph6n tloan chloroform cd' ho4t tfnh g6y doc TB9T mSnh nh6t ron 2 ddng TBLn u,i ui ung no da diy voi gi6 Ei Icso la0'971 Fglml vi l'305 Fclrnl'Trong 4 hqp chit plT.lap ttuqc, hqp ch6t 3-o-debenzoyt"zeytenon ric ch6 ddng TBUTphtii manh nh6t v6i gra ui rcm r,:o pelrt, aong *roi ;rp ;;t;;ilcbn ric ch6 5 aong TBLry khiic: ung thu bidu m0,ung thu gan, ung thu de day, ung thu tl4i ring vi ung thu fpcn ciu.'
K6t tu$n: Ph6n clo4n choloroform c6 hogt tfnh gay tloc t6 bio mgnh nh6t trong 5 ph6n doan r& Bt de tia. D6 ph6nl$p 4 hqp ch6t tu Bt dd th, trong d6 trqp ch6i r-o-debenzoytzeylenon c6 khi ning frc ch6 m4nh 6 dbng TBUT.* Tir kh6a: uvaria grandifrora;'rJvarilactam; 3-o-debenzoylzeylenon; Zeyrenon; pipoxid.
study on chemicol composition and inhibitory activity of cancer cell lines of uvariagranditlora roxb. ex hornern - aflfloflaclaeSummary
In the process of learning knowledge of medicinal plants of Pako, Van Kieu ethnic in euangtri province, uvariagrandiflora has shown in vitro cytotoxic effects of cancir cells in our previous screening test. rnis paper aims to studythe chemical composition and to determine inhibitory activity or calrcei celts of fragm"nis -a i*otuted compounds fromuvaria grandiflora' Materials: Above-ground parts oi uu-iu gt-ainnru harvested in euangtri province.Method" Extraction methods: soaking extraction, liquid - liquid extracti_on, solid - liquid extraction. Isolated bynormal phase' reverse phase column
"hto-atography and Seph'adex LH-zo column chromatography. subscribe toisolation Process by thin-layer chromatography.-Det"*iol"g ril;;; by the method oiou"r"* magnetic resonancesPectroscopy: one - dimensional (IH-NMR, l3e'NMR, DEF[), rwo - dimensional (I{r,fB-, Hsec, cosy, NoEs1.)and electron sPray mass sPectrometry (EsI-Ms). Determinationintriultory activity of cancer cells in in vitro noodels.Resnlts: 4 compounds were isolated frsm uvaria grandiflora including uvarilactam, 3-odebenzoylzeylenon, z*ylenonand Pipoxid' chloroform fragment shows stnongestcanc€r cells cytotoxic activity in'urlt Lcer cel line and gastriccancer cell line with IC5e values arc 0,971pglttl
-T9 r,los pfnrr.'rn i isotatea ,o*pounar, io-aru"n ylzeyrenon hassrongest cytotoxic activity against lung cancir cell line wittr IC;iJb Urrr,, and this compound also inhibis 5 differentcancer cell lines; epithelial cancer, liveicancer, stomach cancer, *lon
"*"r, and leukemia.condusion: choloroform fragrnent has strongest cancer celts cytotoxic activiry among 5 fragments from uvaria
trffiiff?*:Tf,fl*:were isolated from uviria granonora" i"'*ii.r, 3-o-de#;il-"1;on has a strong ab'ity* Key words: uvaria grandiflora; uvarilactam; 3-odebenzoylzeylenon; 7*yrenon;pipoxid.
* Dgi hgc Y Dryc Hud
530
r. DAr vAN otVi€t Nam lA mQt qu6c gia c6 mtc ttQ tla dang sinh hgc cao [], hon nta, cQng ddng cdc d6n tQc ViQt Nam
v5n c6 nhiAu kinh nghiQm ttqc d6o trong sfr dgng ciy cd lim thu6c. Tuy nhi6n, hiQn nay s6 loai tlugc clua vioehi6t xu6t hqp chdt ldm thu6c cbn r6t han ch6 t4l. Do vAy, nghiOn cr?u cdy thudc moi dga r€n tri thuc ban dia
dang h mQt huong tti ttiy tidm ning. Theo k& qui shng lgc ho4t tinh gdy ttQc TBUT in vitro mQt sti cAy
tbu5c cria d6ng bio Pako, Vdn KiAu d Quang Tri mi chfng t6i tld khio sdt, dich chiiSt tohn ph6n ciy Bt de
tia ((lvaria grandiflora -Annonaceae) da th6 hign hoat manh nh6t vd c6 tinh huong tlich tr€n 6 dbng TBUT
thri nghigm [3]. Tuy nhi6n, cho d6n nay d ViQt Nam chua c6 nhiAu nghiOn ctu vA lohi dugc ligu ndy. Vi vay
bi,,i b6o niy nhim: NghiAn cht thinh phht hod hgc theo illnh hrttng ttic dgn{ sinh h\c vd xdc qmh hogttnh .re chd fn(lf crta cdc phdn itogn vd cac hgp chdt ph&n l\p th cdy Brt de fis-
rr. DOI Trlgflc vA PHIIoNG PHAP Ncru,0trt cilU1. D6i tugng nghi€n cni'u
E6i tuqng nghi€n crm li pn6n rOn rnAt dfu c6y Bir dEtia(Ilvaria grandiflora Roxb. ex Homem - Annonaceae).
Mdu thu t+i huy€n Dakr6ng, tinh Quing Tri vdo thr{ng 9 - 2}ll. T0n khoa hgc dup x6c dinh boi TS. Nguy6n
tUti Cuong - ViQn Sinh thdi vi Tii nguyOn sinh vflt - ViQn Han hm Khoa hgc vi c6ng nghQ Vigt Nam. Cdc thu
nghiQm ti6n hanh trOn 6 dbng TBUT: LU-l (ung thu ph6i nguoi), KB (ung thu bi6u m6), MDA-BA-321 (ung
thu vrl), Hep G2 (ung thu gan nguoi), SW-480 (ung thu ruQt k60 vh MKNT (ung thu da dny).
2. Phuong ph6p nghiGn crhr
2.1. Phuong phdp didu ch6 miu thfr
Phin r6n mflt d6t cdy Btr d6 tia clugc phoi, s6y kh6, xay thanh bQt min (5 kg), sau tld chiiit voi methanoi
nryet d6i bang phdp ngi.m d nhiet dO phdng (l hdnnAn x 3 dn). GQp dich chi6t, c6t thu h6i dung m6i duoi 6p
su6t giem thu duo.c cao toin phin (m = 850g).Cao toin phin tluo.c phin trin vio nu6c r6i chitit xu6t phdn
doan bnng phuong phdp chi6t l6ng-l6ng l6n tug vsi c6c dung m6i n - hexan, chlorofornr, ethyl acetal
butanol, thu tlugc dich chi6t c6c phAn tloan vd phin nu6c cdn lai. CAt thu tr6i Aung m6i duoi dp sulit gidm thu
tlugc ci{c cin tuong fmg.
2.2.Phfipthri sinh hgc x6c ilinh tfnh AQc t6 bbo
Phuong ph6p thri tlQ ttQc t6 Uao in vitro [8] ttuo. c ViQn Ung thu Qu6c gia Hoa Ky x,6c nhan h ph€p thri d0
dQc tii biro chuAn nhiim slurg lgc, ph6t hi€n c6c chit c6 khi ndng kim h6m sg ph6t triiin hoac digt TBUT odiAu kiQn in vitro. Ph6p thti ti6n hinh x6c dinh hhm luqng protein tii Uao t6ng si5 dga vdo mit iIQ quang hgc
(OD - Optical Density) do duo. c khi thanh phin protein cta t6 bdo tluo. c nhuQm bdng sulforhodamin B(SRB).Gi6 triOD mdy tlo ilugc ti le thuan voi lugng SRB gin voi ph6n tri protein. KhA ndng ric ch6 ss s6ng
s6t cta ti5 Uao khi tlugc xdc ttinh th6ng qua c6ng thric:
7o s6ng s6t =tOD(chAt thrr) - OD(ngdy 0)l x 100
OD(dOi chfmg dm DMSO l$Vo) - OD(ngiy 0)
% tc ch6,= l00Vo - 7o s6ng s6t
C6c ph6p thi tlugc lip lA I nn. enipticine (Sigma) tlugc sri dpng lim ctrdt di5i chring duong, DMSO 107o
dua.c sri dpng lim A6i ctrung am. Gi6 tr! ICs3 dugc xdc dinh nhd vio phAn mdm mdy tfnh TableCurve. Ctr6t tfuinio c6 ICso < 20 pg/ml (voi chAt chi6t th6, ho{c voi ph6n tlogr h6a hgc) hoflc IC5s <4 ttelntl (voi hopt ch6t tinhttrii5$ ttuqc xem li c6 ho4r tfnh gdy tlQc tti bio vi c6 khi n6ng ric ch6 s1r phrit tri6n hoic di€t TBUT.
2.3. Phuong ph6p nghi€n cr?u vA tro6 hqc t2lPhAn l{p cdc ch6t tinh khi6t bnng sic kf c$t silicagel pha thutmg (Silica gel 60 0,040 - 0,063 mm Qn -
400 mesh ASTM), Merck); silicagel pha ddo YMC (30 - 50 pm, FuJisilisa Chemical Ltd.), Sephadex LH-20.
531
Theo d6i qu6 trinh phtur l0p bang sic ky lop m6ng pha thu&ng, pha tlio (TLC-silicagel 60 F25a Mercls Rp13F25a, Merck). Phit hi$n v6t ch6t bing dEn tu ngogi d hai bu6c s6ng 254 nm vh 366 nm troac atrng ttrutic tnrili dung dich HzSOr l0% trong ethanol tuyet d6i.
Xric tlinh c6u tnic cdc hqp ch6t phin lflp ttuqc dga t€n cdc phuong phdp ph6 bao gdm: ptr6 ttrtii phun mirtli€n tu (ESt - MS) tlo tr€n rnrdy AGILENT 6310 ton Trap; ph6 cQng huong tir het nhdn mQt chi6u 1IH-NMR,ttC-l.[[,IR, DEPT) vDr hai chiAu (HMBC, HSQC, COSY, NoEsY) clo tren mr4y Bruker Avance AM500 FI-NMR t+i Vi$n Ho6 hgc, ViQn Hen lim Khoa hgc vi C6ng nghQ Vigt Nam.
rtr. Ktr euA vA nAn luAx3.1. Tdc dgng rfic cn6 tfUt cria dich chi6t c6c phfrn dognCdc phdn tloan dich chiiit dugc x6c dinh ho4t tinh giy rlOc t6 bao tr6n 2 ddng ti5 bio MDA - BA - 321
(ung thu rnfi) vi MKNT (ung thu da day). K6t qui duo. c rrinh biy nong bang LBang L K6t qui thri host tfnh gdy tlQc t6 blo cta dich chi6t c6c ph6n rloan rrOn 2 dbng d bao MDA-BA-
321vdMKNT
Ndng.lQ
0t9ml)
Phin tlogn H (7o frc chd) Phin dogn C (7o rlc chC) PhAn tlop E (7o rh ch6)
MDA.BA.3zI MKNT MDA.BA-32I MKNT MDA.BA.32I MKNT
100 1o4,47 100,56 99,12 97,89 98,59 98,05
20 93,68 9t,42 97,72 96,92 79,65 80,37
4 37,72 39,09 94,83 74,94 31,32 32,82
0,8 24,12 22,34 36,49 38,13 18,33 21,77
ICso 3,27 3,42 0,971 1,305 8,4 8,08
Ndng dQ(pglrnl)
Phin do4n B (7o irc ch6) Ph6n ttoen N (7o rlc ch6) Ellipticine Vo trc ch|)MDA.BA-
321MKNT MDA.BA-
321MKNT MDA.BA.
321MKNT
100 95,70 97,35 74,91 76,55 90,77 99,78
20 48,16 5L,21 29,47 32,08 59,72 65,72
4 27,L9 30,02 21,84 23,49 19,76 33,33
0,8 21,14 20,46 14,47 r7,82 -0,54 5,76
ICso 22,72 18,77 60,92 52,48 1,249 0,935
* Ghi chil: Ndng ttQ rhri nghiQm cria Ellipticine li l0 pglml; z ltglrnl:O,4 pglml; 0,0g pglrnl.K6t'qud cfro tfrAy,.ngogi trir phdn itoqn nu6c kh6ng c6 khd n6ng fc cfr6 nai dbng t6 bio thri nghiQm,
4 phan do4n cdn lai dAu th6 nien tr{c dpng d cdc mric tlQ kh6c nhau. Trong 116, ph6n doai chloroform tri6 nienhopt tinh t6t ntrAt tron ci hai ddng td uao MDA-BA-321 vir MKNT vctr c6E gia t4 Ic;, Brirqt;g;ti-;;l '305
pg/ml. Do tl6 phAn tloSn chloroform duo. c lga chgn d6 chi6r xuft, phan i6p cic t qp ch6t tinh khtbr.Z.XStqui nghi€n crfru vd ho6 hgc
2.1. Chi6r nr{t va ph6n tgp c6c chdt tinh khi6tPhAn doan chloroform (ki hiQu C, m = 310 g) du-oc chi6t pha r6n t€n cQt silicagel, rua giai Dn lu$ beng
cdc h$ n -hexan l00Eo, n-hexan:aceton 20:1, 10:1, 5:1, l:l vi aceton l007o thu dugc 6 phAn do*n (cl - c6i.PhAn ttoqn C2 tfugc chi6t pha iin, rua giei Bn lu$ beng n-hexan laovo,n - hexan: ihyl acetat 20:1, l0:1, 5:1,l:l vi ethyl acetat 100% thu tlugc 6 phin tto?n (C2A - czn. phan do4n C2B tlugc t6ch Ua"g.$;h.A;;532
hQ methanol: nu6c (3:l) thu tluEc 7 ph6n tlo4a (C2Bl - C2B7). C2B6 dugc phAn lfp titip beng cgt pha thuong
voi h€ n-hexan: ethyl acetat:aceton (20:l:0,5), thu ilugc 5 phin tlo4n (C2B6A - C2B6E). C2B6C tluqc tinh
cnii Ueng cQt SephadexLH-?-[,rua giai bing methanolS0To thu tlugrc hqp ch6t I (UGCI2, m = 25 mg).
PhAn rloan C5 ilugc rich beng cQt silicagel pha thudmg voi hQ chloroform : methanol (40 : l) thu tfugc 4
phdn tlogn (C5A - C5D). Phdn doAn C5A ttugc sic hi lflp lsi bing cQt pha thudng voi h€ n-hexan : aceton
(5 : 1) thu dugc 7 phin ttoan (C5A1 - C5A7). PhAn tloan C5A2 duo.c ti4ch beng cQt pha ttio voi hQ aceton :
nudc : acid formic (3 : I :0,1), thu dugc 4 phAn ttop (C5A2A - C5A2D). C5A2B tluo.c tinh ch6 bang cqt
Sephadex LH-20 voi hQ methanol 807o thu tluo.c 3 phAn tloan (C5A2BI - C5A2B3). Phan doen C5MB2
*uit nien tinh thA hinh kim khi dugc c6 df,c, lec, rira tinh ttrd bing methanol thu tluo. c hqp chfu 2 (UGLE I ,
m=270 mg). Tinh chd phnn doen C5A5 bang cOt Sephadex LH-20, rrla giii bing methanol lNVo thu dugc
hqrp chAt 3 (UGCj, 6 = 167 mg). Phdn doan C5A7 xuAt hi€n tinh thii tring khi tlugc c0 d[c, lgc, rua tinh thd
bing methanol thu dugc hqp chit a Gf higu UGC8, m = 12,5 mg).
2.2,Xicilinh cAu tnfic cic hgp c-h{t di phAn l$p
Th6ng qua viQc phin tfch dt ki€n ptrO mUn I chiAu-2 ehidu vi ptr6 tctrOi MS, c6u tnic cria ci{c ch6t s4ch
I - 4 tluo. c x6c tlinh ti Uv.arilactam (l), (-)-3-O-debenzoylzeylenon (2), 7*ylenon (3) vn Fipoxide (4).
Hqp ch{t I OGC12): BQt miu vimg, tan trong hdn hgp chloroform vi methanol, Rr = 0,30 (n-
hexan/ethyl acetalaceton ?-Ollt},8). Ph6 IH-NMR (DMSO - do) xdc nhin s1r c6 mit cria I nh6m imin 6n
10,76 (s); I nh6m hydroxy tei 6r{ 10,09 (br.s), hQ spin AMX tai 6H 8,62 (lH, d, 8,5); 7,07 (lH, d, 8,5) vi ?,36
(lH, r, 8,5). Ngohi ra cbn c6 2 tfin hiQu singlet cta 2 proton thorn kh6c tai &r 7,84; 7,42 vd proton cria 2 nh6m
rethoxy tai 6n 4,00;4,04.
CAc ph6 t3C-NIr4R vi DEPT cho thfy ph6n tri c6 chria 17 nguyOn tt C gdm 2 nh6m methoxy tai 6c 56,9;
59,8; 5 nh6m methin thom vi 10 C kh6ng li6n k6t H. C6c tfn hi$u t?i 6c 150,5 (C-3); 153,7 (C-8); 154,1 (C4)
rhuQc vi cdc nguy€n trl C cria nhdn thsrn gin voi oxy. Tin hiQu tai 6c 168,3 (s) thuQc vA C cria nh6m -CO-NH-.
Tuong tr{c HMBC cria 3-OMe/C-3, 8-OMe/C-8 cho ph6p ttlnh vi 2 nh6m methoxy vio C-3, C-4. Vi ricna nh6m -CO-NH- duqc khang dlnh qua tuong t6c HMBC cria proton cria nh6m I.IH (6H 10,76) voi C-1, 9,
10, lOa, I l. Cdc ruong t6c vinylic cta cdc proton thsm voi C eta h€ vbng thom H-4C4,10a I l; H-5lC4a"?, 8a; H-6/C-4b, 8; H-7tC-5,8a vi H-9/C-4b, 8, lOa cho ph6p thi6t bp b0 khung phenanthren cria UGC12.
ha vio cr{c dt kiQn ph6, hqp ch6t UGC12 tluo.c x6c dinh ln Uvarilactam
Hqrp chAt 2 (UGLEI): BQt miu vdng nh4t, tan trong c6c dung m6i chloroform, aceton vi methanol, Rs =O,5O (aceton/ nudc/acid formic 3,5lll},l). PhO ESI-MS(-) cria UGLEI cho pic ion gid phAn tu tat nt/2313,4
M-H + 2H2Ol- (IC/|,Vo). K6t trqp voi ph6 '3C, DEPT cho ph€p xr{c ttinh c6ng thric ph6n tu cria UGLEI liCrfipOe (M = 278,3 g/mol).
Ph6 IH-NMR chi ra tfn hiQu cta 5 proton thom thuQc nh6m benzoyloxy tAi 6H 7,46 (2H, ddd,1,0, 8,5, 8,5
Frz);7,60 (lH, fi, 1,0, 8,5 Hz\;7,94 (2H, dd,l,0, 8,5 Hz). Tin hi€u cria 2 cis proton olefinic tai 611 6,07 (lH,dd, 10,5,2,5H2);7,00(lH, dd,lO,5,2,5Hz)vi4protonginvoic6cnguy6ntrlCli€nktitoxytai6r{3,99-469. Ph6 I3C-NMR, DEPT cho bi6t ph6n tri c6 14 nguy€n tt C gdm I nh6m methylen, 9 nh6m methin vh 4
,.ryuydn tu C kh6ng li6n k6t vsi H. Trong tt6 tin hiQu C cria nh6m benzoyloxy t4i 6q 129,5 (C-3',5'); 130,4
€-2',6'); l3l,l (C-l'): 134,3 (C-4') vd 167,2 (C-7'). C cria nh6m Cyl cang huong tAi Ec 197,5 (C-6); 2 C*frnic tai 6c 127,6 (C-5); 153,0 (C-4) vir 4 nguy€n tri C sp3 gin voi oxy tAi 6q 63,3 (C-?); 70,2 (C-3);75,7tC-2); 77,2 (C-l). Cdc tuong t6c COSY vh HMBC cho phdp thitit l6p vdng 6 c4nh voi n6i AOi tai vi trf 4, 5;*6m oxo d v! trf 6 vi chimg 16 nh6m C-7 ni5i v6i C-l vi voi nh6m benzoyloxy.
C.iu hinh tusng d6i cria UGLEI dugc xi{c tlinh dua vio viQc phdn tich hang sii tuong t6c J vi cdc dt kignpm NOESV. Cdc dt kiQn ph6 NMR, MS cta UGLEI chfmg t6 hqp ch6t niy c6 c6u tnic tuong tu voi hqpcra S-O-deUenzoylznylenon ti{ch ra tt loiri (Ivaria purpureatruOc tt6y I l]. Ift6c biQt gita hai hqp chft nhy
533
h d d6u cua ning su6t qlray cuc ([o]o). Theo tii liQu [z], 3-o-debenzoylzeylenon c6 [c]o = +12,5 trong khitt6 hqp ch6t UGLEI c6 [cr]p = - t2,O. Di6u niy chimg t6 UGLEI li mOt diSi quang "ta\-Odebenzoylzeylenon. c6u tn'ic UGLEI dugc x6c tlinh li (-)-3-o.debenzoylzeylenon.
Hqp ch6t 3 (UGCS): BQt miu F6ng, tan rrong aceton, methanol; Rr = 0,40 (hexan/aceto n 2ll hoircmethanoUnudc 3/l). Ph6 rH-MvlR chi ra tfn hiQu cia l0 proton aromaric trong khodng 6 7,51 - 8,0g bao g6m2 doublet (J = 8,0 Hz) vi 2 multiplet tuong ung voi tin hi€u trong vilng 6128,7 - 165,6 trong ph6 t3C-NMR.
EiAu niy chimg t6 sy t6n tai cta 2 nh6m bezoyloxy trong phfur tt. C6c ph6 tD-NMR cung chi ra tin hi6u cta2protoncis-olefinic &87,07 vi6,17(Jr,s= l0,5Hz); I nh6mmethylenoxy6E4,5T vi4,36;2protonctanh6m methinoxy (>CH-O-). Su c6 mit crla I nh6m Cyl trong ph6n nr duo. c thd hiQn 16 qua tin hi€u d &195,1. PhO H,H-COSY chi ra c6c tuong tdc cria H-2fH-3,H-3tH-4,H-4fH-5.PhO I{MBC ctro th6y ff-i to*gt6c voi C-l,C'2,C'6,C-7'chfmg t6 c6 I nh6mbenzoyloxy gin vio v! triC-7. Nh6m benzoyloxy cbn eittinh vi vio C-3 do c6 tuong tr{c gita H-3lC-7-" tr0n ph6 HMBC. Tir cdc ph6n tfch rr6n vi sg phtr hqp vd s6lf,u ph,6 NMR cta UGC5 vdi c6c s6 tleu ph6 cria hqrp ch6t Znylenon rld clugc c6ng b6 [6J cho ptef tningdinh cdu tnic h6a hgc cta UGC5 lh Zeylenon.
ttgp 9!rdt 4 (UGC8): BQt mdu tring, ran ttit trong aceton, Rr = 0,30 (aceton/nu6c Zll),0,4(hexan/aceron5/2). Ph6 tH-Mr4R cria UGCS c6 sg hiQn diQn cria 3 cpm tin hiQu multiplet fng voi l0 proton (thuQc vdngthorn) trong khoing67,49 - 8,10; 2 cis proton olEfinic d 6 5,91 vi 6,14 (J = l0 Hz);2 proron cria nh6mmethylenoxy (-CH2O, d 6 4,91 vh.4,57 voi Jgr, = 12,0 Hz; 3 proton cta ciic nh6m methinoxy (>CH-O-) dkhoing 6 3,66 - 5,63. Ph6 t3c-NMR, DEPT 90, 135 ciing chi ra sg c6 m{r cria 2 nh6m benzoyloxy vst c6cnguy6ntuCcriavdngthomd6129,6-134,5cirngvoi2nguyOntriCnimtrongnh6mcacboxyldD16T,5vd16T,T.TinhiQuctac6cnguyOntriCginvdiOtluocth6ydkhoing655,l-75,8gdmlnh6m-CHzO-d663,7vi3nh6m>CH-O-d655,l,7l,l,75,8vilnguy€ntriCbicfVd66l,0.C6cttacdi6mptrOUUnmgtchidu cta U@S chimg td hqp.ch6t ndy lh mOt din xuit oxy h6a vdng cyclohexene. So sdnh cic dn kiQn ph6cria UGC8 voi cdc dtr ki$n ph6 cria Pipoxid dugc c6ng bti trong Bi liQu [10] cho thAy c6 sU rrirng hqp hoantoin. Do d6 UGCS cluqc x6c tlinh li Pipoxid.'
(-)-3-Odebenzoylzeylenon (2)
Zeylenone (3) Pipoxide (4)
01fr"
\,/",2
reA'
>e%r'"fr-\/
534
Uvarilactam (l)
Hinh l. Gdp tnic h6a hqc c6c hqp ch6t ph6n Bp tu ciy Bt d6 tfa
3. Tdc dgng rfrc cnd fnUt cria c6c hqrp chAt ph6n l$p
Cr{c hqp ch6t ph6n lip duo. c xric dinh khd ning gdy tlQc ti5 bio tr€n dbng TBUT ptrOi t-U-t. Ktit qud thri
nghiQm ttugc thd hiQn trong bing 2.
Bing 2. Kiit qui xrf,c ttinh gi6 d ICso cria cdc hqp chit phdn lip trOn dbng t6 bio LU-l
Ndng dQ Gynt)7o tJc ch6
Hqp ch6t I Hqp ch6t 2 Hqp ch6t 3 Hqrp chdt 4 Ellipticine
ICso 16,66 1,30 37.63 26,19 0,67
Trong 4 hW ctr6t ptrAn lfp dugc, hqp ch6t 2 ((-)-3-O-debenzoylzeylenon) c6 ho4t tinh manh nhAt trOn
dong ti5 bio LU-l voi gi6 tri ICso 1,30 pglml. Do d6 tl6y duo. c xdc clinh li hqtp ch6t c6 tdc dpng irc ch6 su
phdttritin TBUT dbng LU-l. Nh6m nghiOn crnr dE tii5p tUc x6c dinh hoat tfnh cta hqp ch6t ney E€n ? ddng
TBUT kh6c vh tr€n ddng td Uio thu&ng 3T3, k6t qui dugc th.5 hien trong bang 3.
Bdng 3. Hoat tinh gfly ttQc t6 bao cria hqp ctrit (-)-f'O-debenzoylzeylenon
N6ng tlQ(tts/d)
Vo trcch6
KB MDA.BA.23I LNCaP HepG2
UGLEI Ellipticine UGLEl Ellipticine UGLEI Ellipticine UGLEI Ellipticine
ICro 3,71 0,98 17,8 | 0,79 12,63 0,79 3,65 0,86
Ndng dQ
(pe/d)
Vo itc ch€
MKNT sw480 HL-60 3T3( T6 bno thusng)
UGLEI Ellipticine UGLEI Ellipticine UGLEI Ellipticine UGLEI Ellipticine
ICro 2,40 0,94 3,r3 0,78 1,73 0,72 7,19 0,31
K6t qud cho th6y hqp ch6t (-)-3-O-debenzoylzeylenon c6 khi ning ric ch6 nhi€u ddng TBUI kh6c nhau,
ffing qua kh6 ning ric chii 5/7 dbng td beo thri nghi€m (voi c6c gi6 tri ICso < 4ltglml), bao gdm c6c dbng t6
bio: ung thu bi6u m6 KB, ung thu gan HepG2, ung thu da day MKN7, ung thu ruQt klit SW480 vi ung thubgch cAu HL-60. Tr0n ddng t6 bao thubng 3T3, hgrp ch6t nay c6 gi|tri ICso cao hon so vsi Ellipticine di)n23
tfo, d6ng thoi gi6 tri ICso niy nim ngoii ngudng gdy ttQc t6 bao ( > 4 pelrnl). Di6u nhy chfrng t6 tlAy h ho,p
cLit c6 tfnh an toin cao.
Trong 4 hqp ch6t tt6 phAn lip, Uvarilactam mn diu tiOn duo. c tim th6y trong thinh pnan noa hgc loii(Jvaia grandiflora. Uvarilactam li m$t alcaloid thuQc nh6m fuistolactam. MQt sii hqp chdt thuQc nh6n niyc6 khe nnng g6y dQc cric TBUT [5]. Trong khi al6, 3 ho,? chdt Znylenon, (-)-3-O-debenzoylzeylenon vdPipoxid ddu c6 ciu tnfic thuQc nh6m Polyoxygenabed cyclohexen. HiQn nay, nh6m chit niy ttang ifuqc quanrim nghi€n cr?u voi ho4t tinh ric chii TBUT. MQt s6 din chdt Polyoxygenated cyclohexen phin ldp tt chi{Jvaria c6 tr{c dung gAy dQc c6c dbng t6 bno khSi u nguoi nhu t6 bio KB, HCT-8 (ung thu ruQt k6t1, Bel?402 (ung thu gan) vb A 2780 (ung thu bu6ng trung) lgl. Tnylenon cld duo. c chimg minh li mgt ch6t frc ch6v.fo chuydn nucleosid trong ci{c TBUT c6 trucmg Ehrlich t6l. Hqp chdt ney cbn th6 hi€n t6c dpng g6y ttgcnhidu dbng tti bao tltii u nguoi nhu t6 bio HCT-8, BGC-823 (ung thu da diy), Bel7402,MCF-7 vd MCF-?/ADM (ung thu ut), KB vd KBA/CR (ung thu bitiu m6 miQng) t6l. Nhu vgy 4 hqp cnAt pnan Qp duEc c6dfrr tnic thu0c hai nh6m Aristolactam vh Polyoxygenated cyclohexen li nhiing nh6m ch6t c6 tdc dgng tc ch6*i6u dbng TBUT. Dac biet hqp ch6t (-)-3-O-debenzoylzeylenon tld thd hiQn khi ning irc ctr6 mann nniAu&g TBUI voi c6c gi6 tr! IC5s thdp. K6t qui niy md ra tri6n vgng cho viQc nghi€n cuu bdn t6ng hqrp rhu6cco tdc dgng ch5ng kh6i u ttr hqp ch6t (-)-3-OAebenzoylzeylenon.
535
w. KET LU,AN VA rdn NGHI
Trong c6c ph6n doan dich chilit tir cdy Bi d6 tfa, phAn do4n chloroform c6 hopt tinh gdy ttQc tii bio m4nhnhit tr€n 2 dbng TBUT MDA-BA'321, MKNT voi cdc giri tri ICso 0,97 vi 1,30 pg/rnl. Ttr phAn iloanchloroform da ph6n Fp a hqp ch6t Uvarilactanl 3-O-debenzoylzeylenon, Zeylenon vi Pipoxid. Trong tt6,
ho-p chit 3-Odebenzoylzeylenon frc ch6 dbng TBUT LU-l mqnh ntrit voi tCso 1,30 lrg/rnl, tt6ng thoi hqrp
chit niy cdn fc ctr6 S OOng TBUT: KB, HepG2, MKN7, SW480 vi HL-60. Cric k6t qui nghi0n cuu tl5 g6p
phen giai thich kinh nghiQrn dDng thu6c qui gi6 cria ttdng bio dfln tQc vA cho thiy dugc liEu niy cin tluo. c
nghiOn cuu d di$n rQng hon vi mfc alQ sdu hsn vd thanh phdn ho6 hgc vd ho4t tfnh sinh hqc d€ c6 thd dm ra
cric ho4t ch6r c6 uic dung diQt TBUT ttit.
TAI LTEU THAM rcTAO
t. BO Tei nguy€n vi M6i tnrsng (2010), Bdo cdo hiQn trsg mbi tniong quiic gia - ChuyAn di Oa dqng sinh h7c,t.9.
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3. Nguy6n ThiHoii, Trinh ThiEiQp, E6 ThiThdo, Nguy€n Kh6nh Thu! Linh, Nguy6n Bich Hi€n, Hoang ThiDiguHuong (2012), "Sing lgc hoat tinh di€t TBUT mQt s6 cAy rhu6c cta <t6ng bio Pako, Vdn Ki€u d Quring Tn-,Tqp chiDrwc li€u, 17 (2), tr. 95- I 00.
4. ViQn Dugc liQu (2006), Nghi€n cfiu thu6c tv thdodro. c, Nxb l(hoa hgc vi Ki thr.r,it, Ha Ngi, rr.67- 69.
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oxidation product and regioselective Kornblum-Delamare rearangement: Total synthesis of zeylenols and zeylenones",
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I L Yoshifrsa Takeuchi, Qing Wen Shi, Takeyoshi Sugiyama, Takayuki Oritani (2001), '?olyoxygenared Cyclohexenesfrom rhe Chinsse lree, Uvaria purpurea", Bioscience Biotectnology and Biocheml'stry, 66(3), pp. 537-54.
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