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B Y T
TRNG I HC DC H NI
ON NGUYN THNH T
NGHIN CU TNG HP V
TH TC DNG SINH HC MT S
DN CHT CA ACID
3-(BENZIMIDAZOL-2-YL)PROPANOIC
KHA LUN TT NGHIP DC S
H NI-2013
B Y T
TRNG I HC DC H NI
ON NGUYN THNH T
NGHIN CU TNG HP V
TH TC DNG SINH HC MT S
DN CHT CA ACID
3-(BENZIMIDAZOL-2-YL)PROPANOIC
KHA LUN TT NGHIP DC S
Ngi hng dn:
PGS. TS. u n n u n
DS. u n T T an Phc
Ni thc hin:
B mn Cn n i p Dc
Trn i c Dc H i
H NI-2013
LI CM N
Trc ht ti xin by t lng knh trng v bit n su sc n thy PGS. TS.
u n n u n, ngi trc tip hng dn, cho ti nhng li khuyn qu
bu, to mi iu kin gip ti trong qu trnh nghin cu v hon thnh kha
lun tt nghip ny.
Ti xin chn thnh cm n cc thy c trong B mn Cng Nghip Dc,
c bit l DS. Nguyn Vn Giang, DS. Phm Th Hin, DS. Nguyn Th Thanh
Phc v CN. Phan Tin Thnh ca T mn Tng hp Ha dc nhit tnh gip
v to iu kin tt nht ti hon thnh kha lun tt nghip ny.
Trong qu trnh thc hin kha lun ti nhn c s gip ca cc
thy c trng i hc Dc H Ni, cn b Phng Phn tch ph- Vin Ha hc-
Vin Hn lm Khoa hc v Cng ngh Vit Nam, Phng Phn tch hu c- Vin
Hp cht thin nhin- Vin Hn lm Khoa hc v Cng ngh Vit Nam cng th
vin Trng i hc Dc H Ni, ti xin chn thnh cm n.
Cui cng, ti xin gi li cm n n b m v bn b, nhng ngi lun
ng vin, khch l ti trong cuc sng v hc tp.
H Ni, Ngy 21 thng 5 nm 2013.
Sinh vin
on Nguyn Thnh t
MC LC
Trang
Danh mc cc k hiu, cc ch vit tt
Danh mc cc bng
Danh mc cc hnh v, th
T VN ... 1
CHNG 1. TNG QUAN................................................................................... 2
1.1. Khi qut chung v benzimidazol v dn cht. 2
1.2. Tng quan v tc dng sinh hc ca benzimidazol....................................... 3
1.2.1. Tc dng khng nm, khng khun............................................................. 3
1.2.2. Tc dng sinh hc khc.. 9
1.3. Xc nh hng ti nghin cu. 14
CHNG 2. NGUYN LIU, THIT B, NI DUNG V PHNG PHP
NGHIN CU. 15
2.1. Nguyn liu v thit b.. 15
2.1.1. Nguyn liu, dung mi, ha cht.. 15
2.1.2. Thit b, dng c . 16
2.2. Ni dung nghin cu. 17
2.3. Phng php nghin cu............................................................................. 18
CHNG 3. KT QU NGHIN CU.. 20
3.1. Tng hp ha hc. 20
3.1.1. Tng hp acid 3-(benzimidazol-2-yl)propanoic (1).................................... 21
3.1.2. Tng hp ethyl 3-(benzimidazol-2-yl)propanoat (2a). 22
3.1.3. Tng hp methyl 3-(benzimidazol-2-yl)propanoat (2b).. 22
3.1.4. Tng hp 3-(benzimidazol-2-yl)propanhydrazid (3a-1). 23
3.1.5. Tng hp ethyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-yl)propanoat
(3a-2) 24
3.1.6. Tng hp ethyl 3-(1-benzyl-benzimidazol-2-yl)propanoat (3a-3). 25
3.1.7. Tng hp methyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-yl)propanoat
(3b-1)
26
3.2. Kim tra tinh khit ca cc dn cht tng hp c... 27
3.3. Xc nh cu trc ca cc cht tng hp c . 29
3.3.1. Kt qu phn tch ph cc cht trung gian (1, 2a, 2b)............................... 29
3.3.2. Kt qu phn tch ph cc cht sn phm (3a-1, 3a-2, 3a-3, 3b-1). 32
3.4. Th tc dng sinh hc. 35
3.4.1. Th hot tnh khng khun. 35
3.4.2. Th hot tnh khng nm. 38
CHNG 4. BN LUN... 41
4.1. V tng hp ha hc. 41
4.2. V tc dng sinh hc 43
KT LUN V KIN NGH 44
TI LIU THAM KHO
PH LC
DANH MC CC K HIU, CC CH VIT TT
1H-NMR Ph cng hng t ht nhn proton (1H nuclear magnetic
resonance)
AcOH Acid acetic
BuOH n-Butanol
CTCT Cng thc cu to
DMF N,N-dimethylformamid
EC50 Nng tc dng 50% (half maximal effective concentration)
ECA Ethylcloroacetat
EtOH Ethanol
g Gam
h Gi
HEPG2 Dng t bo ung th gan ngi (human hepatocellular liver
carcinoma cell line)
IC50 Nng c ch 50% i tng th (half maximal inhibitory
concentration)
IR Ph hng ngoi (infrared)
MeOH Methanol
MIC Nng c ch ti thiu (minimum inhibitory concentration)
MS Ph khi (mass spectroscopy)
MT Mi trng
PC12 Dng t bo pheochromocytoma thng thn chut (rat adrenal
pheochromocytoma cell line)
Rf H s lu gi (retension of factor)
t0
nc Nhit nng chy
TLC Sc k lp mng (thin layer chromatography)
VSV Vi sinh vt
Dao ng ha tr
DANH MC CC BNG
Trang
Bng 1.1 MIC (g/ml) ca cc cht trong nghin cu ca T. Z. Taher 6
Bng 1.2 Cc dn cht 1H-benzimidazolcarboxamidin 8
Bng 1.3 IC50 (mM) ca cc hp cht trn dng t bo HEPG2 v PC12 10
Bng 1.4 Hot ng chng ng kinh trn chut DBA/2 13
Bng 2.1 Danh mc nguyn liu, dung mi v ha cht 15
Bng 2.2 Danh mc thit b, my mc 16
Bng 2.3 Cng thc ca cc cht tng hp c 17
Bng 3.1 Kt qu tng hp ha hc 27
Bng 3.2 TLC ca cc cht trung gian 1, 2a, 2b 28
Bng 3.3 TLC ca cc cht sn phm 3a-1, 3a-2, 3a-3, 3b-1 28
Bng 3.4 Rf v t0nc ca 7 cht tng hp c 29
Bng 3.5 Kt qu phn tch ph hng ngoi ca 1, 2a, 2b 30
Bng 3.6 Kt qu phn tch ph khi lng ca 1, 2a, 2b 30
Bng 3.7 Kt qu phn tch ph cng hng t ht nhn 1H ca 1, 2a, 2b 31
Bng 3.8 Kt qu phn tch ph hng ngoi ca 3a-1, 3a-2, 3a-3, 3b-1 32
Bng 3.9 Kt qu phn tch ph khi lng ca 3a-1, 3a-2, 3a-3, 3b-1 32
Bng 3.10 Kt qu phn tch ph cng hng t ht nhn ca 3a-1, 3a-2,
3a-3, 3b-1
33
Bng 3.11 Kt qu th tc dng khng khun 38
Bng 3.12 Kt qu th tc dng khng nm 40
DANH MC CC HNH V
Trang
Hnh 1.1 5,6-dimethyl-1-( -D-ribofiranosyl)benzimidazol 2
Hnh 1.2 Cu trc vng benzimidazol 2
Hnh 1.3 Hin tng h bin benzimidazol 3
Hnh 1.4 Mt s dn cht benzimidazol c s dung trong iu tr 3
Hnh 1.5 Cc cht benzimidazol-hydrazid c tng hp 4
Hnh 1.6 Cng thc cu to ca N1-alkyl-2-aryl-5(6)-R-1H-benzimidazol
trong nghin cu ca Hakan Goeker
5
Hnh 1.7 Cng thc cu to cc cht trong nghin cu ca T. Z. Taher 6
Hnh 1.8 Dn cht 3-(2-methylbenzimidazol-1-yl)propanhydrazid 7
Hnh 1.9 Cc dn cht trong nghin cu ca Z. M. Nofal 10
Hnh 1.10 Dn cht 2-methylaminobenzimidazol tng hp bi K. Achar 12
Hnh 1.11 1H-pyrrolo[1,2-a]benzimidazol-1-on tng hp bi A. Chimirri 13
DANH MC CC S
Trang
S 3.1 Quy trnh tng hp chung 20
S 3.2 Tng hp acid 3-(benzimidazol-2-yl)propanoic (1) t o-
phenylendiamin
21
S 3.3 Tng hp ester ethyl 3-(benzimidazol-2-yl)propanoat (2a) 22
S 3.4 Tng hp ester methyl 3-(benzimidazol-2-yl)propanoat (2b) 22
S 3.5 Tng hp 3-(benzimidazol-2-yl)propanhydrazid (3a-1) 23
S 3.6 Tng hp ethyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-
yl)propanoat (3a-2)
24
S 3.7 Tng hp ethyl 3-(1-benzyl-benzimidazol-2-yl)propanoat (3a-3) 25
S 3.8 Tng hp methyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-
yl)propanoat (3b-1)
26
S 4.1 Phn ng ng vng benzimidazol 41
S 4.2 Phn ng trng hp 41
S 4.3 Phn ng ester ha xc tc H+ 42
S 4.4 Phn ng tng hp ester xc tc SOCl2 42
S 4.5 Phn ng alkyl ha 42
S 4.6 Phn ng to amoni bc 4 43
1
T VN
Sc khe lun l mt trong nhng vn c ch trng nht trong mi
quc gia. Vic s dng trc tip cc sn phm t nhin cha tr c t lu v
t c nhiu thnh tu, nhng con ng tm kim thuc mi bng phng php
tng hp lun l cn thit khi m m hnh bnh tt ngy cng phc tp. Vic nghin
cu tm ra thuc mi l kt qu kt hp ca nhiu ngnh khoa hc nh bo ch, ha
dc, vi sinh, dc lm sng, trong tng hp ha dc ng vai tr rt quan
trng. Ngy nay, cc thuc c ngun gc tng hp ha hc ngy cng gi v tr
quan trng trong cng tc chm sc sc khe.
Xu hng nghin cu v pht trin thuc mi thng da vo cu trc ca
cc cht ang c dng lm thuc hoc c trin vng sinh hc tm ra cc hot
cht c th ng dng trong iu tr lm sng.
Nhiu cng trnh nghin cu cho thy dn cht benzimidazol l nhm hp
cht d vng c tc dng sinh hc rt a dng nh: khng khun, khng nm, chng
ung th, chng co git, gim au, chng vim, h huyt p.
gp phn lm phong ph thm cc nghin cu v tng hp v tc dng
sinh hc ca dn cht benzimidazol, chng ti tin hnh thc hin ti: Nghin
cu tng hp v th tc dng sinh hc mt s dn cht ca acid 3-
(benzimidazol-2-yl)propanoic vi cc mc tiu sau:
1. Tng hp v mt s dn cht ester 3-(benzimidazol-2-yl)propanoat vi cc
nhm th khc nhau v tr N1 v tng hp hydrazid
2. Th hot tnh khng khun, khng nm ca hydrazid v mt s dn cht ester
3-(benzimidazol-2-yl)propanoat tng hp c.
2
CHNG 1.
TNG QUAN
1.1. Khi qut chung v benzimidazol v dn cht
Nm 1944, Wooley suy on rng benzimidazol c th hot ng
tng t nh purin th hin tc dng sinh hc [24]. Nm nm sau, Brink
xc nh 5,6-dimethylbenzimidazol l sn phm phn hy ca vitamin B12 v
sau tm thy mt s cc dn xut ca n c hot ng nh vitamin B12 [7].
Hnh 1.1. 5,6-dimethyl-1-( -D-ribofiranosyl)benzimidazol
Cu trc benzimidazol l mt h vng bicyclic m benzen kt hp vi
v tr 4- v 5- ca imidazol. Cc hp cht benzimidazol ni chung, v
benzimidazol carbamat ni ring l nhng cht rn kt tinh vi im nng
chy kh cao v tng i kh tan trong nc [23].
Hnh 1.2. Cu trc vng benzimidazol
Bezimidazol c cha mt nguyn t hydro c gn vi nit 1 v tr
d dng h bin:
3
Hnh 1.3. Hin tng h bin benzimidazol
Hin tng ny cng c tm thy tng t trong imidazol v amidin
[15,25].
1.2. Cc nghin cu v tc dng sinh hc ca benzimidazol
Hnh 1.4. Mt s dn cht benzimidazol c s dng trong iu tr
1.2.1. Tc dng khng nm, khng khun
Nm 2011, Yusuf Ozkay cng cng s tng hp c mt s
benzimidazol-hydrazon (1a 1n); cc dn cht ny th hin tnh khng khun
yu nhng hot tnh khng chng Candida rt ng ch nng khng
c hi [20].
4
Hnh 1.5. Cc cht benzimidazol-hydrazid c tng hp
Cc cht tng hp 1a 1n c th tc dng c ch in vitro vi sinh vt
theo k thut pha long lin tip. Tc dng khng khun c th vi dng vi
khun Gram(+) l: Staphylococcus aureus ATCC (American Type Culture
Collection) 25923, Enterococcus faecalis ATCC 29212, Bacillus subtilis v
Listeria monocytogenes, ng thi th vi vi khun Gram(-) l: Klebsiella
pneumoniae ATCC 13883, Escherichia coli ATCC 35218, E. coli ATCC
25922, Salmonella thyphimurium NRRL (Agricultural Research Service
Culture Collection) B-4420; Proteus vulgaris NRRL B-123 v nm men:
Candida albicans, Candida tropicalis v Candida glabrata ATCC 3658.
Cloramphenicol v ketoconazol c s dng lm thuc i chiu.
Hu ht cc dn cht trn khng c hot tnh chng li cc chng vi
khun, tuy nhin chng vn c hot tnh trung bnh vi E. coli khi so snh vi
cloramphenicol. Tri li vi hot tnh khng khun yu, cc hp cht 1a, 1k,
1l, 1m, 1n c ch mnh vi C. albicans, C. glabrata v C. tropicalis trong
khong MIC= 12,5 400 g/ml. Cht 1k v 1m cho thy hiu lc gp 2 ln
(MIC= 25 g/ml) so vi ketoconazol (MIC= 50 g/ml) i vi tt c cc
chng nm. Bn cnh , hot tnh khng nm vi C. tropicalis ca 1a, 1l, 1n
cng kh cao so vi cc ti liu khc. Cc hp cht khc 1b 1j th hin hot
5
tnh khng nm ngang bng vi ketoconazol. Nghin cu v lin quan cu
trc n tc dng sinh hc (SAR- structure activity relationship) cho thy mt
in t trn vng benzen l yu t quan trng lin quan n tc dng
khng nm ti u. Cc hp cht 1k 1n vi nhm th ht in t (nitro,
triflomethyl, cyano v carboxyl) cho thy tc dng hot tnh khng nm tt
nht [20].
Nm 2009, Hakan Goeker cng cng s tng hp v th cc hot
tnh khng nm Candida in vitro ca mt s N1-alkyl-2-aryl-5(6)-R-1H-
benzimidazol, theo phng php pha long ng. Cc hp cht c th
nghim in vitro cho hot tnh khng nm: C. albicans (ATCC 10231), C.
krusei (ATCC 6258), C. parapsilosis (ATCC 22019), v C. glabrata vi cc
MIC l: 3,12 g/ml; 6,25 g/ml; 12,5 g/ml v 50 g/ml.
R1=-butyl,-propyl,-pentyl
R2=-Cl,-CN,-Br,-F
R3=-Cl,-CN,-F,-Cl,-Br,-NC
R4=-CF3,-F,-Cl,-Br,-I,-CCH,-NO2,-NH2,-
CHO, MeO-N=CH-, CH3CO-, NH2C(N-
OH)CH2-
R5=-Cl,-CN
R5=-F,-OH X,Y,Z= CH, N
Hnh 1.6. Cng thc cu to ca N1-alkyl-2-aryl-5(6)-R-1H-benzimidazol trong
nghin cu ca Hakan Goeker
Kt qu th tc dng sinh hc cho thy cc benzimidazol ny u c
hot tnh tt trn mt s loi Candida. Trong 1-butyl-2-(pyridin-3-yl)-
benzimidazol-5-carbonitril v 1-butyl-2-(pyridin-4-yl)-benzimidazol-5-
carbonitril th hin tc dng ln nht vi gi tr MIC l 6,25 3,125 g/ml
[11].
Nm 2012, T. A. Taher cng cng s tng hp v nh gi kh nng
khng khun, khng nm ca mt s benimidazol-2-yl. Tt c cc cht u
6
c th tc dng in vitro trn cc vi khun Gram(-): Pseudomonas
aeruginosa, Escherichia coli, Salmonella typhi; vi khun Gram(+):
Staphylococcus aureus v nm Candida albicans bng phng php thch
pha long (tobramycin v voriconazol c s dng l thuc i chiu). nh
gi cc hp cht th nghim cho kt qu kh quan, c bit vi vi khun
Gram(-) v Candida albicans vi MIC rt thp (< 0,016 g/ml). Ngc li, vi
khun Gram(+) (Staphylococcus aureus) li c kh nng khng tt c cc cht
th nghim vi MIC > 256 g/ml [22].
Hnh 1.7. Cng thc cu to cc cht trong nghin cu ca T. Z. Taher
Bng 1.1. MIC (g/ml) ca cc cht trong nghin cu ca T. Z. Taher
Hp cht Pseud E.c. Sal. St. Ca.
3a < 0,016 < 0,016 < 0,016 >256 < 0,016
3b < 0,016 < 0,016 < 0,016 >256 < 0,016
3c < 0,016 < 0,016 < 0,016 >256 < 0,016
3d < 0,016 < 0,016 < 0,016 >256 < 0,016
4a < 0,016 < 0,016 < 0,016 >256 < 0,016
7
4b < 0,016 < 0,016 < 0,016 >256 < 0,016
5a < 0,016 < 0,016 < 0,016 >256 < 0,016
5b < 0,016 < 0,016 < 0,016 >256 < 0,016
Tobramycin 1 1,29 - 1 -
Voriconazol - - - - 0,12
Pseud: Pseudomonas aeruginosa, E.c.: Escherichia coli, Sal.: Salmonella typhi,
St.: Staphylococcus aureus, Ca.: Candida albicans.
Nghin cu ca Afaf H. El-masry cng cng s trong vic tng hp v
nh gi tc dng ca cc dn cht 3-(2-methylbenzimidazol-1-
yl)propanhydrazid (nm 2000) trn chng vi khun Gram(+) (Bacillus
cereus), Gram(-) (Escherichia coli), nm men (Saccharomyces cerevisae) v
nm mc (Aspergillus niger), thc hin theo phng php khuch tn a
(gentamycin, ampicillin c dng l cht i chiu) [10].
Hnh 1.8. Dn cht 3-(2-methylbenzimidazol-1-yl)propanhydrazid
Kt qu cho thy 6a l cht c hot ng rt mnh chng li vi khun
Bacillus cereus, nhng c tc dng yu trn Escherichia coli. Cc hp cht
6c, 6d hot ng tt chng li Bacillus cereus, trong khi hp cht 6b, 6g ch
c tc dng yu trn Bacillus cereus. Nhng tt c hp cht c th
nghim cho thy khng c tc dng chng li Saccharomyces cerevisae v
Aspergillus niger [10].
8
Nm 2012, nhn thy t l khng thuc ca cc cu khun Gram(+)
bao gm c Staphylococcus aureus khng methicillin (MRSA), t cu
Coagulas khng methicillin (MR-CNS), Pneumoniea khng penicillin, v
Enterococcus khng vancomycin (VRE) tip tc tng [21], trong ng ch
l S. aureus khi nhiu trng hp chng ny khng li methicillin [12],
Hacer Karatas cng cng s tng hp 1 dy cc cht 1H-
benzimidazolcarboxamidin v nh gi tc dng khng khun, khng nm in
vitro trn Staphylococcus aureus ATTC 25923, Candida albicans ATTC
10231, Candida krusei ATCC 6258, MRSA ATTC 43300 (Staphylococcus
aureus khng methicillin) theo phng php pha long.
Bng 1.2. Cc dn cht 1H-benzimidazolcarboxamidin
CTCT Cht R R
7a H
7b H
7c H
7d n-butyl
7e n-butyl
7f n-butyl
7g p-Cl-benzyl
7h p-Cl-benzyl
9
8a 1-naphtyl NHEt
8b Phenyl NHEt
8c p-Cl-phenyl NHEt
8d p-Cl-phenyl NH-isopropyl
8e p-Cl-phenyl NH(Me)2
9a Cl H
9b Cl Cl
Kt qu th nghim cho thy 8a, 8c, 8d, 8e, 9a, 9b c hot tnh khng
khun tt: chng MRSA (MIC < l g/ml), hp cht 9a, 9b c hot tnh va
phi chng li nm Candida. Cc cht cn li 7a 7h, 8b cho thy tc dng
yu hoc khng c tc dng khng nm, khng khun.
Th nghim cn cho thy nhm amidin thm (trong 9a, 9b) ca 1H-
benizimidazol ng vai tr quan trng trong hot ng trong hot ng khng
khun [16].
1.2.2. Tc dng khc ca benzimidazol
Nm 2011, Z. M. Nofal cng cng s tng hp c 1 dy cc dn
cht ca 1-(1H-benzimidazol-2-yl)-3-R-2-propen-1-on v th hot tnh chng
ung th trn HEPG2 (dng t bo ung th biu m gan), PC12 (t bo ty
thng thn chut). Di y l mt s dn cht thuc nhm ny:
10
Hnh 1.9. Cc dn cht trong nghin cu ca Z. M. Nofal
Kt qu nghin cu tc dng in vitro ca cc hp cht 10 15 c ch s
pht trin ca 2 dng t bo ung th ngi c trnh by bng 1.3. Nghin
cu cho thy, cht 11 c trin vng tc dng cao i vi c PC12 (IC50= 2,4
mM) v HEPG2 (IC50= 0,268 mM), trong khi cht 10a, 10b, 14, 15a, 15b
ch hot ng duy nht trn t bo PC12 v c hot tnh in vitro yu chng li
HEPG2 nng thp. Nghin cu cng ch ra ,-ceton khng no 10a l
yu t tch cc nht chng li dng t bo PC12 (IC50= 0,103 mM), dn xut
isoxazol 11 ca n cho hot ng kp trn c 2 dng t bo ny. Cc dn xut
acetohydrazid c hot tnh r rng trn dng PC12 v hot tnh yu trn
HEPG2, trong 15a cho thy hot tnh r rng nht trn PC12 (IC50= 0,251
mM) do c s hin din ca lu hunh trong chui nh km bn [19].
11
Bng 1.3. IC50 (mM) ca cc hp cht trn dng t bo HEPG2 v PC12
Hp cht IC50 (mM)
HEPG2-24h PC12-24h
10a >30 mM 0,103 mM
10b >30 mM 5mM
11 2,4 mM 0,268 mM
12 >30mM 18 mM
13 >30 mM 3 mM
14 >30 mM 0,954 mM
15a >30 mM 0,251 mM
15b >30 mM 1,5 mM
Nm 1982, trung tm nghin cu dc phm ti Kanebo (Japan)
pht trin 1 dy dn cht 2-(2-pyridinyl)benzimidazol t 55 hp cht v th
tc dng gim au, chng vim. Kt qu cho thy, 2-(5-ethyl-2-
pyridinyl)benzimidazol (KB-1043) l hp cht c hot ng chng vim,
gim au h st tt hn phenylbutazon v tiaramid. Hn na, hp cht ny c
tc dng kch thch tiu ha thp hn v ch s iu tr cao hn 2 3 ln so vi
nhm hp cht cng nghin cu [14].
Nm 2010, mt dy cc dn xut 2-methylaminobenzimidazol c
tng hp bi Kavitha Achar v c sng lc tc dng gim au, chng vim.
12
Hnh 1.10. Dn cht 2-methylaminobenzimidazol c tng hp bi K.
Achar
Tc dng gim au c nh gi theo phng php gy au bng acid
acetic. Gn nh tt c cc hp cht th nghim u cho thy tc dng gim
au mnh khi so snh vi thuc chun nimesulid. Trong , cht 16-g cho
thy tc dng gim au r rt (89%, 100 mg/kg th trng). Cn li hp cht
16-b, 16-d, 16-f, 16-g, 16-h, 16-i, 16-j v 16-k cng cho thy tc dng tt.
Trong h thng vng 2-methylaminobenzimidazol, ngi ta nhn thy rng s
c mt ca -Br, -Cl trn vng benzimidazol v -Br, -Cl, methyl, methoxy trn
phn anilin ti cc v tr khc nhau lm tng cng tc dng gim au. Ngoi
ra cc dn cht trn cng c nghin cu tc dng chng vim. Kt qu cho
thy hp cht 16-a, 16-b c kh nng chng vim tt [6].
Nm 2001, Alba Chimirri cng cng s tng hp mt s dn cht
1H-pyrrolo[1,2-a]benzimidazol-1-on v th c tnh chng co git trn chut
DBA/2 [18]. S liu c tnh ton theo phng php ca Litchfield v
Wilcoxon.
13
Hnh 1.11. 1H-pyrrolo[1,2-a]benzimidazol-1-on tng hp bi A. Chimirri
Bng 1.4. Hot ng chng ng kinh trn chut DBA/2
Cht EC50 (mol/kg)
Pha co git Pha co cng
18 24,1 17,9
19 52,7 27,4
21 23,9 17,0
22 104 79,0
23 46,4 31,7
24 63,8 54,4
Cis-25 63,8 56,9
Trans-25 39,1 25,6
Cis-26 80,3 63,8
Trans-26 43,2 29,1
27 29,2 27,0
28 18,2 9,26
29 27,2 18,5
14
Trong cc hp cht c thay th methyl v tr R2 (18, 19, 21), s c
mt ca fluor v tr C6 (cht 21) khng lm nh hng n tc dng, nhng
nu thay bng clor (cht 19) th li lm gim tc dng. Trong cc hp cht
thay th bng phenyl R2 (22, 23, 24) th hot ng mnh nht l dn cht th
6-Cl (cht 23). Hiu ng co git cng tng ln khi c mt nhm methyl C2
(25 v 26), ng phn trans hot ng mnh hn so vi cis. Cui cng, tc
dng c tng cng mnh khi thay p-tolyl v tr R2 (27, 28, 29), c bit
s kt hp ca nhm th 6-Cl v p-tolyl (cht 28) cho thy hiu qu chng co
git mnh nht [9].
1.3. La chn hng tng hp v th tc dng sinh hc cho ti
T cc ti liu tham kho c chng ti nhn thy rng cc hp cht
ca benzimidazol th v tr C2 v N1 u c nhiu tc dng sinh hc nh:
khng nm, khng khun, chng ung th, virus, chng co git, gim au,
chng vim.[6,9,11,19]. Cc benzimidazol hydrazid cng c cc tc dng
sinh hc ng k, c bit l chng nm [20]. V th hng nghin cu ca
chng ti d kin l tng hp cc dn cht ca benzimidazol-2-propanoat, bao
gm: cc hp cht th v tr N1 v tng hp mt s hydrazid, ng thi th
tc dng khng nm, khng khun ca chng.
15
CHNG 2.
NGUYN LIU, THIT B, NI DUNG
V PHNG PHP NGHIN CU
2.1. Nguyn liu v thit b
Thc hin kha lun ny chng ti s dng mt s ha cht, dung
mi v thit b ca phng th nghim Tng hp Ha dc- B mn Cng
nghip Dc.
2.1.1. u n li u, dun mi v a c t
Bng 2.1. Danh mc nguyn liu, dung mi v ha cht
STT Tn ha cht Xut x
1 Acid acetic 99,8% Merck- c
2 Acid hydrocloric Trung Quc
3 Acid monocloroacetic Trung Quc
4 Acid sulfuric Trung Quc
5 Anhydrid succinic Trung Quc
6 Benzyl clorid Trung Quc
7 Cloroform Trung Quc
8 Dicloromethan Trung Quc
9 Ethanol tuyt i Vit Nam
10 Hydrazin hydrat Trung Quc
11 Kali carbonat khan Trung Quc
12 Methanol Trung Quc
13 Natri acetat Trung Quc
14 n-Butanol Trung Quc
15 Natri carbonat Trung Quc
16 Natri hydroxyd Trung Quc
16
17 N,N-dimethylformamid Trung Quc
18 Natri sulfat khan Trung Quc
19 O-phenylendiamin Merck- c
20 Thionyl clorid Merck- c
21 Xylen Trung Quc
2.1.2. T it b dn c
Bng 2.2. Danh mc thit b, my mc
STT Tn thit b/ dng c Xut x
1 Bn mng Silicagel GF254 70-230 mesh c
2 B lc ht chn khng Buchner Buchi- Thy S
3 Cn k thut XB 320 Precisa- Thy S
4 n soi UV sc k CN6 Vilber Lourmart- c
5 My ct quay chn khng Buchi CH 9230
Model: R-210; No 0700012009
Buchi- Thy S
6 My khuy t gia nhit Wisd Wise stir MSH-20A
Daihansci- Hn Quc
7 My to kh N2 Nitrox UHP N0751
Model UHP N0751E-SIN 9900774
Domnick Hunter Ltd- Anh
8 T hotte Unilab
Model TH 1300
Jinan Unilab Instrusments. Co.
Ltd-Trung quc
9 T lnh Toshiba- Nht
10 T sy Wisd Wiseven
Serial No: 0409300124T002
Daihansci- Hn Quc
11
Bnh cu, bnh sc k, bnh 2 c, bnh nh mc 25 ml,pipet, cc c m, ng nghim, a thy tinh.
Lenz- c
17
- Ph hng ngoi (IR) c ghi trn my Perkin Elmer- USA ti b mn Ha
vt liu- Khoa Ha- Trng i hc Khoa hc t nhin H Ni v my
Impact- 410 Micolet ti Vin Ha hc- Vin Hn lm Khoa hc v Cng
ngh Vit Nam.
- Ph khi lng (MS) c ghi bng my o ph khi 1100 series LC/MSD
Trap Agilent ca Vin Ha hc v Vin Ha hc cc hp cht thin nhin-
Vin Hn lm Khoa hc v Cng ngh Vit Nam.
- Ph cng hng t ht nhn (1H-NMR) c ghi bng my Bruker AV-
500 ti Vin Ha hc- Vin Hn lm Khoa hc v Cng ngh Vit Nam.
2.2. Ni dung nghin cu
- Tng hp cc hp cht theo s sau:
Bng 2.3. Cng thc ca cc cht d kin tng hp
Cng thc Danh php
Acid 3-(benzimidazol-2-
yl)propanoic (1)
18
Ethyl 3-(benzimidazol-2-
yl)propanoat (2a)
Methyl 3-(benzimidazol-2-
yl)propanoat (2b)
3-(benzimidazol-2-
yl)propanhydrazid (3a-1)
Ethyl 3-(1-ethoxy carbonyl
methyl-benzimidazol-2-
yl)propanoat (3a-2)
Ethyl 3-(1-benzyl-benzimidazol-
2-yl)propanoat (3a-3)
Methyl 3-(1-ethoxy carbonyl
methyl-benzimidazol-2-
yl)propanoat (3b-1)
Kim tra tinh khit v xc nh cu trc ca cc cht tng hp c.
Th tc dng khng khun, khng nm ca cht 2a, 2b, 3a-1, 3a-2, 3a-3,
3b-1.
2.3. Phng php nghin cu
2.3.1. Tn p a c v kim tra tin k it
S dng phng php tng hp ha hc, da trn phn ng:
Phn ng ng vng to dn cht ester 3-(benzimidazol-2-
yl)propanoat.
Phn ng tng hp ester bng tc nhn SOCl2.
Phn ng hydrazid ha ethyl 3-(benzimidazol-2-yl)propanoat.
19
Phn ng th N1 ca ester 3-(benzimidazol-2-yl)propanoat.
Dng TLC theo di qu trnh tin trin ca phn ng.
Kim tra tinh khit ca sn phm bng TLC v nhit nng chy.
2.3.2. Xc n cu trc
Cc cht tng hp c c xc nh cu trc bng cc ph: ph hng
ngoi (IR), ph khi (MS), ph cng hng t ht nhn (1H-NMR).
* Ph hng ngoi (IR): ph hng ngoi c ghi ti Khoa Ha, Trng i
hc KHTN H Ni, trn my Perkin Elmer v my Impact- 410 ca Vin Ha
hc- Vin Hn lm Khoa hc v Cng ngh Vit Nam, vi k thut vin nn
KBr trong vng 4000- 400cm-1
. Cc mu rn c phn tn trong KBr sy
kh vi t l khong 1: 200 ri p di dng film mng di p lc cao c
ht chn khng loi b hi m [1,3].
* Ph khi lng (MS): ph khi lng cc cht c ghi ti Vin Ha hc
v Vin Ha hc cc hp cht thin nhin- Vin Hn lm Khoa hc v Cng
ngh Vit Nam, my o LC/MSD Trap Agilent hoc Agilent 6310 vi ch
o ESI hoc LC-MS [5,13].
* Ph cng hng t ht nhn (1H-NMR) c ghi trn my Bruker AV-500
ti Vin Ha hc- Vin Hn lm Khoa hc v Cng ngh Vit Nam dng
DMSO, MeOD lm dung mi [2,17].
2.3.3. T tc dn sin c k n nm, k n k un
Th tc dng sinh hc tin hnh trn 5 vi khun Gram(+), 5 vi khun
Gram(-) v 4 vi nm, thc hin ti b mn Vi sinh- Sinh hc ca Trng i
hc Dc H Ni.
Tt c cc cht tng hp c em th hot tnh theo phng php
khuch tn trn thch v sau o kch thc vng v khun. Chn cht i
chiu l penicillin v streptomycin vi khng khun. Chn intraconazol l
cht i chiu vi phng php khng nm.
20
CHNG 3
THC NGHIM, KT QU NGHIN CU
3.1. Tng hp ha hc:
Sau khi tham kho cc ti liu v da vo iu kin phng th nghim,
cc ha cht cho php, chng ti tin hnh tng hp mt s dn cht ester
3-(benzimidazol-2-yl)propanoat v tng hp hydrazid theo s sau:
S 3.1. Quy trnh tng hp chung
Tc n n v iu ki n: a) anhydrid succinic, xylen, 1400C, 3h; b) EtOH, SOCl2,
800C, 2h; c) MeOH, SOCl2, 80
0C, 2h; d) hydrazin hydrat, EtOH, 90
0C, 4h; e) ECA,
DMF, K2CO3, 250C, 12h; f) Benzyl clorid, DMF, K2CO3, 25
0C, 2h; g) ECA, DMF,
K2CO3, 250C, 4h.
21
3.1.1. Tng hp acid 3-(benzoimidazol-2-yl)propanoic (1)
- Cht acid 3-(benzimidazol-2-yl)propanoic c tng hp theo quy trnh:
S 3.2. Tng hp 3-(benzimidazol-2-yl)propanoic (1) t o-phenylendiamin
Ha tan hn hp gm: 5,4g (0,05mol) 1,2-phenylen diamin v 30ml
xylen trong bnh cu 250ml. Thm tip 10g (0,1mol) anhydrid succinic.
Lp sinh hn v un hi lu, kt hp khuy t trong 3h.
Kim tra phn ng bng TLC vi pha ng n-BuOH: AcOH: H2O = 9:
2: 2,5.
X l: hn hp phn ng sau khi kt thc c lm lnh, gn ly ta.
Ha tan ta thu c trong 60ml HCl 1M ( 600C). Lm lnh, lc ly
dch qua phu lc Buchner. Dch lc c thm 1g than hot, dng a
thy tinh khuy 600C trong 10 pht. Lc qua phu lc Buchner, ra 2
ln vi 10ml nc nng. Dch lc c vo cc c m, lm lnh v
trung ha dch lc bng natri acetat n pH= 6 th xut hin sn phm
(1) mu trng. trong 12h ta ht sn phm. Lc qua phu lc
Buchner, ra ta 3 ln vi 10ml nc lnh, thu c 6,65g sn phm
th. Kt tinh li sn phm trong EtOH 700, thu c tinh th trng, hnh
kim, nh.
Kt qu:
Hiu sut: 70% (6,65g).
Cm quan: tinh th mu trng, hnh kim, nh.
T0nc: 229-2300C
Rf = 0,67 (n-BuOH: AcOH: H2O = 9: 2: 2,5)
22
3.1.2. Tng hp ethyl 3-(benzimidazol-2-yl)propanoat (2a)
- Quy trnh tng hp:
S 3.3. Tng hp ester ethyl 3-(benzimidazol-2-yl)propanoat (2a)
Thm 30ml EtOH tuyt i vo bnh cu 2 c 100ml, lm lnh n 50C,
va khuy va thm t t 0,36ml (4,9mmol) SOCl2, sau thm tip
1g (4,9mmol) acid 3-(benzimidazol-2-yl)propanoic. Lp sinh hn, un
hi lu trong 2h. Sc kh N2 trong sut qu trnh phn ng.
Phn ng c theo di bng TLC vi pha ng n-BuOH: AcOH: H2O
= 9: 2: 2,5.
X l: hn hp phn ng sau khi kt thc c c thu hi dung mi
di p sut gim. Lm lnh bng , thm 30ml nc lnh v trung
ha bng Na2CO3 n pH= 6 th s xut hin sn phm (2a) mu vng
nht. Lc thu ta qua phu Buchner. Ra ta 2 ln vi 10ml nc, ht
kit v sy kh.
Kt qu:
Hiu sut: 70,6% (0,81g)
Cm quan: cht rn mu vng nht.
T0nc= 136-138oC
Rf = 0,3 (CHCl3: MeOH= 20: 1)
3.1.3. Tng hp methyl 3-(benzimidazol-2-yl)propanoat (2b)
- Quy trnh tng hp:
S 3.4. Tng hp ester methyl 3-(benzimidazol-2-yl)propanoat (2b)
23
Thm 30ml MeOH vo bnh cu 2 c 100ml, lm lnh n 50C, va
khuy va thm t t 0,36ml (4,9mmol) SOCl2, sau thm tip 1g
(4,9mmol) acid 3-(benzimidazol-2-yl)propanoic. Lp sinh hn, un hi
lu trong 2h. Sc kh N2 trong sut qu trnh phn ng.
Phn ng c theo di bng TLC vi pha ng n-BuOH: AcOH: H2O
= 9: 2: 2,5.
X l: hn hp phn ng sau khi kt thc c c thu hi dung mi
di p sut gim. Lm lnh bng , thm 30ml nc lnh v trung
ha bng Na2CO3 n pH= 6 th s xut hin sn phm (2b) mu trng.
Lc thu ta qua phu Buchner. Ra ta 2 ln vi 10ml nc, ht kit v
sy kh.
Kt qu:
Hiu sut: 62% (0,66g)
Cm quan: cht rn mu trng lp lnh.
T0nc= 144-1460C
Rf = 0,24 (CHCl3: MeOH= 20: 1)
3.1.4. Tng hp 3-(benzimidazol-2-yl)propanhydrazid (3a-1)
- Quy trnh tng hp:
S 3.5. Tng hp 3-(benzimidazol-2-yl)propanhydrazid (3a-1)
Ha tan hn hp gm: 1g (4,58mmol) ethyl 3-(benzimidazol-2-
yl)propanoat v 17ml EtOH tuyt i trong bnh cu 100ml. Sau
thm 8ml (82,4mmol) hydrazin monohydrat 50%. Lp sinh hn, un
hi lu v khuy trong 4h.
Phn ng c theo di bng TLC vi pha ng CHCl3: MeOH = 9: 1
24
X l: lnh trong 15 pht s xut hin ta sn phm (3a-1) mu
trng, lc qua phu Buchner, ra ta 2 ln vi nc lnh.
- Kt qu:
Hiu sut: 77% (0,73g).
Cm quan: cht rn mu trng.
T0nc= 265-2680C.
Rf = 0,16 (CHCl3: MeOH = 9: 1)
3.1.5. Tng hp ethyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-
yl)propanoat (3a-2)
- Quy trnh tng hp:
S 3.6. Tng hp ethyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-
yl)propanoat (3a-2)
Ha tan hn hp gm: 0,2g (0,9mmol) ethyl 3-(benzimidazol-2-
yl)propanoat v 7ml DMF khan trong bnh cu 2 c 50ml. Thm tip
0,13g (0,9mmol) K2CO3 khan, cui cng thm t t 0,11ml (1,0mmol)
ECA vo hn hp trn. Lp dng c lm khan (c gn CaO), sc kh N2
trong sut qu trnh phn ng, khuy nhit phng trong 12h.
Phn ng c theo di bng TLC vi pha ng CHCl3: MeOH= 20: 1
X l: hn hp phn ng sau khi kt thc c lc loi b mui v c,
ra bng CH2Cl2, thm 20ml nc lnh ri chit 2 ln vi CH2Cl2. Gp
dch chit v lc dch chit vi dung dch NaOH loi sn phm b
thy phn. Dch CH2Cl2 c lm khan v ct thu hi dung mi p
sut gim. Cht rn (3a-2) c kt tinh li trong nc, trong t lnh
1 ngy. Lc thu ta qua phu Buchner.
- Kt qu:
25
Hiu sut: 72% (0,2g)
Cm quan: cht bt mu trng.
T0nc= 65-670C.
Rf = 0,7 (CHCl3: MeOH= 20: 1)
3.1.6. Tng hp ethyl 3-(1-benzyl-benzimidazol-2-yl)propanoat (3a-3)
- Quy trnh tng hp:
S 3.7. Tng hp ethyl-3-(1-benzyl-benzimidazol-2-yl)propanoat (3a-3)
Ha tan hn hp gm: 0,25g (1,15mmol) ethyl 3-(benzimidazol-2-
yl)propanoat v 7ml DMF khan trong bnh cu 2 c 50ml. Thm tip
0,16g (1,15mmol) K2CO3 khan, cui cng thm t t 0,13ml (1,2mmol)
benzyl clorid vo hn hp trn. Lp dng c lm khan (c gn CaO),
sc kh N2 trong sut qu trnh phn ng, khuy nhit phng trong
1h30 pht.
Phn ng c theo di bng TLC vi pha ng CHCl3: MeOH= 20: 1
X l: hn hp phn ng sau khi kt thc c lc loi b mui v c,
ra bng CH2Cl2, thm 20ml nc lnh ri chit 2 ln vi CH2Cl2. Gp
dch chit dicloromethan, lc dch chit vi dung dch NaOH 10%
loi sn phm b thy phn. Dch CH2Cl2 c lm khan bng Na2SO4
khan v ct thu hi dung mi p sut gim. Cht rn (3a-3) c kt
tinh li trong nc, trong t lnh 1 ngy. Lc thu ta qua phu
Buchner.
- Kt qu:
Hiu sut: 46,0% (0,16g)
Cm quan: cht rn mu nu.
T0nc= 85-880C.
26
Rf= 0,68 (CHCl3: MeOH= 20: 1)
3.1.7. Tng hp methyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-
yl)propanoat (3b-1)
- Quy trnh tng hp:
S 3.8. Tng hp methyl 3-(1-ethoxy carbonyl methyl-benzimidazol-2-
yl)propanoat (3b-1)
Ha tan hn hp gm: 0,2g (0,98mmol) methyl 3-(benzimidazol-2-
yl)propanoat v 7ml DMF khan trong bnh cu 2 c 50ml. Thm tip
0,14g (0,98mmol) K2CO3 khan, cui cng thm t t 0,13ml
(1,18mmol) ECA vo hn hp trn. Lp dng c lm khan (c gn
CaO), sc kh N2 trong sut qu trnh phn ng, khuy nhit
phng trong 4h.
Phn ng c theo di bng TLC vi pha ng CHCl3: MeOH= 20: 1
X l: hn hp phn ng sau khi kt thc c lc loi b mui v c,
ra bng CH2Cl2, thm 20ml nc lnh ri chit 2 ln vi CH2Cl2. Gp
dch chit dicloromethan, lc dch chit vi dung dch NaOH 10%
loi sn phm b thy phn. Dch CH2Cl2 c ra, lm khan v ct thu
hi dung mi p sut gim. Cht rn (3b-1) c kt tinh li trong
nc, trong t lnh 4 ngy. Lc thu ta qua phu Buchner.
- Kt qu:
Hiu sut: 64,2% (0,18g)
Cm quan: tinh th mu trng, hnh kim, nh.
T0nc= 103-1040C
Rf= 0,48 (CHCl3: MeOH= 20: 1)
27
Nh vy, chng ti tng hp c 7 cht. Sau y l bng tm tt kt qu
tng hp ha hc:
Bng 3.1. Kt qu tng hp ha hc
STT Cht CTCT Cm quan T0nc
Hiu
sut
(%)
Rf
1 1
Tinh th mu
trng, hnh
kim.
229-
2300C
70% 0,67
2 2a
Cht rn mu
vng nht.
136-
1380C
70,6% 0,30
3 2b
Cht rn mu
trng lp lnh.
144-
1460C
62% 0,24
4 3a-1
cht rn mu
trng.
265-
2680C
77% 0,16
5 3a-2
Cht rn mu
trng.
65-
670C
72% 0,69
6 3a-3
Cht rn mu
nu
85-
880C
46% 0,68
7 3b-1
Tinh th mu
trng, hnh
kim, nh
103-
1040C
64% 0,48
3.2. Kim tra tinh khit
Chng ti kim tra tinh khit ca cc cht tng hp c bng
sc k lp mng v nhit nng chy nh sau:
- TLC c tin hnh trn bn nhm trng sn silicagel GF254 70-230 mesh,
quan st di n t ngoi bc sng 254 nm.
28
- Dung mi ha tan: EtOH 960.
- H dung mi: n-BuOH: AcOH: H2O (9: 2: 2,5); cloroform: methanol (20: 1)
v cloroform: methanol (9: 1).
Bng 3.2. TLC ca cc cht trung gian (1, 2a, 2b)
TLC
Vt o-ph 1 1 2a 1 2b
H dung mi
n-BuOH: AcOH:
H2O (9: 2: 2,5)
CHCl3: MeOH (20: 1) CHCl3: MeOH (20: 1)
Ghi ch: o-ph: o-phenylendiamin
Bng 3.3. TLC ca cc cht sn phm (3a-1, 3a-2, 3a-3, 3b-1)
TLC
Vt 2a 3a-1 2a 3a-2 2a 3a-3 2b 3b-1
Dung
mi
CHCl3: MeOH
(9: 1)
CHCl3: MeOH
(20: 1)
CHCl3: MeOH
(20: 1)
CHCl3: MeOH
(20: 1)
29
- o nhit nng chy cc cht.
Gi tr Rf v nhit nng chy (t0
nc) ca cc cht c tm tt trong bng
Bng 3.4. Rf v t0nc ca 7 cht tng hp c
Cht H dung mi khai trin Rf T0nc(
0C)
1 n-BuOH: AcOH: H2O (9: 2: 2,5) 0,67 229-230
2a CHCl3: MeOH (20: 1) 0,30 136-138
2b CHCl3: MeOH (20: 1) 0,24 144-146
3a-1 CHCl3: MeOH (9: 1) 0,16 265-268
3a-2 CHCl3: MeOH (20: 1) 0,70 65-67
3a-3 CHCl3: MeOH (20: 1) 0,68 85-88
3b-1 CHCl3: MeOH (20: 1) 0,48 103-104
Nhn xt:
- Kt qu TLC trn c 2 h dung mi thu c 1 vt sn phm, gn, r
di nh sng n t ngoi bc sng 254nm, khng c vt l.
- Nhit nng chy ca cc cht dao ng khong hp t 2 40C.
S b kt lun cc cht tng hp c l tinh khit.
Tin hnh o ph xc nh cu trc cc cht tng hp c.
3.3. Xc nh cu trc ca cc cht tng hp c
3.3.1. Kt qu phn tch ph cc cht trung gian (acid 1 v ester 2a, 2b)
a. Ph hng ngoi (IR)
30
Bng 3.5. Kt qu phn tch ph hng ngoi ca 1, 2a, 2b
Cht CTCT S sng (cm-1) ng vi cc dao ng
maxNH maxC=O maxC=C maxC-N
1
3177 1636 1557 1218
2a
3443 1728 1545 1272
2b
3149 1728 1537 1204
Nhn xt: Cc cht u c s sng ng vi cc dao ng ca cc nhm
lin kt, trong dao ng ca nhm C=O l dao ng mnh. 2a, 2b nhm
C=O r v cao hn acid 1.
b. Ph khi lng (MS)
Bng 3.6. Kt qu phn tch ph khi lng ca 1, 2a, 2b
Cht Cng thc cu to m/z
1
M=190,07 189,0 [M-H]
-
2a
M=218,11 218 [M]
+
2b
M=204,09 205,06 [M+H]
+
Nhn xt: T cc d liu phn tch ph bng 3.4 v ph (ph lc
8 10) chng ti u thy c pic phn t c s khi ng bng s khi d
kin vi cng mnh.
c. Ph cng hng t ht nhn proton (1H-NMR)
31
Bng 3.7. Kt qu phn tch ph cng hng t ht nhn 1H ca 1,2a,2b
Cht Cng thc cu to S liu phn tch 1H-NMR
1
1H-NMR (500MHz, DMSO, ppm):
2,80 (2H, t, C2H2); 3,04 (2H, t, C3H2);
7,11 (2H, d, C5H + C6H); 7,46 (2H, d,
C4H + C7H)
2a
1H-NMR (500MHz, DMSO, ppm):
1,21 (3H, t, CH3); 2,91 (2H, t, C2H2);
3,19 (2H, t, C3H2); 4,12 (2H, q,-CH2-);
7,20 (2H, dd, C5H + C6H); 7,50 (2H,
dd, C4H + C7H)
2b
1H-NMR (500MHz, DMSO, ppm):
2,88 (2H, t, C2H2); 2,06 (2H, t, C3H2);
3,59 (3H, s, O-CH3); 7,07-7,13 (2H, m,
C5H + C6H); 7,41 (1H, d, C4H); 7,51
(1H, d, C7H); 12,21 (1H, s, NH)
Ghi ch: : chuyn dch ha hc (ppm); s: singlet; d: doublet; t: triplet; q: quartet;
m: multiplet
Nhn xt: da vo kt qu phn tch ph bng 3.5 v cn c vo ph
(ph lc 15 20) chng ti nhn thy:
- S lng proton, dch chuyn, bi ca tn hiu l ph hp vi cng
thc cu to ca 3 cht. Cc cht u c cc proton ca khung benzimidazol
nm trong vng dch chuyn t 7,1 7,5. Cht 1, 2b u c proton ca NH
vng 12; ring cht 2a khng hin proton ca NH l do o trong dung mi
MeOD.
- Kt hp vi kt qu ph IR, ph MS c th khng nh 3 cht 1, 2a, 2b
chng ti tng hp c c cng thc cu to nh d kin.
32
3.3.1. Kt qu phn tch ph cc cht sn phm (3a-1; 3a-2; 3a-3; 3b-1)
a. Ph hng ngoi (IR)
Bng 3.8. Kt qu phn tch ph hng ngoi ca 3a-1, 3a-2, 3a-3, 3b-1
Cht CTCT
S sng (cm-1) ng vi cc dao ng
max
C-H
max
C-N
max
C=O
max
C=C
max nhm
c trng
khc
3a-1
2853 1175 1655 1534 3274 ( NH)
3a-2
2993
2937 1174 1733 1519
3a-3
2926
2863 1176 1728 1509
3b-1
2959
2886 1170 1736 1515
Nhn xt: Cc cht u c s sng ng vi cc dao ng ca cc nhm
lin kt. Trong dao ng ca nhm NH l dao ng mnh, c trng cho
3a-1. Dao ng ca nhm C=O c trng cho cc sn phm 3a-2, 3a-3, 3b-1.
b. Ph khi lng (MS)
Bng 3.9. Kt qu phn tch ph khi lng ca 3a-1, 3a-2, 3a-3, 3b-1
Cht Cng thc cu to m/z
3a-1
M= 204,10 204 [M]
+
33
3a-2
M= 304,14
305,13 [M+H]+
3a-3
M= 308,15
309,13 [M+H]+
3b-1
M= 290,13
291,09 [M+H]+
Nhn xt: T cc d liu phn tch ph bng 3.7 v ph (ph lc
11 14) chng ti u thy pic phn t c s khi ng bng s khi d kin
vi cng mnh.
c. Ph cng hng t ht nhn proton (1H-NMR)
Bng 3.10. Kt qu phn tch ph cng hng t ht nhn 3a-1, 3a-2, 3a-3, 3b-1
Cht Cng thc cu to S liu phn tch 1H-NMR
3a-1
1H-NMR (500MHz, DMSO, ppm):
2,55 (2H, t, C3H2); 3,01 (2H, t, C2H2);
4,16 (2H, s, NH2); 7,05-7,11 (2H, m,
C5H + C6H); 7,37 (1H, d, C4H);
7,48 (1H, d, C7H); 9,06 (1H, s, O-
NH); 12,15 (1H, s, NH)
3a-2
1H-NMR (500MHz, DMSO, ppm):
1,17 (3H, t, CH3); 1,22 (3H, t, CH3);
2,86 (2H, t, C2H2); 3,03 (2H, t, C3H2);
4,07 (2H, q, O-CH2); 4,18 (2H, q, O-
CH2); 5,21 (2H, s, N-CH2); 7,14-7,19
(2H, m, C5H + C6H); 7,47 (1H, d,
C4H); 7,55 (1H, d, C7H)
34
3a-3
1H-NMR (500MHz, DMSO, ppm):
1,15 (3H, t, CH3); 2,87 (2H, t, C2H2);
3,08 (2H, t, C3H2); 4,05 (2H, q, O-
CH2); 5,50 (2H, s, N-CH2); 7,12-7,15
(4H, m, 4CH); 7,26-7,28 (1H, m, CH);
7,31-7,33 (2H, m, 2CH); 7,44-7,46
(1H, m, CH); 7,56-7,58 (1H, m, CH)
3b-1
1H-NMR (500MHz, DMSO, ppm):
1,22 (3H, t, CH3); 2,89 (2H, t, C2H2);
3,03 (2H, d, C3H2); 3,61 (3H, s,
OCH3); 4,18 (2H, q, OCH2); 5,12 (2H,
s, N-CH2); 7,14-7,19 (2H, m, C5H +
C6H); 7,47 (1H, d, C4H); 7,56 (1H,
d, C7H)
Ghi ch: : chuyn dch ha hc (ppm); s: singlet; d: doublet; t: triplet; q: quartet;
m: multiplet
Nhn xt: da vo kt qu phn tch ph bng 3.8 v cn c vo ph
(ph lc 21 28) chng ti nhn thy:
- S lng proton, dch chuyn, bi ca tn hiu l ph hp vi
cng thc cu to ca 4 cht d kin. Cc cht u c cc proton ca khung
benzimidazol nm trong vng dch chuyn t 7,1 7,6. Cht 3a-1 vn c 1
proton ca NH v tr 12 do khng th vo v tr N1 ging nh 3a-2, 3a-3,
3b-1. Cht 3a-3 c 9 proton vng dch chuyn t 7,1 7,6 do trong phn t
c 2 vng benzen.
- Kt hp vi kt qu ph IR, ph MS c th khng nh 4 cht 3a-1,
3a-2, 3a-3, 3b-1 chng ti tng hp c c cng thc cu to nh d kin.
35
3.4. Th tc dng sinh hc
Vi mc ch tm kim cc cht c hot tnh sinh hc cao, c trin vng
trong s dng lm thuc, chng ti tin hnh th tc dng sinh hc ca cc
dn cht tng hp c. Da vo mt s kt qu nghin cu v hot tnh sinh
hc ca benzimidazol-2-yl v benzimidazol hydrazid nh: khng nm, khng
khun, chng ung th, virus, chng co git, gim au, chng vim.
[6,9,11,19], chng ti nh hng thm d tc dng khng khun, khng
nm ca cht 2a, 2b, 3a-1, 3a-2, 3a-3, 3b-1.
3.4.1. Nguyn tc, cch tin hnh th hot tnh khng khun
3.4.1.1. Cc vi sinh vt s dng trong nghin cu
Th tc dng khng khun c thc hin i vi 5 vi khun Gram(+)
v 5 vi khun Gram(-) sau:
Vi khun Gram(+) gm: Bacillus cereus ATCC 9946 (Bc),
Bacillus pumilus ATCC 10241 (Bp),
Bacillus subtilis ATCC 6633 (Bs),
Sarcina lutea ATCC 9341 (SL),
Staphylococcus aureus ATCC 1128 (Sta).
Vi khun Gram(-) gm: Escherichia coli ATCC 25922 (EC),
Proteus mirabilis BV 108 (Pro),
Pseudomonas aeruginosa VM 201 (Pseu),
Salmonella typhi DT 220 (Sal),
Shigella flexneri DT 112 (Shi).
Khng sinh i chng: Streptomycin: 14 g/ml
Penicillin: 27 UI/ml
3.4.1.2. Mi trng th nghim
Mi trng th nghim trong nghin cu c thnh phn nh sau:
36
Thnh
. phn
MT
NaCl
(%)
Cao
tht
(%)
Pepton
(%)
Thch
(%) Nc pH
Canh thang
nui cy VSV 0,5 0,3 0,5 0 Va
100ml
7,0 7,4
Thch thng 0,5 0,3 0,5 1,6
3.4.1.3. Nguyn tc
Th tc dng khng khun tin hnh theo phng php khuch tn trn
thch. Mu th (c cha hot cht th) c t ln lp thch dinh dng
cy vi sinh vt kim nh, hot cht t mu th khuch tn vo mi trng
thch s c ch s pht trin ca vi sinh vt kim nh to thnh vng v
khun.
Php th c thc hin ti b mn Vi sinh- Sinh hc, Trng i hc
Dc H Ni.
3.4.1.4. Tin hnh
- Mu th c pha vo dung mi thch hp (EtOH) vi nng l 100
g/ml. Cc khoanh giy lc v trng v c sy kh c tm 3 ln vi
dung dch mu th gm: mu cht 2a, 2b, 3a-1, 3a-2, 3a-3, 3b-1, sau mi ln
tm cc khoanh giy lc c cha mu th u c sy nhit thp hn
600C n kh ht dung mi.
- Chun b mi trng v cy VSV kim nh:
VSV kim nh c cy vo mi trng canh thang, ri nui cy cho
pht trin trong t m 370C trong thi gian 18 24 gi n nng 107 t
bo/ml (kim tra bng pha long v dy dch chun). Mi trng thch
37
thng v trng (tit trng 1180C/30 pht) c lm lnh v 45 500C v
c cy ging VSV kim nh vo vi t l 2,5 ml/100 ml. Lc trn VSV
kim nh phn tn u trong mi trng thch thng, ri vo a petri v
trng vi th tch 20 ml/a v cho thch ng li.
- t khoanh giy lc: khoanh giy lc c tm cht th v x l nh trn
c t ln b mt mi trng thch thng cha VSV kim nh theo s
nh sn.
- cc a petri c mu th c t nh trn trong t m t0 = 370C trong
18 24h, ri sau ly ra c kt qu, o ng knh vng v khun nu c
(s dng thc kp Panmer chnh xc 0,02 mm).
- nh gi kt qu: da trn ng knh vng v khun v c nh gi theo
cng thc:
2
1 1
( )
1
n n
i i
i i
D D D
D sn n
D : ng knh trung bnh vng v khun
iD : ng knh vng v khun th i
s: lch thc nghim chun c hiu chnh
n: s th nghim lm song song (n = 3).
3.2.2.4. Kt qu th nghim
38
Bng 3.11. Kt qu th tc dng khng khun
Mu ng knh vng v khun [mm] ( D , s)
Bc Bs Bp SL Sal EC Shi Pro Pseu Sta
2a - - 12,05
0,69
10,29
1,25
9,13
0,60 - - - -
10,12
0,46
2b - - 11,34
0,14 - -
12,11
1,17 -
9,42
0,66 - -
3a-1 15,42
0,79 - - - - - - - - -
3a-2 - - - - 10,97
0,68
9,59
0,46 - -
10,59
0,84 -
3a-3 - - 13,78
0,37 - -
10,47
0,72
11,03
0,45
11,10
0,52 -
10,91
0,34
3b-1 12,10
0,43 - - - - - - - - -
Peni 19,38
0,37
20,47
0,70 -
12,91
0,47 - - - - -
11,42
0,67
Strep 21,50
0,66
20,16
1,21 -
12,73
0,41
13,73
1,50
13,20
1,05
12,07
0,89
15,53
0,81
17,37
0,67
17,37
0,12
Ghi ch: Peni: penicillin; Strep: streptomycin
Kt lun:
- Mu 3a-1, 3b-1 c tc dng vi 1 vi khun th.
- Mu 2b, 3a-2 c tc dng vi 3 vi khun th.
- Mu 2a c tc dng vi 4 vi khun th.
- Mu 3a-3 c tc dng vi 5 vi khun th.
3.4.2. Nguyn tc, cch tin hnh th hot tnh khng nm
3.4.2.1. Cc vi sinh vt s dng trong nghin cu
39
Vi sinh vt kim nh: + Candida albicans (Ca)
+ Aspergillus niger (As)
+ Saccaromyces ceravitiae (Sa)
+ Penicilium sp. (Pe)
Cht i chng: Intraconazol 50 g/ml
3.4.2.2. Mi trng th nghim
Mi trng th nghim trong nghin cu c thnh phn nh sau:
Thnh
. phn
MT
Glucose
(%)
Pepton
(%)
Thch
(%) Nc pH
Sabouraud
nui cy 2,0 1,0 0
Va
100ml 7,0 7,4
Sabouraud
th nghim 2,0 1,0 1,6
3.4.2.3. Nguyn tc
Nguyn tc tng t php th khng khun.
3.4.2.4. Tin hnh
- Tin hnh tng t vi phng php khng khun. Mt s ch trong php
th khng nm:
+ Mu th c pha vo EtOH vi nng 100 g/ml.
+ Nm kim nh nui cy trong t m 28 300C, thi gian 18 24h.
+ Cc a petri c mu th c t trong t m t0= 28 300C, trong
24 48h, ri sau ly ra c kt qu, o ng knh vng v nm nu c.
- nh gi kt qu: da trn ng knh vng v nm v c nh gi theo
cng thc:
40
2
1 1
( )
1
n n
i i
i i
D D D
D sn n
D : ng knh trung bnh vng v nm
iD : ng knh vng v nm th i
s: lch thc nghim chun c hiu chnh
n: s th nghim lm song song (n=4).
3.2.2.4. Kt qu th nghim
Bng 3.12. Kt qu th tc dng khng nm
Mu ng knh vng v nm [mm] ( D , s)
Ca As Sa Pe
2a 9,73
0,31 -
11,16
0,61 -
2b - - - -
3a-1 - - - -
3a-2 Ht mu
3a-3 - 10,05
0,59
8,67
0,68 -
3b-1 8,15
0,76 -
10,21
0,46 -
Intra 16,33
2,01
15,26
0,41
9,73
0,43
12,39
0,75
Ghi ch: Intra: intraconazol
Kt lun:
- Cc cht 2a, 3a-3, 3b-1 c tc dng vi 2 vi nm th, khng c cht
no c tc dng vi Penicilium sp.
- Cht 2b, 3a-1 khng c tc dng vi tt c vi nm th.
41
CHNG 4.
BN LUN
4.1. V tng hp ha hc
Qua vic tng hp cc cht, chng ti c nhn xt sau
- Phn ng ng vng benzimidazol: gia o-phelylendiamin v anhydrid
succinic thc hin theo phng php ca Bibhucharan Chatterjee [8], xy ra
vi hiu sut kh cao. Phn ng c th xy ra 2 giai on nh sau:
S 4.1. Phn ng ng vng benzimidazol
Mt s yu t nh hng n hiu sut phn ng l:
+ Nhit phn ng: chng ti thy nhit ti u ca phn ng l
140 1450C (nhit xylen bt u si, to s hi lu). Nu nhit thp,
phn ng xy ra chm v khng hon ton. Nu nhit cao qu s to ra
nhiu tp do s trng hp hoc amin b oxy ha. Cc giai on s trng hp
nh sau:
S 4.2. Phn ng trng hp
+ Lng HCl 1M dng loi acid succinic. Nu dng t th s khng
loi b c ht acid succinic, cn khi dng nhiu qu th s lm sn phm
kh kt tinh. Chng ti nhn thy ch cn thm t t HCl 1M v un nng n
khi ta tan ht, lnh l loi c acid succinic.
42
- Phn ng tng hp cc ester: phn ng to ester gia acid carboxylic v
EtOH; MeOH xc tc H+
(H2SO4, HCl kh) c hiu sut khng cao do phn
ng thun nghch (H= 46%). C ch phn ng nh sau:
S 4.3. Phn ng ester ha xc tc H+
Chng ti nhn thy khi s dng tc nhn SOCl2 to ra cht trung
gian l acyl clorid th thu c ester vi hiu sut cao nht, phn ng xy ra
hon ton. Tuy nhin khi s dng SOCl2 cn phi lm khan dng c [2].
S 4.4. Phn ng tng hp ester tc nhn SOCl2
- Phn ng alkyl ha : to dn cht th v tr N1 thc hin theo phng
php gin on trong gio trnh K thut sn xut dc phm tp I, xy ra
theo c ch SN [4]:
S 4.5. Phn ng alkyl ha
Hiu sut phn ng thng thp. Cc iu kin phn ng sau cn c ch :
+ Nguyn liu l cc ester, trong mi trng base (K2CO3) d b thy
phn. V th, trnh thy phn ester phn ng cn lm khan tuyt i: sy
43
dng c trc khi dng, lm khan dung mi DMF v K2CO3, lp b dng c
lm khan (c CaO) loi hi m. Ngoi ra phn ng trnh un nng, ch cn
khuy nhit phng.
+ Dung mi s dng l DMF, kh loi khi x l v th lm sn phm
kh kt tinh, lm gim hiu sut.
+ T l mol gia benzimidazol v tc nhn th. Nhn thy t l mol
benzimidazol: tc nhn th = 1:1,1 l tt nht. Nu dng lng tc nhn th t
hn th phn ng khng hon ton; nu dng nhiu qu th c th to ra mui
amoni bc 4 ti N3 nh sau:
S 4.6. Phn ng to amoni bc 4
- Phn ng hydrazid: gia ethyl ester v hydrin hydrat c hiu sut kh cao.
Phn ng nhanh, x l n gin nn d dng thu c sn phm.
4.2. V tc dng sinh hc
- Cc cht ester: ethyl ester (2a) c tc dng tt (th hin trn 4 vi khun:
Bacillus pumilus, Sarcina lutea, Salmonella typhi, Staphylococcus aureus v
2 vi nm: Candida albicans, Saccaromyces ceravitiae); trong khi dn cht
methyl ester (2b) c hot tnh yu hn (3 vi khun: Bacillus pumilus,
Escherichia coli, Proteus mirabilis, khng c tc dng trn cc vi nm th).
- Cc dn cht th N1: dn cht th vi benzyl clorid (3a-3) cho thy tc dng
tt nht (c vng v khun trn 5 vi khun: Bacillus pumilus, Escherichia
coli, Shigella flexneri, Proteus mirabilis, Staphylococcus aureus v 2 vi nm:
Aspergillus niger, Saccaromyces ceravitiae). Trong cc dn cht th vi
ECA: cht 3a-2 tc dng tt trn vi khun (Salmonella typhi, Escherichia coli,
44
Pseudomonas aeruginosa), 3b-1 tc dng tt trn vi nm (Candida albicans,
Saccaromyces ceravitiae).
- Dn cht hydrazid cho thy tc dng yu: khng c hot tnh trn vi nm v
ch c tc dng trn 1 vi khun th (Bacillus cereus).
T kt qu th hot tnh ta thy: khi chuyn ester (2a, 2b) thnh
hydrazid (3a-1b) th mt tc dng trn tt c vi khun th v vi nm, nhng
li xut hin hot tnh trn chng Bacillus cereus. Khi gn thm cc nhm
akyl vo N1 th c th mt hot tnh trn 1 s chng, nhng li xut hin hot
tnh trn cc chng khc; c bit nhm benzyl (3a-3) cho thy tc dng trn
nhiu chng vi khun, vi nm.
45
KT LUN V KIN NGH
Kt lun
1. tng hp c cc cht nh d kin, trong cc hp cht 3a-2, 3a-3, 3b-1
cha thy cng b trong cc ti liu tham kho c. Bao gm:
- Acid 3-(benzimidazol-2-yl)propanoic (1)
- Ethyl 3-(benzimidazol-2-yl)propanoat (2a)
- Methyl 3-(benzimidazol-2-yl)propanoat (2b)
- 3-(benzimidazol-2-yl)propanhydrazid (3a-1)
- Ethyl 3-(1-methoxy carbonyl methyl-benzimidazol-2-yl)propanoat (3a-2)
- Ethyl 3-(1-benzyl-benzimidazol-2-yl)propanoat (3a-3)
- Methyl 3-(1-methoxy carbonyl methyl-benzimidazol-2-yl)propanoat (3b-1)
Da trn c s phn tch TLC, o nhit nng chy, d liu phn tch ph IR,
ph MS v ph cng hng t proton 1H-NMR chng ti xc nh c cu
trc ca cc cht tng hp ph hp vi cu trc d kin.
2. th tc dng khng nm, khng khun ca 6 cht: 2a, 2b, 3a-1, 3a-2, 3a-3,
3b-1. Cc cht u c hot tnh khng khun hoc khng nm nhng tc dng
khng cao.
xut
tip tc pht trin kt qu ca kha lun ny, chng ti a ra mt s xut
1. Tip tc nghin cu phn ng tng hp dn cht ca ester 3-(benzimidazol-2-
yl)propanoat vi nhm th v tr N1 hoc tng hp lm di thm mch ester.
2. Tip tc th nghim cc hot tnh sinh hc trin vng vi cc dn cht mi ca
ester 3-(benzimidazol-2yl)propanoat nh: tc dng chng ung th, tc dng
gim au chng vim, tc dng h huyt p, gp phn vo vic tm ra nhm
dn cht mi c tc dng sinh hc trin vng.
TI LIU THAM KHO
Ting Vit
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Ha Hu C, Trng i Hc Dc H Ni, trang 4-29.
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3. Nguyn Hu nh, Trn Th (1999), ng dng mt s phng php ph
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4. T Minh Kong (2007), K thut sn xut dc phm tp I, Nh xut bn Y
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5. Nguyn nh Triu (2005), Cc phng php phn tch vt l v ha l tp
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19. Nofal Z. M., et al. (2011), "Novel benzimidazole derivatives as expected
anticancer agents", Acta poloniae pharmaceutica, 68(4), pp. 519-534.
20. zkay Yusuf, Tunal Yamur, Karaca Hlya, et al. (2011), "Antimicrobial
activity of a new series of benzimidazole derivatives", Archives of
pharmacal research, 34(9), pp. 1427-1435.
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based surveillance of current antimicrobial resistance patterns and trends
among Staphylococcus aureus: 2005 status in the United States", Annals of
clinical microbiology and antimicrobials, 5(1), pp. 2.
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Antimicrobial and Cytotoxic Agents", Chemical and Pharmaceutical
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and their reversal by purines", Journal of Biological Chemistry, 152(2), pp.
225-232.
25. Wright John B. (1951), "The Chemistry of the Benzimidazoles", Chemical
Reviews, 48(3), pp. 397-541.
PH LC
Ph lc 1: Ph hng ngoi IR ca cht 1
Ph lc 2: Ph hng ngoi IR ca cht 2a
Ph lc 3: Ph hng ngoi IR ca cht 2b
Ph lc 4: Ph hng ngoi IR ca cht 3a-1
Ph lc 5: Ph hng ngoi IR ca cht 3a-2
Ph lc 6: Ph hng ngoi IR ca cht 3a-3
Ph lc 7: Ph hng ngoi IR ca cht 3b-1
Ph lc 8: Ph khi lng MS ca cht 1
Ph lc 9: Ph khi lng MS ca cht 2a
Ph lc 10: Ph khi lng MS ca cht 2b
Ph lc 11: Ph khi lng MS ca cht 3a-1
Ph lc 12: Ph khi lng MS ca cht 3a-2
Ph lc 13: Ph khi lng MS ca cht 3a-3
Ph lc 14: Ph khi lng MS ca cht 3b-1
Ph lc 15: Ph 1H-NMR ca cht 1
Ph lc 16: Ph 1H-NMR m rng ca cht 1
Ph lc 17: Ph 1H-NMR ca cht 2a
Ph lc 18: Ph 1H-NMR m rng ca cht 2a
Ph lc 19: Ph 1H-NMR ca cht 2b
Ph lc 20: Ph 1H-NMR m rng ca cht 2b
Ph lc 21: Ph 1H-NMR ca cht 3a-1
Ph lc 22: Ph 1H-NMR m rng ca cht 3a-1
Ph lc 23: Ph 1H-NMR ca cht 3a-2
Ph lc 24: Ph 1H-NMR m rng ca cht 3a-2
Ph lc 25: Ph 1H-NMR ca cht 3a-3
Ph lc 26: Ph 1H-NMR m rng ca cht 3a-3
Ph lc 27: Ph 1H-NMR ca cht 3b-1
Ph lc 28: Ph 1H-NMR m rng ca cht 3b-1
Ph lc 1: Ph hng ngoi IR ca cht 1
Ten may: GX-PerkinElmer-USA Resolution: 4cm-1
BO MON HOA VAT LIEU-KHOA HOA-TRUONG DHKHTN
Nguoi do: Phan Thi Tuyet MaiTen mau: F-esteDate: 4/5/2013
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600.0
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
3443
3058
2981
2903
2846
2741
2687
2635
1728
1623
1592
1545
1451
1428
1375
1353
1333
1272 1189
1152
1116
1054
1031
936
887
791
745
618
Ph lc 2: Ph hng ngoi IR ca cht 2a
Ph lc 3: Ph hng ngoi IR ca cht 2b
Ten may: GX-PerkinElmer-USA Resolution: 4cm-1
BO MON HOA VAT LIEU-KHOA HOA-TRUONG DHKHTN
Nguoi do: Phan Thi Tuyet MaiTen mau: HydrazitDate: 4/5/2013
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600.0
0.0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100.1
cm-1
%T
3327
3274
3081
3051
3016
2974
2939
2853
2737
2661
1768
1655
1602
1534
1481
1451
1432
1366
1326
1312
1273
1243
1223
1175
1158
1109
1022
1000
929
888
778
765
748
737
698
666
616
Ph lc 4: Ph hng ngoi IR ca cht 3a-1
Ph lc 5: Ph hng ngoi IR ca cht 3a-2
Ph lc 6: Ph hng ngoi IR ca cht 3a-3
Ph lc 7: Ph hng ngoi IR ca cht 3b-1
Ph lc 8: Ph khi lng MS ca cht 1
Ph lc 9: Ph khi lng MS ca cht 2a
Ph lc 10: Ph khi lng MS ca cht 2b
Ph lc 11: Ph khi lng MS ca cht 3a-1
M= 204
Ph lc 12: Ph khi lng MS ca cht 3a-2
Ph lc 13: Ph khi lng MS ca cht 3a-3
Ph lc 14: Ph khi lng MS ca cht 3b-1
Ph lc 15: Ph 1H-NMR ca cht 1
Ph lc 16: Ph 1H-NMR ca cht 1 (m rng)
Ph lc 17: Ph 1H-NMR ca cht 2a
Ph lc 18: Ph 1H-NMR ca cht 2a (m rng)
Ph lc 19: Ph 1H-NMR ca cht 2b
Ph lc 20: Ph 1H-NMR ca cht 2b (m rng)
Ph lc 21: Ph 1H-NMR ca cht 3a-1
Ph lc 22: Ph 1H-NMR ca cht 3a-1 (m rng)
Ph lc 23: Ph 1H-NMR ca cht 3a-2
Ph lc 24: Ph 1H-NMR ca cht 3a-2 (m rng)
Ph lc 25: Ph 1H-NMR ca cht 3a-3
Ph lc 26: Ph 1H-NMR ca cht 3a-3 (m rng)
Ph lc 27: Ph 1H-NMR ca cht 3b-1
Ph lc 28: Ph 1H-NMR ca cht 3b-1 (m rng)