You are on page 1of 12

Nghin cu cc phng php phn lp

Zerumbone c cht lng cao t thn r cy


gng gi (zingiber zerumbert sm.) v
chuyn ha Zerumbone thnh cc hp cht
c hot tnh sinh hc
L Th Thy
Trng i hc Khoa hc T nhin, Khoa Ha hc
Lun vn ThS Chuyn ngnh : Ha hu c; M s: 60 44 27
Ngi hng dn: PGS.TS. Vn Ngc Hng
Nm bo v: 2011

Abstract: Khi qut v chi Gng (Zingiber), cy Gng gi (Zingiber


zerumbet. Sm), cng dng v cc hot cht sinh hc ca Gng gi
(Zingiber zerumbet Sm). Trnh by cc phng php thc nghim:
phng php phn lp zerumbone t thn r cy gng gi; iu ch cc
dn xut ca Zerumbone; kho st hot tnh chng ung th ca cc dn
xut ca Zerumbone. a ra kt qu nghin cu v tho lun: iu ch
Zerumbone cht lng cao (> 99%);tTng hp mt s dn xut ca
zerumbone v kho st hot tnh chng ung th ca chng; kho st
hot tnh chng ung th ca 3-isopropylaminozerumbone.

Keywords: Ha hu c; Hot tnh sinh hc; Ha hc; Cy gng


Content
Gng gi (Zingiber zerumbet Sm) l cy thuc dn tc ni ting t lu i
khng ch nc ta m cn rt nhiu nc ng Nam nh Indonexia, Thi
Lan, Min in
Gng gi l cy mc hoang di ph bin nc ta t Bc ti Nam, c bit
l cc tnh min ni nc ta.
Hp cht c hot tnh sinh hc chnh ca gng gi l Zerumbone- n l cht
c hot tnh phng nga v chng ung th mnh trn 10 loi ung th khc nhau,

c bit l ung th gan v ung th v. V vy, gn 20 nm tr li y, Zerumbone


nhn c s quan tm ca rt nhiu nh khoa hc trn th gii, c bit l cc
nh khoa hc Nht Bn v M. Ngoi ra nhng nghin cu gn y ca cc nh
khoa hc chng minh rng dn xut ca zerumbone cng c hot tnh phng
nga v chng ung th mnh. Chnh v vy m chng ti c bit quan tm ti
vic iu ch zerumbone c cht lng cao t cy gng gi chuyn ha
zerumbone thnh cc cht khc nhau nhm to ra c nhng hp cht c hot
tnh sinh hc phc v cho ngnh y dc. y cng chnh l l do m chng ti
chn ti: Nghin cu cc phng php phn lp zerumbone c cht lng cao
t thn r cy gng gi( Zingiber zerumbet Sm) v chuyn ha zerumbone thnh
cc hp cht c hot tnh sinh hc.
Gng gi (Zingiber zerumbet Sm) l cy thuc dn tc, mc hoang v ph
bin nc ta. Nh trong tng quan trnh by, trong 20 nm tr li y cc
kt qu nghin cu ca trong v ngoi nc v hot tnh chng ung th ca
Zerumbone khng nh rng: Zerumbone l mt tc nhn chng ung th
mnh, n c ch s pht trin ca 10 loi ung th khc nhau. C ch chng ung
th ca Zerumbone c chng minh r rng, n thc y qu trnh t cht
(apoptosis) ca t bo ung th lm cho ung th khng pht trin c, n c ch
NF- kB hot ng nn ngn nga s to thnh ung th v pht trin ung th.
Zerumbone li khng c, nguyn liu cho iu ch zerumbone l cy gng gi
(Zingiber zerumbet Sm)- nc ta li c nhiu v cht lng tt, hn na gn y
ngi ta pht hin ra rng mt s dn xut ca zerumbone c hot tnh chng ung
th ti mt mnh hn zerumbone nhiu. Chnh v vy m iu ch Zerumbone c
cht lng cao tng hp cc dn xut ca zerumbone c hot tnh sinh hc l
mc tiu v ni dung ca ti ny.
1. iu ch zerumbone c cht lng cao
ti ca chng ti l mt phn ca ti nghin cu khoa hc quc gia
m s CNHD- T- 018/10-11 thuc chng trnh KHCN trng im quc gia
pht trin CN ha dc n nm 2020. Trong ti ny ngi ta xc nh
nguyn liu cho sn xut zerumbone l c gng gi (rhizomer of Zingiber

zerumbet Sm) thu mua ti Tam o tnh Vnh Phc vo cui thng 11 dng
lch, do ThS Nguyn Quc Bnh thu mua, phn loi, gim nh v cung cp.
iu ch zerumbone c th i t c gng gi ti hay c gng gi kh.
Kho st sinh l c gng gi ca chng ti cho thy t lc thu hoch cui thng
11 u thng 12 dng lch, bo qun n thng 3 thng 4 nm sau th c gng
gi mc mm v bin cht, hm lng Zerumbone rt thp. Chnh v l do ny
nn chng ti nghin cu iu ch zerumbone t bt c gng gi kh, hiu
sut chuyn i c t c kh l 14,28%, hn na nguyn liu kh cho php
sn xut c quanh nm nn thun tin cho vic cng nghip ha qu trnh sn
xut.
thu c phn khng phn cc ny chng ti chit tng cc cht trong
bt c gng gi kh bng EtOH 96%, loi bt EtOH cn 1/5 th tch, ri pha
thm 4/5 nc mui 10% v chit bng n-hexan, sau lm kh v loi n-hexan
c phn khng phn cc hiu sut t 4% tnh theo nguyn liu kh.
T phn khng phn cc tin hnh iu ch zerumbone tinh khit bng 3
phng php: Sc k ct (CC), sc k lp mng iu ch (PTLC) v phng
php kt tinh phn on (FCr). So snh cht lng v hiu sut zerumbone ca 3
phng php ny c s liu trn bng 3.1.
Bng 3.1: Hiu sut zerumbone thu c t 3 phng php phn lp khc
nhau
Phng php

CC

PTLC

FCr

Hiu sut (%)

0,10

0,08

0,35

im chy

66-67%

66-67%

66-67%

Rf

0,75

0,75

0,75

Kt qu trn cho thy phng php ti u iu ch Zer tinh khit t thn r


gng gi l kt tinh phn on nhit thp.
T cc kt qu trn chng ti xut quy trnh cng ngh sn xut
zerumbone tinh khit nh s 3.1. Thc hin qu trnh ny mt cch nghim
chnh chng ti thu c zerumbone 99,5% t hiu sut 0,35% tnh theo nguyn
liu ti. Trong khi cng bng cch chit phn khng phn cc ri qua

phng php sc k ct Abdul v cng s trong bng pht minh ca mnh s


US2009/0239953 A1 cng b Sep.24,2009 ch thu c zerumbone vi hiu sut
0,062% tnh theo nguyn liu ti. Trong cc ti liu khc chng ti cng cha
tm thy quy trnh iu ch zerumbone t c gng gi c hiu sut cao. S trnh
by trn cho thy quy trnh iu ch zerumbone 99,5% t c gng gi vng Tam
o c nhiu u im v cho hiu sut cao, tuy nhin hiu sut sn phm cn
ph thuc cht lng nguyn liu.

Bt c gng gi kh
1. Chit bng EtOH 96%
2. Loi bt EtOH cn 1/5V
3. Thm 4/5V nc mui,
chit bng n-hexan
4. Lm kh, loi n-hexan

Cng on I
iu ch du Zerumbone

Du Zerumbone

1. Kt tinh chm nhit thp


2. Lc qua phu thy tinh xp G4

Cng on II
iu ch Zerumbone th

Zerumbone th

1. Kt tinh phn on nhit


thp v dung mi thch hp
2. Lc butne qua giy lc

Cng on III
iu ch Zerumbone tinh
khit

Zerumbone tinh
khit
4

S 3.1 : Quy trnh cng ngh sn xut Zerumbon tinh khit t bt c


gng gi kh
tinh khit ca sn phm c kim tra bng UPLC- MS trn my sc ph ch
cho 1 pic duy nht (ph lc 1 trang 62)
Cu trc ca Zerumbone c chng minh bng cc phng php ph :
+ Ph khi ca Zerumbone cho pic M+H =219,19 ngha l phn t lng
Zer-1 l 218 v (xem ph lc 2).
+ Ph IR cho thy trong phn t Zerumbone c lin kt i 2 nhm th dng
trans c H = 965,18cm-1 v lin kt i ba nhm th c H = 831,33cm-1 (xem ph
lc 3 trang 63).
+ Ph 1H-NMR cho thy phn t Zer-1 c 2H ca lin kt i loi E dng pic
AB, dd 5,96ppm v 5,88ppm, J = 16,40Hz. Hai nhm metyl c H = 6,00; J =
11,12Hz v H = 5,25 v J = 10,63, mt nhm metylen c H = 1,90; J = 12,72.
Mt nhm gemdimetyl c H = 1,07 v 1,20ppm dng singlet v 2 metyl singlet
khc c H = 1,79 v 1,54ppm (xem ph lc 4).
+ Ph 13C-NMR cho thy trong phn t Zerumbone c 15 nguyn t cacbon,
trong c mt nhm >C=O, C = 204,33 ppm. 6 cacbon olefin c chuyn dch
ha hc t 124ppm n 160ppm v 8 cacbon vng trng mnh c C t 11ppm
n 42ppm (ph lc 5).
+ Ph DEPT cho thy phn t Zerumbone c 4 nhm CH3, 3 nhm CH2, v 4
nhm CH (ph lc 6).
Da trn nhng d liu ph, cng thc phn t ca Zerumbone c xc
nh l C15H22O , cng thc cu to nh trong hnh 3.1 v cc d liu ph c
th hin trong bng 3.2.

Hnh 3.1: Cng thc cu to ca Zerumbone


Bng 3.2: S liu ph NMR ca Zerumbone
13

C-NMR

H-NMR

S C

C ppm

H ppm

S H

bi

J.Hz

42.38

2.45; 1.90

2H

d,d

11.39;
12.72

127.14

2.50

1H

10.60

136.26

6.01

39.42

2.34

2H

12.55

29.61

2.22

2H

12.00

148.80

6.01

1H

11.32

137.93

204.33

124.97

5.97

1H

16.37

10

160.72

5.87

1H

16.40

11

37.84

12

15.19

1.54

3H

13

11.76

1.79

3H

14

24.38

1.07

3H

15

24.17

1.20

3H

2. Tng hp mt s dn xut ca zerumbone v kho st hot tnh chng


ung th ca chng
Zerumbone l mt serquyterpen xeton vng ln, , khng no, c cng
thc chung C15H22O, cu to phn t hnh 3.1, tn gi ha hc l 2,6,9,9tetramethyl- (E,E,E)- 2,6,10- cycloundecatrien-1- on. Hp cht cacbonyl ,
khng no trong t nhin l loi hp cht c hot tnh sinh hc rt l th v
c cc nh khoa hc trn th gii c bit quan tm. Zerumbone l mt v d
in hnh v c trnh by trong phn tng quan, hot tnh sinh hc ni bt
ca zerumbone l c ch mnh s pht trin ca t bo ung th theo c ch
apoptosis, loi tr NF- kB hot ng. Nhm chc sinh hc ca zerumbone l
nhm cacbonyl , khng no.
Vo nm 2010 Uraiwan Songsiang v cng s ch ra rng dn xut 3amino, 3- amino th c hot tnh chng s pht trin ca 5 dng ung th ti mt
khc nhau nng IC50 75 M trong khi nng ny zerumbone khng
c tc dng- y l mt kt qu th v [42]. Theo hng ny ngi ta tng hp
rt nhiu dn xut ca zerumbone nhm nng cao hiu qu ca zerumbone. Cu
to phn t ca Zerumbone cho thy c nhiu kh nng tng hp cc dn xut
ca Zerumbone nh: dn xut ca nhm cacbonyl, dn xut ca lin kt i lin
hp vi nhm C=O, dn xut ca lin kt i c lp (C6=C7) v cc dn xut
ca oix ho m vng. ng u trong chuyn ho Zerumbone l cc nh khoa
hc Nht Bn [25].
Theo hng chuyn ha Zerumbone thnh cc hp cht c hot tnh sinh hc
chng ti amin ha zerumbone bng amoniac v isopropylamin.

3. Kho st hot tnh chng ung th ca 3-isopropylamino-Zerumbone


Nhm lm r hiu qu v hot tnh chng cc ung th: ung th gan HepG2; ung th phi- Lu v ung th c tim RD ca dn xut 3-isopropylaminozerumbone so vi zerumbone chng ti kho st hot tnh chng ung th ca 2
hp cht ny i vi 3 dng ung th k trn.Kt qu c th hin trong bng sau:
7

Bng 1: Kh nng gy c t bo ca 3-isopropylamino-Zer v Zer


K hiu mu

IC50 (g/mL)
HepG2

Lu

RD

Zerumbone

0,31

1,00

0,36

3-isopropylamino-Zerumbone

2,23

>5

>5

Kt qu trn cho thy hot tnh chng ung th ca dn xut 3-isopropylaminozerumbone yu hn nhiu so vi Zerumbone i vi 3 dng ung th th. iu
ny cng cho thy nu h thng lin kt i lin hp vi nhm cacbonyl trong
phn t zerumbone b gim th hot tnh chng ung th ca zerumbone i vi 3
dng ung th HepG2,Lu v RD cng b gim.

References
TING VIT

1. Bo co tng kt ti QGTD0309-2006 thng 2


2. Bo co k nim 45 nm thnh lp Vin Dc liu TW 9-5-200
3. L Trn c (1997), Cy thuc vit nam. Trng hi ch bin tr bnh ban u,
tr 291-292, NXB nng nghip, H ni
4. V vn Chi (1997), T in cy thuc Vit nam, tr 536, NXB Y hc.
5. V Vn Chi, Dng c Tin (1987), Phn loi thc vt. Thc vt bc cao, tr
461-464, NXB i hc v Trung hc chuyn nghip, H ni
6. Trnh nh Chnh (1995), Nghin cu thnh phn ha hc ca tinh du mt s
cy thuc h Gng (Zingiberaceae) Vit nam, Lun n PTS Khoa hc v
Ha hc, Trng H S phm H ni I.
7. Phm Hong H (1999), Cy c Vit nam, quyn 3, tr 553, NXB Tr H Ni.
8. Tt Li (2005), Nhng cy thuc v v thuc Vit nam, in ln th mi ba, tr
368- 369, NXB Y hc H Ni.

9. L nh Mi, Lu m C, Trn Minh Hi, Trn Huy Thi, Ninh Khc Bn


(2002), Ti nguyn thc vt c tinh du Vit nam, tp 2, tr 94-119, NXB
Khoa hc v K thut, H Ni.
10. Nguyn Quang Thch (2009), C s cng ngh sinh hc tp 3, Cng ngh sinh
hc t bo, trang 343-344, Nh xut bn gio dc Vit Nam.
11. Vn Ngc Hng, Th Thanh Thy (2006), Gng gi (Zingiber zerumbet.
Sm) vng Tam o, mt nguyn liu qu cho iu ch Zerumbone, Tp ch
Khoa hc v cng ngh, tp 44, s 3, trang 65-69.
TING ANH

12. A. Hoffman, L. M. Spetner and M. Burke (2002), Redox-regulated


mechanism may account for zerumbones ability to suppress cancer-cell
proliferation, Carcinogenesis 23 (11): 1961-1962.
13. Abdul, A.B.H., A.S. Al-Zubairi, N.D. Tailan, S.I.A. Wahab, Z.N.M. Zain, S.
Ruslay and M.M. Syam (2008), Anticancer activity of natural compound
(zerumbone) extracted from Zingiber zerumbet in human HeLa cervical
cancer cells, International Journal of Pharmacology 4, (3), 160--168
14. Ahmad Bustanmam Abdul (2009), US2009/0239953
15. Bokyung Sung, Sonia Jhurani, Kwang Seok Ahn, Yoichi Mastuo, Tingfang Yi,
Sushovan Guha, Mingyao Liu, and Bharat B. Aggarwal (2008),
Zerumbone Down-regulates Chemokine Receptor CXCR4 Expression
Leading to Inhibition of CXCL12-Induced Invasion of Breast and
Pancreatic Tumor Cells, Cancer Res, 8938-8944.
16. Buckingham J (1982), Dictionary of Organic compounds, New YorkLondon-Toronto, Chapman and Hall, 5, 5763.
17. Dai.J.R., Cadellina II.J.H., Mc Mahon.J.B., and Boyd.M.R (1997),
Zerumbone, an HIV-inhibitory and cytotoxic sesquiterpene of Zingiber
aromaticum and Z. zerumbet, Natl. Prod. Lett, 10, 115-118.
18. Dae Sik Jang, Ah-Reum Han, Gowooni Park, Gil-Ja Jhon, Eun-Kyoung Seo
(2004), Flavonoids and aromatic compounds from the Rhizomes of
Zingiber zerumbet, Arch Pharm Res 27, (4), 386-389.

19. Dai J.R. et al (1997), Nat. Prod. Lett 10, 115-118.


20. Dev S. (1960), Tetrahedron 8, 171-180.
21. Dung.N.X., chinh.T.D., Piet A.Leclereq (1995), Chemical investigation of the
acrial part of Zingiber zerumbet (L.) Sm. from Vietnam, Jour. Essent. Oil
Res.(USA), 7 (2), 153-157
22. Huang.G.C., Chien.T.Y., Chen.L.G., Wang.C.C (2005), Antitumor effects of
zerumbone from gingiber zerumbet in P-388D1 cells in vitro and in vivo,
Planta Med, 71 (3), 219-224
23. Kirana C, McIntosh GH, Record IR, Jones GP. (2003), Antitumor activity of
extract of Zingiber aromaticum and its bioactive sesquiterpenoid
zerumbone, Nutr Cancer.45(2):218-25.
24. Matthes.H.W.D., Luu.B., Ourisson.G (1980), Cytotoxic components of Zingiber
zerumbet, curcuma zedoaria and C. domestica, Phytochemistry, 19, 2643-2650.
25. Masuda.T., Jitoe.A., Kato.S., Nakatani.N (1991), Acetylated flavonol glycosides
from Zingiber zerumbet, Phytochemistry, 30, 2391-1392.
26. M. Golam KADER, M. Rowshanul HABIB, Farjana NIKKON, Tanzima
YEASMIN, Mohammad A. RASHID, M. Mukhlesur RAHMAN, Simon
GIBBONS (2010), Zederone from rhizomes of Zingiber zerumbet and
staphylococus activity, Boletn Latinoamericano y del Caribe de Plantas
Medicinales y Aromticas, 9 (1), 63 68.
27. Mohamed Yousif Ibrahim , Ahmad Bustamam Hj Abdul, Tengku Azmi
Tengku Ibrahim, Siddig Ibrahim AbdelWahab, Manal Mohamed Elhassan
and Syam Mohan (2010), Attenuation of cis platin- induced nephrotoxity
in rats using Zerumbone, African Journal of biotechnology Vol 9, 44344444.
28. Murakami.A, Takahashi.M., Jiwajinda.S., Koshimizu.K., Ohigashi (1999),
Identification of zerumbon in Zingiber zerumbet Smith as Potent inhibitor
of

12-O-tetradecanoylphorbol-13-acetate

induced

Epstein-Barr

virus

activation, Biosci Biotechnol Biochem, 63 (10), 1811-2.


29.

Murakami.A.,

Takahashi.D.,

Kinoshita.T.,

Koshimizu.K.,

Kim.

H.W.,

Yoshihiro.A., Kakamura.Y., Jiwajindo.S., Terao.J., Ohigashi.H (2002),

10

Zerumbone, a southeast Asian ginger sesquiterpene, markedly suppresses free


radical generation, proinflammatory protein production, and cancer cell
proliferation accompanied by apoptosis the alpha, beta-unsatureted carbonyl
group is a prerequisite, Carcinogenesis, 35 (5), 795-802
30. Nakamura.Y., Yoshida.C., Murakami.A., Ohigashi.H., Osawa.T., Uchida.K
(2004), Zerumbone, a tropical ginger sesquiterpene, activates phase II
drug metabolizing enzymes, FEBS Letters, 13, 572 (1-3), 245-250
31. N.P. Damodaram and Sukh Dew (1965), Stereochemistry of Zerumbone,
Tetrahedron Letters, 24, 1977-1981.
32. Ozaki.Y., Kawahara.N., and Harada.M (1991), Anti-inflammatory effect of
Zingiber cassumunar.Roxb. and its active principles, Chem.Pharm. Bull,
39, 2353-2356.
33. Samir Kumar Sadhu, Amina Khatun, Takashi Ohtsuki, Masami Ishibashi
(2007), First isolation of sesquiterpenes and flavonoids from Zingiber
spectabile and identification of Zerumbone as the major cell groth
inhibitory component, Natural product research, 21, (14), 1242-1247.
34. Sal S.R. et al (1981), Aust J..Chem. 34, 243-247.
35. SA Sharifah Sakinah, S Tri Handayani and LP Azimahtol Hawariah (2007),
Zerumbone induced apoptosis in liver cancer cells via modulation of
Bax/Bcl-2 ratio, Cancer Cell International 7, 1186.
36. Takahashi.S et al (2002), Japanese Journal of cancer and chemotherapy, 27, 70.
37. Takashi Kitayama et al (2001), Chemistry of Zerumbone, part 2 Regulation
of Ring bond Cleavage and unique antibacterial activities of Zerumbone
derivatives, Biosci, Biotechnol. Biochem. 65, 2193-2199.
38. Takashi Kitayama et al (2003), The chemistry of Zerumbone part 5
Structural transformation of the dimethylamine derivatives, Tetrahedron
59, 4857-4866.
39. Takashi Kitayama et al (2002), The chemistry of Zerumbone IV. Asymetric
Synthesis of Zerumbol, Journal of Molecular Catalysis 17, 75-79.

11

40. Takashi Kitayama et al (2007), Synthesis of a novel inhibitor against MRSA


and VRE: preparation from Zerumbone ring opening material showing
histidine-kinase inhibition, Bioorganic and Medicinal chemistry Letter 17,
1098-1101.
41. Takada Y, Murakami A, Aggarwal BB. (2005), Zerumbone abolishes NFkappaB and IkappaBalpha kinase activation leading to suppression of
antiapoptotic and metastatic gene expression, upregulation of apoptosis,
and downregulation of invasion.
42. Uraiwan Songsiang et al. (2010), Cytotoxicities against cholagiocarcinoma
cell lines of Zerumbone derivatives, European Journal of Medicinal
Chemistry 45, 3794-3802.
43. Vimala.S., Norhanom.A.W., Yada.M (1999), Anti-tumour promoter activity
in Malaysian ginger rhizobia used in traditional medicine, British Journal
of cancer, 80, 110-116.

12

You might also like