Nghiên cứu tổng hợp và thử hoạt tính sinh học của một số dẫn chất thiazolidin 2,4 dion

B Y T
I HI




NGUYN QUC THNG

U TNG H
HO HC CA MT S
DN CHT THIAZOLIDIN-2,4-DION

N TT NGHI








 NI - 2013



B Y T
I HI

NGUYN QUC THNG

U TNG H
HOC CA MT S
DN CHT THIAZOLIDIN-2,4-DION

N TT NGHI


ng dn:

1. TS. Phan Th 
2.  

c hin:

B c

i hi


I - 2013

LI CM 
 vi tt c s   n thy
PGS. TS. Nguyn Hi Nam    TS. Phan Th  
thc ting d bo tn
.
i li cn   , 
k thu i hc D o
u kin tt nhi gian thc hin.
i li c  i h
N tdy d, ch bo  sut c va qua.
 c s  c  Vin
Hp ch- Vin  h Vi 
t liu   i hc Khoa hc t  i hc Quc gia
i.
Cui li c c bi m i
ng lc cho trong cuc sc tp.


i,  



Nguyn Quc Thng


 2
 2
 2
1.1.2. T 5
 12
 14
-2,4-dion 14

N
2 15
 15

 17
 17
 17
 19
 19
 19
 20
 20
 21
 22
 22
 32
 33



 38

 39
 39
 41
 43
 
 





DANH MCH VIT TT:
13
C-NMR
Ph cng t h
13
C (
13
C nuclear magnetic resonance)
1
H-NMR
Ph cng t hproton (
1
H nuclear magnetic resonance)
CTCT
c cu to
CTPT

DCM
Dicloromethan

DMF
N,N-dimethylformamid
DMSO
Dimethyl sunfoxid
EtOH
Ethanol
HAT
Histon acetyltranferase
HDAC
Histon deacetylase
IR
Ph hng ngoi (Infrared spectroscopy)
HepG2
T 
MeOH
Methanol
MS
Ph khng (Mass spectroscopy)
PPAR-
Peroxisom proliferator activated receptor gamma
MTT
Thuc nhum 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium
bromid
PTP 1B
Protein tyrosin phosphat 1B
SAHA
Acid suberoylanilid
STT
S th t
SWL68

T ng
SW620
T 
TZD
Thiazolidin-2,4-dion
TLC
Sp mng (Thin Layer Chromatography)
R
f
H s  (Retention factor)






DANH MNG
STT
ng
Trang
Bng 1.1
Kt qu th  p cht TZD
3
Bng 1.2
 IC
50
c logP
4
Bng 3.1
Hiu su s n phc tng

hp
32
Bng 3.2
 R
f

o
nc cn phm cui 4a-4d
33
Bng 3.3
Kt qu  khng ct trung gian
34
Bng 3.4
Kt qu  hng ngoi ct
34
Bng 3.5
Kt qu  khng ct sn phm
cui
35
Bng 3.6
Kt qu 
1
H-NMR cn phm cui
36
Bng 3.7
Kt qu 
13
C-NMR cn phm cui
37
Bng 3.8


39
Bng 3.9

42













DANH M 

STT

Trang

ng gii vi s

6

Cu tng ca HDAC
7


Ct s hot cht c ch HDAC
8

Ca mt s 
8

Cng ca HDAC
9


10

Zn
2+

trong protein
10

 
11

 i gian
12


12

ng c-
13


Mt s TZD c ch PTP1B mu
14








`STT

Trang
3.1
ng ht
22
3.2
Tng hp thiazolidin-2,4-dion
22
3.3
Tng ht 2a-d
23
3.4
Tng ht 3a-d
26
3.5
Tng ht 4a-d
29

1



T V
Thiazolidindion (TZD) n cht  c bing chng
  ng nh ho   th peroxisom proliferator activated receptor
gamma c ch protein tyrosin phosphat 1B [13,14]. Mt s thu u tr 
p 2 dc s d 
Pioglitazon [2, 17].
p chng
 n [6]m [7,8], ch [18].
c bit trong thi gian g    u v dn cht ca TZD ng
ng ti ho    t  ph
  mt s   [12,26,28].  t k 
thc mi ca TZD        c ch enzym histon
deacetylase (mt enzym quan tr )
[26]. Kt qu thu c rt kh i mt s cht chn
.
T t qu nhm m n cht
thiazolidindion trin vng, c hi u tng h

 ng sinh hc ca mt s dn cht thiazolidin-2,4-dion 
sau:
1. Tng hp N-(2-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)ethyl)
benzensulfonamid t s dn cht.
2. Tht tng hc.
2




1.1. 
Thiazolidindion c gn cha gc
thiazolidin-2,4-dion:

 c chu ng sinh h
ng n [6,7], chng nm [7,8], ch [18], c t 
chng ung th [12,26,28], ho   th PPAR-, c ch protein PTP1B
[8,12,14].
Hin nay,   u v ho  a     
nhiu  , c ch HDAC, hoPPAR- c ch PTP1B.
1.1.1.
  Patil  cng s    ng hp mt s dn cht
 (I)  in vitro   m:
Hop62 i), PC3 n tin lit), MCF7 G2
  ch cu), Gurav  
hng). N t th nghim trong khong t 10
-4
ti 10
-7
M, ti

song song vi 2 mu th, mt m dng cht c ch t mu s dng
cht c ch  [29]. Sau 48 gi n ca mi
 t   t n ca t  c ch
50% (GI
50
).  logarit n ti GI
50
  hin cho s nhy
cm c p ch logGI
50
< -t


Y=R-NH Ib,Ic,Ie
Y=R Ia,Id
I
3



Bng 1.1: Kt qu th  p cht TZD I
Hp
cht
R
logGI
50

K562
MCF7
HepG2

Hop62
PC3
Gurav
KB
Ia

>-4,0
-4,53
>-4,0
-6,72
-4,53
>-4,0
>-4,0
Ib

>-4,0
>-4,0
>-4,0
-6,73
>-4,0
>-4,0
>-4,0
Ic

-5,65
>-4,0
>-4,0
-4,51
-5,65
-6,71

>-4,0
Id

>-4,0
-4,53
>-4,0
-5,64
-4,53
>-4,0
>-4,0
Ie

-6,72
-6,71
>-4,0
>-4,0
-5,60
-6,73
-5,65
Doxorubicin
-5,59
-6,88
<-7,0
-6,97
-6,96
-6,97
<-7,0
GI
50
: n c ch 50% s n ca t 

Nh kt qu u cho thy, hp cht Ie ng c ch t
nh nhc tng hp. Chc ch 5 trong s 7
 c th nghi3, Gurav ht Ic
cho thng yc ch c 4 trong s  
. Hp cht Ia Id  
  cht Ib ch c ch c duy nh 
Hop62. Tt c  
(HepG2) enzym amidase trong t  
p cht, khi hing [29].
Tip tu . ng s n
ng hp cha khung TZD (II)  
4



      i     t cung), HT-   c
i), MCF- c ch n c
24 gi c ghi nh IC
50
 [7]. S dng phn mm
 th hin s ng gic) c
hp cht tng hc, t  suy luc kh 


Bng 1.2:  IC
50
c logP
Cht
Ar
 IC

50
(M)
HeLa
HT29
A549
MCF-7
logP
IIa

72
88
90
100
3,05
IIb

38
30
42
48
3,86
IIc

34
32
36
38
3,22
IId

68
72
78
100
3,50
IIe

48
48
55
58
3,08
IIf

30
32
36
30
2,68
II
5



Cht
Ar
 IC
50
(M)
HeLa

HT29
A549
MCF-7
logP
IIg

62
78
80
90
2,05
IIh

60
80
72
84
1,88
IIi

80
94
76
90
2,54
IC
50
: n   cht hoc c ch
T kt qu u  nhn thy hp cht v benzen
(IIa) benzen (IId) cho  p v IC

50
cao t 60-
100   brom (IIb) hoc flo
(IIc) ho v IC
50
c trong khong 40-50 
Hp ch  ng mnh nh  -trimethoxylbenzen (IIe) v 
IC
50
thp nht t 30-. p cht d a Nit (IIg, IIh
oxy (IIi IC
50
 mc cao t 60-90 .
 logP cn chbenzen (IIa-IIfn
ch  (IIg-IIi), chng t n du cp cht IIa-IIf
p cht IIg-IIin du tht trong nh
  [7].
t qu y n cht mang khung TZD 
ng trong vic c ch s n c 
1.1.
1.1.2.1. iston deacetylase
Histon deacetylase i b 
acetyl t -N-acetyl lysin amino acid ca phn histon [16]. Loi   
pha histon, dn gin
gia histon vi ADN. u, nhim sc th th
ng hp protein dic biu hi
6

trng. Trong khi enzym histon acetyltranferase 
dc li. Trong t l
 nh s biu hin gen [4].

.1: ng gii vi s 
     ra rng, s c ch     p ca
 ph bin t bii trong c
cht nhim s   ng, dn ti s
i u. c ch hong cng ca nhiu
thuc ch n hin nay [16,22].
V        ng nhau,
u gn sau :
+ Ion Zn
2+
 a HDAC nm      
t mnh nht vi pht ph
t c ch nh vi Zn
2+
ng c
ch  nh [10].
+ t Van der Walls v
chc cu to bc bi
 i
 p vn ng dea[20].
Trt hot
Trng
7

: Cu tng ca HDAC
1.1.2.2. Mt s cht c ch 
T nh ng c ch HDAC ca natri butyrat 
 n trong vit c ch 
nhiu cht c ch ng mnh nhc
bin hi - mt sn pha Streptomycesc sn
xun n thi [25]n tht c
ch  c FDA cp p        d u tr u
lympho t i da: SAHA (V - chn gc tng
hp, Romidepsin  - chp t Chromobacterium violaceum
[32]ng 15 cht c ch u th nghim
  u tr .


SAHA
Romidepsin
8





TSA
Acid valproic
1.3: Ct s hot cht c ch HDAC
ng hp ca b  c Ti h
ng hp mt s dn chng c ch HDAC d
ca SAHA. Ct qu c v kh c ch 
t s   t kh quan [5,27].

n1:1-3
n2:1-2
n:1-6
1.4: C ca mt s 
Theo mt s  u tham kho, hiu qu  ng c  t c ch
HDAC d t ca 3 yu t [10] :
-  t vi Zn
2+
tng c
HDAC 
- Cu nch hydrocarbon, n
- 
ng n mt ca enzym.
D   u bit v c    t c ch HDAC, R.
 cng s t k mt s hp cht ca TZD
(IIIa IIIb) nhm c ch p
cht d s do phc vi
ion Zn
2+
tng c[26].
9





1.5: Cu ng ca HDAC

a
IIIa-b
b

1.6:

qua docking       MOE (Molecular Operating





Chu
hp

kt (acid
hydroxamic)
 mt
protein
Khong trng
ng
10

Environment). Kt qu docking cho th lc cao ca hai chc thit k i
vi HDAC, th hin s p v c


ng sinh hc ca 2 hp cht mi tng hqua 
t ng c ch enzym HDAC.
 a hai hp chc tng hp ti
   ng (WRL68). Hai
 ng ph thudng ph thui gian.
S d ng c 
ng. Kt qu  
Bi kt qu cho thy  n  
HepG2: cht IIIa   IIIb  
   a hai cht g     IIIa  
(IIIbt hi   chn lc
 
11





  sau 48h
m ti c ch t a 2 cht tm
48 gi, 72 gi, 96 gi . Ti thm k, cht IIIa c ch
82,98% t  t IIIb ng
ch t IIIa cho thy hoc ch mnh
i gian so vi cht IIIbm hn ch 
  

 i gian
Tip t c ch HDAC in vitro cc tng
h      i chi    t qu vi hai cht c ch
% t t theo liu ca IIIa
% t t
% t t theo liu ca IIIb
% t t
% t t theo thi gian ca IIIb
% t t
% t t theo thi gian ca IIIa
% t t
12



HDAC m       t qu     bng
1.10.


Cht IIIb    t qu c ch HDAC t   i cht IIIa
(42,11%), chi SAHA
(57,89%) [26].
               
--           
--
enzym.
thiazolidindion
ng ho-

t ch vn chn l th PPAR-
qua ho-u chnh s biu hin ca mt s gen ma
PPAR-a t -
s  tr  d t  dng
c ch  c ch s tng hp glucose  
ng s oxy LDL (low density lipoprotein)  i th
i thin s nhy cm vi insulin ca t m n 
gi chng m[13,7].


SB SAHA 3a 3b
13




ng c-
   i thu        
dng ph i th
ng. Mt thu ngng s d
mch, hin ti ch c s dng. u
tr ting cho thy nhin.
ng c ch PTP1B
t phosphotyrosin phosphatase, enzym th
g        n   
phosphotyrosin ca enzym kinase ho       
gin truyn ca th th  th leptin  c
ch   nh  ng c  i th th, d n
gim n [9,30]c chng minh
t m quan trn cu tr

ng. Trong mt s u gt s dn chc
chng c ch enzym PTP1B [9].


2: Mt s TZD c ch PTP1B mu
Tin t 
T  nhy cm insulin
T  

bi
n s
chuy
a
b
IVa-b
14




 -2,4-dion
1.2.1.1. Phn a thioure vi acid monocloacetic
E. ng s ng h-2,4-dion theo
 [23]:

Phn ng xn:
- n 1: to pseudothiodohydantoin:

- n 2: thng ca HCl sinh ra trong
n mt:

c s dng nhiu su ti
gin.
1.2.1.2. Phn t acid hydrocloric
. Joshi (1968) [20] n n
u sut thc s dng.

15




N
2
1.2.2.1. Phn ng to dn xut sulfonamid
Phn ng gia dn xuc 1 x
S
N
2 [1]:


1.2.2.2. Phn ng to ether t dn xun xut phenol

Phn ng th gia dn xun t t
  S
N
2 [1]. 

phenol.


-2,4-dion  ng cao
      
ng:
chuyn tip
16





 [3]. 
n-2,4-dion 

