tổng hợp và thử tác dụng sinh học một số dẫn chất benzensulflonamid mang khung...
Post on 16-Dec-2015
33 Views
Preview:
DESCRIPTION
TRANSCRIPT
-
B Y T
TRNG I HC DC H NI
NGUYN HUY PHONG
TNG HP V TH TC DNG SINH
HC MT S DN CHT
BENZENSULFONAMID MANG
KHUNG THIAZOLIDIN-2,4-DION
KHA LUN TT NGHIP DC S
H NI-2014
-
B Y T
TRNG I HC DC H NI
NGUYN HUY PHONG
TNG HP V TH TC DNG SINH
HC MT S DN CHT
BENZENSULFONAMID MANG
KHUNG THIAZOLIDIN-2,4-DION
KHA LUN TT NGHIP DC S
Ngi hng dn:
1. TS. Phan Th Phng Dung
2. DS. Th Mai Dung
Ni thc hin:
B mn Ha Dc
Trng i hc Dc H Ni
-
LI CM N
Trong thi gian nghin cu v lm kha lun, em xin chn thnh cm
n TS. Phan Th Phng Dung, PGS.TS Nguyn Hi Nam, Dc s
Th Mai Dung gip em hon thnh tt ti kha lun tt nghip.
Bn cnh , em cng xin cm n cc anh ch k thut vin b mn Ha
Dc, Trng i hc Dc H Ni gip v to iu kin tt nht cho
em trong thi gian thc hin kha lun. Em xin cm n s gip ca cc
cn b phng Phn tch hu c - Vin Hp cht thin nhin - Vin Hn lm
Khoa hc v Cng ngh Vit Nam v Phng Ha vt liu - Khoa Ha -
Trng i hc Khoa hc t nhin - i hc Quc gia.
Em cng xin chn thnh cm n cc thy c gio trong trng i hc
Dc H Ni ging dy v trang b cho em nhng kin thc c bn trong
hc tp nghin cu kho lun cng nh trong cng vic sau ny.
Nhng li ng vin, khch l t gia nh, s chia s, hc hi t bn b
cng gp phn rt nhiu cho kha lun tt nghip ca em t kt qu tt
hn.
H Ni, ngy 11 thng 5 nm 2014
Ngi vit
Nguyn Huy Phong
-
MC LC
DANH MC CC K HIU,CC CH VIT TT
DANH MC CC BNG
DANH MC CC HNH V
DANH MC CC S
T VN
CHNG 1.TNG QUAN
1.1. MT S MC TIU PHN T CA DN CHT THIAZOLIDIN-
2,4-DION ............................................................................................................. 2
1.1.1. Tc dng c ch enzym histon deacetylase (HDAC) ................................... 2
1.1.2. Tc dng hot ha PPAR- ca TZD trong iu tr i tho ng ............. 5
1.1.3. Tc dng c ch PTP1B ............................................................................... 7
1.2. MT S TC DNG KHC CA CC DN CHT THIAZOLIDIN-
2,4-DION ............................................................................................................... 9
1.2.1. c tnh t bo ............................................................................................. 9
1.2.2. Tc dng khng khun, khng nm ............................................................. 11
CHNG 2. NGUYN LIU, THIT B, NI DUNG V PHNG
PHP NGHIN CU ........................................................................................ 15
2.1. NGUYN VT LIU, THIT B ................................................................. 15
2.1.1. Ha cht ..................................................................................................... 15
2.1.2. Thit b, dng c ......................................................................................... 15
2.2. NI DUNG NGHIN CU .......................................................................... 16
2.2.1. Tng hp ha hc ....................................................................................... 16
2.2.2. Th c t nh t b o in vitro v nh gi s b t nh ging thuc ca mt
s n cht tng hp c .................................................................................... 16
2.3. PHNG PHP NGHIN CU.................................................................. 16
2.3.1. Tng hp ha hc v kim tra tinh khit ................................................ 16
-
2.3.2. Xc nh cu trc ........................................................................................ 17
2.3.3. Th tc dng c t nh t b o in vitro .......................................................... 17
2.3. . nh gi mc ging thuc ca cc n cht tng hp c .................. 18
CHNG 3. THC NGHIM, KT QU V BN LUN .......................... 20
3.1. HA HC ..................................................................................................... 20
3.1.1. Tng hp ha hc ....................................................................................... 20
3.1.2. Kim tra tinh khit ................................................................................. 28
3.1.3. Xc nh cu trc ........................................................................................ 29
3.2. TH HOT TNH SINH HC ..................................................................... 34
3.3. BN LUN .................................................................................................. 34
3.3.1. Tng hp ha hc ....................................................................................... 35
3.3.2. Tc dng sinh hc ....................................................................................... 38
KT LUN V KIN NGH ............................................................................ 40
Kt lun ................................................................................................................ 40
Kin ngh.............................................................................................................. 40
TI LIU THAM KHO
PH LC
-
DANH MC CC K HIU,CC CH VIT TT
13C-NMR Ph cng hng t ht nhn
13C (
13C nuclear magnetic resonance
)
DCM Dicloromethan
DMF N,N-dimethylformamid
DMSO Dimethyl sunfoxid
EtOH Ethanol
HAT Histon acetyltranferas
HDAC Histon deacetylase
HepG2 T b o ung th gan
1H-NMR Ph cng hng t ht nhn proton (
1H nuclear magnetic
resonance)
IR Ph hng ngoi (Infrared spectroscopy)
MeOH Methanol
MIC Minium inhibitory concentration
MS Ph khi lng (Mass spectroscopy)
MTT Thuc nhum 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium bromid
PPAR- Peroxisom proliferator activated receptor gamma
PTP1B Protein tyrosin phosphatase 1B
SAHA Acid suberoylanilid
SWL68 T b o gan bnh thng
SW620 T b o ung th i trng
TLC Sc k lp mng (Thin Layer Chromatography)
TZD Thiazolidin-2,4-dion
-
DANH MC CC BNG
STT Tn bng Trang
1 Bng 3.1: Hiu sut v cc ch s ha l ca cc sn phm
c tng hp
28
2 Bng 3.2: Gi tr Rf v tonc ca cc sn phm cui 4a-4d 29
3 Bng 3.3: Kt qu phn tch ph hng ngoi ca cc cht 29
4 Bng 3.4: Kt qu phn tch ph khi lng ca cc cht sn
phm cui
30
5 Bng 3.5: Kt qu phn tch ph 1H-NMR ca cc sn phm
cui
31
6 Bng 3.6: Kt qu phn tch ph 13
C-NMR ca cc sn phm
cui
32
7 Bng 3.7: Tc ng hng t b o ung th i tr ng (SW620) 34
8 Bng 3.8: nh gi mc ging thuc theo quy tc
Lipinsky
38
-
DANH MC CC HNH
STT Tn hnh Trang
1 Hnh 1.1: Vai tr ca HDAC 2
2 Hnh 1.2: Cu trc khung ca SAHA 3
3 H nh 1.3: Cu trc 2 hp cht c ch HD C 4
4 Hnh 1.4: Kt qu c ch HDAC ca cc cht sau 8h 5
5 Hnh 1.5: S c ch hot ng ca th th PPAR 6
6 Hnh 1.6: Cc d n cht TZD vi tc dng h ng huyt 7
7 Hnh 1.7: Cc TZD c ch PTP1B mi c nghin cu 8
8 Hnh 1.8: Cc d n cht TZD theo nghin cu ca Vijay Patil 9
9 Hnh 1.9: Cc d n cht c tng hp trong nghin cu ca
Avupati
10
10 Hnh 1.10: Cc d n cht TZD theo nghin cu ca Alegaon
Shankar
11
11 Hnh 1.11: Cc d n cht 3-benzyl-5-(4-cloro-2-piperidin-1-
yl-thiazon-5-yl-methylen) thiazol-2,4-dion
12
12 Hnh 1.12: Cc d n cht ca pyrazolyl-2, 4-thiazolidindion 12
13 Hnh 1.13: Dy cht A1, A2, A3, A4, A5, B1, B2, C1, C2,
D1
13
14 Hnh 3.1: C ch phn ng tng hp cc cht 2a-d 35
15 Hnh 3.2: C ch phn ng tng hp cc cht 3a-d 36
16 Hnh 3.3: Giai on cng hp 37
17 Hnh 3.4: Giai on ngng t 38
-
DANH MC CC S
STT Tn s Trang
1 S 3.1: Quy trnh tng hp chung 20
2 S 3.2: Quy trnh tng hp thiazolidin-2,4-dion 20
3 S 3.3: Quy trnh tng hp cht 2a 22
4 S 3.4: Quy trnh tng hp cht 3a 23
5 S 3.5: Quy trnh tng hp cht 4a 24
-
1
T VN
Cc d n cht ca thiazolidindion (TZD) c bit n vi rt nhiu tc
dng [9,11,13,18], trong ni bt nht l tc dng h ng huyt nh hot
ha th th peroxisom proliferator activated receptor gamma (PPAR-). T
TZD thc y s sao chp iu ha gen, gip tng hp mt s protein cn
cho cc t b o p ng vi insulin nn l m tng hiu nng insulin. Mt trong
nhng d n cht TZD in hnh c s dng trong i tho ng typ II l
Pioglitazon [28].
Bn cnh , trong thi gian gn y, TZD c cc nh khoa hc
tip cn theo hng nghin cu khc, l h nng c ch cc dng t bo
ung th [20,22]. Nm 2011, trong cng b ca Jung M., cc TZD c
chng minh l nhng cht c kh nng c ch enzym histon deacetylase
(HDAC), l mt enzym c s biu hin qu mc trong qu trnh hnh thnh
cc t b o ung th [20]. Hng nghin cu n y c cc nh khoa hc rt
quan tm v ch . Nhm nghin cu tng hp ha c, b mn Ha c,
trong thi gian 3 nm tr li y cng thit k 1 s dy d n cht ca TZD
v th hot tnh ca chng theo hng tip cn ny [4]. Tuy nhin kt qu th
c ch HDAC v c tnh t bo trn cc t b o ung th li cha my kh
quan, trn c s thay i mch cacbon s d n n thay i tng tc ca cht
vi mc tiu phn t , nhm mc ch tm im cc d n cht thiazolidindion
trin vng trong tng lai, chng ti thc hin t i Tng hp v th tc
dng sinh hc mt s dn cht benzensulfonamid mang khung
thiazolidin-2,4-dion vi cc mc tiu sau:
1. Tng hp N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)
propyl)benzensulfonamid v mt s d n cht.
2. Th c t nh t b o ca cc cht tng hp c.
-
2
CHNG 1. TNG U N
1.1. MT S MC TIU PHN T CA DN CHT THIAZOLIDIN -
2,4-DION
1.1.1. Tc dng c ch enzyme histon deacetylase (HDAC)
1.1.1.1. Khi nim v vai tr ca enzym histon deacetylase trong s to ung
th
Histon deacetylase (HDAC) l mt nhm cc enzym xc tc qu trnh
loi b nhm acetyl t -N-acetyl lysin amino acid ca phn histon [10,19].
u amin ca histon mang in ng nn tng tc mnh vi phn
phosphat mang in m trn phn t ADN to nn cu trc nucleosom v cu
trc bc cao hn ca nhim sc th. HDAC xc tc cho qu trnh deacetyl ha
l m tng s t ch in ng trn u N ca histon d n n lin kt cht hn
vi u phosphat ca ND gy ng xon NST c ch qu trnh phin m.
Cc sai lch trong qu trnh phin m l mt trong nhng nguyn nhn d n
n s hnh thnh khi u [19].
Trong khi enzym histon acetyltranferase (HAT) xc tc qu trnh acetyl
ha c tc dng ngc li. H T v HD C l 2 enzym iu ha qu trnh
acetyl ha t quyt nh s biu hin gen.
Hnh1.1: Vai tr ca HDAC v HAT
Mt s thng k gn y cho thy cc HD C c lin quan n nhiu
giai on iu ha c bn ca qu trnh sinh hc trong t b o ung th nh chu
Ngng t cc nhim sc th
ngn cn phin m
Ko dn cc nhim sc th
kch thch phin m
-
3
trnh t bo, s bit ha, s cht t b o theo chng trnh, c s di chuyn,
s xm ln v s to mch. V vy, c ch hot ng ca enzym HDAC l
ch tc ng ca nhiu thuc chng ung th ang c nghin cu trong giai
on hin nay [10,20,31,32].
1.1.1.2. Cu trc ca cc cht c ch HDAC
Hiu qu tc dng ca cc cht c ch HDAC da trn s c mt ca 3
yu t chnh [15] :
- Nhm kt thc (zinc-binding group-ZBD): l nhm lin kt vi Zn2+
ti trung tm hot ng ca cc enzym HD C, thng c cu trc acid
hydroxamic, thiol, benzamid, mecaptoceton...
- Cu ni (lin er): thng l cc mch hydrocarbon, nm trong lng
enzym.
- Nhm kho hot ng (capping group): thng l vng thm hoc
pepti vng, thng nm trn b mt ca enzym.
Hin nay, c nhiu ti liu cng b v hot tnh c ch HDAC ca
nhiu d n cht vi cc cu trc khc nhau [31,32]. c bit n nhiu l
SAHA (Vorinostat, Zolina), mt c cht c cu trc amid-alkyl-
hydroxamic, c FDA cp php lu h nh trn th trng c tc dng iu tr
u lympho t b o T i da [10,19]. Mc d cc cht c ch HDAC c cu
trc hc nhau, nhng c bn gm 3 phn chnh: nhm kha hot ng lin
quan n hiu lc v t nh c hiu (thng l aryl), phn cu ni (akyl) v
nhm kt thc gn kt vi ion Zn2+ (acid hydroxamic) [15] (hnh 1.2).
Hnh 1.2: Cu trc khung ca SAHA
Nhm kha hot ng Cu ni Nhm kt thc
-
4
Cho n nay c nhiu cng trnh trn th gii khi nghin cu lin quan
cu trc tc dng ca cc cht c ch HD C tp trung vo vic thay i
cu ni [33,34] hoc thay i nhm kha hot ng [35].
1.1.1.3.Kh nng c ch HDAC ca TZD
Thi gian gn y, trong mt s nghin cu v TZD c t i liu cng
b v hot tnh c ch HDAC ca chng. Nm 2011, Rhea Mohan v cng s
thit k 2 hp cht ca thiazolidindion l : N-(6-(2,4-dioxothiazolidin-3-
yl)hexyl)benzamid (SRR1) v N-(6-(2,4-dioxothiazolidin-3-y)hexyl) benzen
sulfonamid (SSR2) vi nh hng c ch HD C trn ng t b o ung th
gan (hnh 1.3). y, tc gi s dng khung TZD l nhm to phc vi ion
Zn2+
ti trung tm hot ng ca enzym [22].
C
O
N
NS
O
O
H
N
NS
O
O
H
S
OO
SSR1 SSR2
H nh 1.3:
nh gi c tnh t bo ca hai hp cht c tng hp c tin
hnh trn 2 dng: t b o ung th gan (HepG2) v t bo gan bnh thng
(WRL68). Hai tiu ch c nh gi l tc ng ph thuc vo liu v tc
dng ph thuc vo thi gian. Nghin cu chng minh kh nng gy c
ca 2 hp cht ny trn 2 dng t bo, tuy nhin v n c s chn lc hn trn
t b o ung th (HepG2). Bn cnh cht SSR1 cho thy hot tnh c ch
mnh hn theo thi gian so vi cht SSR2. Tuy nhin li c im hn ch khi
tc dng trn dng t b o bnh thng (WRL68) li mnh hn trn ng t
b o ung th (HepG2).
-
5
Tip tc nh gi h nng c ch HDAC invitro ca cc cht c
tng hp trn dng t b o ung th gan (HepG2), i chiu v so snh kt qu
vi hai cht c ch HDAC mnh l SB (sodium butyrat) v SAHA (hnh1.4).
Hnh 1.4: K t qu c ch HDAC ca cc cht sau 8h
Cht SSR2 lc ny cho kt qu c ch HDAC tt hn so vi cht SSR1
(42,11%), cht so snh SB (52,32%) v c hot tnh gn tng ng vi
SAHA (57,89%).
T t qu nghin cu ca nhm tc gi Rhea Mohan, c th thy rng
cc hp cht ca thiazoli in-2, - ion c tim nng ln trong vic c ch
enzym HD C. Nhm thiazoli in-2, - ion ng vai tr l nhm tc ng, c
ch trung tm hot ng ca enzym.
1.1.2. Tc dng hot ha PPAR- ca TZD trong iu tr i tho ng
1.1.2.1. Tc dng ca cc dn cht TZD ln th th PPAR-
Peroxisom proliferator (PPAR) gm 3 loi: PPAR-, PPAR- v PP R-
. Trong peroxisom proliferator activate gamma (PP R-) l mt th th
mng nhn t bo, c chc nng nh nhng yu t phin m iu chnh s
biu hin ca gen. PPAR- c mt trong cc t bo ni m v cc t b o c
trn mch mu [24].
Khi c gn vi cc cht ch vn (nh cc TZD), PP R- chuyn
thnh dng hot ng. PPAR- hot ng to dimer vi receptor X
% HDAC ca HepG2 b c ch sau 8h
-
6
retinoid alpha (RXR-) ( c gn vi cht ch vn ni sinh 9-cis retinoic
acid) to thnh phc hp proliferator proxisom (PPRE).
Hnh 1.5: S ho ng ca th th PPAR
Phc hp ny gn vi ND l m iu ha qu trnh phin m, dch m
ca gen trong nhn t bo, lm cho kch thch d tr v s dng acid bo,
triglycerid t bo m, kch thch s dng glucose v c ch oxi ha acid bo
c, c ch s tng hp glucose gan, ngo i ra cn tng cng s oxi ha
cc LDL i thc bo, t ci thin s nhy cm vi insulin ca t bo,
gim nng glucose trong mu, gim acid bo trong mu, chng x va
ng mch v cao huyt p [8,13].
1.1.2.2. V tr ca cc dn cht TZD trong iu tr i tho ng hin nay
Thiazolidindion c dng trong iu tr i tho ng type 2 k t
cui nhng nm 1990. Tuy nhin troglitazon b rt khi th trng mt vi
nm sau o nhim c gan.Tip , nm 2010, o tng nguy c bin c
tim mch, c quan y t chu u ngh tm nh ch s dng rosiglitazon. Do
ch cn pioglitazon l thiazolidindion v n c s dng iu tr i tho
ng type 2, nhng vic s dng pioglitazon phi c kim sot cht ch
do gp mt s tc dng ph trn gan, xng, tim v c th gy ung th b ng
quang. iu ny, cng vi s pht trin ca cc chin lc iu tr c ph
-
7
duyt trong vi nm qua, t ra cu hi l c nn s dng thiazolidindion
(c th l pioglitazon) trong iu tr i tho ng type 2 hay khng [28]?.
V o nm 2009, Pattan Shashikant R v cng s tng hp 1 lot cc
d n cht thiazolidin-2,4-dion c cu trc nh sau [25,26]:
S
O
O
RS
NH
O
O
Hnh 1.6: Cc dn cht TZD vi tc dng h ng huy t
Cc cht n y c em th nghim trn chut bch, o nng ng
huyt trong mu cc thi im 0 gi, 1 gi, 3 gi, 6 gi, dng Glibenclamid
lm cht chun so snh. Kt qu thu c 6 cht c tc dng h ng
huyt ng .
Gn y nht, nm 2011, Mehen ale-Munj v cng s [36] tin hnh
tng hp cc d n cht 5-arylidenthiazolidin-2,4-dion, t th nghim tc
dng h ng huyt v kim sot m mu. Kt qu l c 2 d n cht (Z)-5-(2-
4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)acetyl)-2-hydroxy
benzamid (IIIa) v (Z)-2-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)-
N-(5-nitro-thiazol-2-yl)acetami (IIIb) u cho tc dng kim sot ng
huyt, cholesterol (CHL) v triglyceride (TG) trn chut c Sprague
Dawley c ch n giu m sau 14 ngy s dng.
Nh vy, c th thy rng TZD v n cn cha ng nhiu tim nng
trong iu tr i tho ng type 2.
1.1.3. Tc dng c ch PTP1B
1.1.3.1. Khi nim v tc dng ca PTP1B
Protein tyrosin phosphatase 1B (PTP1B) l mt phosphotyrosin
phosphatase, enzym thy phn nhm phosphat gn trn acid amin tyrosin.
-
8
PTP1B tham gia phn ng dephosphoryl ha phosphotyrosin ca enzym
kinase hot ha insulin v leptin. PTP1B lm gim thng tin d n truyn ca
th th insulin v th th leptin m m, gan, c. c ch PTP1B l m tng
nhy cm, p ng ca insulin v leptin ti th th, d n n gim nng
glucose, lipid trong mu [14,17].
V vy, c ch PTP1B s l mt chin lc hiu qu iu tr bnh
tiu ng loi II v bnh bo ph.
1.1.3.2. Tc dng c ch PTP1B ca cc dn cht TZD
Trong mt s nghin cu gn y, cc n cht TZD cn c chng
minh c tc dng c ch enzym PTP1B. Mt s d n cht ca TZD, c th l
benzyliden-2,4-thiazolidindion vi cc nhm th v tr ortho v para
c tng hp v nghin cu tc dng trn ch PTP1B [14,29]. Kt qu
tm c hai d n cht c hot tnh sinh hc mnh nht l a v b (hnh 1.7).
S
NH
O
O
O
CF3
Br
O
SO
O
CH3
a b
Hnh 1.7: Cc TZD c ch PTP1B mi c nghin c u
Th nghim invitro cho thy, cc d n cht n y ng thi c hai tc
dng: c ch enzym PTP1B v hot ha PPAR-. Hot tnh c ch PTP1B
ca b IC50 = 1.3m, cn a tc dng yu hn vi IC50 = 5m. S c mt ca
nhm sulfonyl ca hp cht b c vai tr quan trng trong vic to tng tc
vi c 2 th th PTP1B v PPAR-.
Trn invivo, a v b c tc dng nh l cht h ng huyt, ci thin
-
9
ng dung np glucose. Hai hp cht n y cng c ch ng s tng cn,
chng bo ph do c tc dng h triglicerid, cholesterol huyt [14].
Nghin cu ny m ra hng pht trin cc thiazolidindion mi vi tc
dng h ng huyt mnh ng thi c th ng nh nhng cht iu ha
lipid mu, chng bo ph v gim thiu cc bnh v tim mch trong tng lai.
1.2. MT S TC DNG KHC CA CC DN CHT THIAZOLIDIN-
2,4-DION
1.2.1. c tnh t bo
Nm 2010, Vijay Patil v cng s [16] tin hnh tng hp mt s d n
cht mang khung TZD (hnh 1.8).
SNH
O
O
OY
O
Y=R-NH-(1b,1c, 1d)
Y=R-(1a,1d)
Cht 1a 1b 1c 1d 1e
R
Hnh 1.8: Cc dn cht TZD theo nghin c u ca Vijay Patil
Sau th c tnh invitro trn 7 dng t b o ung th bao gm: ung
th phi (HOP62), ung th tuyn tin lit (PC3), ung th v (MCF7), ung
th gan (HEPG), ung th bch cu (K562), ung th ming (GUR V), ung th
vm hng (KB).
Kt qu nghin cu cho thy, hp cht 1e c tc dng c ch t bo ung
th mnh nht trong cc cht c tng hp. Cht ny c ch 5 trong s 7
-
10
dng t b o ung th c th nghim l K562, MCF7, PC3, GUARV v KB.
Cht 1c c ch c 4 trong s 7 dng t bo: K562, HOP62, PC3 v
GUARV. Hp cht 1a v 1d cng cho thy tc dng trn 3 dng t bo ung
th l MCF7, HOP62 v PC3. Ring cht 1b ch c ch c duy nht 1 dng
t bo l HOP62.
Tip , vupati v cng s [37] tng hp cc d n cht 5-aryl v th
tc dng c t bo. Cc d n cht c cng thc cu to i y:
S
NHO
OR
O
Hnh 1.9: Cc d n cht c tng hp trong nghin cu ca Avupati
Kt qu cho thy: trong cc d n cht tng hp c th hp cht ((Z)-5-
(4-((E)-3-oxo-3-(thiophen-2-yl)prop-1-enyl)benzyliden)-1,3-thiazolidin-2,4-
dion) c tc dng c t bo tt nht vi gi tr ED50 l 4.000.25g/ml.
Avupati cn phn tch mi lin quan gia cu trc v tc dng c t bo ca
cc d n cht TZD. Tc gi thy rng tc dng c ch t b o c lin quan n
nhn thiazolidin-2,4-dion v cc nhm th hc nhau l m tng cng hoc
gim bt tc dng c ch t bo, c th:
- Nhm th halogen ti v tr meta hoc para trn vng bn l m tng tc
dng c ch t bo.
- Tc dng c ch t bo b gim khi thay th phenyl bng naphatalen
hoc khi c thm nhm hot ng nh hy roxyl trn vng phenyl.
- V iu th v nht l hot ng c ch t b o tng rt mnh khi thay
th phenyl bng thiophen-2-yl (c 2 nhm carbonyl v tr , ).
Gn y nht v o thng 8 nm 2013, nhm tc gi Alegaon Shankar
cng c 1 bo co v cc d n cht TZD trn kh nng gy c tnh t bo
[12].
-
11
3a: Ar = 2-Cl C6H4 3f: Ar = 2-OH-5-NO2 C6H3
3b: Ar = 2,3-di-Cl C6H3 3g: Ar = 3,5-di-Br-4-OH C6H2
3c: Ar = 2-Cl-5-NO2 C6H3 3h: Ar = 4-Br-2-F C6H3
3d: Ar = 4-Cl-3-NO2 C6H3 3i: Ar = 4-Br-3-F C6H3
3e: Ar = 2,3-di-OH C6H3 3j: Ar = 2,4-di-NO2 C6H3
3k: Ar = 2-furyl 3m: Ar = 1-methylpyrol
3l: Ar = 2-thienyl
Hnh 1.10: Cc dn cht TZD theo nghin c u ca Alegaon Shankar
Cc cht n y c nh gi c tnh trn cc dng t b o ung th
ngi: ung th c t cung (HeLa), ung th i trc trng (HT-29), ung th
phi (A549), ung th v (MCF-7). Kt qu cho thy hu ht cc cht u c
kh nng chng li ung th phi ( 5 9) v ung th v (MCF-7). Cc cht 3a,
3c, 3e, 3f, 3g, 3j, 3l, v 3m cho thy tc dng ng vi IC50 nm trong
khong 40-50 M. Cc cht 3a, 3b, 3g, 3k, 3l c kh nng chng li A549
vi IC50 ln lt l 38, 37, 45, 36, 30 M.
Cc kt qu nghin cu trn phn no cho thy cc d n cht mang
khung TZD c tc dng trong vic c ch s pht trin ca cc t bo ung
th, m ra hng mi trong pht trin cc thuc iu tr ung th trong tng
lai.
1.2.2. Tc dng khng khun, khng nm
Vi hot tnh khng khun, khng nm tim nng, cc n cht ca TZD
v ang thu ht c s ch ca cc nh tng hp ha c. Mt s d n
cht ca TZD c hot tnh sinh hc tt c cc nh khoa hc cng b trn
cc tp ch quc t [21,23].
Nm 2006, M.C.Unlusoy v cng s tng hp thnh cng d n cht
ca 3-benzyl-5-(4-cloro-2-piperidin-1-yl-thiazon-5-yl-methylen)thiazol-2,4-
dion.
Cu trc ca chng nh sau:
S
N
Ar- O
O
O
OH
-
12
N
S
S
N
O
O
Cl
R1
R2
N
R1= H, Cl
R2=H, Br, Cl, F, NO2
Hnh 1.11: Cc dn cht 3-benzyl-5-(4-cloro-2-piperidin-1-yl-thiazon-5-yl-
methylen)thiazol-2,4-dion.
Kt qu th hot tnh sinh hc cho thy chng c tc dng kh tt trn
cc chng vi khun Gram (+): Bacillus subtillis, Staphylococus aureus v vi
khun Gram (-): E.coli [30].
Nm 2011, trong mt bo co ca nhm tc gi Aneja D.K v hot tnh
khng khun, khng nm ca 24 d n cht pyrazolyl-2,4-thiazolidindion, mt
ln na cho thy tim nng ca cc d n cht TZD [38].
NN
S
NO
O
R2
R1
O
O
4a-h: R2=C2H5
5a-h: R2=CH3
6a-h: R2=H
4-5-6 a b c d e f g h
R1 H Me OMe Cl F Br OH NO2
Hnh 1.12: Cc dn cht ca pyrazolyl-2,4-thiazolidindion
24 cht n y c em th nghim hot tnh khng nm ca 2 loi nm
l: Aspergillus niger v Aspergillus flavus. Kt qu thu c rt kh quan khi
c 24 cht u c tc dng khng nm, c bit 2 cht mnh nht 4b v 4e c
tc dng khng nm A.niger (70%) v A.flavus (67,7%). 11 cht 4d, 4e, 4g,
5a, 5h, 6a, 6b, 6d, 6e, 6f, v 6h cho thy tc dng khng nm A.flavus trn
-
13
60% so vi 77,7% ca Fluconazol. Cn vi kh nng hng nm A.niger, cc
cht khng trn 60% l 4b, 4d, 4e, 4h, 5c, 5d, 6a, 6b, 6d, 6e, 6f.
Th kh nng hng hun: cht 4a, 4h, 5h, c tc dng khng
Staphylococus aureus mnh nht (MIC = 6 g/mL), 2 cht 6a v 6h c tc
dng khng Bacillus subtillis (MIC = 6 g/mL).
Vi mong mun tm ra thm cc cht khng khun, khng nm khc
mang hung TZD, nm 2012, Shital L. Nawale v Avinash S. Dhae tng
hp nn 10 d n cht khc nhau ca thiazolidin-2,4-dion: A1, A2, A3, A4,
A5, B1, B2, C1, C2, D1 [23] (hnh 1.13).
SNH
O
O
OEtO
O
B1
SNH
O
O
O
OCH3
EtO
O
A1
SNH
O
O
O
EtO
O
B2
SNH
O
O
O
OCH3
NH
O
NHS
NH2A2
SNH
O
O
NHNHH N
NH NH
2
C1
SNH
O
O
O
OCH3
NH
O
NHO
NH2A3
SNH
O
O
NH
NHH2N
SC2
SNH
O
O
O
OCH3
EtO
OA4
SNH
O
O
O
Cl
D1
SNH
O
O
HO
OCH3
NHNH
S
NH2
A5
Hnh 1.13: Dy cht A1, A2, A3, A4, A5, B1, B2, C1, C2, D1
10 cht c xc nh nng c ch ti thiu trn 3 chng vi sinh vt
l Bacillus subtilis, Pseudomonas aerugenosa v Staphylococcus aureus. Cht
chun c so snh l Streptomycin. Kt qu cho thy A2, A5 l c hot
tnh khng khun mnh vi gi tr MIC i vi c 3 vi khun u l 31.25(g)
-
14
T nhng nghin cu trn ta thy ngy nay vi s gia tng sc
khng ca vi khun, nm th vic pht trin cc d n xut 2,4-thiazolidindion
l cn thit tm ra cc cht mi trong vic khng vi khun v nm.
Vi cc kt qu nghin cu khoa hc c cng b trn y, cho
thy tip cn cc d n cht TZD l mt hng nghin cu y tim nng
thit k cc c cht c hot tnh sinh hc trn i tho ng, ung th cng
nh khng khun, khng nm. Chng ti hy vng, kha lun s gp phn lm
phong ph hn cc n cht TZD v tm ra c nhng c cht c tc dng
sinh hc tt c th ng dng c trong lm sng.
-
15
CHNG 2. NGUYN LIU, THIT B, NI DUNG V
PHNG PHP NGHIN CU
2.1. NGUYN VT LIU, THIT B
2.1.1. Ha cht
Cc ha cht, dung mi dng trong qu trnh thc nghim l loi dng
trong tng hp c nhp t cng ty Merck hoc Sigma-Aldrich. Cc ha
cht ny c s dng trc tip khng qua tinh ch thm. Bao gm:
+) Benzensulfoclorid
+) 4-Clorobenzensulfoclorid
+) 4-Methylbenzensulfoclorid
+) 4-Methoxybenzensulfoclorid
+) 3-Bromopropanamin
+) 4-Hydroxybenzaldehyd
+) Acid cloroacetic
+) Thioure
+) Acid acetic bng
+) Piperazin
Cc ha cht v dung mi khc: DMF, aceton,dicloromethan, ethanol,
methanol, NaOH, HCl, nc ct, N,N-dimethylformamid.
2.1.2. Thit b, dng c
- Dng c thy tinh: bnh cu y trn ung t ch 50 ml c nt m i, sinh
hn hi lu, pipet, bnh chit, phu thy tinh, bnh chy sc k lp mng
(TLC).
- My khuy t gia nhit.
- My ct quay chn khng Buchi R-210.
- Phu lc Buchner
- Cn phn tch, cn k thut Shimazu.
-
16
- T lnh, t sy, my siu m.
- Bn mng silicagel Merck 70-230 mesh chy sc k lp mng.
- My o nhit nng chy nhit in (Electrothermal igital) xc
nh nhit nng chy.
- My Per in Elmer xc nh ph IR.
- My khi ph HP 5989B-MS ghi ph MS.
- My cng hng t Bruker AV-500 ghi ph 1H-NMR, 13C-NMR.
2.2. NI DUNG NGHIN CU
2.2.1. Tng hp ha hc:
Tng hp cc hp cht sau:
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)
benzensulfonamid (4a).
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)-4-
methylbenzensulfonamid (4b).
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)-4-
methoxy benzensulfonamid (4c).
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)-4-
chlorobenzensulfonamid (4d).
2.2.2. Th c t nh t o in itro nh gi s t nh ging thuc c
t s n cht tng h c
2.3. PHNG PHP NGHIN CU
2.3.1. Tng hp ha hc v ki tr tinh khit
- Nghin cu tng hp 4 cht trong mc tiu bng phng php ha
hc.
- Dng TLC theo di tin trnh ca phn ng.
- Kim tra tinh khit ca sn phm bng TLC v o nhit nng
chy.
-
17
2.3.2. Xc nh cu trc
Cc cht tng hp c xc nh cu trc bng cc loi ph sau: ph
hng ngoi (IR), ph hi lng (MS), ph cng hng t proton (1H-NMR),
ph cng hng t carbon (13C-NMR).
Ph hng ngoi (IR): ph hng ngoi c ghi ti Khoa Ha - Trng i
hc Khoa hc T nhin (KHTN) H Ni i hc Quc Gia H Ni, trn
my Per in Elmer vi thut vin nn KBr trong vng 4000-600cm-1. Cc
m u rn c phn tn trong KBr sy h vi t l 1:200 ri p i ng
film mng i p lc cao c ht chn hng loi b hi m.
Ph hi (MS): ph hi lng cc cht c ghi ti Khoa Ha, Trng
i hc KHTN H Ni vi ch o EI v Vin Ha hc cc hp cht thin
nhin Vin Hn lm Khoa hc v Cng ngh Vit Nam vi ch o ESI.
Ph cng hng t ht nhn (1H-NMR, 13C-NMR) c ghi trn my
Bru er V-500 ti Vin H n lm Khoa hc v Cng ngh Vit Nam ng
DMSO l m ung mi, cht chun ni l tetramethylsilan (TMS).
2.3.3. Th tc dng c t nh t o in itro
Th hot t nh hng t b o ung th (c t nh t b o) in vitro c thc
hin ti hoa Dc, Trng i hc uc gia Chungbu , Cheongju, H n
uc theo phng php MTT v gi tr IC50 c t nh trn phn mm
GraphPad Prism.
Dng t b o th nghim: t b o ung th i tr ng (SW620). Dng t
b o ung th c s dng t Ngn h ng t b o ung th ca Vin nghin cu
sinh hc v cng ngh sinh hc H n uc (KRIBB) v c nui cy trong
mi trng DMEM (Dulbecco s mo ifie ) hoc RPMI b sung 10 FBS
(huyt thanh b o thai b).
c t nh t b o ca cc cht c th bng phng php MTT theo cc
bc sau:
-
18
c hu n
Cc t b o pha logarit c tripsin ha v phn tn v o hn ch n
t b o trong mi trng DMEM hoc RPMI b sung 10 FBS. iu chnh
nng hong 1,5x104 n 3,5x104 t b o, sau chia u v o cc ging ca
a 96 ging, mi ging 200 L, cc a sau c 37oC trong iu in
5% CO2.
Sau 2 gi , cc m u th c chun b trong 20 L mi trng
DMEM/RPMI b sung 10 FBS t ung ch gc 10 mg/mL trong dimethyl
sulfoxid (DMSO) ri thm 2 L m u th v o cc ging nhiu nng hc
nhau. Cc a n y sau c thm 8 gi. Tt c cc m u c chun b
sau cho nng cui cng ca DMSO hng qu 0,1%.
c Tin h nh th
Sau 8 gi , thm v o mi ging 20 L thuc nhum MTT (nng
MTT 5 mg/ mL trong mui m phosphat PBS). Cc a c thm 3 gi
37oC trong iu in 5% CO2. Tip theo mi ging c cho 100 L dung
dch DMSO, 5 pht cho MTT formazan c ha tan. hp th c
c bc sng 510 nm.
Gi tr IC50 l nng ca m u th m , hp th gim i 50 so
vi nhm chng (trng m t nh l ging ch thm mi trng nui cy). Kt
qu cui cng l gi tr trung bnh ca ln o c lp vi gi tr hp th
khc nhau hng qu 5 . Gi tr IC50 c t nh a trn phn mm
GraphPa Prism. Trong phng php th n y, IC50 20 g/mL c coi l c
hot t nh.
2.3. . nh gi c ging thuc c cc n cht tng h c.
T nh gi tr logP ca cc n cht tng hp c bng phn mmEPIsuite.
uy trnh bao gm v cu trc 2D, to Smiles notation bng phn mm
Chems etch .0 ca CD labs sau a Smile notation v o phm mm
-
19
EPIsuite t nh cc gi tr logP.
nh gi mc ging thuc ca cc n cht a trn quy tc Lipins y
- Khi lng phn t ca cht phi nh hn 500 g/mol.
- S trung tm nhn lin t hy ro (N, O) phi nh hn 10.
- S trung tm cho lin t hy ro (NH, OH) phi nh hn 5.
- Cn bng gia t nh thn nc/ u: logP
-
20
CHNG 3: THC NGHIM, KT QU V BN LUN
3.1. HA HC
3.1.1. Tng hp ha hc
Qu trnh tng hp 4 cht trong kha lun n y c tin h nh theo s
3.1
OH
OCl
+NH2 NH2
S
iv
SNH
O
NH
H+
SNH
O
O
R
SCl
OO
i
R
SNH
OO
Br
R
SNH
OO
O
CHO
ii
R
SNH
OO
O
S
NH
O
Oiii
S 3.1: Quy trnh tng hp chung
Tc nhn v iu kin: i) 2-bromoethanamin, DMF, TEA, 25oC, 24h; ii) p-hydroxy
benzaldehyd, MeOH, K2CO3, 80oC, 24h; iii) thiazolidin-2,4-dion, EtOH, piperazin,
80oC, 24h; iv) HCl, H2O, 100
oC, 2h.
3.1.1.1. Tng hp thiazolidin-2,4-dion
S phn ng:
OH
OCl
+NH2 NH2
S
iv
SNH
O
NH
H+
SNH
O
O
S 3.2: Quy trnh tng hp thiazolidin-2,4-dion
Tc nhn v iu kin :iv) HCl, H2O, 100oC, 2h.
Tin hnh phn ng:
- Cho 9.45g (0.1 mol) acid monocloroacetic; 7.6g thioure (0.1 mol)
-
21
thioure vo bnh cu 500 ml. Thm tip 100 ml nc ct, lp sinh h n, un
hi lu bng bp in c o bc.
- Sau khi dung dch si khong 1h, rt tip 10 ml HCl 10% qua
sinh hn vo bnh cu. Tip tc hi lu thm 2h na.
- Kim tra phn ng bng TLC vi pha ng DCM:MeOH:AcOH
= 90:10:1.
X l hn h thu c:
- Rt hn hp phn ng vo cc c m, vo t lnh, s xut
hin cc tinh th.
- kt tinh lnh trong 24h.
- Lc ta qua phu lc Buchner, r a 2 ln vi 50ml nc ct
lnh.
- Ty mu: cht rn sau hi thu c, c ha tan trong 50ml
nc nng, thm 50 mg than hot, ng a thy tinh khy nng 10 pht. Lc
qua phu lc Buchner, r a 2 ln vi 10ml nc nng. Dch lc thu c
vo cc c m, vo t lnh trong 24h. Sn phm s kt tinh, mu trng nh
vng. Lc dung dch qua phu lc Buchner, r a ta 3 ln vi 10ml nc lnh.
Kt qu:
Hiu sut: 72% (8.4g).
Cm quan: tinh th mu trng .
Tonc: 124.5-126.2oC.
3.1.1.2.Tng hp (Z)-N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)
phenoxy) propyl)benzensulfonamid (4a)
tng hp cht 4a chng ti tin hnh quy trnh phn ng gm 4
giai on tng t trong s phng php chung.
a. Tng h p cht 2a
-
22
SCl
OO
SNH
OO
Br
NH2Br+
i
S 3.3: Quy trnh tng hp cht 2a
Tc nhn v iu kin : i) DMF, TEA, 25oC, 24h.
Tin hnh phn ng
- Cho 0.3ml d= 1.184g/ml (2mmol) benzensulfoclorid (1a), 2ml
DMF vo bnh cu dung tch 50ml, lm lnh dung dch bng nc .
- Thm 482 mg (2.2 mmol) 3-bromopropanamin hydrobromid vo
bnh cu.
- Tip tc thm t t 0.6ml TEA, lc u trong hn hp nc
sao cho mu ca dung dch hng c m ln.
- Phn ng c tin hnh trong nhit phng, kt hp khuy t,
thi gian 24h.
- Kim tra phn ng bng TLC vi pha ng DCM:MeOH = 40:1.
X l hn h thu c:
- Thm dn 10ml dung dch nc lnh, tip tc acid ha vi 10ml
dung dch HCl 5% (kim tra bng giy qu vn nng).
- Chit thu sn phm 3 ln bng dung mi dicloromethan , mi ln
5-10ml dung mi. Thu lp dung mi, lm khan bng Na2SO4. Gp dch chit
ca 3 ln.
- Ct quay thu hi dung mi bng my ct quay chn khng
40oC n hi thu c sn phm.
- Sy sn phm 60oC trong 4h.
Kt qu:
Hiu sut: 75% ( 0.41g)
Cm quan: tinh th vng
-
23
Tonc : 123,2-125,4oC.
b. Tng h p cht 3a
SNH
OO
BrS
NH
OO
O
CHO
ii
OH
CHO
+
S 3.4: Quy trnh tng hp cht 3a
Tc nhn v iu kin: ii) MeOH, Na2CO3, 80oC, 24h.
Tin hnh:
- Cn hn hp phn ng gm 240mg (2 mmol)p-hydroxyl
benzaldehyd cng 220mg K2CO3 vo trong bnh cu 50ml.
- Thm tip 10ml dung mi MeOH v gia nhit ln 80oC. Qu
trnh hot ha din ra trong vng 30-60 pht.
- Ha tan 1 mmol (0,28g) tinh th N-(3-bromopropyl)benzen
sulfonamide (2a) trong 10ml ung mi MeOH, ng pipet ht v a t t
vo dung dch c hot ha. Gi nhit 80oC, khy t, hi lu trong
khong 24h.
- Kim tra phn ng bng TLC vi pha ng DCM:MeOH=
40:1.
X l hn hp phn ng:
- Ct thu hi bt dung mi bng my ct quay nhit 40oC.
- Thm t t 20ml dung dch aci HCl 5 c lm lnh vo
hn hp phn ng, ng a thy tinh khuy u, xut hin ta mu trng
ng.
- Lc ta qua phu thy tinh, r a 3 ln vi 15ml nc ct nng
n khi dch lc trung tnh (th vi giy qu vn nng). Sy kh ta 60oC
trong khong 24h.
Kt qu:
-
24
Hiu sut: 60% (0.41mg).
Cm quan: tinh th trng ng.
Tonc: 188,0-190,9oC.
c.Tng h p cht 4a
SNH
OO
O
CHO
SNH
OO
O
S
NH
O
O
iiiS
NH
O
O
+
S 3.5: Quy trnh tng hp cht 4a
Tc nhn v iu kin: iii) EtOH, piperazin, 80oC, 24h.
Tin hnh:
- Cn 200mg cht 3a cng vi 120mg thiazolidin-2,4-dion vo
bnh cu 50ml.
- Thm tip 5ml EtOH khan v 0.12ml piperazin vo binh cu.
- Gia nhit 80oC, khuy t, lp sinh hn, hi lu trong 24h.
- Kim tra ph ng bng TLC vi pha ng DCM: MeOH:AcOH=
90:5:1.
X l hn hp phn ng:
- Ct thu hi bt dung mi bng my ct quay nhit 40oC.
- hn hp phn ng ra cc c m c sn 10ml HCl 5 c
lm lnh, to ta mu vng.
- Lc ta, r a ta bng nc lnh ti khi dch lc trung tnh.
- Sn phm c kt tinh li trong dung mi EtOH. Tinh th thu
c em sy kh 60oC trong 48 gi.
Kt qu:
Hiu sut: 91% (0.38g).
Cm quan: tinh th mu vng .
Tonc : 230,6-231,9oC
-
25
D liu ph c trnh by bng 3.3, 3.4, 3.5, 3.6.
3.1.1.3.Tng hp (Z)-N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)
phenoxy)propyl)-4-methylbenzenesulfonamid (4b)
a. Tng h p cht 2b
Quy trnh tin h nh tng t vi quy trnh tng hp cht 2a, i t 0.40g
(2mmol) cht 4-methylbenzen-1-sulfonyl chlorid (1b). Kt qu nh sau:
Hiu sut: 79% (0.46g).
Cm quan: tinh th mu trng nh vng
Tonc: 135,6-137,1oC.
b. Tng h p cht 3b
Cht N-(3-(4-formylphenoxy)propyl)-4-methylbenzensulfonamid (3b)
c tng hp t 0.30g (1mmol) N-(3-bromopropyl)-4-methylbenzen
sulfonamid (2b) vi p-hy roxybenzal ehy theo phng php tng t nh
tng hp cht 3a. Kt qu nh sau:
Hiu sut: 61% (0.20g).
Cm quan: tinh th mu trng .
Tonc: 199,1-201,6oC
c. Tng h p cht 4b
Cht N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)-2-methyl
phenoxy) propyl)-4-methylbenzensulfonamid (4b) c tng hp t 0.32g
(1mmol) cht N-(3-(4-formylphenoxy)propyl)-4-methylbenzensulfonamid
(3b). Qu trnh tng hp tng t nh tng hp cht 4a. Kt qu nh sau:
Hiu sut: 90% (0.38g).
Cm quan: tinh th mu vng.
Tonc : 250,6-251,3 oC.
D liu ph c trnh by bng 3.3, 3.4, 3.5, 3.6.
3.1.1.4. Tng hp (Z)-N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)
-
26
phenoxy)propyl)-4-methoxylbenzensulfonamid (4c)
a. Tng h p cht 2c
Quy trnh tin h nh tng t vi quy trnh tng hp cht 2a, i t 0.42g
(2mmol) cht 4-methoxybenzen-1-sulfonylchlorid (1c). Kt qu nh sau:
Hiu sut: 81% (0.49g).
Cm quan: tinh th mu vng.
Tonc : 140,1-142,7 oC.
b. Tng h p cht 3c
Cht N-(3-(3-formylphenoxy)propyl)-4-methoxybenzensulfonamid (3c)
c tng hp t 0.31g (1mmol) N-(3-bromopropyl)-4-methoxybenzen
sulfonamid (2b) v p-hy roxybenzal ehy . Phng php tng hp tng t
nh vi cht 3a. Kt qu nh sau:
Hiu sut: 62% (0.21g).
Cm quan: tinh th mu trng ng.
Tonc : 205,6-207,8oC
c. Tng h p cht 4c
Cht(Z)-N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)
propyl)-4-methoxybenzensulfonamid (4c) c tng hp t 0.34g (1mmol)
cht N-(3-(4-formylphenoxy)propyl)-4-methoxybenzensulfonamid (3c) v
thiazolidin-2,4-dion. Quy trnh tng hp tng t nh vi cht 4a. Kt qu
nh sau:
Hiu sut: 89% (0.39g).
Cm quan: tinh th mu vng nht.
Tonc: 260,6-261,7oC
D liu ph c trnh by bng 3.3, 3.4, 3.5, 3.6.
3.1.1.5. Tng hp (Z)-N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)
phenoxy) propyl)-4-clorobenzensulfonamid (4d)
-
27
a. Tng h p cht 2d
Quy trnh tin h nh tng t vi quy trnh tng hp cht 2a, i t 0.42g
(2mmol) cht 4-clorobenzen-1-sulfonylchlorid (1d). Kt qu nh sau:
Hiu sut: 70% (0.43g).
Cm quan: tinh th mu vng.
Tonc: 143,5-145,7oC.
b. Tng h p cht 3d
Qu trnh tng hp cht N-(3-(4-formylphenoxy)propyl)-4-cloro-
benzensulfonamid (3d) i t 0.31g (1mmol) cht N-(3-bromopropyl)-4-
chlorobenzensulfonamid (2d) cng vi 4-hydroxy benzaldehyd. Qu trnh
tng hp tng t nh vi cht 3a. Kt qu nh sau:
Hiu sut: 59% (0.2g).
Cm quan: tinh th mu vng nht.
Tonc: 202,5-203,9oC.
c. Tng h p cht 4d
Cht N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl) -
4-cloro-benzensulfonamid (4d) c tng hp t 0.34g (1mmol) cht N-(3-
(4-formylphenoxy)propyl)-4-clorobenzensulfonamid (3d) v thiazolidin-2,4-
dion. Qu trnh tng hp din ra tng t nh tng hp cht 4a. Kt qu nh
sau:
Hiu sut: 88% (0.39g).
Cm quan: tinh th m u v ng m.
Tonc: 232,7-233,6oC.
D liu ph c trnh by bng 3.3, 3.4, 3.5, 3.6.
Di y l bng tng hp cc ch s l ha v hiu sut tng hp ca
cc cht 4a-d:
-
28
Bng 3.1: Hiu sut v cc ch s ha l ca cc sn phm c tng h p
R
SNH
OO
O
S
NH
O
O
CHT R MU SC HIU SUT(%)
4a H Vng xn 91
4b 4-CH3 Vng nht 90
4c 4-OCH3 Vng nht 89
4d 4Cl V ng m 88
3.1.2. Ki tr tinh khit
tinh khit ca cc cht 4a-4b-4c-4d c kim tra bng sc k TLC
v o nhit nng chy (Tonc) ca chng.
Sc k lp mng (TLC): c tin hnh trn bn mng nhm trng silicagel
Merck 70-230 mesh, quan st vt sc i n t ngoi bc sng 254 nm.
Dung mi ha tan cc cht l aceton. Kt qu thu c trn sc u cho
thy mt vt gn, r hi soi i nh sng n t ngoi.
o n i nng chy: cc cht sau hi c tinh ch bng cch kt tinh
li u c dng tinh th rn c o im nng chy bng my o nhit
nng chy nhit in (Electrothermal digital). Nhit nng chy ca cc
cht dao dng trong khong 1-2oC.
Nh vy t kt qu chy sc k lp mng v o nhit nng chy ca 4
d n cht 4a-d, chng ti khng nh cc cht tng hp c l tinh khit v
c th s dng o ph IR, MS, 1H-NMR, 13C-NMR
-
29
Bng 3.2: Gi tr Rf v tonc ca cc sn phm cui 4a-4d
Cht R Ph ng Rf To
nc(oC)
4a H DCM:MeOH= 9:1 0.40 230,6-231,9
4b 4-CH3 DCM:MeOH= 9:1 0.40 250,6-251,3
4c 4-CH3O DCM:MeOH= 9:1 0.39 260,6-261,7
4d 4-Cl DCM:MeOH= 9:1 0.41 232,7-233,6
3.1.3. Xc nh cu trc
khng nh cu trc ca cc cht tng hp c chng ti tin hnh
ghi ph khi lng (MS) ca d n cht aldehyd trung gian v ghi ph hng
ngoi (IR), ph khi lng (MS), ph cng hng t proton (1H-NMR), ph
cng hng t carbon (13C-NMR) ca cc sn phm cui. Kt qu ph ca
cc cht c trnh by c th phn phn tch ph v ph lc.
Kt qu phn tch ph cc sn phm cui 4a-d:
a) Ph hng ngoi IR
Bng 3.3: Kt qu phn tch ph hng ngoi ca cc cht 4a-d
R
SNH
OO
O
S
NH
O
O
CHT R S sng (cm-1) ng vi cc o ng
C-O C=C(Ar) C=O C-H(Ar) CH(alken) NH
4a H 1264 1604
1569;1514
1742
1694 2937 3243 3519
4b 4-CH3 1266 1594
1568;1513
1735
1691 3121 3291 3445
4c 4-OCH3 1269 1569
1588;1566
1736
1690 3970 3121 3293
-
30
4d 4-Cl 1254 1596
1571;1510
1741
1695 3036 3274 3446
Ghi ch: l dao ng ha tr
Nhn xt: T d liu ph IR bng 3.4 v ph (ph lc 5-8) cho thy
cc cht u c s sng c trng vi cc ao ng ca cc nhm lin kt.
Trong c nh hp th c trng ca nhm C=O v nhm NH ca d vng
ami , nh hp th ca nhm ete C-O-C, nh hp th ca C-H benzen v
nh hp th ca C-H alken, cho thy sn phm c cu trc ph hp vi cng
thc cu to d kin.
b ) Ph MS
Bng 3.4: Kt qu phn tch ph khi lng ca cc cht sn phm cui
R
SNH
OO
O
S
NH
O
O
CHT R KLPT m/z
4a H 418 417,15[M-H]-
4b 4-CH3 432 430,5 [M-H]-
4c 4-OCH3 448 446,6 [M-H]-
4d 4-Cl 452 450,5 [M-H]-
Nhn xt: Qua kt qu phn tch ph bng 3.4 v ph ca 4 cht
4a-d (ph lc 1- 4), chng ti thy trn tt c cc ph u c pc phn t
c s khi ng bng s khi d kin ca cht tng ng vi cng mnh.
Ph t c pc phn mnh, cc pc phn mnh c cng nh. V vy c
th khng nh ph l ph hp vi cng thc ca 4 cht d kin 4a-d.
c) Ph cng hng t ht nhn 1H (
1H-NMR)
-
31
Bng 3.5: Kt qu phn tch ph 1H-NMR ca cc sn phm cui
R
SNH
OO
O
S
NH
O
O
6
23
45
1
1'
2'3'
4'
5'
6'
7'8'
9'
10'
STT CTCT S liu phn tch ph 1H-NMR
4a R= H
1H-NMR (500MHz, DMSO, ppm): 1,829 (2H, m, CH2);
2,92 (2H, t, J = 7.5Hz, CH2); 4,010 (2H, t, J = 6.0 Hz,
CH2) ; 7,01 (2H, d, J = 8.5Hz, H-2 ,6 ); 7,515 (2H, d, J =
9.0Hz, H-3 ,5 ); 7,572 (2H, t, J = 7.5Hz, H-3,5); 7,632
(1H, t, J = 1.0Hz, H-4) ; 7,696 (1H, s, H-7 ); 7,730 (1H, s,
NH); 7,797 (2H, t, J = 7.5Hz, H-2,6); 12,500 (1H, s, NH)
4b R= CH3
1H-NMR (500MHz, DMSO, ppm): 1,827 (2H, m,
CH2) ; 2,341 (3H, s, CH3); 2,900 (2H, t, J = 6.5 Hz , CH2)
; 3,988 (2H, t, J = 6.0Hz, CH2); 6,995 (2H, t, J = 9Hz, H-
2 ,6 ); 7,3 6 (2H, d, J = 8Hz, H-3 ,5 ); 7,513 (2H, d, J =
9Hz, H-3,5); 7,610 (1H, s, H-7 ); 7,666 (2H, d, J = 8Hz,
H-2,6); 7,729 (1H, s, NH).
4c R=OCH3
1H-NMR (500MHz, DMSO, ppm): 1,822(2H, m, CH2);
2,886 (2H, t, J = 6.5Hz, CH2); 3,799 (3H, s, CH3); 4,016
(2H, t, J = 6Hz, CH2); 6,997 (2H, d, J = 4Hz, H-2 ,6 );
7,059-7,077 (2H, m, H-3,5); 7,523 (2H, d, J = 5.5Hz, H-
3 ,5 ); 7,5 3 (1H, s, H-7 ) ; 7,705 (1H, s, NH); 7,726 (2H,
d, J = 3Hz, H-2,6).
-
32
STT CTCT S liu phn tch ph 1H-NMR
4d R= Cl
1H-NMR (500MHz, DMSO, ppm): 1,829 (2H, m, CH2);
2,94 (2H, t, J = 6Hz, CH2); 3,991 (2H, t, J = 6Hz, CH2);
6,994 (2H, d, J = 8.5Hz, H-2 ,6 ); 7,512 (2H, d, J = 8.5Hz,
H-3 ,5 ); 7,616 (2H, d, J = 8Hz, H-3,5); 7,723 (1H, s, H-
7 ); 7,786 (2H, d, J = 8.5 Hz, H-2,6); 7,819 (1H, s, H-NH).
Ghi ch: : chuyn dch ha hc (ppm); s: singlet; d: doublet; t: triplet;
m: multiplet a vch.
Nhn t: T ph 1H-NMR ca cc cht tng hp c c th thy s
lng proton, dch chuyn, bi tn hiu l ph hp vi cng thc cu
to d kin ca c 4 cht 4a-d. Cc ph u c tn hiu proton ca nhm NH,
proton ca 2 vng benzen nm trong vng dch chuyn t 7,0-8,0 ppm v 4
proton ca cu ni nm trong vng dch chuyn t 1,0-4,0 ppm. Tn hiu ca
proton nhm NH trong vng thiazolidin xut hin dng vch n trn ph
ca cht 4a, nhng ph ca cht 4b-d khng c tn hiu ny. C th do
proton ca nhm NH linh ng d b trao i nn cho tn hiu rt yu hoc
khng cho tn hiu trn ph . Cc d n cht c nhm th methyl v methoxy
u xut hin tn hiu proton c trng.
d) Ph cng hng t ht nhn 13
C (13
C-NMR)
Bng 3.6: Kt qu phn tch ph 13C-NMR ca cc sn phm cui
R
SNH
OO
O
S
NH
O
O
6
23
45
1
1'
2'3'
4'
5'
6'
7'8'
9'
10'
STT Cng thc
cu to
S liu phn tch ph 13
C-NMR
-
33
4a R= H
13C-NMR (125MHz, DMSO, ppm): 167,903 (C-
10 ); 167,377 (C-9 ); 160,150 (C-8 ); 1 0,37 (C-
1 ); 132,326 (C-1) ; 132,000 (C-2, C-6); 131,812
(C-4); 129,182 (C-3, C-5); 126,405 (C-3 , C-5 );
125,461 (C-4 ); 120,266 (C-7
); 115,291 (C-2 , C-
6 ); 6 ,945 (CH2); 39,499 (CH2); 28,700 (CH2).
4b R= CH3
13C-NMR (125MHz, DMSO, ppm): 167,938 (C-
10 ); 167, 53 (C-9 ); 160,149 (C-8 ); 1 2,551 (C-
1 ); 137,512 (C-1);131,990 (C-2, C-6); 131,779
(C-4); 129.585 (C-3, C-5); 126.476 (C-3 , C-5 );
125,456 (C- ); 120,302 (C-7 ); 115,277 (C-2 , C-
6 ); 6 ,920 (CH2); 39,333 (CH2); 28,653 (CH2),
20,918 (CH3).
4c R= OCH3
13C-NMR (125MHz, DMSO, ppm): 167,921 (C-
10 ); 167, 22 (C-9 ); 162,052 (C-8 ); 160,155 (C-
1 ); 131,986 (C-2, C-6); 131,782 (C-1); 128,595
(C-3, C-5); 125,447 (C-4); 120,268 (C-4 ); 115,265
(C-3 , C-5
); 114,831 (C-7
); 114,274 (C-2
, C-6
);
64,941 (CH2); 55,544 (CH2); 39,500 (CH2);
28,623 (CH3).
4d R= Cl
13C-NMR (125MHz, DMSO, ppm): 167,98 (C-
10 ); 167,561 (C-9 ); 160,07 (C-8 ); 139,261 (C-
1 ); 137,220 (C-1) ; 131,971 (C-2, C-6); 131,691
(C-4); 129,295 (C-3, C-5) ; 128,365 (C-3 , C-5
);
125,499 (C-4 ); 120,410 (C-7
); 115,234 (C-2
, C-
6 ); 64,793 (CH2); 39,497 (CH2); 28,621 (CH2).
Ghi ch: : chuyn dch ha hc (ppm).
-
34
Nhn xt: Qua kt qu phn tch ph bng 3.6 v ph cc cht
phn ph lc (ph lc 21-32) chng ti nhn thy :
S lng carbon, dch chuyn ha hc ca cc pic l ph hp vi
cng thc cu to d kin. Tt c cc cht u c 2 pic ca nhm C=O, 6 pic
ca vng benzen v 3 pic ca nhm CH2. D n cht methyl v methoxy u
xut hin pic ring bit ca nhm th.
Nh vy kt hp vi ph IR, MS, 1H-NMR c th khng nh cc cht
chng ti tng hp c cng thc cu to ng nh kin. Trn c s cc
cht ny tip tc c em i th hot tnh sinh hc.
3.2. TH HOT TNH SINH HC
Cc cht 4a-4d c tin h nh th tc ng in vitro trn ng t b o
ung th i tr ng (S 620) ti hoa Dc, Trng i hc uc gia
Chungbuk Cheongju, H n uc theo phng php MTT v gi tr IC50 c
t nh trn phn mm GraphPa Prism. Kt qu c trnh b y bng 3.7.
Bng 3.7: Tc ng hng t b o ung th i tr ng (SW620)
R
SNH
OO
O
S
NH
O
O
CHT R c tnh t bo IC50(g/ml) 4a H >30
4b 4-CH3 >30
4c 4-OCH3 >30
4d 4-Cl 10,89 hi ch : IC50: nn (g/m in a o n
Nhn xt: Ch c cht 4d c tc dng c ch pht trin t b o ung th,
gi tr IC50 =10,89 g/mL. Cc cht cn li khng c tc dng c ch pht
trin t b o ung th.
3.3. BN LUN
-
35
3.3.1. Tng hp ha hc
* Vi phn ng tng hp thiazolidin-2,4-dion:
- Phn ng xy ra d dng v hiu sut cao.
* Vi phn ng tng hp cc cht 2a-d:
- Phn ng gia d n xut benzensulfoclorid v amin bc 1 xy ra theo
c ch SN2 :
Hnh 3.1: phn ng tng h p cc cht 2a-d
- Xc tc phn ng n y thng l cc amin bc 3 nh triethylamin
(TEA), pyridin, quinolin, 4-dimethylaminopyridin. Qua qu trnh thc
nghim, TE c la chn lm xc tc. TEA c tc dng hp th HCl
sinh ra trong qu trnh (trnh cho phn ng chy theo chiu ngc li). Lng
TEA tha c loi i bng dung dch HCl long. Cn ch l phn ng din
ra nhit thng, khi nh TEA phi nh t t v va (0,6ml) vo hn
hp ngm trong nc , phn ng khng sinh nhit v mi trng phn
ng khng qu base, khng to ra cc sn phm khng mong mun trong qu
trnh phn ng.
- Tc nhn acyl ha dng trong phn ng ny l cc sulfonyl clorid, cht
ny d b phn hy bi nc nn phn ng phi c tin hnh trong mi
trng han nc (s dng dung mi DMF khan, cc dng c v ha cht
u phi c sy h trc khi tin hnh phn ng).
- Trong 2 thnh phn chnh tham gia phn ng, thc t thng ng
2-bromoethanamin do tc nhn ny d loi b bng dung dch HCl.
* Vi phn ng tng hp cc cht 3a-d:
-
36
- Phn ng th gia d n xut halogen v mt tc nhn base c cp in
t t o hay gi u electron nh cc alcol, phenol hay ester cng theo c ch
SN2 .
Hnh 3.2: phn ng tng h p cc cht 3a-d
- S ng K2CO3 chuyn nhm phenol th nh phenolat, nhm tng h
nng phn ng vi cc d n xut brom trong dy. p-hydroxybenzaldehyd c
ha tan trong ung mi l methanol sau thm K2CO3 ng bt, gia nhit
v huy trn trong 30-60 pht nhm hot ha al ehy . Giai on ny rt
quan trng v nu hot ha khng tt adehyd s khng phn ng sinh ra
sn phm.
- Bn cnh , trong qu trnh thc hin cc phn ng to dy cht 3a-d,
i hi cn -hydroxybenzaldehyd v cn phi loi b cht ny trong qu
trnh tinh ch (ha tan ta vi dung mi DCM, lc vi nc NaHCO3 bo ha
ha tan benzal ehy , sau chit ly phn dung mi v tin hnh ging
phn x l). Do trong cng thc ca 4-hydroxybenzaldehyd cn cha nhm
cbonyl nn nu cn cht ny, s c th xy ra phn ng ngng t aldol
vi thiazolidin-2,4-dion trong phn ng tip theo to dy cht 4a-d, nn nh
hng n s tinh khit ca cc sn phm cui. V vy cn cho 4-hydroxy
benzaldehyd vi lng va trnh phn ng ph giai on tip theo.
* Vi cc phn ng tng hp cc cht 4a-d:
- y l phn ng ngng t aldol gia cc aldehyd vi nhm methylen
ca vng thiazoldidin-2,4- ion, c tin h nh trong mi trng ethanol vi
-
37
xc tc l piperazin. Vng thiazolidin-2,4- ion c hai nhm carbonyl ht in
mnh cng thm xc tc piperazin c t nh base o l m tng h nng phn
ng ca nhm methylen v tr s 5. Phn ng ngng t cho hiu sut cao
(trung bnh xp x 90%).
- C ch phn ng xy ra theo hai giai on:
+ Giai on cng hp: (xem hnh 3.3) nhm methylen v tr s 5 c
cc hy ro linh ng c th ng tch hi nguyn t cacbon hi c mt
xc tc base, y s dng xc tc l piperazin hnh th nh tc nhn i nhn
mnh. Anion va hnh th nh tn cng v o cacbon mang in t ch ng ca
nhm cacbonyl trong al ehy to th nh sn phm cng hp. Giai on din ra
theo s :
Hnh 3.3: Giai on cng h p
+ Giai on ngng t: (xem hnh 3. ) i tc ng ca proton, mt
phn t nc c loi ra to th nh lin t i, qu trnh xy ra ng o
sn phm to th nh c mc nng lng thp nh h ni i lin hp. Giai
on din ra theo s sau:
-
38
Hnh 3.4: Giai on n n
3.3.2. Tc dng sinh hc
nh gi mc ging thuc theo 5 quy tc Lipinsky c trnh by
trong bng 3.8.
Bng 3.8: nh gi mc ging thuc theo quy tc Lipinsky
R
SNH
OO
O
S
NH
O
O
CHT R logP MW S NH, OH S N,O
4a H 2,81 418 2 7
4b 4-CH3 3,36 432 2 7
4c 4-OCH3 2,89 448 2 7
4d 4-Cl 3,46 452 2 7
Nhn xt: Tt c cc cht u tha mn c 5 yu cu ca quy tc
Lipinsky v ging thuc.
Gi tr IC50 cao i vi ba cht 4a, 4b v 4c (IC50>30 g/mL) nn 3 cht
ny khng c hot tnh, cht 4d cho gi tr IC50 =10,89 g/ mL, gi tr ny
mc trung bnh. So snh vi cht c nhm th 4-Cl cng b trc y
-
39
trong lun vn thc s c hc ca Lm Th Ha th 4d cng c c tnh trn
dng t b o S 620 cao hn (IC50 = 10,89 g/mL vi 12,3 g/mL). Tuy
nhin, khi so snh vi cht c nhm th 2-Cl v 3-Cl, th c tnh li thp
hn. C th nhn thy n xut ca nhm th clo cho hot t nh tt hn cc
n cht hng c nhm th.
-
40
KT LUN V KIN NGH
1. KT LUN
T cc kt qu nghin cu trnh b y c th rt ra 1 s kt lun sau
1.1. V tng hp v khng nh cu trc
tng hp c cc cht nh in, cc cht u cha c cng
b trong bt t i liu n o, bao gm:
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)
benzensulfonamid
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)-4-
methyl benzensulfonamid.
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)-4-
methoxy benzensulfonamid.
- N-(3-(4-((2,4-dioxothiazolidin-5-yliden)methyl)phenoxy)propyl)-4-
clorobenzensulfonamid.
* Cu trc ca cc sn phm c xc nh bng cch phn tch cc
ph MS, IR, 1H-NMR,
13C-NMR.
1.2. V hot tnh sinh hc
Bc u th c hot t nh c ch t b o ung th i tr ng ca cc
cht tng hp c. Kt qu cho thy:
- Tc ng c ch t b o ung th: ch c cht 4d th hin tc ng c
ch ng t b o ung th S 620, 3 cht cn li khng c hot tnh.
2. KIN NGH
- Tin hnh th hot tnh khng t b o ung th in vitro ca cc cht
tng hp c trn cc dng t b o ung th hc nh: MCF-7: ung th v,
PC3: ung th tuyn tin lit, sPC: ung th tuyn ty, tm ra cht c tc dng
mnh l m c s cho vic th tc dng in vivo.
- Tin h nh th hot t nh ca cc cht tng hp c trn mt s ch
-
41
tc ng hc c p ng vi nhm thiazoli in-2, - ion c a ra trong
cc nghin cu: HDAC, PPAR-, PTP1B.
- Tin h nh tng hp cc n cht thiazoli in-2,4-dion vi cc nhm th
hc nhau, c th thay Cl bng Br, I, F hoc thay i v tr ca nhm th Cl.
-
TI LIU THAM KHO
Ting Vit
1. Trn Mnh Bnh, uang t (2007), Ha h p I, NXB Y hc.
2. Nguyn Hi Nam (2011), Lin quan cu trc v tc dng sinh hc, Ti liu
a i hc, i hc Dc H Ni.
3. V Th Thanh Tm (2013), Tng hp mt s aci hy roxamic hng c
ch histondeacetylase, Kha lun tt nghi c s, Th vin i hc Dc
H Ni, H Ni.
4. Lm Th Ha (2013), Nghin cu tng hp v th tc dng sinh hc ca 1
s d n cht thiazolidindion, Lun vn d c hc, Th vin i hc
Dc H Ni,H Ni.
5. Phm Khnh Phong Lan (2007), n q an x nh
cu trc h p cht h , Quyn 1, Ti liu sau i hc, i hc y c
thnh ph H Ch Minh.
6. Phm Khnh Phong Lan(2007), n q an x nh
cu trc h p cht h , Quyn 2, Ti liu sau i hc, i hc y c
thnh ph H Ch Minh.
7. PGS.TS Trn T An (2007), Ha phn tch tp 1, NXB Y hc.
Ting Anh
8. Chan ra V., (2008), Structure of the intact PP R-gamma-RXR-nuclear
receptor complex on ND, National Institutes of Health, 456(7220), 350-
356.
9. Blanquicett C., Roman J., Hart C.M. (2008), "Thiazolidinediones as anti-
cancer agents", Cancer therapy, 6(A), 25.
10. Cress W.D., Seto E. (2000), "Histone deacetylases, transcriptional control,
and cancer", Journal of cellular physiology, 184(1), 1-16.
-
11. Faine A.L. et al, (2011), "Anti-inflammatory and antioxidant properties of
a new aryliden-thiazolidinedione in macrophages", Current medicinal
chemistry, 18(22), 3351-3360.
12. Alegaon S.G., Alagawadi K.R. (2013), " Synthesis, characterization, and
biological evaluation of thiazolidine-2,4-dione derivatives ", Medicinal
Chemistry Research, 987-994.
13. Lee H., et al., (2005), Epitope analysis of PP R gamma monoclonal
antibo y Pg 8.3 an its application for screening PP R gamma ligan s,
Journal of Immunological Methods, 296, 125-134
14. Bhattarai B.R et al, (2009), "Thiazolidinedione derivatives as PTP1B
inhibitors with antihyperglycemic and antiobesity effects", Bioorganic &
medicinal chemistry letters, 19(21), 6161-6165.
15. Bieliauskas A.V, Weerasinghe Sujith VW, Pflum Mary Kay H (2007),
"Structural requirements of HDAC inhibitors: SAHA analogs functionalized
adjacent to the hydroxamic acid", Bioorganic & medicinal chemistry letters,
17(8), 2216-2219.
16. Patil V. et al, (2010), "Synthesis and primary cytotoxicity evaluation of
new 5-benzyliden-2, 4-thiazolidinedione derivatives", European journal of
medicinal chemistry, 45(10), 4539-4544.
17. Seely B.L. et al, (1996), "Protein tyrosine phosphatase 1B interacts with
the activated insulin receptor", Diabetes, 45(10), 1379-1385.
18. Shul'gina MV. et al, (1996), "Mechanisms of the antibacterial activity of
some thiazolidinedione derivatives", Pharmaceutical Chemistry Journal,
30(6), 355-358.
19. Johnstone R.W. (2002), "Histone-deacetylase inhibitors: novel drugs for
the treatment of cancer", Nature Reviews Drug Discovery, 1(4), 287-299.
-
20. Jung M. (2001), "Inhibitors of histone deacetylase as new anticancer
agents", Current medicinal chemistry, 8(12), pp, 1505-1511.
21. McIntyre R.S. et al., (2007), "Thiazolidinediones: From antioxidant to
neurotherapeutic?", Medical hypotheses, 69(4), 773-777.
22. Mohan R. et al., (2012), "Design, synthesis, and biological evaluation of
novel 2, 4-thiazolidinedione derivatives as histone deacetylase inhibitors
targeting liver cancer ell line", Medicinal Chemistry Research, 21(7), 1156-
1165
23. Shital L. Nawale and Avinash S. Dhake (2012)," Synthesis and
evaluation of novel thiazolidinedione derivatives for antibacterial activity",
Der Pharma Chemica, 4(6),2270-2277.
24. Schoonjans Kristina, Staels Bart, Auwerx Johan (1996), "Role of the
peroxisome proliferator-activated receptor (PPAR) in mediating the effects of
fibrates and fatty acids on gene expression", Journal of lipid research, 37(5),
pp. 907-925.
25. Pattan Shashikant R, Kekare Prajact, NS Dighe, SB Bhawar, Nikalje Ana,
Pati Ashwini and MB Hole (2009)," Synthesis and Evaluation of Some New
Thiazolidinedione Derivatives for Their Antidiabetic Activities ",Asian J.
Research Chem,123-126.
26. Pattana Shashikant R, Kekareb Prajact, Ashwini Patilc, Ana Nikaljec, BS
Kitturd (2009)," Studies on the Synthesis of Novel 2,4-Thiazolidinedione
Derivatives with Antidiabetic Activity" Iranian Journal of Pharmaceutical
Sciences, 5(4): 225-230
27. Bashir A. Bhat,Shashikanth Ponnala, Devi Prasad Sahu, Priti Tiwari,
Brajendra K.Tripathi and Arvind K.Srivastava (2004),"Synthesis and
antihyperglycemic activity profiles of novel thiazolidinedione derivatives",
Bioorganic & Medicinal Chemistry, 58575864.
-
28. A. Consoli & G. Formoso (2013), "Do thiazolidinediones still have a role
in treatment of type 2 diabetes mellitus?" Diabetes, Obesity and Metabolism,
967977.
29. Bharat Raj Bhattarai, Bhooshan Kafle, Ji-Sun Hwang, Seung Wook Ham,
Keun-Hyeung Lee, Hwangseo Park, Inn-Oc Han, Hyeongjin Cho (2010),
"Novel thiazolidinedione derivatives with anti-obesity effects: Dual action as
PTP1B inhibitors and PPAR-c activators", Bioorganic & Medicinal
Chemistry Letters, 67586763
30. M.C.nsoy, O.B, Dndar, N.Atlantar, R.Ertan(2006), "Synthesis and
Antimicrobial Activity of some new 3-substituted benzyl-5-(4-cloro-2-
piperidin-1-yl-thiazole-5-yl-methylene)thiazolidin-2,4-dion Derivaties ",
Turk, J.Chem, (30), 355-360.
31. Nam N.H. et al. (2011), "Benzothiazole -containing hydroxamic acids as
histone deacetylase inhibitors and antitumor agents", Bioorganic & Medicinal
Chemistry Letters, 21(24), pp. 75097512.
32. Nam N.H., et al., (2011), Novel Hy roxamic aci s having histone
deacetylase inhibiting activity and pharmaceutical composition for
treating cancer comprising the same as active ingre ient, Korean patent No
10-2011-0050864.
33. Kim D., et al., (2003), Synthesis an biological evaluation of 3-(4-
substitutedphenyl)-N-hydroxy-2-propenamides, a new chass of histone
deacetylse inhibitors, J.Med.Chem., 46, p. 4745-5751.
34. n rianov V., et al., (2009), Novel ami e erivatives as inhibitors
of histone deacetylase: Design, synthesis and S R, European Journal
of Medicinal Chemistry, 44(3), p. 1067-1085.
-
35. Dow G.S., et al., (2008), ntimalarial activity of phenylthiazol-
bearing hydroxamate-base histone eacetylase inhibitors, Antimicrobial
agents and chemotherapy, 52, p. 3467-3477.
36. Mehen ale M., et al., (2012), Synthesis an evaluation of the
hypoglycemic and hypolipidemic activity of 5-benzylidene-2,4-
thiazolidinedione analogs in a type 2 iabetes mo el, Med Chem Res,20,
642-647.
37. Avupati V.R et al,(2012), Sythesis, characterization an biological
evaluation of some novel 2,4-thiazolidindions as potential cyttoxic,
antimicrobial ans antihyperglycemic agents, Bioorganic and Medicinal
Chemistry Letters, 22, 6442-450.
38. Deepak K Aneja et al, (2011), Synthesis of new pyrazolyl-2,4-thiazolidin
iones as antibacterial an antifungal agent, Organic and Medicinal
Chemistry Letters, 1-15
-
PH LC
Ph lc 1: Ph khi lng ca cht 4a
Ph lc 2: Ph khi lng ca cht 4b
Ph lc 3: Ph khi lng ca cht 4c
Ph lc 4: Ph khi lng ca cht 4d
Ph lc 5: Ph hng ngoi ca cht 4a
Ph lc 6: Ph hng ngoi ca cht 4b
Ph lc 7: Ph hng ngoi ca cht 4c
Ph lc 8: Ph hng ngoi ca cht 4d
Ph lc 9: Ph H1-NMR ca cht 4a
Ph lc 10: Ph H1-NMR m rng ca cht 4a
Ph lc 11: Ph H1-NMR m rng ca cht 4a
Ph lc 12: Ph H1-NMR ca cht 4b
Ph lc 13: Ph H1-NMR m rng ca cht 4b
Ph lc 14: Ph H1-NMR m rng ca cht 4b
Ph lc 15: Ph H1-NMR ca cht 4c
Ph lc 16: Ph H1-NMR m rng ca cht 4c
Ph lc 17: Ph H1-NMR m rng ca cht 4c
Ph lc 18: Ph H1-NMR ca cht 4d
Ph lc 19: Ph H1-NMR m rng ca cht 4d
Ph lc 20: Ph H1-NMR m rng ca cht 4d
Ph lc 21: Ph C13-NMR ca cht 4a
Ph lc 22: Ph C13-NMR m rng ca cht 4a
Ph lc 23: Ph C13-NMR m rng ca cht 4a
Ph lc 24: Ph C13
-NMR ca cht 4b
Ph lc 25: Ph C13
-NMR m rng ca cht 4b
Ph lc 26: Ph C13
-NMR m rng ca cht 4b
-
Ph lc 27: Ph C13
-NMR ca cht 4c
Ph lc 28: Ph C13
-NMR m rng ca cht 4c
Ph lc 29: Ph C13-NMR m rng ca cht 4c
Ph lc 30: Ph C13-NMR ca cht 4d
Ph lc 31: Ph C13-NMR m rng ca cht 4d
Ph lc 32: Ph C13-NMR m rng ca cht 4d
-
Ph lc 1: Ph khi lng ca cht 4a
C:\LTQ Orbitrap\...\Dung_2_120824160948 8/31/2012 3:00:11 PM Dung_2.4a
Dung_2_120824160948 #1511 RT: 9.34 AV: 1 NL: 3.42E4
T: ITMS - c ESI Full ms [50.00-800.00]
50 100 150 200 250 300 350 400 450
m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rela
tive A
bundance
417.1546
149.1159 346.2292
325.3794
167.1257 311.3499285.1730183.1214
358.269991.0456 257.1882108.1116 447.2584229.200764.9858 205.3339
SNH
OO
O
S
NH
O
O
-
Ph lc 2: Ph khi lng ca cht 4b
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 3: Ph khi lng ca cht 4c
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 4: Ph khi lng ca cht 4d
SNH
OO
O
S
NH
O
O
Cl
-
Ph lc 5: Ph hng ngoi ca cht 4a
Ten may: GX-PerkinElmer-USA Resolution: 4cm-1
BO MON HOA VAT LIEU-KHOA HOA-TRUONG DHKHTN
Nguoi do: Phan Thi Tuyet MaiTen mau:12aDate: 10/15/2012
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
0.0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100.0
cm-1
%T
3519
3243
2937
1742
1694
1604
1569
1514
1479
1447
1424
1399
1321
1308
1264
1181
1157
1092
1071
1050
1014
944
901
824
744
724
689
644
607
589
563
533
405
SNH
OO
O
S
NH
O
O
-
Ph lc 6: Ph hng ngoi ca cht 4b
Ten may: GX-PerkinElmer-USA Resolution: 4cm-1
BO MON HOA VAT LIEU-KHOA HOA-TRUONG DHKHTN
Nguoi do: Phan Thi Tuyet MaiTen mau: 12dDate: 10/15/2012
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
0.0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
97.7
cm-1
%T
3445
3291
3121
2970
2875
2788
1735
1691
1594
1568
1513
1464
1425
1332
1266
1225
1182
1156
1124
1092
1078
1024
972
910
836
824
814
691
662
642
608
576
541
490
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 7: Ph hng ngoi ca cht 4c
Ten may: GX-PerkinElmer-USA Resolution: 4cm-1
BO MON HOA VAT LIEU-KHOA HOA-TRUONG DHKHTN
Nguoi do: Phan Thi Tuyet MaiTen mau: 12eDate: 10/15/2012
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
0.0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
97.3
cm-1
%T
3293
3121
2970
2941
2839
2794
1763
1736
1690
1596
1588
1566
1514
1496
1460
1426
1334
1314
1300
1269
1261
1230
1183
1154
1109
1098
1079
1020
972
908
856
837
817
718
691
666
641
608
580
556
540532
491
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 8: Ph hng ngoi ca cht 4d
Ten may: GX-PerkinElmer-USA Resolution: 4cm-1
BO MON HOA VAT LIEU-KHOA HOA-TRUONG DHKHTN
Nguoi do: Phan Thi Tuyet MaiTen mau:12bDate: 10/15/2012
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
0.0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
97.0
cm-1
%T
3446
3274
3141
3036
2929
2768
1741
1695
1596
1571
1510
1474
1421
1394
1328
1254
1227
1180
1156
1091
1068
1013
968
904
826
755
705
688
640
618
608
568
541
532
483
SNH
OO
O
S
NH
O
O
Cl
-
Ph lc 9: Ph H1-NMR ca cht 4a
SNH
OO
O
S
NH
O
O
-
Ph lc 10: Ph H1-NMR m rng ca cht 4a
SNH
OO
O
S
NH
O
O
-
Ph lc 11: Ph H1-NMR m rng ca cht 4a
SNH
OO
O
S
NH
O
O
-
Ph lc 12: Ph H1-NMR ca cht 4b
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 13: Ph H1-NMR m rng ca cht 4b
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 14: Ph H1-NMR m rng ca cht 4b
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 15: Ph H1-NMR ca cht 4c
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 16: Ph H1-NMR m rng ca cht 4c
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 17: Ph H1-NMR m rng ca cht 4c
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 18: Ph H1-NMR ca cht 4d
SNH
OO
O
S
NH
O
O
Cl
-
Ph lc 19: Ph H1-NMR m rng ca cht 4d
SNH
OO
O
S
NH
O
O
Cl
-
Ph lc 20: Ph H1-NMR m rng ca cht 4d
SNH
OO
O
S
NH
O
O
Cl
-
Ph lc 21: Ph C13
-NMR ca cht 4a
SNH
OO
O
S
NH
O
O
-
Ph lc 22: Ph C13-NMR m rng ca cht 4a
SNH
OO
O
S
NH
O
O
-
Ph lc 23: Ph C13-NMR m rng ca cht 4a
SNH
OO
O
S
NH
O
O
-
Ph lc 24: Ph C13-NMR ca cht 4b
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 25: Ph C13-NMR m rng ca cht 4b
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 26: Ph C13
-NMR m rng ca cht 4b
SNH
OO
O
S
NH
O
O
CH3
-
Ph lc 27: Ph C13-NMR ca cht 4c
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 28: Ph C13-NMR m rng ca cht 4c
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 29: Ph C13-NMR m rng ca cht 4c
SNH
OO
O
S
NH
O
O
OCH3
-
Ph lc 30: Ph C13-NMR ca cht 4d
SNH
OO
O
S
NH
O
O
Cl
-
Ph lc 31: Ph C13-NMR m rng ca cht 4d
SNH
OO
O
S
NH
O
O
Cl
-
Ph lc 32: Ph C13-NMR m rng ca cht 4d
SNH
OO
O
S
NH
O
O
Cl
top related