GII THIU LUN N

1. Tnh cp thit ca lun n

Nghin cu, tm kim cc hp cht c ngun gc thin nhin nh
sesquitecpen lacton c hot tnh sinh hc cao ng dng trong y hc,
nng nghip v cc mc ch khc trong i sng con ngi l mt trong
nhng nhim v quan trng v ang c cc nh khoa hc trong v
ngoi nc ht sc quan tm.
Vi s pht hin ra nhiu cht c hot tnh sinh hc c gi tr t thin
nhin, cc nh khoa hc c nhng ng gp ng k trong vic to ra
cc loi thuc iu tr nhng bnh nhit i v bnh him ngho nh:
penicillin (1941); artemisinin (nhng nm 1970); ...., ko di tui th v
nng cao cht lng cuc sng ca con ngi. Thin nhin khng ch l
ngun nguyn liu cung cp cc hot cht qu him to ra cc bit dc
m cn cung cp cc cht dn ng tng hp ra cc loi thuc mi. T
nhng tin cht c phn lp t thin nhin, cc nh khoa hc chuyn
ho chng thnh nhng hot cht c kh nng tr bnh rt cao.
Sesquitecpen lacton c bit n nh mt nhm ln cc sn phm
t nhin c phn lp ch yu t h Asteraceae, Artemisia Lp cht
ny c cu trc v hot tnh sinh hc rt a dng v phong ph. Mt trong
s sesquitecpen lacton thin nhin c quan tm nghin cu l hp cht
parthenin.
Cy Parthenium hysterophorus vi hot cht chnh l parthenin
thuc khung pseudoguainolide c bit n vi cc hot tnh sinh hc
nh khng l amit, chng st rt, khng ung th do chng ti chn
parthenin lm nguyn liu u nghin cu chuyn ha mt s dn xut
ca parthenin nhm tm kim nhng hot tnh sinh hc ca cc dn xut
tng hp.
2. Mc ch ca lun n
- Nghin cu chit tch v xc nh cu trc ca parthenin;
- Tng hp cc dn xut ca parthenin;
- Xc nh cu trc ho hc ca cc dn xut chuyn ha c;
- Thm d hot tnh sinh hc ca chng
3. Nhng ng gp mi ca lun n
ti nghin cu hon ton mi khng trng lp vi cc cng trnh
lun n cng b vi nhng im ng gp mi sau:
- Nghin cu v chit tch, xc nh cu trc ca parthenin t hoa
ca cy Parthenium hysterophorus v tng hp dn xut mi ca
parthenin, chuyn ha c sesquitecpen lacton khung pseudoguainolid
1

vi h vng -lacton v -metyl ca dihydroisoparthenin;

- Tng hp c 35 dn xut ca parthenin trong c 27 dn xut
mi v thm d hot tnh HIV v gy c t bo ca 09 dn xut.
- Tng hp 03 dn xut bng phng php kh ha v tm cc iu
kin ti u kh ni i v tr C-13 to h vng -lacton v -metyl
ca dihydroisoparthenin 220. Tip tc kh ha thu c hn hp ng
phn ca (4R, 4S)-hydroxy-dihydroisoparthenin 221 v 222;
- T Parthenin 6 kh ha thu c 05 dn xut gm Hymenolin 235,
(11S)-dihydrohymenolin 236, (11R)-dihydrohymenolin 237, (11R-axetyl)dihydrohymenolin 238 v (11S-axetyl)-dihydrohymenolin 239. Cc dn
xut 235 - 239 c xc nh cu trc bng cc phng php ph v so
snh vi kt qu nghin cu ca cc cht c cng b.
- tng hp c 09 dn xut este mi 223-231 t (4R)-hydroxy
dihydroisoparthenin v (4S)-hydroxy-dihydroisoparthenin.
- tng hp c 03 dn xut mi 232-234 c cha lin kt N-O
v 06 dn xut mi 240-245 c cha lin kt N-N ca parthenin.
- tng hp c 09 dn xut 246-254 ca khung pseudoguainolid
vi h vng -lacton v -metyl khng lin hp ca parthenin trong c
07 dn xut mi v 02 dn xut c cng b trong ti liu tham kho.
- Th hot tnh khng HIV v gy c t bo ca 09 dn xut ca
parthenin v so snh vi hot tnh ca Stavudin (D4T) v Zidovudin
(AZT). Kt qu th hot tnh sinh hc khng HIV ca 09 hp cht thp
hn D4T v AZT nhng t c hn D4T v AZT.
4. B cc ca lun n
Lun n bao gm 99 trang vi 03 bng, 45 s , 32 hnh v bao
gm cc phn: M u (2 trang); Tng quan (24 trang); Thc nghim (33
trang); Kt qu v tho lun (39 trang); Kt lun (2 trang); Danh mc cc
cng trnh cng b ca tc gi; 85 ti liu tham kho; Phn ph lc.
NI DUNG CHNH CA LUN N
Chng 1: TNG QUAN
Trn c s nghin cu ti liu, phn tng quan lun n cp n:
1.1. Vi nt v sesquiterpen lacton
1.2. Hot tnh sinh hc ca sesquitecpen lacton
1.3. Parthenin, dn xut ca parthenin v hot tnh sinh hc
Chng 2-THC NGHIM
2.1. Ha cht v trang thit b
2.2. Chit tch, tinh ch v xc nh cu trc ca parthenin t hoa ca cy
Parthenium hysterophorus.
2

2.3. Tng hp mt s dn xut ca parthenin.

2.4. Th hot tnh khng HIV v gy c t bo ca 09 dn xut ca
parthenin v so snh vi hot tnh ca Stavudin (D4T) v Zidovudin
(AZT).
Chng 3- KT QU V THO LUN
3.1. Tch v xc nh cu trc ha hc ca Parthenin
3.1.1. Quy trnh tch parthenin
T Hoa ca Parthenium hysterophorus (5 kg) c sy kh, xay nh
sau chit vi MeOH (5 L) trong thit b chit soxhlet. Dch chit th
MeOH c c c thu c (97 g); Cn chit MeOH c tch sc k ct
trn ct silica gel vi h dung mi ra gii chloroform: MeOH (99:1 n
65:35) theo t l tng dn thu c parthenin 6 (2,1 g, hiu sut 0,3%) l
cht rn mu trng c nhit nng chy 155-156 oC.
3.1.2 Xc nh cu trc ca parthenin:
Ph 1H-NMR xut hin cc tn hiu ca 17 proton, cc tn hiu c
trng cho nhm metyl (ppm) 1,13 (3H) ca C-15; 1,29 (3H) ca C-14;
proton c trng ca nhm CH2 ti C-8, C-9 v nhm CH ti C-10 c
xc nh ti 1,67-2,36 (5H, m); proton v tr C-7 ti 3,51 (1H, m);
Da vo hng s tng tc gia cc tn hiu v ph HSQC xc nh
c cc proton tng tc vi nhau v tr cacbon tng ng: 5,01 (1H ,
d, J = 7,9 Hz, H-6); 5,60 (1H, d, J = 2,6 Hz, H-13); 6,19 (1H, d, J = 5,7
Hz, H-3); 6,28 (1H, d, J = 2,6 Hz, H-13); 7,52 (1H, d, J = 5,7 Hz, H-2).

15

HO

2
3

10

1
5

9
8

6
14

7
11
12

13

Hnh 3.1. Ph 1H-NMR ca hp cht 6

3

Ph 13C-NMR gm 15 tn hiu c trng cho 15 nguyn t cacbon

khung sesquitecpen lacton, kt hp vi ph HSQC v HMBC xc nh
c v tr ca cc nguyn t cacbon c trng cho hp cht parthenin:
17,4 (C-15); 18,3 (C-14); 28,3 (C-8); 29,8 (C-9); 40,6 (C-10); 44,2 (C-7);
59,1 (C-5); 78,7 (C-6); 84,4 (C-1); 121,6 (C-13); 131,7 (C-3); 140,4 (C11); 163,0 (C-2); 170,7 (C-12) c trng cho cc bon tr C-12 lin kt
lacton; 210,5 (C-4) c trng cho cc bon v tr C-4 lin kt C=O.
Ph HSQC cho thy s tng tc trc tip H-C ca 15 nguyn t cc
bon v proton tng ng vi cc v tr, ph HMBC cho bit cc tng tc
xa khng nh thm cu trc ca parthenin. T kt qu nghin cu ph
1
H-NMR, 13C-NMR, HSQC, HMBC, EI-MS v so snh vi ti liu tham
kho chng minh hp cht nhn c l parthenin.
3.2. Kh ha parthenin to thnh cc dn xut.
HO

HO

HO

NaBH4
O

3
4

221

220

OH
10

1
5

H2
Pd/C
t=8 h

9
8

6
14

222

11
12

HO

HO

NaBH4

MeOH/THF
HO

H
O

13

HO

H2
Pd/C
t=20 h

235

236

HO

237
HO

HO

+
O

t=10 h

15

HO

HO

H2
Pd/C

MeOH/THF
HO

O
O

O
O

220

246

247

S 3.1. S kh ha parthenin bng cc tc nhn khc nhau

3.2.1. Phn ng kh ha parthenin v chuyn v ni i
Parthenin 6 c kh ha di tc dng ca H2 v Pd/C 10% trong
MeOH thu c dihydroisoparthenin 220 vi hiu sut 78%. Ph 1H-NMR
ca hp cht xut hin tn hiu 1,83 (3H, s) c xc nh ca nhm metyl
ti C-13, proton v tr C-7 ti 3,51 (1H, m) ca parthenin sau khi b kh
ha khng xut hin trong ph.
Ph 13C-NMR c s chuyn dch ca cc v tr cc bon C-7 t 44,2
ppm xung trng thp 162,7 ppm. Nhm metylen b kh ha thnh nhm
4

metyl v tr C-13, tn hiu trn ph cc bon cho thy s chuyn dch ha

hc t 121,6 ppm ln trng cao 11,1 ppm. Nh vy, parthenin sau khi
kh ha bng H2 trn Pd/C 10% trong MeOH thu c sn phm ch yu
l kh ni i C1=C2 thnh hai nhm CH2 v c s chuyn dch ni i ca
C-13 vo pha trong to lin kt i ti C7=C11.
v tr C-2 v C-3 khng cn tn hiu c trng cho ni i ca
parthenin m chuyn dch v pha trng cao c gn cho nhm CH 2.
S chuyn dch chuyn dch ha hc cc v tr ph hp vi cu trc ca
hp cht 220 v c so snh vi ti liu tham kho.
HO

HO

HO

NaBH4, MeOH

t = 16 h
to = 25 oC

HO

220

HO

221 (52%)

O
O

222 (28%)

S 3.2: S chuyn ha cc dn xut 221 v 222

Cht 220 c kh ha bi tc nhn NaBH4, xc tc DMAP trong
MeOH thu c hn hp ng phn ca (4S)-hydroxydihydroisoparthenin
(221) v (4R)-hydroxy-dihydroisoparthenin (222) (s 3.3) vi hiu sut
tng ng l 52% v 28%.

Hnh 3.2. Ph 1H-NMR ca hp cht 221

5

Ph 1H-NMR xut hin cc tn hiu ti (ppm) 5,55 (1H, s) c

trng cho proton H-6; 4,80 (1H, m) c gn cho proton H-4; 2,69-1,31
(9H, m, H-2, H-3, H-8, H-9, H-10); Cc nhm metyl ca hp cht 221
c xc nh 1,81 (3H, s, Me-13); 1,11(3H, d, J = 7,5Hz, Me-15), 0,65
(3H, s,Me-14).

15

HO

2
3

HO

10

1
5

9
8

6
14

7
11
12

13

Hnh 3.3. Ph ESI-MS ca hp cht 221

Ph khi lng ion ha bi in t (ESI-MS) c pic gi phn t
[M+1]+ = 267,1 tng ng vi cng thc phn t C15H22O4.
Ph DEPT ca hp cht 221 xut hin tn hiu ti (ppm): 86,15
c trng cho nhm CH v tr C-4 sau khi kh ha nhm C=O chuyn
dch t pha trng thp t 215,5 ppm v pha trng cao.
Ph 13C-NMR ca hp cht 221 xut hin tn hiu 164,17 ppm c
trng cho cacbon v tr C-7 sau khi c s chuyn dch ni i t ngoi
vo trong vng ca parthenin.
xc nh c nhm hydroxy v tr C-4 vi cu hnh hay
da vo ph NOESY. Ph NOESY ca hp cht 221 c tn hiu tng tc
ca H-4, H-6 v nhm hydroxy v tr C-1 nn nhm hydroxy v tr C-4
c xc nh v tr .
Ngc li, hp cht 222 trn ph NOESY proton H-4 khng c
tng tc vi H-6 v nhm OH v tr C-1 nhng li c tng tc vi Me14 v Me-15. Do , hp cht 222 c xc nh nhm hydroxy v tr .

15

HO

2
3

HO

10

1
5

9
8

6
14

7
11
12

13

Hnh 3.4. Ph NOESY ca hp cht 221

Hnh 3.5. Ph 1H-NMR ca hp cht 222

7

3.2.2. Kh ha parthenin to thnh hymenolin, dihydrohymenolin v dn

xut axetat.
Parthenin 6 c kh ha di tc dng ca H2 vi s tham gia ca
Pd/C thu c Hymenolin 235, tip tc kh ha di tc dng ca NaBH4
trong hn hp dung mi MeOH/THF thu c hn hp ng phn (11S)dihydrohymenolin 236 (55%) v (11R)-dihydrohymenolin 237 (32%). Dn
xut ca hn hp ng phn dihydrohymenolin c axetyl ha thu c
(11R-axetyl)-dihydrohymenolin 238 (85%) v (11S-axetyl)-dihydrohyme
-nolin 239 (82%).
Hp cht 235: Ph 1H-NMR c tn hiu ca cc nhm metyl H
(ppm) 1,13 (3 H, d, J = 7,6 Hz, Me-15); 1,31 (3 H, s, Me-14); 1,31 (3 H, d,
J = 7,6 Hz, Me-13); Nhm metylen v tr C-13 b kh ha v metyl cho
tn hiu duplet vi J = 7,6 Hz. Ph 13C-NMR xut hin tn hiu C (ppm)
41,6 c trng cho C-11 chuyn dch v pha trng cao; 210,4 c trng
cho cacbon v tr C-4.
HO

HO

HO

MeOH/THF (2/1)
+
O

NaBH4, 0 oC, 1 h
O

HO

HO

235

236 (55%)

237 (32%)

S 3.3: Kh ha hymenolin to thnh dihydrohymenolin

Hnh 3.6. Ph 1H-NMR ca hp cht 239

8

Hp cht 236: Ph 13C-NMR c tn hiu C (ppm) 84,92 c trng

cho cacbon C-4 chuyn dch v pha trng cao.
xc nh c nhm hydroxy v tr C-4 vi cu hnh hay
da vo ph NOESY ca dn xut sau khi axyl ha.
HO

HO

(CH3CO)2O
Py, 25 oC, 6 h
HO

AcO

238 (82%)

237

HO

HO

(CH3CO)2O
Py, 25 oC, 6 h

HO

AcO

O
O

236

239 (85%)

S 3.4: Phn ng axyl ha dihydrohymenolin

Hnh 3.7. Ph 1H-NMR ca hp cht 238

9

OH

H
AcO

H
O
O

Hnh 3.8. Ph NOESY ca hp cht 239

Ph NOESY ca hp cht 239 c tn hiu tng tc ca H-4, H-6, H13 v nhm hydroxy v tr C-1 nn nhm hydroxy ca hp cht 236 v
tr C-4 c xc nh l v tr .

Hnh 3.9. Ph NOESY ca hp cht 238

10

Ngc li, ph NOESY ca hp cht 238 proton H-4 khng c tng

tc vi H-6, H-13 v nhm OH v tr C-1 nhng li c tng tc vi Me14 v Me-15. Do , hp cht 237 c xc nh nhm hydroxy v tr .
3.2.3. Kh ha parthenin v to thnh hp cht 246 v 247
HO

HO

HO

HO

H2, Pd/C
MeOH, 20 h, 25 oC
O

O
O

O
O

220 (75%)

O
O

246 (7%)

O
O

247 (3%)

S 3.5: S kh ha parthenin to thnh cc hp cht 246 v 247

Parthenin 6 c kh ha bng H2 vi xc tc Pd/C thu c hp
cht 246 (7%) v 247 (3%). Ph 1H-NMR ca hp cht 246 xut hin
nhng tn hiu c trng cho s kh ha ti v tr C-13. Proton H-13 ca
hp cht 6 c s chuyn dch ha hc v pha trng cao 1,2 (3H, d, J= 7,5
Hz, H-13). Ph 13C-NMR c cc tn hiu c trng cho s chuyn dch ca
C-7 t 162,7 ppm v pha trng cao 48,19 ppm v C-12 t 175,13 ppm v
pha trng cao 42,7 ppm.

HO

O
O

Hnh 3.10. Ph 1H-NMR ca hp cht 246

11

Xc nh nhm metyl ti v tr C-13 bng ph NOESY. Proton H-6

khng c tng tc vi cc nhm metyl ti cc v tr C-15, C-14 v C-13.
Cc nhm metyl ti v tr C-14 v C-15 c xc nh cu hnh cng
pha v c tng tc vi nhm metyl ti v tr C-13 nn nhm metyl ti v
tr C-13 c xc nh v tr
Tng t hp cht 246, Ph 1H-NMR ca hp cht 247 xut hin
nhng tn hiu c trng cho s kh ha ti v tr C-13.
Ph APT c cc tn hiu c trng cho s chuyn dch ca C-7 t
162,7 ppm v pha trng cao 46,36 ppm v C-12 t 175,13 ppm v pha
trng cao 42,4 ppm.
Sau khi kh ha parthenin to thnh cc hp cht sesquitecpen lacton
thuc khung pseudoguainolid vi h vng -lacton v -metyl tin hnh
tng hp cc dn xut nhm tm kim nhng hp cht c hot tnh sinh
hc l th. Tng hp cc dn xut este, dn xut amit v azit ti v tr C-4
ca parthenin.

HO

O
O

Hnh 3.11. Ph NOESY ca hp cht 246

12

3.2.4. Chuyn ha cc este ca parthenin

3.2.4.1. Chuyn ha cc este ca (4S)- hydroxy-dihydroisoparthenin 221
O H

OH

O
H
O
O

223 (70%)
E3N, DMAP
CH2Cl2, 25 oC, 4 h

O H

OH

OH

OH

H
E3N, DMAP
CH2Cl2, 25 oC, 2 h

O
O

224 (75%)

O
E3N, DMAP
CH2Cl2, 25 oC, 2 h

HO

H
H

Cl

221

E3N, DMAP
CH2Cl2, 25 oC, 8 h

225 (75%)

E3N, DMAP
CH2Cl2, 25 oC, 2 h
E3N, DMAP
CH2Cl2, 25 oC, 8 h

O H

OH

O H

O H

OH

O
H

OH

Br

MeO
O

228 (63%)

OMe

227 (62%)

226 (72%)

S 3.6: S chuyn ha (4S)- hydroxy-dihydroisoparthenin 221

Tng hp cc dn xut este ca (4S)-hydroxy-dihydroisoparthenin
221 bng phn ng vi anhydric axit tng ng trong mi trng kim
Et3N v xc tc DMAP thu c cc dn xut 223-228.
Nhn vo ph 1H-NMR ca cc hp cht 223 cho thy r s chuyn
dch ca proton H-4 ca hp cht 221 ti H 4,80 ppm v pha trng
thp. Khung ca hp cht 221, kt hp vi d liu ph ca hp cht 223:
Ph 1H-NMR xut hin tn hiu ca nhm axetyl ti H (ppm) 2,07 (3H, s,
CH3-C=O ); proton H-4 chuyn dch t 4,80 ppm v pha trng thp 5,70
ppm cho thy s lin kt ca nhm axetyl vi oxi ca nhm hydroxy ti v
tr C-4.

13

OH

HO

+
H

OH

E3N, DMAP

H
O

CH2Cl2, 25 oC, 2 h

221

224 (75%)

S 3.7: S chuyn ha hp cht 221 to thnh 224

Hp cht 224: Ph 1H-NMR xut hin tn hiu ca nhm isobutyl ti
H (ppm) 1,23-1,17 (6H, dd, 2xCH3); proton H-4 chuyn dch t 4,80 ppm
v pha trng thp 5,67 ppm cho thy s lin kt ca nhm isobutyl vi
nhm hydroxy ti v tr C-4.
OH

HO

O
Cl

+
H

OH

CH2Cl2, 25 oC, 2 h

Cl

E3N, DMAP

H
O
O

Cl

221

225 (75%)

S 3.8: S chuyn ha hp cht 221 to thnh 225

Hnh 3.12. Ph 1H-NMR ca hp cht 225

Hp cht 225: Ph 1H-NMR xut hin tn hiu ca vng thm th
14

1, 4 ti H (ppm) 7,87 (2H, d, J = 9 Hz, H-2 v H-6); 7,56 (2H, d, J = 9

Hz, H-3 v H-5); Proton H-4 chuyn dch t 4,80 ppm v pha trng
thp 5,77 ppm cho thy s th ca nhm th 4-clobenzoyl vo nhm
hydroxy ti v tr C-4.
OH

HO

O
Cl

+
H

OH

CH2Cl2, 25 oC, 2 h

Br

E3N, DMAP

Br

221

226 (72%)

S 3.9: S chuyn ha hp cht 221 to thnh 226

Hp cht 226: Ph 1H-NMR xut hin tn hiu ca vng thm th 1,
4 ti H (ppm) 7,87 (2H, d, J = 9 Hz, H-2 v H-6); 7,55 (2H, d, J = 9 Hz,
H-3 v H-5); proton H-4 chuyn dch t 4,80 ppm v pha trng thp
5,76-5,79 ppm cho thy s th ca nhm th 4-bromobenzoyl vo nhm
hydroxy ti v tr C-4.
HO

OH

CH2Cl2, 25 oC, 8 h

OMe

OMe

221

OH

E3N, DMAP

Cl

+
H

O H

227 (62%)

S 3.10: S chuyn ha hp cht 221 to thnh 227

Hp cht 227: Ph 1H-NMR xut hin tn hiu ca vng thm th 1,
3 ti H (ppm) 7,61 (1H, dd,, J = 7,5 v 2,5 Hz, H-6); 7,55 (1H, dd J = 1,5
v 2,5 Hz, H-2); 7,32 (1H, t, J = 7,5 Hz, H-5); 7,07 (1H, ddd, J = 2,5; 1,5
v 7,5 Hz, H-4); 5,76 (1H, dd, J = 6 v 9,5 Hz, H-4); proton H-4 chuyn
dch t 4,80 ppm v pha trng thp 5,75-5,78 ppm cho thy s lin kt
ca nhm th 3-metoxybenzoyl vi nhm hydroxy ti v tr C-4.
HO

OH

MeO

E3N, DMAP

Cl

+
H

O H

OH

O
H

CH2Cl2, 25 oC, 8 h
MeO

O
O

221

228 (63%)

S 3.11: S chuyn ha hp cht 221 to thnh 228

Hp cht 228: Ph 1H-NMR xut hin tn hiu ca vng thm th 1,
4 ti H (ppm) 7,97 (2H, d, J = 9 Hz, H-2 v H-6); 6,90 (2H, d, J = 9 Hz,
15

H-3 v H-5); proton H-4 chuyn dch t 4,80 ppm v pha trng thp
5,74 (1H, dd, J = 6 v 9,5 Hz, H-4) cho thy s lin kt ca nhm th 4metoxybenzoyl vi nhm hydroxy ti v tr C-4, nhm metoxy gn vng
benzen c xc nh ti 3,84 (3H, s, OCH3).
3.2.5. Chuyn ha cc este ca (4R)- hydroxy-dihydroisoparthenin 222:
O H

OH

O
H
O
O

229

O H

OH

OH

OH

HO
H

H
H

Br

O
O

Cl

231

230

222

S 3.12. S chuyn ha (4R)- hydroxy-dihydroisoparthenin 222

Hp cht (4R)-hydroxy-dihydroisoparthenin 222 vi nhm hydroxy
v tr C-4 c cu hnh cng c phn ng vi mt s anhydric v dn
xut clorua thu c hp cht 229-231.
OH
HO

+
H

OH

O
O

Et3N, DMAP
CH2Cl2, 25 oC, 6 h

H
O
O
O

229 (70%)

222

S 3.13: S chuyn ha hp cht 222 to thnh 229

Nhn vo ph 1H-NMR ca cc hp cht 229 xut hin nhng tn
hiu c trng cho cu trc ca sn phm.
Ph 1H-NMR xut hin tn hiu ca nhm axetyl ti H (ppm) 2,09
(3H, s, CH3-C=O); proton H-4 chuyn dch t 4,80 ppm v pha trng
thp 4,92 ppm cho thy s lin kt ca nhm axetyl vi nhm hydroxy ti
v tr C-4.
Hp cht 230: Ph 1H-NMR xut hin tn hiu ca nhm 4clobenzoyl ti H (ppm) 7,91 (2H, d, J = 9 Hz, H-4 v H6); 7,39 (2H, d,
J = 9 Hz, H-3 v H-7); 5,77(1H, bs, H-6 ); proton H-4 chuyn dch t
4,80 ppm v pha trng thp 5,10 (1H, dd, J = 6 v 9,5 Hz, H-4) cho thy
s lin kt ca nhm 4-clobenzoyl vi nhm hydroxy ti v tr C-4.
16

OH
HO

Cl
O

+
H

OH

H
O

Et3N, DMAP

CH2Cl2, 25 oC, 8 h

O
O

Br
Br

222

231 (72%)

S 3.14: S chuyn ha hp cht 222 to thnh 231

Hp cht 231: Ph 1H-NMR xut hin tn hiu ca vng thm th 1,
4 ti H (ppm) 7,91 (2H, d, J = 9 Hz, H-4 v H-6); 7,39 (2H, d, J = 9 Hz,
H-3 v H-7); proton H-4 chuyn dch t 4,80 ppm v pha trng thp
5,09 ppm cho thy s lin kt ca nhm 4-brombenzoyl vi nhm hydroxy
ti v tr C-4.

Hnh 3.13. Ph 1H-NMR ca hp cht 231

3.2.6. Tng hp (4)-oxim-dihydroisoparthenin 232 v cc dn xut
HO

HO

HO
NH2OH. HCl
EtOH, Et3N, 80 oC, 2 h
O

Et3N, 25 oC, 8 h
HON

R
O

O N

O
O

220

232 (91%)

233: R=Me (78%)

234: R=Ph (81%)

S 3.15: S tng hp (4)-oxim-dihydroisoparthenin 232, dn xut

233 v 234
17

Dn xut cha mt ni t ca dihydroisoparthenin c chuyn ha

vi s tham gia phn ng ca hydroxylamin trong mi trng Et 3N thu
c (4)-oxim-dihydroisoparthenin 232 vi hiu sut cao 91%. Hp cht
trung gian 232 c este ha thu c hp cht 233 (78%) v 234 (81%).

Hnh 3.14. Ph 1H-NMR ca hp cht 233

Hp cht 232: Ph 1H-NMR c tn hiu H (ppm) 5,4 (1H, s, H-6)
c trng cho proton v tr C-6; 1,89 (3H, s, Me-13) c gn cho nhm
metyl v tr C-13 v 1,08 (3H, d, J = 8,0Hz, Me-15) xc nh nhm metyl
v tr C-15 v 0,70 (3H, s, Me-14) c gn cho metyl v tr C-14. Ph
13
C-NMR c tn hiu C (ppm) 215,17 c gn cho cc bon v tr C-4.
Hp cht 233: Ph 1H-NMR c tn hiu H (ppm) 5,52 (1H, s, H-6)
c trng cho proton v tr C-6; 2,26 (3H, s,Ac) c xc nh ca nhm
axetyl; 1,82 (3H, s, Me-13) c gn cho nhm metyl v tr C-13; v
1,05 (3H, d, J = 8,0Hz, Me-15) xc nh nhm metyl v tr C-15; v 0,97
(3H, s, Me-14) c gn cho metyl v tr C-14.
Hp cht 234: Ph 1H-NMR c tn hiu H (ppm) 8,01-7,41(5H, Bz)
c trng cho vng benzen; H (ppm) 1,82; 1,05; 0,97 c xc nh ca
nhm metyl; 1,82 (3H, s, Me-13) c gn cho nhm metyl v tr C-13;
v 1,05 (3H, d, J = 8,0Hz, Me-15) xc nh nhm metyl v tr C-15; v
0,97 (3H, s, Me-14) c gn cho metyl v tr C-14.

18

Hnh 3.15. Ph 1H-NMR ca hp cht 234

3.2.7. Tng hp (4)-hydrazin-dihydroisoparthenin 240 v cc dn xut
Phn ng ca dihydroisoparthenin 220 vi hyrazin trong EtOH d
dng thu c hp cht 240 vi hiu sut cao 94%. Nguyn liu trung gian
240 cha nhm amin phn ng vi anhydrit axit tng ng thu c dn
xut 241 (80%), 242 (72%) v phn ng vi mt s dn xut benzandehit
thu c dn xut 243 (71%), 244 (70%) v 245 (90%) (s 3.18).
OH

Boc

H
N N

(Boc)2O
O

OH

OH

N
NH2

Py, 50 C, 4 h

N
NH
O

242 (72%)

240
O

245 (90%)

Py, 90 oC, 2 h
HOOC

Py, 25oC, 16 h

O
OH

N
NH
O
O

241 (80%)

S 3.16: S tng hp cc hp cht 241-245

19

Hp cht 241: Ph 1H-NMR c tn hiu H (ppm) 2,32 (3H, s,

COCH3) c xc nh gn vi nhm (NH-).
Hp cht 242: Ph 1H-NMR c tn hiu H (ppm) 1,24 (6H, dd, J = 4
v 8 Hz, 2 x Me-3) c trng cho hai nhm metyl ca hp cht 242.
CHO

OH

N
NH2

OH

N
N

R
Toluen, 90 oC, 2 h

243-244

240

243: R=p-metoxy (71%)

244: R=p-axetyl (70%)
S 3.17: S tng hp cc hp cht 243-244
Hp cht 243: Ph 1H-NMR c tn hiu H (ppm) 8,37 (1H, s, H-1)
c trng cho nhm CH gn vi vng thm, 7,98 (2H, d, J = 9 Hz, H-3 v
H-5); 6,91 (2H, d, J = 9 Hz, H-2 v H-6) c trng cho th 1,4 vng
benzen, 3,83 (3H, s, OCH3) c gn cho nhm metoxy gn vng benzen.
Hp cht 244: Ph 1H-NMR c tn hiu H (ppm) 8,33 (1H, s, H-1)
c trng cho nhm CH gn vi vng thm, 7,70 (2H, d, J = 9 Hz, H-3 v
H-5); 7,83 (2H, d, J = 9 Hz, H-2 v H-6) c trng cho th 1,4 vng
benzen, 2,62 (3H, s, COCH3) c gn cho nhm axetyl gn vng benzen.
Hp cht 245: Ph 1H-NMR c tn hiu H (ppm) 1,52 (9H, s, Boc)
c trng cho nhm Boc. Ph khi lng ESI-MS vi pic ion phn t m/z
= 379. Ph 13C-NMR cho thy s chuyn dch ha hc v tr C-4 ca
dihydroisoparthenin C (ppm) 215,5 (C=O) v 167,8 c trng cho nhm
(N=C).
3.2.8. Tng hp cc dn xut ca 246
HO

HO
NH2OH.HCl

Py, EtOH, 70 oC, 4 h

O
O

HOHN

O
O

246

248 (76%)

S 3.18: S tng hp cht 248

T hp cht 246 v hydroxylamin clorua trong mi trng kim Py,
phn ng c thc hin iu kin nhit 70 oC trong thi gian 4 gi
20

thu c hp cht 248 vi hiu sut (76%). Ph 1H-NMR ca hp cht cho

thy cc tn hiu (ppm) 5,08 (1H, d, J=8,5 Hz); 2,73 (1H, q, J= 10 Hz);
2,59-2,51 (2H, m); 2,39-2,33 (2H, m); 2,11-2,06 (2H, m); 1,88-1,81 (2H,
m); 1,58-1,52 (2H, m); 1,26 (3H, d, J= 7 Hz, H-13); 1,2 (3H, s, H-14); 1,14
(3H, d, J = 7,6 Hz, H-15); c trng ca hp cht 248.
Tng hp dn xut ca hp cht 248 phn ng vi clorua axitvi s
tham gia ca xc tc trietylamin trong dung mi diclometan thu c dn
xut 250 v 251.
HO

HO
O
HOHN

Et3N, CH2Cl2,
Cl

25 C, 2 h

O
248

O N
O

O
O

250: R = C6H5 (58%)

251: R = CH3 (65%)

S 3.19: S tng hp cc cht 250 v 251

T nguyn liu u 248 v benzylclorua trong mi trng kim
trietylamin, phn ng c thc hin nhit 25oC trong 2 gi thu c
hp cht 250. Cu trc ca hp cht 250 c xc nh bng ph 1H-NMR
vi cc tn hiu xc nh vng thm C6H5 (ppm) 8,03 (2H, dd, J = 1,2
Hz, 8,2 Hz); 7,56 (1H, td, J = 1,2 v 7 Hz); 7,43 (2H, t, J = 8 Hz); tn hiu
5,26 (1H, s, H-6); 1,35 (3H, s, H-14); 1,30 (3H, d, J= 7 Hz, H-13); 1,18
(3H, d, J = 7,5 Hz, H-15).
Hp cht 251: Cu trc ca hp cht 251 c xc nh bng ph
1
H-NMR vi cc tn hiu xc nh (ppm) 5,14 (1H, s, H-6); nhm axetyl
2,21 (3H, s, Ac); v cc nhm meyl 1,27 (3H, s, H-14); 1,16 (3H, d, J = 7
Hz, H-15);
3.2.9. Tng hp cc dn xut 247
T hp cht 247 v hydroxylamin clorua phn ng trong mi trng
baz Py thu c 249 (hiu sut 73%). Cu trc ca sn phm c xc
nh bng ph 1H-NMR vi nhm hydroxy lin kt vi NH ti 8,2 (1H, s);
proton H-6 ti 4,89 (1H, d, J = 5,6 Hz); cc proton thuc khung parthenin
2,91-2,86 (2H, m); 2,82 (1H, q, J = 10 Hz); 2,61 (1H, dd, J = 9 Hz); 2,542,47 (1H, m); 2,15-2,11 (1H, m); 2,08-2,02 (1H, m); 1,67-1,62 (2H, m);
1,53-1,49 (2H, m); 1,25 (3H, s, H-14); 1,16 (6H, d, J = 7 Hz, H-15 v H13);

21

HO

HO

NH2OH.HCl
O

Py, EtOH, 70 oC, 4 h

HOHN

247

249 (73%)

S 3.20: S tng hp cht 249

HO

HO

HO

Ac2O
Py, 25 oC, 4 h

NH2NH2
O

EtOH, Py H2N N
80 oC, 4 h

HN N

O
O

252 (96%)

249

O
O
253(84%)

Toluen
80 oC

HO
O
HO

N N
HO

O
O

254 (60%)

S 3.21: S tng hp cc hp cht 252- 254

T nguyn liu u 249 tng hp cht trung gian 252 bng cch tc
dng vi hyrazin trong EtOH vi hiu sut cao 96%. Sn phm trung gian
252 cha nhm amin bc 1 d phn ng vi axetic anhydric thu c dn
xut 253 vi hiu sut 84%.
Cu trc ca sn phm 253 c xc nh bng ph 1H-NMR
(ppm) 8,53 (CH3NH-); 5,0 (1H, d, J = 8 Hz, H-6); 2,55-2,45 (2H, m); 2,382,31 (2H, m); 2,28 (3H, s); 2,15-2,05 (2H, m); 1,94-1,87 (1H, m); 1,781,73 (1H, m); 1,63-1,58 (2H, m); 1,30 (3H, d, J = 7 Hz, H-13); 1,12 (3H, s,
H-14); 1,16 (3H, d, J = 7 Hz, H-15); v ph 13C-NMR (ppm) 180 (C-1);
174 (C-12); 165 (C-4); 85,8 (C-1); 82,5 (C-6); 56 (C-5); 48,5 (C-7); 42,8
(C-11); 42,2 (C-10); 34,9 (C-3); 30 (C-9); 25,6 (C-2); 22,5 (C-2); 20 (C2); 18,6 (C-13); 18,2 (C-14); 16 (C-15).
Nhm tm kim dn xut c cha vng thm, hp cht 252 cho tc
dng vi p-hydroxybenzandehyt thu c hp cht 254 vi hiu sut 60%.
Cu trc ca sn phm c xc nh bng ph 1H-NMR (ppm) 8,82
(1H, -CH=); 7,6 (2H, d, J = 7,5 Hz ); 6,8 (2H, d, J = 7,5 Hz ); 5,17 (1H, d,
22

J = 8 Hz); 2,88-2,82 (1H, m); 2,65-2,56 (2H, m); 2,55-2,46 (1H, m); 2,392,34 (1H, m); 2,15-2,09 (2H, m); 1,96-1,91 (1H, m); 1,74-1,7 (1H, m); 1,61,5 (4H, m); 1,33 (3H, s); 1,27 (3H, s); 1,17 (3H, s).
3.3. Hot tnh khng HIV v kh nng gy c t bo
Hin nay, nhm cht quan trng nht ca cc cht chng HIV/AIDS,
gm c zidovudin (AZT), stavudin (d4T), didanosin (ddI), lamivudin
(3TC)... Cc nucleozit ny c phosphat ho nh enzym kinase to thnh
5-triphosphat v c gn vo mch ADN ca virus nh enzym phin m
ngc ca HIV.
Cc hp cht este ca cc thuc (cc tin cht) c nghin cu rt
nhiu bi v chng c nhng c tnh l tng do chng c bn hp l
trong in vitro, d dng c thy phn nh qu trnh trao i cht. iu
cho php to ra nhng thuc c thi gian bn hu thch hp nn c nh
hng nghin cu chuyn ha cc dn xut este mi ca parthenin.
Hot tnh khng HIV ca cc dn xut c xc nh theo phng
php so mu MTT (the tetrazolium-based colorimetric). Cc php th hot
tnh ny c thc hin ti Vin nghin cu virut Tibotec, Vng quc B.
Hot tnh khng HIV c th trn t bo MT-4 gy nhim bi chng
HIV-1 IIIB. Kh nng gy c t bo c th nghim trn dng t bo
MT4-CMV-EGFP vi cc cht chun l cc thuc chng HIV/AIDS
bit v cht khng HIV phin m ngc AZT v D4T.
3.4. Hot tnh sinh hc ca cc dn xut tng hp t parthenin
Parthenin v cc dn xut ca n c xc nh c nhng hot tnh
sinh hc l th nh khng l amit, st rt, ung thNhm tm kim nhng
hot tnh sinh hc ca cc dn xut tng hp c t parthenin, chng ti
th hot tnh sinh hc khng HIV trn dng IIIB v hot tnh gy c t
bo trn dng t bo MT4-CMV-EGFP ca 09 dn xut tng hp c.
3.5. Kt qu th hot tnh khng HIV
Kt qu th hot tnh khng HIV cho thy gi tr EC50 ca hp cht
243 cao nht 97 M/ml v cc hp cht khc cho gi tr > 197 M/ml
th hin hot tnh khng HIV ng ch .
Kt qu th hot tnh sinh hc khng HIV ca 09 hp cht thp hn
D4T v AZT nhng t c hn D4T v AZT.
KT LUN
Trong qu trnh thc hin lun n ny, nghin cu sinh thu c
cc kt qu nghin cu v chit tch, xc nh cu trc ca parthenin t
hoa ca cy Parthenium hysterophorus v tng hp dn xut mi ca
23

parthenin, chuyn ha c sesquitecpen lacton khung pseudoguainolid

vi h vng -lacton v -metyl ca dihydroisoparthenin.
T nguyn liu u parthenin tng hp c 35 dn xut ca
parthenin trong c 27 dn xut mi, thm d hot tnh HIV v gy c
t bo ca 09 dn xut.
Tng hp 03 dn xut bng phng php kh ha v tm cc iu
kin ti u kh ni i v tr C-13 to h vng -lacton v -metyl
ca dihydroisoparthenin 220. Tip tc kh ha thu c hn hp ng
phn ca (4R, 4S)-hydroxy-dihydroisoparthenin 221 v 222 dng tng
hp cc dn xut ca parthenin:
- tng hp c 09 dn xut este mi 223-231 t (4R)-hydroxydihydroisoparthenin v (4S)-hydroxydihydroisoparthenin;
- tng hp c 03 dn xut c cha lin kt N-O mi ca
parthenin: 4-oxim-dihydroisoparthenin 232; (4)-dihydroisoparthenin(1)axetyl oxime 233; (4)-dihydroisoparthenin-(1)phenoxy oxime 234.
T Parthenin 6 kh ha thu c 05 dn xut gm Hymenolin 235,
(11S)-dihydrohymenolin 236, (11R)-dihydrohymenolin 237, (11R-axetyl)dihydrohymenolin 238 v (11S-axetyl)-dihydrohymenolin 239. Cc dn
xut 235 - 239 c xc nh cu trc bng cc phng php ph v so
snh vi kt qu nghin cu ca cc cht c cng b.
tng hp c 06 dn xut c cha lin kt N-N mi ca
parthenin: (4)-hydrazin-dihydro isoparthenin 240; Acetohydrazindihydroisoparthenin
241;
3,3-dimethyl-4-oxobutanoic
hydrazin
dihydroisoparthenin 242; 4-methoxylbenzyl-hydrazin-dihydroisoparthenin
243;
4-axetylbenzyl-hydrazin-dihydroisoparthenin
244;
tbutyloxicarbonyl-hydrazin-dihydroisoparthenin 245.
tng hp c 09 dn xut 246-254 ca khung pseudoguainolide
vi h vng -lacton v -metyl khng lin hp ca parthenin trong c
07 dn xut mi v 02 dn xut c cng b trong ti liu tham kho.
Th hot tnh khng HIV v gy c t bo ca 09 dn xut mi ca
parthenin v so snh vi hot tnh ca Stavudin (D4T) v Zidovudin
(AZT). Kt qu cho thy hot tnh khng HIV thp hn D4T v AZT
nhng t c tnh hn D4T v AZT.
24