Nghiên cứu phương pháp tổng hợp Stavuđin, một loại thuốc chống HIV có hiệu quả hiện nay

DAI HOC QUOC GIA HA
IMOI
TRUdNG
DAI HOC KHOA HOC TU NHIEN
BAG CAO KET QUA
THUC
HIEN
D^
TAI DAC BIET
CAP DAI HOC QUOC GIA HA NOI 2001-2003
Ten de tdi:
NGHIEN
CIJU PHirONG
PHAP TONG HOP STAVUDIN
MOI
LOAI THUOC CHONG HIV CO HIEU QUA HIEN NAY
Mflso.QGOl-04,
•
'••
r
Chd nhiem
de tdi: GS.
TSKH.
Nguyen
Dhih
Trieu
T^LlNGTA*^i•!1(•.'
.!iN
\' v.
iirK
mJs^

HA NOI, 2004
MUC LUC
I.
Ma
dau.
II.
Tom
tat
ket qua chfnh.
_L
Ket qua ve mat khoa hoc.
2.
Ket qua phuc vu thuc te.
.*
3.
Ket qua dao tao.
4.
Ket qua nang cao tiem
luc
khoa hoc.
5.
Tinh hinh
su dung kinh
phi.
Abstract
in.
Phan noi dung chfnh.
1.
Dat van
d^

2.
Tong quan vi cac van de nghien
cifu
2.1.
Vong dai
ciia
HIV.
;
2.2.
Cac loai thudc chong HIV.
2.3.
Phiro'ng
phap chung
t6ng
hop stavudin (D4T)
va/idovudin
(AZT) 1
2.4. Phuo'ng phap long hap thymidin. 1
3.
i^huang
phap
nghien cu'u
va
kci
qua
3.1
Tong hap Stavudin (D4T) va Zidovudin (AZT) I
3.2
Tdng
hop thymin va uraxin 2

3.3 Tdng hap
2'-hidroxithvmidin
2
3.4 Tdng hap mot sd hap chat di vong co hoat tfnh sinh hoc. 2
3.5
Thir
hoat tfnh khang
khuan
khang nam. 2
IV.
KclluAn.
3
V.
KiC'M
nghi.
3
Tai lieu
tham khao 3
Phieu dang ky
ket
qua
nghien ciru
khoa hoc
'W'nu lal
c;ic
cdng
liliih nghien ciiii
khoa hoc
ciia
ca nhan

Phil liic
Phu luc
1:
Phd hdng ngoai
Phu luc 2: Phd cdng hirdng
lu"
hat nhan
I.
M6
DAU.
Ten de tai:
"Nghien
ctru phirang phap tdng hap stavudin, mot
loai
thudc chdng
HIV
CO
hieu qua hien
nay".
Ma sd:QG
01-01
Caquan:
Dai hoc Qudc gia Ha noi
Chu
tri:
GS. TSKH.
Nguyen Dinh
Trieu
Dan vi: Trudng Dai hoc Khoa
hpc

Tu nhien
Cac thanh vien:
1.
GS. TSKH.
Nguyfin Dinh
Trieu
2.
Thac
sT
Doan Duy Tien
3.
Thac
sT
Tran Thi Thanh Van
Thdi gian
ihirc
hien: 2 nam (tu thang
5-2001)
H.
TOM TAT KET QUA CHINH.
Di tai:
"Nghien
cuu phuang phap tdng
hgp
stavudin, mot
loai
thudc chdng HIV co hieu
quii hicn nay".
Chfnh thuc dugc phe duyet
la

de tai dac biet cap Dai hgc Qudc gia Ha noi vao thang
nam 2001 vdi thdi han 24 thang va tdng kinh phf
60
trieu ddng. Den nay de lai da
dm
hoan thanh va dat dugc nhung ket qua nhu sau:
1.
Kel qua ve mat khoa hoc.
• Da nghien cuu long hgp dugc
.V-ZO-lhymin
cho
2\3'-(hdeoxy-2\3
didchidrothymin (Stavudin, D4T). Phan ung dugc tdng hgp
theo
hai budc. buc
dau
lien
khda nhdm chiic -OH d vi trf
5'-
ciia
Deoxyriboza bing Uic nhan
RSO2C
sau do tach loai nudc
bang
lac nhan RONa irong dung moi ancol de lhu
due
stavudin (D4T).
• Da nghien cuu tdng hgp dugc chat
l-(3-azido-2J-dideoxy-P-[
crythropentofuianosyl)

thymin (AZT). Qua trinh phan ung dugc thuc hien theo
h;
budc,
budc dau tien can phai khda nhdm chuc -OH d
C-5'
cua vdn
pcnlofuranoza bAng mot tac nhan khda Z-Y (nhu trityl
(C(^U<,);^CC
iriankylsunfonylclorua
RSO2CI ).
Budc hai
tij'san
pham nhan dugc ciia phan un
Ircn
cho
phixn
ung
liep
vdi nalriazit cho san pham cudi cung la AZT.
• Da
nghien
cuu tdng hgp dugc thymin,
nguyen lieu
dfiu tdng hgp cac chat thuc
chdng HIV, di tu axil oxalic va clanol dat hieu suat cao (han 80%).
• Da
iij'liieii ciiii
ldng hgp
ihrgc
3'-hidroxilhymin

cpia
phan ung
ciia
Ihymin vdi
thii
ngan clorua cho
N-mccuaclorualhymin.
rdi sau dd cho phan ling licp vc
ieIiaa\eiyl-P-riboriiiano/o'.
•
loii!'
hop
(liuir
mol
st"i
(ian
xufii eiia
o\a(Ii;i/ol.
o\a(liia/.ol
va
Iria/ol co
hoat lin
khang khuan va
kliaiig
nam.
• Cau
true
ctia cac san phiim tdng hgp dugc da dugc xac dinh qua cac phuang pha
phd hdng ngoai, cong hudng
tu'

hat nhan va phd khdi lugng.
• Dfl
Iham
dd llur
hoai li'nh
sinh
h()c ciia
mol sd
chai
di vdng tdng hgp
ductc
va
tha
chung cd uic dung khang khuan. khang nam.
• Ciic cdng
irinh
khoa hoc
dfi
dugc cdng bd :
1/
Phan
Dinh
Long,
NguyCm Dinh
Trieu.
Tdng hgp
vii diic
trung phd cua mot sd
dSn xuiYl
furan

Tiip
chl Hda hoc.
T.39,
sd 1, 27-30 (2001).
2/
Nguyen Dinh
Trieu.
Doiin
Duy Tien. Tnrang Quang Trung
Tdng hgp Thymin.
Iloahoc
ung
dung,
sd
9.
16(2003).
3/
Nguyen
Dhih
Trieu.
Tain
Thi Phuang Thiio
Tdng
h(;p.
xiic
dinh
ci'iu
tnic
vii
ihii"

hoiit linh
sinh hoc
ciia
mot sd diln
xuii't
the
(
\'l trf
sd5
eua
2-innino-l
,3.4-lhiazol Vii
1.2-diammo-l
,3.4"lria/.ol.
Biio Ciio d Hdi nghi khoa
hc;)c vii
cong nghe Hda huu ca
loiin
qudc
htn
thu
;
(2{)()1).
Tuyeu
Ulp
e;ie edng Irinh hdi nghi khoa hoe
\';i ediig
nghe Hda
lurii ecr
tr 9'

(2001).
4/
Nguydn
Dinh Trieu, Truang Quang Trung
Tdng hgp mot
sddiln xuiit
day
2-furyIidenxiniimoyl
hidiiizon
Biio
Ciio
d Hgi nghi Hda hgc va Cdng nghe Hda Huu co
lOiin
quoc
Iiin
thu
(2001).
5/
Doiin
Miinh Chfit,
Nguyen Dinh
Trieu.
•
Tdng hgp
vii
tfnh toan mot sd thdng sd CiYu
tiio ciia
5-iU"yl-2-mecapto-l,3,'
oxadiazol.
Tuyen tap cac cdng trinh khoa hoc

ki
niem 45
nilm
ngay thanh lap Khoa
He
hgc,
Truc^ng
Dili hgc Tdng hgp
Hii
ndi
(2001)
6/ Pham Van Chiic, Nguyen Thiiy
HAng,
Nguyen
Dinh
Trieu
Tdng hgp. nghien
ciVu
cau tao
ciia
mgl sd
dSn xuiYl aryliden-]
tolylsunfonylhidrazon vii fomiizan
chua nhdm
arylsunfonyl.
Biio Ciio d hgi nghi khoa
hocTrudng
Dili hoc Khoa hgc Tu
nhien,
DHQG

H
ngi,
ngiinh
hcxi
hgc 2002.
2.
Kci
<|u;i
phuc
vii
thuc te.
De
liii dii
ldng hop
Ihiinh
cdng chi'it stavudin (D4T)
vii
zidovudin (AZT)
lii loiii
thuc
duc;c dung phd bien hien
niiy Iren ihe
gidi va trong nudc de
dieu
tri benh HIV-AIDS. T
kel quii
niiy,
neu cd dieu kien tiep tuc nghien cuu quii trinh cdng nghe
vii
thu nghiem

lAr
siing Iren
benh
nhfin
cd the siin
xuitt thucic
stavudin (D4T)
vii
zidovudin (AZT) trong nud
ihay
cho nhap ngoai
riil diil
lien.
3.
Ket qua dao lao.
DUii Viio
ngudn kinh
phi ciia dC liii dfi
dao
tiio
dugc 02 thiic sy va 07 cu
nhfin
hda hgc :
• Nguyen Tluiy
Hiuig.
biio ve
Ihiic
sy
niim
2002

cV
hoi ddng Trudng Dili hgc Kho
hoe
T'lr
nhien.
Hii
noi.
•
\)i)l\\\
Duy Tien.
Ix'm
ve
Ihae
sy nam 2003 d hoi ddng Tnrdng Diu hoe Khoa hoe
Ti
nhien.
Hii
ngi.
• Ciic sinh vien
dii
hoan
[\vdvh
khdii
luan tdt nghiep : Tnrong Quang Trung,
TiTin
Th
Phuang
Thiio,
Dang The Anh, Nguyen Hii:
Doiin,

NguySn
Viln
Hii,
Phimg
Thi
Ti
Anh. Nguyen Viln Hieu.
•
Dii huc)'ng
diln mot sd sinh vien lam nghien
cii'u
khoa hgc.
4.
Kc'l
<|ua nang cao
licni hic
khoa hoc.
Tren ea sd
Ihue hien
de
uii dii diio liio
dirge 02
lluie
sy irong dd cd thac sy
Doiin
Du^
Ticn,
Iil
ngudi thuc hien chfnh
cila

de tai, da thi diU hgc bdng cua Bg giiio
due.
gui di dac
lao Tien Sy d Cong hda Lien bang
Due.
Pang ngudn kinh
plif dC
lai da
muii Ihem
dung eu
vii
hda chitt dd thue hien
66
tai
v;
diio liio Ihiic
sy
Vil
cu
nhiln
hda hoc.
5.
Tinh
hinli sii
dung kinh
phi.
Tdng kmh
i^hf
diroe cilp
2

lulm hi
60 trieu (mdi nam la 30 Iricu ddng). Kinh phf
chi
yeu
dilng miui
dung cu hda chill
vil
thuc mudn nhiln edng.
Den
nay da
quvel loiin
dugc
khoang 47 6S0 000 ddng, cdn
hii
se quyei
loiin
sau khi nghiem lhu.
ABSTRACT
Acquired immunodeficiency syndrome (AIDS) is he resull of an infection by
humii
immunodeficiency virus(es) (HIV). This retrvivus shows a specific tropism
fc^r
the
heipe
inducr T cells leading to their depiction. The resultant immunosuppression
predispose
HIV patients to
life-ihrealcning
opportunistic infections.
The present project relates to a process for the synthesis of

2\3'-didcoxy-2\3
didehydro-P-D-erythropentofuranosylthymin,
also
known
iis
2\3'-dideoxy-2\3
didehydrothymidine or stavudin (D4T), a drug anti HIV-AIDS.
In this project the study on the synthesis of
3'-iizido-3'-deoxythymidine
or AZT
drug
anli
HIV-AIDS is also realized.
In
iiddilion,
some derivatives of
1,3,4-oxiidiiizol,
1,3,4-oxathiiizol,
1.3,4-triiizc
andlhcir biological
iiclivitics
will be studied.
The struclure of
all
producls
will
identified by the modern spectroscopy.
Xac nhan cua Trirdng Dili
ht)c
hien

,
HlL'.
TRUONG
lla
noi,
ni^ayX^thang
3
ndm
2004
Chu nhiem de tai
GS.
TSKH.
Nguyen Dinh
Trieu
GS.TS.
J'Adu
,'hi/ii.
Xiic nhan cua Dai hoc Qudc gia
H;i
nc)i
HI.
FHAN
N()I
DUNCi CHINH.
Ten
de
tiii:
^'Nghien
curu phuang phap tdng hop stavudin, mot loai thuoc chdng
HIV cd hieu qua hien

nay"'.
MilscK
"
Q(i
01-01
Chii
nhiem de tai: GS.TSKH. Nguyen Dinh Trieu
Thdi gian thuc hien: 24 thang
(5/2001-5/2003)
Kinh phf: 60 trieu dong
1.
Diit
van de.
HIV-AIDS hicn nay
hi
mgl
ciln
benh (he
ki,
hilng
m;l'y
ehiic
Iricu ngucri
iren
the gic
da bi nhiem va nhieu ngudi bi cudp di sinh menh. 0 nudc la cung
vily,
tren lilt cii
ciic tin
Ihilnh

phd trong Cii nudc diu cd ngudi bi
nhi^m
HIV, mot sd da chuyen sang AIDS va
che
vl Ciln benh
niiy
1I-7J.
Vice
dieu tri ddi hdi phai su dung ciic
loiii
thudc diil
li6n,
tdn ker
vl giii Ihudc rill diit do dd sd ngudi dugc dieu tri bilng cac
loiii
lhuoc dac
hieu
rill
hiin
che
nirc'yc hi
hien nay.
Vice lir
siin
xiull
ra ciic
loiii
thudc
uily
irong nuac cd the hit giil thiinh,

nuck
la hien
cilng
cd
vili
xf nghiep siin
xuitt
nhirng
chii
yeu
hi
gia cdng ddng gdi, cd
lhuoc
vAn
nhap do
66
giii thanh
vAn
cao. Neu nhu nghien
ciiu
dugc cdng nghe san xui
dugc Ciic
loiii
thudc tren trong nudc
chad
chan giii ihanh thudc se ha han giii
nhilp
ngoai.
Trong sd' ciic thudc dieu
Iri

HIV-AIDS hicn nay duac
sir
dung r()ng
ifii dieu
tri
ben
IIIV-A IDS iren the gicri
va d
Viel
nam
lii
cap AZT (Zidovudin)
vii
D4T (stavudin)
[7-I6J.
Muc lieu
ciia
66 lai la
butifc
diiu
tim hieu
cdng nghe siin
xuAl
AZT
vil
D4T
vil
lien
hilni
ldng hop Irong phdng

ihf
nghiem. Cho den nay cdng nghe siin xuat hai
loai
thudc tren
vili
dirc;c
ciic nuac siin
xufil
giu
kin,
iren
ciic
piitent chi
neu
ciie
nguyen
tfic
chung do dd
chiin;
ldi sau khi tham khiio
liii
lieu phiii tien hiinh ciic thf nghiem
thilm
dd,
tim phuung
phiip
ch
hieu
suiil idt
nhill.

2.
Tdng quan ve cac ViVn de nghien
ciiu.
2.1.
Vong dot
ciia
HIV[1-7J.
HIV
(hiunan immimodeficicne
virus) la mot retro virut cd vat lieu di truyen la RN/
(axil nbonuclcic). HIV ed khii nilng nhiem nhieu te bao
limpoT,
te bao mono,
diii
thuc ba(
LimpoT hd trg la loai hay bi virut lily
nhi^m
nhat vl chiing cd chua chat chi diem
CD^
ircn
mill le bao. Trong quii trinh nhiln len
ciia
HIV cd mdt sd chiing
chii
chdl:
•
'InriVc
lien
vinii lllV phiu
gan rdi

hdii va'i
lap
niiliig lij")il ciia
le
biio
limpoT hd
Irg
ihdng
qua
iuang
hie giua glycoprotein gp 120 d lap vd cua HIV va receptor DC4
trei
mill le bao T hd trg (le bao CD4).
• Tu
miieh
dan RNA
ciia
virut hinh thanh
miich
kep DNA nhd vai trd
ciia
enzim
phiei
mil nguc/c
reverse
Iranscripliise,
•
Miiu
DNA
niiy

dugc hda nhiip vao bd miiy nhiem
silc ciia
te bao
vilt chii
nhd
enziiT
integiiise.
• DNA hda nhap
niiy
(lien virut) qua ca che phien ma binh
Ihirdng cila
le bao.
tiio
ri
nhieu RNA mdi
ciia
virut
vii nigl
sd protein khiic
ciia
virut nhu gp 41. gp 120, p 24
p
17 Vil
mgl sd enzim. Nhung protein nay dugc xu ly che bien bdi enzim proteaza.
Md
Ihe
he
mc)l vinil
sau khi
ihiuin

phuc dugc giiii phdng. Mdi chang khiic nhau
ciia
qui
trinh
nhan Ien cua HIV nhat
la
cac enzim phien ma ngugc deu
la
cac dfch de thudc c
Ih^
tac dong.
2.2,
Cdc loai thudc chdng HIV.
Nhung nghien
ciiu
vi quii trinh
di6u
tri HIV-AIDS cho thiiy mot sd hgp chill thuc
loai
nuclcosit tu
nhien
nhu ihymidin,
uiiixin,
xiticiin.
iidcnosin
cd khii
uilng
uc che s
phiit trien
ciia

virut HIV[l-7].
Ciic
loiii
thudc tren dugc phfin theo tiic dung dieu tri nhu sau:
2.2.1
Thudc ngan chan su biim dfnh va xam nhap cua le
biio
vat
chii
TCD4 cua virut HIV.
Khi virut HIV
vilo
dugc ca thd ngudi, trudc tien ciic protein bi milt GP 120
(glucoz
protein)
ciia
virut HIV tien sat vao be mat
CLia
te bao vat
chii
TCD4 cua ngudi
Vil
thiiy dt
hinh
diing. Trong khi bien diing chiing de
Id
khiic udn
V^
cd khii nang chui
niio

te
biic
Dua Vilo yeu
id
khdng giiin
niiy
co the
sir
dung mot sd hda chill dd uc che su
xilm
nhap
cii
virulHIV.
Loiii
thudc tieu
bieu
cho tac dung nay
la
CD4 (ten hda hgc:
2\3'-didcoxy-2',3'
didehidrocytidin).
Cdng thirc cau
tiio
NH-
N
HO
O
N
,0.
^

">
^
liude lie
che
ICMI
men
pliilu
giiii
diim ciia virul
HIV.
Khi virut HIV vao dugc
be
bao TCD4 cila ngudi cluing
sir
dung
loiii
men phan giii
diim
(enzym
proleaza)
dugc dung cho viec long hgp ra protein
ciui
tnic loi cua virut HIN
giiij) eliiiiig
sinh
InrcVng iiliaiih
chdng. Mdt sd hap
chAl
sau cd khii nilng lie che enzyn
proleii/a ciia

virul HIV :
a.
Indiiiiivia
(Indinavir) :
Ten hoa hoe : |
1 (1
S.
1 R) SS|^2.3.5-lndeo\i-N(2.3 (lihidro-2-hicIr()\i-1-II-indcn-1
yl)-5 (2[(l.l-dimclylclyl)
anionoj cacbonyL4(3-pyridinylmetyl)-l-pii)erazinyI)-2
phenylmelyl]-D-erylhropcnloamil.
Cdng
Ihiic
philn
111
C^.H^nNsOf,
l„
=
L53-
I54"C
Tiic dung : lhuoc
Idp
hidroxiaminopentan
iimit
mdi Uong cac thudc
lic che
proteazi
ciia HIV I Iil loiii
thudc chdng virul.
b.

Rilonavia (Ritonavir)
Ten hda hoe :
|5S-(3\8R\lOR\l lR^)]10-hidroxi-2-metyl-5-(I-metyIetyI)-4
lhiazolyl)-3,6-dioxo-8,l
i-bis(phenylmetyl)-2,4,7,l2-lclraiizalridecan-l
3-dioxo-
cS.l
1- bis
(phcnyimctyl)-2.4.7.12-telraiizatridecan-13-oie-thiazolylmelyl
este.
Cdng
ihii'c
philn
lu
C.7H.,xN(,OsS2
Tiic dung : chill kieu peplit,
li'c
che proleaza
ciia HIVi hi loai
thudc chdng virul.
2.2,3
Thudc uc che men sao chep ngugc (reversetran)
Uc che enzim nay dd ngan
chrin
su tao thanh DNA tu RNA
ciia
virul va do dd
ngiln
HIV khdng gia nhap dugc vao DNA ciia vat chii
vii

n^m
tiem
An
trong nhan te bao. Thudc
di1u lien
ciia
loai
niiy hi
zidovudin (AZT,
Retrovir),
lamivudin (Epivi, 3TC). stavudin
(D4T).
Trong
10 nfim
qua Ciic thudc nhdm nay dugc
sir
dung
nhieu
nlull,
diic biel
hi
AZT.
a. Zidovudin (AZT, Retrovir).
La diln xuat
cila
thymidin, cd nhdm
N^
d vi trf 3'.
Ten hda hgc:
l-(3-azido-2,3-didcoxi-P-D-erylhropcntofuranosyI)-thymih.

(ling ihue
philn
lu:
C'|,,l
I,
iN.,()|.
Cdng
ihirc
ciTu
liio:
:()
Tiic dung : cd
hoiU
tfnh chdng virut HIV rill miinh, tiVn cdng virut khi nd
ihoiit
ra
khdi lc bao
nhidm
benh de nhiem triing Ciic tc bao khiic.
h.
SlaviicJin
(D4T,
Zeril)
Ten hda hoc :
2'.3'-didchidro-3'-deoxithymidin
Cdng thiic philn lu:
CinHpN^Oj.
C
Viii!'
lliii'e

e;lii
lao;
O
N
CH
x>
O
H
HO—I
.0
t,,^
:
165-166"C
(dang tinh the khdng mau)
Tlie dung : chdng virut, uc che enzim phien
mil
nguc;c.
Dily Iil
loiii
thude ed hieu quit
dic^u
tri
HIV
cao, nhill
hi
khi
sii"
dung ket hgp
D4T/3TC/hidinavir.
2.2.4. Mdt sd chill cd

Uie
dung ehdng
HIV
mdi.
Theo mol sd
liii
lieu gfin dily
[16-21]
mdt sd hgp chill day
oxadiiizol,
iriiizol
hoiic
disunfua cd
hoiil
tfnh chdng HIV nhu :
o
II
CH^—C-NH
N-N
Ho'
S-CFL-CO-NHAr
CO-N
/
C.H
6'^5
CO~N
\
CH2
CH2
Q1H5

2.3.
Phuong phdp chung tdng hop stavudin (D4T) vd zidovudin (AZT).
Plurang phiip cluing de tdng hgp D4T
vil
AZT, theo ciic
tiii
lieu cdng bd
ihi deu xuitt
phiii
lir
ihymidin. Phiin ung
diui
ticn
lii
khda nhdm chuc -OH d vi trf 5'-
ciia
vdng
p-iiboIluiinozc:f
bilng ciie
Uic
nhiln
kluie nhiui
; rdi sau dd chuyen hda ihilnh
chuig
(C) cd
Ciiu
etc:
O
ON
110-

Oil
I livinidiii
(/\)
/•IL
/ x
o
;A
zo-
N
O
N'
.0,
Oil
(B)
/
n
:);i/tt
O
IN
O
N-
zo-
0.
/
(C)
Cl
Tiic nhiln khda nhdm chuc Z-X d dily cd the la:
Axil pholphoric
H^PO.j.
- Trilyl

(C^J-iO^CCl.
AnkyI-
vil arylsunfonylclorua
RSO2CI.
Trimetylsilanclorua
(CH3)^SiCI
f)ieii
che AZT bilng
Ciich
cho hgp chill C phiin ung vdi
NaN^,
cdn didu che stavudin cho
nd phiin ling
vc)i
mgl chill
biiza liich loiii
ILO:
MN
O
o
N
C-l
I,
O
HN
O
CM3
x\^\
iO-
>\'^

VJ
N'
ZO-
\
.0.
)
/J-
N,
Zidovucliii
(AZT)
O
HN'
Cl
O^N
10-
,0.
Stavudin
{D4T)
Tiic nhiln
B'
ed the
hi
nalrianeolal
RO*"'Nii'^*
hay mudi
telraiinkyUimonihalogenua
R4N'^'X.
2.4. Phuo'ng phdp tdng hop thymidin.
Thymidin
chii

yeu thu dugc
til"
su thuy philn
nucleosit
biing
xilc
tiic enzim, luy nhien
ve mill ldng hgp cung
dii
dugc nghien cuu di
lir
thymin
vil f5-D-ribofuranozcr
nhu sau:
-I CJhiiyCn
xylose (I) thiinh
1,2-di-0-axelyl-3,-*S-di-()-benzoylofuninose
(lla) hoac
1,2.3,5-letra-O-iixelyIfuranosc
(lib).
HO-
0
T
T
OH
(I)
(a) Ac : axetyl
R : b enzoyl
RO
OH

O
OR
OA-
OAc
( II )
(b) Ac, R: axetyl
+
Chuyen ihymin
(III)
thiinh 0,0-bis(trimetylsilyl)thymin (IV):
^ 0Si(CH3)3
HN
>^
(CH.^.SiCl HN
CH
O
N"
H
hviniM
(CH3)3SiO
N
0.0-his(lriiiiclylsilylj!hymin
(IV)
II
+
Cho
1,2,3,5-tetra-O-axetyIfuranose
phan ung vdi 0,0-bis(lrimetyIsilyl)thymin
ch(
2'-hidroxithymidin (V):

O
/CH,
OSi(CH3)3 |I[SJ
CH,
IO-
.()-
WMc/-'OAc ^
H
OA^
{
II
)
(Cll3)3SiO
N'
DCIICf,,
18h,
t
2)Na/CIl3C)ll
!0
O N
on /
OH
2'-hidr'o.\ilhyniidin
(V)
-f-
Tir
(V) chuyen thiinh thymidin qua Ciic phiin ii'ng sau:
O
HN'^
O^N

HO^^O.
Koii
HL
(CrJ-L),CO,
O
N
CH,
O
i
I
N
Oil
O
N Piridin
/ N
lIBi-
Na.CO,
llO-n
\o
OH
(CFLCOMf AcO-n\o
(V)
Oil
(V»)
O
OAC
(VH)
MN
J
CH,

HN
O
lIUi
O
N
HT/NJRancy
O N
AcO-
.O.
MOn
,o.
7
AcO ^'
CM
Thvmidin
3.
Phirong phiip nghien ci'ru va ket
quii.
3.1.
long hop Stavudin (D4T) vd Zidovudin (AZT).
Dua
iheo
eae
lai lieu
nghien euu ehiing ldi
dii ihue
hien dieu che D4T
vil
AZT theo
Ciie

biroe siiu
:
3.1.1
Tdng hgp
5'-aiylsunfonyloxilhymidin
vii
3\:'*-diaiylsunfonyllhyinidin.
Vc)-i
mue dfch
khdii
nhdm
ehiic
-OH d vi trf
5'-oxi
ciia
ihymidin chiing
tdi dii
chon tiic
nhiln
iuylsunlonNleloruii
eho pluin ling vdi ihymidin:
12
o
HN
O
O
N
AiO->S0
HN
CH3

O^
N
HO-i
O,
LJ
ArSO.Cl
OH
(1)
Of
o
O N
HN
A1O2SO-]
o^
O
0S02Ar
(2)
CH,
CH
Phan ung nilyed
ihd
eho
hdn
hop
(1) vil
(2)
hoile
chi eho ( !)
hoilc
(2)

lily
dieu kien lien
lulnh phan ung. Cluing ldi
dii ihiiy
ddi dieu kien de nhan
dirc;e
(I)
hoilc
(2). hieu
suiit
phan
ling eao
(kluxing
90^,'V').
dang tinh khiei. Siin phfim
dii
dugc chirng minh
cilli Uio
qua
[)huoiig
phiip phd hdng
ngoiii.
edng hudng tu hiU
nluln
prolon
vil
'•'C.
Vf du phd 'H-NMR cua chat
5'-iirylsunfonyloxithymidin
(I):

0 ppm
1.78
2,02
3.SO
4.20
5,20 Vil 6.20
7.20
7.20-7.80
11.40
Nhdm
CH,
CH.
(2')
CIKL)
CH
(4^)
CH(5')
=
CH
Ar
NH
O
4II ^ CIL
'INi
AiO.SO-i^
^O.
4'
/
O
jN

(>
H
3'
2
OH
13
Phd '
t-NMR
ciia
chitt
5'-arylsunfonyIoxithymidin
(1)
5 ppm
21,061
38,323
69.896
70.108
83,171
83,965
109.752
127.501
130.142
132,095
135,838
145,100 •
150,138 C=0
163,597 C=0
Phd
hdn^
ngoai

eiia
5'-aiylsiinfonyloxilhymidin
(I):
Nhdm
CH,
CH2
(5')
CII
(3^)
CH2(2')
CH
(4')
CHd')
=
CH
-
CH Vil
= C-
vdng
ihcfm
N
f'
M
'()!
Nil
V
V,.
,•
V(^-^^(l)enzen)
^'si)2

3370 cm
'
3172
cm"'
172!
em'
1659
cm''
1596,
1477
em-'
1359.
1272.
1 176 cm
o
CW^
4
-IN3
2
CH
AiO.SO
N
('
11
0,
NLJ
3-
2'
OH
Dai

hoc
Ouoc
gta
Tosydliymidinc
Tue Jan 06 22
50
11 2004
9£
'JG-
94
92-
90-
BO-
86-
84-
82-
(ID-
7U-
76-
74-
72-
70-
G8'
66-
G4-
(">2
GO-
2000
Woveruiiiibi'is
(LIII

1)
1000
Pho hdng
ngoiii ciia
S'-arylsunfonyloxithymidhi
(1):
14
-
Dai
hoc Ouoc
gia
- Bis-tosyllhymrdine
- Tue
Jan
06 22 57 44 2004
88
86
84-
82-^
80^
78-^
76-j
74-;
72-|
70-i
G8-i
66-_
QA-
G2-
60-;

50-
2000
WavcnuiTibers (cni-1)
Phd hdng
ngcnii
cua 3',
S'-diarylsunfonyloxithyinidin
(2)
Phd hdng ngoai
cuii
3\5'-diarylsunfonyIoxilhymidin
(2) khdng cdn cd mat nhdm
-Olid
vung .34()0-32()0em'.
Phd 'H-NMR cho
cik dinh
dac trung d
vising
7,2-7,8 ppm, so vdi chat
5'-arylsunfonyIoxithymidin
thi sddinh
nhieu han chiing
id cii
2 nhdm -OH da bi khda.
3.1.2'long
hc)|) Sia\'uclin (D4'r)
Di
til'
5'-arylsunfonyloxilhymidin,
thuc hien phiin ung tiich

loiii H2O ti^r
nhdm -OH d vj
trf 3'- va H
a
vi
irf
2'
khi cd mat cua
biiza
huu ca
uiilriiaicolat
va
tclraiinkylaminium
halo);einia
R.|N'X
Iheo scy
dd
siui:
O
HN"
CH
H
ArOoSO^
O,
w
B-
O
O 'N
HN
CH3

HO-
3'
.0.
OH
5'-;ir>'lsunroinloxilhvniidin
kJ
Stavudin
Nhiel
do ndng chay
l„^j
165
"C.
hieu
suiit:
30-40%
Phd edng hudng
lu
nhiln 'H-NMR
vil
' C-NMR (kel hap giiii phd
2D-C0SY,
HMBC,
IMC^Oeuasiavudin (D4T)
Pho 'H-NMR:
5
ppm
1,72
3.66
4.78
.'i.lO

5,91
6,39
6.81
7,65
•
i'l
5
ppm
12.13
62,25
87,28
88.87
125.90
136.74
150.77
163,84
Nhom proton
-CII,
-CH,
(2')
-CH(4')
-
OH (5')
-CHd')
=
CH(2')
= CH(3')
=
CH (6)
o

"C-NMk:
Nhom
caebon
-
CH,
(5)
-
CH2
(5')
-
CH (4')
=
CH(3')
•
-
CH(l')
= CH(2-)
c=o
c=o
o
4U .
Ch,
HN
6
110-
02
N 11
.0.
•Y,
^

O
110-
41
L
t
HN
^
Ay
^K
CH3
6
H
r
N^
.V
^ritr\r'
85-
DO
75-
60-
513-
50-
4 5-"
4 0
-
3000
2000
Wavenumbers (cm-l)
1000
Phd hong ngoai

ciia
stavudin (D4T)
16
D4T-DMSO-1H
"'"S"
-V''
r^""
'tl/!
J
.'
V\.
T'.!
(,i
Phd
CHTNH-'H ciia
stavudin
(D4T)
D4T-Dr.SO-
C13CPD
i I
!
m'
M«i>**w^wii«f
|i|»M««<ww»M»i«wiiN«*w»iw«w»w*wW»^<MMiN|W»»yw^
i«Wk|Mi^^taaMMhiAwMftMnM« *ttm»*tKm»mm»ttmlt<mmmf^mm
Phd
CHTNH-'^C ciia
stavudin
(U4T)
17

'^''^^^V^'
ilUJtliJ,'.
"lU
Mi
[;,"
•:
DT/^^^
•^w,
., /
3.1.3 Tong hcrp Zidovudin (AZT)
AZT duac ldng hap qua phan ii'ng
true
tiep cua
5'-arylsunfonyIoxithymidin
vdi
natriazit
NaN^
trong dung mdi thich hop:
O o
.CH3 U CH
HN
O^N
H
ArO.SO-
O
NaN.
O"
N
HN
10

i\u7
.0
/
OH
N,
Siin phrlm la
tinh thd khong mau, ket tinh lai trong nudc:
t.,,:117-119''C
hieu suat: 70%
Phd hdng ngoai chi ra su cd mat cua nhdm azit
N-,
d
2115,
2083 cm'', vdi cudng do
manh.
Dai tine Quuc gia
A7T.TICH
I .>'.>
•I
!
. I
'.
.j
:
I
. i.l
•
!,, •!
IV
11

"ffl
N<
06
04
62
m
Gfl
OG
5-1
52
0
302
>
r^j
1
OS'S
211
in
in
CO
0
r^
T
CM
to
•o
C-J
1280.
0
2000

Wavcnumt:Pt5 (cm-lj
Phd hong ngoai cua zidovudin (AZT)
^
:•]
18
Pho cdng hudng
tir
nhan
'H
va phd COSY cua hap chat tdng
hgp
ra phu hap vdi cau
tao
CLia
AZT dudi day;
5 ppm
1,8
2,4
3,6
4,4
5,2
6,2
7,8
11,3
Nhom
CH,
CH^iT)
CHA51
CH(3')
CH (4')

CH(I')
= CH (6)
NH
HN
O
4A
3
,CH.
HO-
0"2
N
H
.0.
K?
N3
• Pho
c6ng
huong
tir
nhan
'^C-NMR
cho cac tfn hieu
phd
cua chat tong hop ra
('^C-
NMR va DEPT):
5 ppm
12
36
60

61
84
85
135
150
16.^
Nhom
CH,
CH^CV)
CH(3')
CH2
(2')
CH (4')
CHd')
=
CH (6)
C=0
C=0
o
HN
O^N
CH
H
HO-
.0.
h_J
N.
Phd hong ngoai cua
hgp
chat cho

ca'c
dinh hap thu dac
irifng
nhu sau:
VNM
Von
Veil
VNI
Vc=o
Vc-N
3462,87
cm-'
3367,95
cm''
3021,00
cnr'
2900,
2800 cm-'
2115,50;
2085,51
cm"'
1692,74; 1660 cm-'
1280,23 cm-'
O
HN
3
1
0"'2
^N
HO-

5'
O.
V_7
N-^
CH^
H
19
TIEN-AZT,
DMf^O
,
III
'"Sv
^^;;-'ii^*^^*
Phd CHTNH-'H cua zidovudin (AZT)
TTKN-AZT,
DMSO,
Cl3CPD
Phd
CHTNH-'^C ciia
zidovudin (AZT)
20
3.2.
Tong hpp thymin vd uraxin
Chiing
toi da nghien
cu:u
ldng hap thymin va uraxin
la
cac
nguyen lieu

66 ldng
hc;p
thymidin va
uridin.
3.2.1
Tdng
hgp
thymin: di
tij
axit oxalic, axit axetic, etanol
theo
sa dd sau:
HOOC-COOH
^^^g^g^'^*
C2H5OCO-COOC2H5 COOC2H3
JJ^
^'^^^°^^''^
' ' \c2H,0Na C = 0
NaBH^HN-"^
-^'*-'
C2H,C00H ^gg^"> C2H3COOC2H3 / CH-CH3
QA^.
axil propionic
COOC2H5 |.j
Thymin
San pham
la
tinh
the mau
tring,

t^/,.:
330"C
(tai
lieu
335'*C),
phd
hdng ngoai,
phd
CHTN-'H
va
'^C
da
xac nhan
cau tao cua nd la
thymin:
• Phd
hdng ngoai:
Vr^ii
3060; 3062
cm'
(lien
hgp
_js[_]-j
o=C-
)
c^o
Vf
C
-C
1744,90;

1677,51
cm"
1610
cm'
V( N 1245,70
va
1215,01 cm
l^hd
cong hucVng
ti^r
hat
nhan
'H
(5, ppm)
5
a,,
1^733
d.a.
7.247
Of,,,
10.570
;
10.981
^lid
cdng
hucVng lir
hat nhan
'^C
(ppm)
5,11,

'1-773
dc.M 107,667
3,,,
137,698
6c.o
151,482
5c-o 164.911
3.2.2 Tdng hcrp uraxin:
Uraxin dugc tdng hgp
t|ua
phan
u/ig
ciia ure
va axil malic dudi dAy:
o=c'^
\
UIC
COOH
I
Nil:
CHOH
+
CH.
NH.
I
^
COOH
axil malic
21
O

oleum
Uraxin
San pham la tinh the mau trang, nhiet do ndng chay
t,,
=
332"C,
cau tao cua nd dugc
xac minh qua cac phuang phap phd hdng ngoai, phd cong hudng
ti^
hat nhan 'H va
'''C
nhu
sau:
• Phd hdng ngoai
(v,
cm')
Nil
c=o
C=C
C-N
3112
1716,32; 1671,36
1650
1237,17
PhdCirn-lN-'H(6,
ppm)
'^-CH
(5)
J,,,=
15Hz

"NH
5,455
;
5,440
7,391
;
7,376
10,804;
10,974
o
IN^4
O
W^
PhdCHTHN-'^C(6,ppm)
dc(^^
100,226
8
142,170
151,511
164,338
'(•(6)
h
C=0(2)
2000
WavenumbOfs icm-i)
Phd hdng ngoai
ciia
thymin
22
3 000

2000
Wavenumbers
(cm-1)
Phd hong ngoai
ciia
uraxin
}.}
.
ronj;
hup
2'-hidn)xilhyinidin
Nham
ihuc
hien ldng
hqp
Ihymidin chung ldi da thuc hicn mdt
sc")
buxrc phan ung sau:
o
o
.ni}
II
C-H3
(1)
HN
O
N'
11
llgCl.
1-1N

O^
N'
HgCl
(I)
(2)
llO-n
(3)
.O,
ll(>-OH
OH
O
N
CH3
(CHiCO).O
O^
'N
HizCI
AcO-i
AcO-i
O
AcoT^OAc
OAc
Ac=COCH3
(H)
N
O
HO-
O
^N'
C)

K!y
OH
(ill
Hgp chat (I) cd
t„,
=
330-33l"C,
phd hdng ngoai cho
Vc=o
165,10.
v,.^,-
^^96,
v^,,
3555,42 ,
Vf,.,,^
481,84 cm'.
Ilgp chai (II)
cd
l,„
=
82"C,
hieu sua't 44%, phd hdng ngoai cho cac dinh hn'p lhu OH d
3300-3400
cm-',
cho
Vc=o ^
1749,08 cm''.
Hgp chat (III) tinh the mau trang, phd hdng ngoai cho cac dinh hap thu
v^j^
d 3486,7

cm-',
cho
Vc=oCf
1738,82 cm-',
Vc^ova
VC.N
d 1373,19 va 1202,96
cm-'.
Nhiet do ndng chay
183-184"C.
Hieu suat thap.
01
t
ran
^
lUO
05
90
65
80
75
70
65
60
50
nft
15
40
?iC^
.III

1
1
•
Oitl
)ir><:
Uuoc (ii.t
TieNHIOROXY
I
1
I.
\
•1
\
1
1
\
•
/
,'
"•""
'
" •;
'
,
,
y

/
/
/

/ ^
/
/
1 1
1 '
1
1-
1
•
fij
:
•.
«;
i
-

-v^
V
j
1 •
r-4
TO
,
I
'i 1
, 1
1
i
1 1
1

1
1
1
I
V
.
/ \
1
1
1
1
i
•
^
1
'•
"
;«
o
'^-
i
M
1
ir
/
1 '1
'
I'"
i
'

1
)•
!
f
1
"•
' 'i'l
i
Wavenumbers
(cm-1)
Phd hong ngoai cua 2'-hidroxithymidin
Bang 1: Ket qua tdng hgp stavudin, zidovudin va cac chat dau
TT
1
3
4
5
6
7
Ten chat
Stavudin (D4T)
Zidovudin (AZT)
Monoarylsunfonyllhymidin
Thymin
Uraxin
Tclraaxelyl-l^-ribofurannoza
5-Melyl
uridin
tnc
("C)

165
117-119
172
326-330
332
82
183-184
Hieu suat
(%)
30-40
60-70
96
40
-
44
thap
Phd chung minh
IR,
'H-,
'^C-NMR,
COSY,
HETCOSY, HMQC
IR,
'H-,
'^C-NMR,
COSY,
HETCOSY, HMQC
IR,'H-,''C-NMR
IR,
'H-,

'^C-NMR
IR, 'H-,
"C-NMR
IR
IR
24
3,4. Tdng hap mpt
sdhap
chdt di vdng co hoat tinh sinh hpc.
Nhd kinh
phi
cua de tai, chiing tdi da nghien cuu tdng hgp chat di vdng cd hoat
tinh
sinh hoc nhu khang khuan, khang nam, khang
HIV.
So 66 tdng hgp nhu sau:
X~^,
Ji^COOH
X-<
>-CH=N-NH-Z
O.N-^
V-CH=NOH
SO.CI
X-^
ii^COCl
(I)
O
^.HsOH
Z: H.
CONH2-

CSNH2
S02Cr,H4CH3
O
il.NNHCSNH.
X-^
J>-CONHNH2CSNH2
(IV)
^
II,SO,,
(V)
o S
II.NNI
1
/ v
N-N
X-OK
>-NI
IVI) O
N^
NIN
O.N-^
J)—CH=N-OCH2COOC2H5
(VII)
^
i-I.NNH.
(II
J O
Z:li.S02CV,ll4<'ih
(v,„,
^^

O.N^
V-CH=N-OCH.
CS
KOIl
r-—.
N-N
X; H. Br.
NO^
DCSyKOM
2)
H.NNH.
0-C N11NH.
fl~\
'^"^
0:N-^
J^CH^N-OCH:-^
3~SI
() N
I
NH.
(I\)
'^s