Nghiên cứu tổng hợp và thử tác dụng sinh học một số dẫn chất 2-Arylbenzimidazol.pdf

B Y T

TRNG I HC DC H NI

NG TH KHNH

NGHIN CU TNG HP V TH

TC DNG SINH HC MT S DN

CHT 2-ARYLBENZIMIDAZOL

KHA LUN TT NGHIP DC S

H NI 2013

B Y T

TRNG I HC DC H NI

NG TH KHNH

NGHIN CU TNG HP V TH

TC DNG SINH HC MT S DN

CHT 2-ARYLBENZIMIDAZOL

KHA LUN TT NGHIP DC S

Ngi hng dn:

1. PGS. TS. Nguyn nh Luyn

2. DS. Phm Th Hin

Ni thc hin:

B mn Cng nghip Dc

Trng i hc Dc H Ni

H NI 2013

LI CM N

Sau mt thi gian lm vic khn trng c s gip tn tnh ca cc

thy c gio, gia nh cng bn b, ti hon thnh Kha lun tt nghip Nghin

cu tng hp v th tc dng sinh hc mt s dn cht 2-arylbenzimidazol.

Vi tt c s knh trng, trc tin ti xin by t lng bit n su sc n

thy gio PGS. TS Nguyn nh Luyn trc tip hng dn, ch bo tn tnh v

to mi iu kin gip ti nghin cu thc hin kha lun ny.

Ti cng xin gi li cm n chn thnh ti thy gio DS. Nguyn Vn

Giang, DS. Phm Th Hin v CN. Phan Tin Thnh ca T mn Tng hp Ha

dc - B mn Cng nghip dc hng dn, gip v to mi iu kin tt

nht cho ti trong sut thi gian thc hin kha lun va qua.

Ti cng xin c gi li cm n n cc thy, c gio thuc B mn Cng

nghip Dc, cng nh cc thy c trong Trng i hc Dc H Ni dy bo

ti tn tnh trong sut nm nm hc v to iu kin thun li cho ti hon thnh

kha lun tt nghip ny.

Cui cng, ti xin gi li bit n su sc n gia nh, bn b nhng ngi

ng vin, gip ti sut thi gian i hc v trong sut qu trnh thc hin

ti Kha lun tt nghip.

H Ni, ngy 21 thng 05 nm 2013

Sinh vin

ng Th Khnh

MC LC

T VN ............................................................................................................. 1

CHNG I: TNG QUAN ....................................................................................... 2

1.1. TNG QUAN V TC DNG SINH HC CA MT S DN CHT 2-

ARYL BENZIMIDAZOL ............................................................................... 2

1.1.1. Tc dng khng nm, khng khun ............................................................ 2

1.1.2. Tc dng chng ung th ............................................................................. 3

1.1.3. Tc dng chng tng huyt p .................................................................... 8

1.1.4. Tc dng chng virus ................................................................................. 8

1.2. CC PHNG PHP TNG HP MT S DN CHT 2-ARYL

BENZIMIDAZOL ........................................................................................... 9

1.2.1. ng vng to nhn 2-aryl-1H-benzimidazol t o-phenylendiamin hoc

dn cht ca o-phenylendiamin .................................................................. 9

1.2.1.1. Phn ng vi acid ................................................................................. 9

1.2.1.2. Phn ng vi aldehyd ......................................................................... 10

1.2.2. To cc dn cht trn nhn 2-aryl-1H-benzimidazol ............................... 12

1.2.2.1. Phn ng bromo ha ........................................................................... 12

1.2.2.2. Phn ng alkyl ha ............................................................................. 13

1.2.3. Phn ng th vo v tr N-1 trn nhn 2-aryl-1H-benzimidazol ............... 15

CHNG II: NGUYN LIU, THIT B, NI DUNG V PHNG PHP

NGHIN CU .......................................................................................................... 16

2.1. NGUYN LIU V THIT B .................................................................... 16

2.1.1. Ha cht v dung mi .............................................................................. 16

2.1.2. Thit b, dng c ...................................................................................... 17

2.2. NI DUNG NGHIN CU ......................................................................... 18

2.3. PHNG PHP NGHIN CU .................................................................. 19

2.3.1. Tng hp ha hc v kim tra tinh khit ............................................ 19

2.3.2. Xc nh cu trc ..................................................................................... 19

2.3.3. Th tc dng sinh hc .............................................................................. 20

CHNG III: THC NGHIM, KT QU V BN LUN .............................. 21

3.1. HA HC ...................................................................................................... 21

3.1.1. Tng hp ha hc ..................................................................................... 21

3.1.1.1. Tng hp 4-(1H-benzimidazol-2-yl)-2-methoxyphenol (I) ............... 21

3.1.1.2. Tng hp 4-(1H-benzimidazol-2-yl)-2-bromo-6-methoxyphenol (II)

............................................................................................................ 22

3.1.1.3. Tng hp 2-(4-benzyloxy)-3-bromo-5-methoxyphenyl)-1H-

benzimidazol (III) ............................................................................... 24

3.1.1.4. Tng hp ethyl 2-[2-(4-benzyloxy-3-bromo-5-methoxyphenyl)-1H-

benzimidazol-1-yl] acetat (IV) ........................................................... 25

3.1.2. Kim tra tinh khit .............................................................................. 26

3.1.3. Xc nh cu trc ..................................................................................... 28

3.1.3.1. Kt qu ph hng ngoi (IR) .............................................................. 28

3.1.3.2. Kt qu ph khi lng (MS) ............................................................. 28

3.1.3.3. Kt qu ph cng hng t ht nhn proton (1H-NMR) .................... 29

3.2. TH TC DNG SINH HC ...................................................................... 31

3.2.1. Th hot tnh khng khun ...................................................................... 31

3.2.1.1. Ging vi sinh vt kim nh ............................................................... 31

3.2.1.2. Mi trng th nghim ...................................................................... 32

3.2.1.3. Nguyn tc .......................................................................................... 32

3.2.1.4. Tin hnh ............................................................................................ 32

3.2.1.5. Kt qu th nghim ............................................................................ 33

3.2.2. Th hot tnh khng nm ......................................................................... 34

3.2.2.1. Ging vi sinh vt kim nh ............................................................... 34

3.2.2.2. Mi trng th nghim ...................................................................... 34

3.2.2.3. Nguyn tc .......................................................................................... 35

3.2.2.4. Tin hnh ............................................................................................ 35

3.2.2.5. Kt qu th nghim ............................................................................ 36

3.2.3. Th hot tnh gy c t bo .................................................................... 37

3.2.3.1. Nguyn tc .......................................................................................... 37

3.2.3.2. Cch tin hnh .................................................................................... 37

3.2.3.3. Kt qu ................................................................................................ 38

3.3. BN LUN ................................................................................................... 39

3.3.1. V tng hp ha hc ................................................................................ 39

3.3.1.1. Phn ng ng vng to nhn benzimidazol v phn ng th trn

nhn thm ........................................................................................... 39

3.3.1.2. Phn ng th v tr N-1 trn nhn benzimidazol ............................. 40

3.3.2. V tc dng sinh hc ................................................................................ 40

3.3.2.1. V tc dng khng khun ................................................................... 40

3.3.2.2. V tc dng khng nm ...................................................................... 41

3.3.2.3. V tc dng gy c t bo ................................................................. 41

KT LUN V XUT ...................................................................................... 42

TI LIU THAM KHO .............................................................................................

PH LC ......................................................................................................................

CH GII CH VIT TT

1H-NMR

Ph cng hng t ht nhn proton (1H Nuclear Magnetic

Resonance proton)

ADN Acid 2-deoxynucleic

ATCC American Type Culture Collection

BrdU 5-bromo-2'-deoxyuridin

BVDV Virus gy tiu chy b (Bovine Viral Diarhoea)

CVB Coxsackie B virus

DMF N,N-dimethylformamid

DMSO Dimethyl sulfoxid

EC50 Nng 50% tc dng ti a (Effective Concentration of 50%)

EtOH Ethanol

HIV Virus suy gim mim dch ngi (Human immunodeficiency

virus)

HSV-1 Herpes Simplex Virus

IC50 Nng c ch ti thiu 50% s pht trin ca t bo (Inhibitory

concentration of 50%)

IR Ph hng ngoi (Infrared Spectroscopy)

LM Nng di micromol (Low Micromol)

MIC Nng c ch ti thiu (Minimum Inhibitory Concentration)

MMC Mitomycin C

MRSA T cu vng khng methicillin (Methicillin Resistant

Staphylococus aureus)

MS Ph khi lng (Mass Spectroscopy)

MTT Thuc nhum 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl

tetrazolium bromid

MW Vi sng (Micro Wave)

NTCC National Type Culture Collection

Rf H s lu gi (Retention factor)

RSKK Refik Saydam National Type Culture Collection

SKLM Sc k lp mng

SNU T bo ung th d dy ngi

THF Tetrahyrdofuran

to

Nhit

tonc Nhit nng chy

VSV Vesicular stomatitis virus

VV Vaccinia virus

YFV Virus gy st vng da (Yellow Fever Virus)

DANH MC CC BNG

STT Tn bng Trang

1 Bng 1.1: Gi tr IC50 ca cc dn cht benzimidazol-4,7-dion trn 3

dng t bo ung th bng kho nghim MTT 5

2 Bng 1.2: Kt qu tng hp mt s dn cht 2-arylbenzimidazol 11

3 Bng 2.1: Danh mc cc dung mi, ha cht 16

4 Bng 2.2: Danh mc cc thit b 17

5 Bng 2.3: Danh mc cc dng c 17

6 Bng 3.1: Kt qu tng hp ha hc 26

7 Bng 3.2: Gi tr Rf v nhit nng chy (to

nc) ca cc cht 27

8 Bng 3.3: Kt qu ph hng ngoi ca cc cht I-IV 28

9 Bng 3.4: Kt qu ph khi lng ca cc cht I-V 28

10 Bng 3.5: Kt qu ph cng hng t ht nhn proton ca cc cht

I-V 29

11 Bng 3.6: Kt qu th tc dng khng khun 34

12 Bng 3.7: Kt qu th tc dng khng nm 36

13 Bng 3.8: Kt qu th hot tnh gy c t bo ca 2 cht III, IV 39

DANH MC CC HNH V, TH

STT Tn hnh v, th Trang

1 Hnh 1.1: Mt s dn cht th v tr 2, 5 ca benzimidazol v

benzoxazol 2

2 Hnh 1.2: Cng thc cu to mt s dn cht 4-methoxyphenyl-

1H-benzimidazol 4

3 Hnh 1.3: Cng thc cu to ca mt s dn cht 2-aryl-1H-

benzimidazol 7

DANH MC CC S

STT Tn s Trang

1 S 1.1: o-phenylendiamin ngng t vi acid carboxylic thm 9

2 S 1.2: 4-cloro-o-phenylenediamin ngng t vi acid 2-amino

benzoic

10

3 S 1.3: Tng hp 2-alkyl v 2-aryl-benzimidazol theo phng

php vi sng

10

4 S 1.4: o-phenylendiamin ng vng vi aldehyd 10

5 S 1.5: o-phenylendiamin ngng t vi aldehyd 11

6 S 1.6: Tng hp dn cht 4-(benzimidazol-2-yl)phenol 12

7 S 1.7: o-phenylendiamin ng vng vi aldehyd thm bng cng

ngh vi sng

12

8 S 1.8: Phn ng th N-1 ca dn cht 2-aryl-1H-benzimidazol 15

9 S 3.1: Cc giai on tng hp cc dn cht 2-phenyl

benzimidazol

21

10 S 3.2: Tng hp 4-(1H-benzimidazol-2-yl)-3-methoxyphenol (I) 22

11 S 3.3: Tng hp bromo vanillin 22

12 S 3.4: Tng hp 4-(1H-benzimidazol-2-yl)-2-bromo-6-

methoxyphenol (II)

23

13 S 3.5: Tng hp 4-benzyloxy-3-bromo-5-methoxybenzaldehyd 24

14 S 3.6: Tng hp 2-(4-benzyloxy)-3-bromo-5-methoxyphenyl)-

1H-benzimidazol (III).

25

15 S 3.7: Tng hp ethyl 2-[2-(4-benzyloxy-3-bromo-5-

methoxyphenyl)-1H-benzimidazol-1-yl] acetat (IV)

25

1

T VN

Ngy nay, tnh trng lm dng thuc din ra ph bin trn th gii c bit l

cc nc ang hoc km pht trin, m Vit Nam nm trong s . in hnh l

vic s dng thuc khng sinh khng hp l to ra rt nhiu chng vi khun

khng v a khng thuc, khin cho vic iu tr tr nn ngy cng kh khn. Mt

khc, nhim mi trng, ch n ung, sinh hot khng hp l lm xut

hin thm nhiu bnh him ngho nh bnh ung th, bnh tim mch, bnh tiu

ng, bnh ni tit. Thc t ny i hi con ngi cn tm ra nhiu loi thuc mi

c tc dng iu tr hiu qu.

Nhiu cng trnh nghin cu gn y cho thy cc dn cht 2-

arylbenzimidazol l nhm hp cht d vng c tc dng sinh hc rt a dng nh:

khng nm, khng khun, khng t bo ung th, khng vim, h huyt pVic

nghin cu tng hp v nh gi tc dng sinh hc ca chng ha hn rt nhiu

trin vng.

Chnh v vy, chng ti thc hin ti: Nghin cu tng hp v th tc

dng sinh hc ca mt s dn cht 2-arylbenzimidazol vi cc mc tiu sau:

1. Tng hp mt s dn cht 2-arylbenzimidazol vi cc nhm th khc nhau

trn nhn thm v v tr N-1.

2. Th tc dng gy c t bo v khng nm, khng khun ca mt s dn

cht 2-arylbenzimidazol tng hp c.

2

CHNG I: TNG QUAN

1.1. TNG QUAN V TC DNG SINH HC CA MT S DN CHT

2-ARYL BENZIMIDAZOL

1.1.1. Tc dng khng nm, khng khun

Nm 1997, Sener v cng s tng hp mt s dn cht th v tr 2, 5 ca

benzimidazol v benzoxazol, ng thi th tc dng c ch vi sinh vt in vitro theo

k thut pha long 2 ln lin tip. Tc dng khng khun c th vi 3 dng vi

khun Gram(+) l: Staphylococcus aureus ATCC 6538, Streptococcus faecalis

ATCC 10541, Bacillus subtilis ATCC 6033 v 3 dng vi khun Gram() l:

Escherichia coli ATCC 10536, Klepsiella pneumoniae NTCC 52211, Pseudomonas

aeruginosa RSKK 355. Kt qu cho thy cc cht tng hp c c tc dng khng

khun yu trn S. aureus, S. feacalis vi MIC nm trong khong 25-50 g/ml.

Ring 5-nitro-2-cyclohexylmethylbenzimidazol (X=NH, Y=CH2, R=NO2) c tc

dng tt hn trn S. aureus vi MIC = 12,5 g/ml.

X= O, NH; R= H, Cl, NO2, CH3; Y= CH2 hoc khng c.

Hnh 1.1: Mt s dn cht th v tr 2, 5 ca benzimidazol v benzoxazol

i vi tc dng khng nm, hu ht cc cht tng hp c u c tc

dng tt trn Candida albicans RSKK 628 vi MIC trong khong 12,5-50,0 g/ml

[21].

Nm 2005, G. Hakan cng cng s tin hnh tng hp v th tc dng

khng khun in vitro trn cc chng vi khun S. aureus v S. aureus khng

methicilin (MRSA), E. coli, E. faecalis, khng nm C. albicans ca 22 dn cht 1,2

ca 1H-benzimidazol-N-alkyl-5-carboxamidin. Kt qu cho thy hp cht 1-(2,4-

diclorobenzyl)-N-(2-diethylaminoethyl)-1H-benzimidazol-5-carboxamidin v hp

cht 3,4-diclorophenyl v tr C-2 (1) c hot tnh khng khun kh mnh, vi MIC

3

= 3,12 mg/ml i vi S. aureus v MRSA, c tc dng tt trn E. coli, E. faecalis,

C. albican [13].

1

Nm 2006, Ayhan Kilchigil v cng s th tc dng chng nm Candida

albicans, Candida glabrata v Candida krusei ca mt s dn cht benzimidazol.

Kt qu th tc dng in vitro hp cht 2-(p-fluorophenyl)-5-nitro-1-

propylbenzimidazol (2) (MIC=12,5 g/ml) c hot tnh i vi C. albicans tng t

nh fluconazol. Tuy nhin cha c hp cht no tng hp c c tc dng tt vi

c ba loi nm nh cc cht i chiu (Fluconazol, Metronidazol) [14].

2

1.1.2. Tc dng chng ung th

ADN topoisomerase l enzym xc tc cho qu trnh tho xon ca ADN siu

xon c Wong pht hin nm 1971. ADN topoisomerase c vai tr quan trng

trong hu ht cc giai on ca chu trnh t bo, n gn vi ADN to thnh phc

hp thun nghch enzym-ADN. Da vo c ch xc tc c hai loi ADN

topoisomerase l ADN topoisomerase I v II. n nm 1984, Tewey v cng s

nghin cu v cng nhn vai tr ca ADN topoisomerase nh l mc tiu phn t

ca cc thuc chng ung th. Camptothecin l thuc u tin gy c vi

topoisomerase I (c chit tch t Camptotheca acuminata) [23]. Sau nm

1993, Chen v cng s nghin cu v tm ra nhm cht mi cha vng

benzimidazol c tc dng gy c vi ADN topoisomerase I, l Hoechst 33258

4

v Hoechst 33342 thuc dn cht bis-benzimidazol. Bng cch gn vo ADN,

nhng cht ny n nh phc hp thun nghch enzym-ADN, gy mt hot tnh

enzym v cui cng gy cht t bo [7,9,27,28].

Nm 1996, Kim Jung Sun v cng s tng hp mt dy cc dn cht 4-

methoxyphenyl-1H-benzimidazol v th tc dng c ch topoisomerase I. Cht 5-

nitro-2-(4-methoxyphenyl)-1H-benzimidazol th hin tc dng c ch ln nht vi

topoisomerase I. Tri ngc vi cc dn cht bis-benzimidazol trn, nhng dn

cht trong nghin cu c rt t hoc khng c i lc gn vi ADN. Trong th

nghim gy c t bo, nhm cht ny khng c s khng cho vi dng t bo

khng topoisomerase I ca camptothecin. Tip tc, nghin cu mi lin quan cu

trc - tc dng ca 10 cht trong bng di y, cho thy tc dng gy c vi

topoisomerase I hiu qu hn khi nhm th v tr s 5 l cht nhn lin kt hydro

[15].

Hnh 1.2: Cng thc cu to mt s dn cht 4-methoxyphenyl-1H-benzimidazol

Nm 2000, Ahn Chan Mug v cng s nghin cu tc dng chng ung th

trn 3 dng t bo (dng t bo ung th bch cu chut P388, cc dng t bo ung

th biu m d dy ngi SNU-1 v SNU-16) ca mt dy cc dn cht 2-alkyl, 2-

X CN CHO CH2OH CH2NH2 CONH2 COOH H Br NO2 H

Y H H H H H H H H H NO2

5

aryl, v 2-piperazinylbenzimidazol. Cc hp cht ny c kh nng gy c mnh

chng li tt c ba dng t bo th nghim [6].

Bng 1.1: Gi tr IC50 ca cc dn cht benzimidazol-4,7-dion trn 3 dng t

bo ung th bng kho nghim MTT

Cht IC50 (g/ml)

P388 SNU-1 SNU-2

6-aziridinyl-1,2,5-trimethylbenzimidazol-4,7(1H)-

dion

0,36 0,34 0,18

6-aziridinyl-2-ethy1-1,5-dimethylbenzimidazol-4,7

(1H)-dion

1,20 1,00 0,44

6-aziridinyl-l,5-dimethyl-3-propylbenzimidazol-4,7

(1H)-dion

1.36 1,05 0,37

6-aziridinyl-2-isopropyl-1,5-dimethylbenzimidazol-

4,7(1H)-dion

1,27 3,34 0,30

6-aziridinyl-2-butyl-l,5-dimethylbenzimidazol-

4,7(1H)-dion

2,05 0,95 0,34

6-aziridinyl-2-cyclohexyl-l,5-

dimethylbenzimidazol-

4,7(1H)-dion

1,46 1,43 0,46

2-(m-amino)phenyl-6-aziridinyl-l,5-

dimethylbenzimi

dazol-4,7(1H)-dion

0,21 0,32 0,40

2-(acetamidophenyl)-6-aziridinyl-1,5-

dimethylbenzimidazol-4,7(1H)-dion

0,38 0,26 0,30

6-aziridinyl-l,5-dimethyl-2-(4-methyl-l-piperazinyl)

benzimidazol-4,7(1H)-dion

0,21 0,30 0,09

MMC 0,43 0,40 0,86

6

Nm 2008, Emine v cng s tip tc nghin cu hot tnh c ch ADN

topoisomerase I, II ca t bo nhn tht vi mt vi dn cht th v tr 2, 5 ca

benzoxazol v benzimidazol. Mi tm dn cht c th hot tnh tho xon ca

ADN topoisomerase I, II trong h thng t bo t do (pBR322 plasmid ADN).

Trong 2-(4-methoxyphenyl)-5-nitro-1H-benzimidazol (3) gy c ch c hai loi

ADN topoisomerase. ng thi vi IC50=248 M, cht ny c hiu lc gy c

ADN topoisomerase I tt hn cht i chiu l camptothecin [18].

3

Nm 2011, Romero Castro cng cng s tng hp v nh gi s b kh

nng chng ung th ca su dn cht 2-aryl-5-nitro-1H-benzimidazol. c tnh t

bo c nh gi trn by dng t bo ung th ngi bng cch s dng kho

nghim MTT. Hp cht 2-(4-cloro-3-nitrophenyl)-5-nitro-1H-benzimidazol (4) c

tc dng tt nht trn dng t bo A549 (dng t bo ung th phi). Hp cht ny

c kh nng gy cht t bo theo chu trnh trn dng t bo K562 (dng t ung th

bch cu), c ch hot ng ca poly-(ADP-ribose)-polymerase (PARP). Nhng kt

qu nghin cu trn cho thy hp cht 4 l mt tc nhn chng ung th mnh [19].

4

Nm 2012, Balram Soni v cng s tng hp v nh gi hot tnh gy c

t bo ca 11 dn xut 2-aryl-1H-benzimidazol chng li cc dng t bo ung th

ngi. Nghin cu c th trn hai dng t bo K-562 (bnh ung th bch cu

nguyn bo ty) v HL-60 (bnh bch cu promyelocytic). Trong s cc hp cht

th nghim vi dng t bo K-562, cc hp cht 1a, 7a v 10a cho thy hot ng

mnh v hp cht 5a tc dng tt c ch s pht trin trn dng t bo HL-60 [8].

7

R R1

1a H H

2a H -2NO2

3a H -3NO2

4a H -2Cl

5a H -4Cl

6a -2NO2 -2NO2

7a -2NO2 -3NO2

8a -3NO2 -2NO2

9a -3NO2 -3NO2

10a -2Cl -2NO2

11 -2Cl -3NO2

Hnh 1.3: Cng thc cu to ca mt s dn cht 2-aryl-1H-benzimidazol

Nm 2012, Ju-Fang Liu v cng s thc hin nghin cu vi hp cht 1-

benzyl-2-phenyl-benzimidazol (BPB) (5) cho thy BPB c tc dng chng ung th

trn t bo sn ngi. BPB gy cht t bo theo chu trnh c nh gi bng kho

nghim MTT v phn tch dng cytometry. Trong nghin cu ny, ta thy BPB gy

cht t bo theo chu trnh trong dng t bo sn (JJ012 v SW 1353 ). Trong nghin

cu trn ng vt cho thy gim 40% khi lng khi u sau 21 ngy iu tr [16].

5

Nm 2013, Yurttas cng cng s nghin cu tng hp cc dn cht 1-(2-aryl-

2-oxoethyl)-2-[(morpholin-4-yl)thioxomethyl]benzimidazol v th tc dng chng

ung th ca chng. Tc dng chng ung th ca cc hp cht ny c nh gi

bng kho nghim MTT, phng php BrdU trn dng t bo A549 (dng t bo

ung th phi). Hu ht cc hp cht tng hp c tc dng chn lc ng k trn

dng t bo ny [26].

8

1.1.3. Tc dng chng tng huyt p

Nm 2006, Samuel Estrada-Soto v cc cng s tin hnh nghin cu tc

dng ca ca cc dn cht v tr o-, p-ca nhn phenyl gn vo C2 trn nhn

benzimidazol vi cc nhm th (-H , -CH3 , -NO2 , -CF3) trn ng mch ch ca

chut. Cc dn cht 2-(5-nitro-1H-benzimidazol-2-yl)phenol (6) v 2-(4-

methoxyphenyl)-5-nitro-1H-benzimidazol (3) c tc dng gin mch, chng tng

huyt p [12].

6

1.1.4. Tc dng chng virus

Nm 2007, Vasquez cng cng s nghin cu tng hp cc dn xut ca 1-

benzoyl-2-(2-nitrophenyl)-1H-benzimidazol v th tc dng c ch s sao chp

ngc ca virus HIV-1. Trong , hp cht 1-(2,6-diclorobenzoyl)-2-(2-

nitrophenyl)-1H-benzimidazol (7) cho hot ng c ch chng li RT-HIV l tt

nht [24].

7

Nm 2010, Mechele Tonelli v cng s th tc dng chng virus ca 76 dn

cht 2-phenylbenzimidazol. Kt qu cho thy tc dng chng virus theo th t gim

dn CVB-2, BVDV, HSV-1 v YFV, trong khi khng c tc dng trn HIV-1,

VSV v RSV, VV ch nhy cm vi 1 s hp cht. Hp cht 5,6-dicloro-2-(4-

nitrophenyl)benzimidazol (8) c hot tnh cao v chn lc chng li VV (EC50 = 0,1

9

LM), kt qu cho thy c tc dng mnh hn 18 hoc 110 ln so vi cc thuc tham

chiu l acid mycophenolic v 6-azauridin (EC50 = 1,8 LM v 11 LM) v gim c

tc ng c hi ln cc t bo th [29].

8

1.2. CC PHNG PHP TNG HP MT S DN CHT 2-ARYL

BENZIMIDAZOL

1.2.1. ng vng to nhn 2-aryl-1H-benzimidazol t o-phenylendiamin hoc

dn cht ca o-phenylendiamin

1.2.1.1. Phn ng vi acid

Thng thng, o-phenylendiamin c ngng t vi cc acid mono

carboxylic, trong iu kin phn ng loi nc, s dng cc acid nh acid

polyphosphoric (PPA), acid hydrocloric, acid boric hoc acid p-toluensulfonic to

ra cc nhm th khc nhau v tr s 2 trn nhn benzimidazol. Tuy nhin vic s

dng tc nhn yu hn c bit l acid Lewis, mui v c, acid v c ci thin c

hiu sut v tinh khit ca sn phm [11].

S 1.1: o-phenylendiamin ngng t vi acid carboxylic thm

Trong mt s trng hp, phng php Phillips dng HCl 4N v un hi lu

cho php thc hin phn ng nhit thp hn v tng hiu sut phn ng. Vi

cc acid thm nh acid benzoic th hiu sut phn ng thng thp, tng hiu

sut cn thc hin phn ng trong ng kn vi nhit 180oC-185oC [25].

Theo phng php ca Sharma v cng s, tng hp nhn benzimidazol i

t 4-cloro-o-phenylendiamin c ngng t vi acid 2-aminobenzoic s dng acid

polyphosphoric (PPA) theo s sau [22]:

10

S 1.2: 4-cloro-o-phenylenediamin ngng t vi acid 2-amino benzoic

T nm 1995, nhiu dn cht benzimidazol c tng hp theo cng ngh

vi sng. Nm 2007, Dubey v Moorthy tng hp cc cht 2-alkyl v 2-

arylbenzimidazol theo phng php ny. Thi gian phn ng gim n pht, hiu

sut tng 10-50% so vi phng php trc y [11].

R= H, CH3, C6H5, 4-NH2C6H4, 4-ClC6H4

S 1.3: Tng hp 2-alkyl v 2-arylbenzimidazol theo phng php vi

sng

1.2.1.2. Phn ng vi aldehyd

Phn ng ca o-phenylendiamin vi aldehyd cn c cht oxy ha to nhn

benzimidazol. Nhng cht oxy ha c s dng nh: nitrobenzen, benzoquinon,

natri metabisulfit, thy ngn (I) oxyd, ch (IV) tetra acetat, iod, ng (II) acetat,

indium perfluorooctan sulfonat[11].

S 1.4: o-phenylendiamin ng vng vi aldehyd thm

Weidenhagen l ngi u tin thc hin phn ng gia diamin v aldehyd

s dng ng acetat l cht oxy ha trong dung mi nc hoc alcol. Theo phng

php ca Weidenhagen, nhiu dn cht benzimidazol th v tr s 2 c tng

hp [25].

Nm 2006, Nagawade v cng s nghin cu tng hp mt s dn cht

benzimidazol t o-phenylendiamin v dn cht vi aldehyd s dng cht xc tc

BF3.OEt2 cho hiu sut rt cao. Phng php ny, c tc dng rt tt vi cc

Cht oxy ha

11

aldehyd thm, khng no hoc bo [17]. Di y l mt s dn cht 2-

arylbenzimidazol c tng hp theo phng php ny:

Bng 1.2: Kt qu tng hp mt s dn cht 2-arylbenzimidazol

Diamin Aldehyd Sn phm

Hiu

sut

%

tonc (

oC)

90 288-290

89 292-293

84 310-311

90 198-200

86 220-221

Nm 2012, Chen v cng s nghin cu tng hp cc dn cht v tr C2

trn nhn benzimidazol s dng cht xc tc FeCl3/Al2O3. Kt qu cho thy phn

ng din ra iu kin nh nhng hn, thi gian phn ng ngn hn, hiu sut cao

hn, cht xc tc khng c v r tin [10].

S 1.5: o-phenylendiamin ngng t vi aldehyd

12

Nm 2009, Savall v cng s nghin cu tng hp dn cht 2-

arylbenzimidazol s dng cht xc tc l natri metabisulfit (Na2S2O5) cho hiu sut

kh cao trn 85% [20].

S 1.6: Tng hp dn cht 4-(benzimidazol-2-yl)phenol

Gn y, Navarrtete v cng s s dng cng ngh vi sng trong phn

ng ng vng ca o-phenylenediamin vi aldehyd trong tng hp dn cht 2-

(alkyloxyaryl)-1H-benzimidazol vi hiu sut cao. Phn ng c 3 thnh phn v

khng cn dung mi [11].

S 1.7: o-phenylendiamin ng vng vi aldehyd thm bng cng ngh vi sng

1.2.2. To cc dn cht trn nhn 2-aryl-1H-benzimidazol

1.2.2.1. Phn ng bromo ha

a. Phn ng bromo ha

Phn ng bromo ha l phn ng ha hc nhm a 1 hay nhiu nguyn t

brom vo hp cht hu c.

C th to dn xut brom theo 2 phng php chnh nh sau:

- Th hydro hay nhm nguyn t khc bng brom.

- Cng hp brom vo lin kt kp ca phn t hu c.

b. Tc nhn bromo ha

Brom lng

- Cht lng mu nu sm, d bay hi.

- Thng s dng bromo ha cc hp cht thm theo c ch i in t hoc

cng hp v lin kt bi ca hydrocarbon khng no.

13

Acid hydrobromic (HBr)

- ng dng cng hp v lin kt bi ca hydocarbon khng no theo c ch phn

ng cng hp, hoc th nhm OH alcol bng brom theo c ch i nhn.

Mui ca brom vi kim loi kim (NaBr)

- Thng dng th cc dn cht clo

Cc tc nhn khc

N-bromosuccinimid (NBS) l mt cht cung cp brom hay c

s dng. Cc phn ng bromo ha dng NBS l: cng brom vo

alken, bromo ha dn xut carbonyl v tr alpha (), bromo ha

hp cht thm [1,3].

1.2.2.2. Phn ng alkyl ha

a. Phn ng alkyl ha

Alkyl ha l qu trnh thay th mt hoc nhiu nguyn t hydro ca hp cht

hu c bng mt hoc nhiu nhm alkyl.

Nhm alkyl c th lin kt trc tip vi carbon, oxy, nit hoc lu hunh

tng ng ta c cc qu trnh C-alkyl ha, O-alkyl ha, N-alkyl ha hoc S-alkyl

ha,

b. Cc tc nhn alkyl ha

Cc alcol (R-OH)

Methanol v ethanol l nhng tc nhn alkyl ha quan trng. Hu nh trong

mi trng hp alkyl ha, phi s dng acid v c lm xc tc.

14

Alcol thng dng alkyl ha cc amin hoc alcol khc. Sn phm l cc

amin hoc ester.

Cc alkyl halogenid (R-X)

Cc alkyl halogenid mch carbon ngn (C=1-3) l cht lng c si thp.

Tc nhn hay dng l methyl halogen v ethyl halogen. Alkyl halogenid c s

dng alkyl ha cc amin.

Cc arakyl halogenid (Ar-(CH2)nX)

Benzyl clorid hoc benzyl bromid c s dng nhiu. N thng c

dng bo v nhm -OH ca ng hoc celluose trong ha hc cc

hydratcarbon.

Cc ester ca acid v c cha oxy

Gm cc dialkyl sulfat, alkyl nitrat, alkyl phosphat, dialkyl carbonat. Tc

nhn c s dng nhiu nht l dimethyl sulfat. Tc nhn ny c th alkyl ha

nhm OH phenol trong mi trng kim, nc, nc-alcol, thm ch c trong mi

trng acid. Cc alkyl nitrat rt t khi s dng v d gy n.

Cc ester ca acid sulfonic

Cc ester hu c ca acid sulfonic thm c ngha trong thc t. Vic s

dng n ging vi dialkyl-sulfat. Nhm tc nhn ny thng c dng alkyl

ha nhm OH phenol hoc NH amin. Mt s tc nhn in hnh l methyl-

benzensulfonat, benzyl-benzensulfonat hoc methyl, benzyl ester ca acid p-toluen

sulfonic.

Cc mui amoni bc 4

Trong phenyl-trimethyl-amoni clorid l tc nhn hay c s dng. Tc

nhn ny methyl ha chn lc nhm OH phenol trong mi trng kim.

Cc tc nhn alkyl ha khc

Cc epoxid: c s dng iu ch cc polyether-alcol, cht c tc dng

lm sc cng b mt.

Cc hp cht c kim: Hp cht c magnesi dng iu ch cc ether hn

tp [4].

15

1.2.3. Phn ng th vo v tr N-1 trn nhn 2-aryl-1H-benzimidazol

Cc dn cht 2-aryl-1H-benzimidazol thng c th vi tc nhn R-X (X

l halogen) to ra cc nhm th khc nhau v tr N-1 trn nhn 2-

arylbenzimidazol. Phn ng thng c thc hin trong cc dung mi nh: aceton,

DMF, THF Vi cc cht xc tc base: natri hydrid, natri hydroxid, kali carbonat

khan

S 1.8: Phn ng th N-1 ca dn cht 2-aryl-1H-benzimidazol

16

CHNG II: NGUYN LIU, THIT B, NI DUNG V PHNG

PHP NGHIN CU

2.1. NGUYN LIU V THIT B

Kha lun s dng mt s ha cht, dung mi v thit b ca phng th

nghim Tng hp Ha Dc B mn Cng Nghip Dc Trng i hc Dc

H Ni. Bao gm:

2.1.1. Ha cht v dung mi

Bng 2.1: Danh mc cc dung mi, ha cht

STT Dung mi, ha cht Ngun gc

1 Aceton Trung Quc

2 Acid acetic bng Merck

3 Acid hydrobromic Trung Quc

4 Benzyl clorid Trung Quc

5 Cloroform Merck

6 Dicloroethan Trung Quc

7 DMF Trung Quc

8 Ethanol Trung Quc

9 Ethyl acetat Trung Quc

10 Ethyl cloroacetat Trung Quc

11 Kali bromat Trung Quc

12 Kali carbonat khan Trung Quc

13 Kali iodid Trung Quc

14 Methanol Trung Quc

15 Methyl cloroacetat Trung Quc

16 Natri metabisulfit Trung Quc

17 Natri sulfat khan Trung Quc

18 n-hexan Trung Quc

19 o-phenylendiamin Merck

17

STT Dung mi, ha cht Ngun gc

20 Vanillin Vit Nam

2.1.2. Thit b, dng c

Bng 2.2: Danh mc cc thit b

STT Tn thit b, my mc Ngun gc

1 Cn k thut Sartorius BP2001S c

2 Cn phn tch Mettler Toledo AB204S Thy S

3 My ct quay Buchi B491 Thy S

4 My o nhit nng chy EZ-Melt M

5

My o ph cng hng t ht nhn proton 1H-NMR

Brucker AV-500MHz (Vin Hn lm Khoa hc v Cng

ngh Vit Nam)

M

6 My o ph hng ngoi Impact-410 (Vin Hn lm Khoa

hc v Cng ngh Vit Nam) M

7 My o ph khi lng 1100 Series LC/MSD Trap (Vin

Hn lm Khoa hc v Cng ngh Vit Nam) c

8

My o ph khi lng phn gii cao FT-ICR-MS-Varian

910MS (Vin Hn lm Khoa hc v Cng ngh Vit

Nam)

c

9 My khuy t gia nhit IKA c

10 T sy Memmert c

Bng 2.3: Danh mc cc dng c

STT Tn dng c Ngun gc

1 Bn mng Silicagel GF254 70-230 mesh c

2 Bnh cu 2 c 100ml c

3 Bnh cu y trn loi 50ml, 100ml c

Cc c m 100ml, 250ml, 500ml Trung Quc

18

STT Tn dng c Ngun gc

4 a thy tinh, pipet paster Trung Quc

5 Nhit k thy ngn Trung Quc

6 Phu lc thy tinh Trung Quc

7 Phu lc Buchner Trung Quc

8 Pipet chnh xc 1ml, 5ml, 10ml Trung Quc

9 Nhit k thy ngn Trung Quc

10 Phu lc thy tinh Trung Quc

11 Phu lc Buchner Trung Quc

12 Pipet chnh xc 1ml, 5ml, 10ml Trung Quc

2.2. NI DUNG NGHIN CU

- Tng hp ha dc: Tng hp 4 dn cht 2-phenylbenzimidazol, c cng thc

cu to nh sau:

Cht I

Cht II

Cht III Cht IV

- Kim tra tinh khit ca 4 cht I-IV bng SKLM v o nhit nng chy,

xc nh cng thc cu trc cc cht tng hp c bng ph hng ngoi IR, ph

khi lng MS, ph cng hng t ht nhn 1H-NMR.

19

- Th tc dng khng nm, khng khun vi cc cht I, II, III, IV.

- Th tc dng chng ung th vi cht III, IV.

2.3. PHNG PHP NGHIN CU

2.3.1. Tng hp ha hc v kim tra tinh khit

S dng phng php tng hp ha hc, da trn 4 phn ng:

- Phn ng ng vng to dn cht th 2-phenylbenzimidazol theo phng php

mc 1.2.1.

- Phn ng bromo ha to dn cht bromo theo phng php mc 1.2.2.

- Phn ng alkyl ha theo phng php mc 1.2.2

- Phn ng th vo v tr N-1 trn nhn 2-phenylbenzimidazol theo phng php

mc 1.2.3.

Dng sc k lp mng theo di tin trin ca phn ng vi 2 h dung mi

khai trin ethylacetat: n-hexan (7: 3) hoc cloroform: methanol (9: 1), hin mu

bng n t ngoi bc sng 254nm.

Kim tra tinh khit ca sn phm bng SKLM c tin hnh trn bn

nhm trng sn silicagel GF254 Merck 70-230 mesh, quan st di n t ngoi

bc sng 254nm, dung mi ha tan: ethanol 96o, h dung mi khai trin

ethylacetat: n-hexan (7: 3) hoc cloroform: methanol (9: 1). o nhit nng chy

bng my o nhit nng chy EZ-Melt.

2.3.2. Xc nh cu trc

Xc nh cu trc ca cc cht tng hp c da trn kt qu phn tch ph

hng ngoi (IR), ph khi lng (MS), ph cng hng t ht nhn proton (1H-

NMR).

Ti phng phn tch cu trc phn t Vin ha hc - Vin Hn lm Khoa hc

v Cng ngh Vit Nam, cc cht I-IV c ghi ph hng ngoi (IR) trn my

vi k thut vin nn KBr ghi trong vng 4000-400 cm-1.

Ti phng phn tch khi ph ca Vin Ha hc - Vin Hn lm Khoa hc v

Cng ngh Vit Nam cc cht I, IV c ghi ph khi lng trn my 1100 Series

20

LC/MSD Trap - Agilent, cht III c ghi ph khi lng phn gii cao FT-ICR-

MS-Varian 910MS. Ti phng Ha vt liu -Khoa Ha - i hc Khoa hc T

nhin - i hc quc gia H Ni ph MS ca cht II c ghi trn my LTQ

Orbitrap XL theo phng php EI.

Ti phng phn tch ph - Vin ha hc - Vin Hn lm Khoa hc v Cng

ngh Vit Nam, ph cng hng t ht nhn proton (1H-NMR) c ghi trn my

Brucker AV-500MHz, s dng dung mi l DMSO, ly mc l pic ca cht chun

ni tetramethylsilan (TMS).

2.3.3. Th tc dng sinh hc

Th tc dng khng nm, khng khun ti b mn Vi sinh v sinh hc -

Trng i hc Dc H Ni bng phng php khuch tn trn thch.

Th hot tnh gy c t bo ti Phng Sinh hc thc nghim - Vin ha hc

cc hp cht thin nhin - Vin Hn lm Khoa hc v Cng ngh Vit Nam theo

phng php ca Skehan cng cng s (1990) v Likiwitayawuid cng cng s

(1993) hin ang c p dng ti Vin nghin cu ung th Quc gia ca M

(NCI) v Trng i hc Dc, i hc tng hp Illinois, Chicago, M.

21

CHNG III: THC NGHIM, KT QU V BN LUN

3.1. HA HC

3.1.1. Tng hp ha hc

Sau khi tham kho cc ti liu v da vo iu kin phng th nghim, cc

ha cht cho php, cc dn cht 2-phenylbezimidazol c tng hp theo s sau:

S 3.1: Cc giai on tng hp cc dn cht 2-phenylbenzimidazol

3.1.1.1. Tng hp 4-(1H-benzimidazol-2-yl)-2-methoxyphenol (I)

S phn ng:

22

S 3.2: Tng hp 4-(1H-benzimidazol-2-yl)-2-methoxyphenol (I)

Tin hnh:

Thm 2,7 g (2,5 mmol) o-phenylendiamin v 3,8 g (2,5 mmol) vanillin vo

bnh cu cha 45 ml DMF, khuy cho n khi tan hon ton. Sau , cho t t 4,75

g (2,5 mmol) Na2S2O5 vo v un hn hp phn ng 900C trong 2 gi.

Sau khi kt thc phn ng, rt t t ton b hn hp vo cc c m cha

150ml nc lnh, khuy mnh trong 30 pht. Lc ly ta, sau em kt tinh li

trong ethanol 40o, sy kh 60-70oC.

Kt qu:

- Cm quan: tinh th mu trng xm.

- Rf = 0,51 (h dung mi khai trin ethylacetat:n-hexan = 7:3)

- tonc= 219oC 221oC.

- Khi lng: m = 5,41 g.

- Hiu sut phn ng t 90,2%.

3.1.1.2. Tng hp 4-(1H-benzimidazol-2-yl)-2-bromo-6-methoxyphenol (II)

a. Tng hp Bromovanillin

S phn ng:

S 3.3: Tng hp bromovanillin

Tin hnh:

Thm 1,0 g (6,6 mmol) vanillin v 0,4 g (2,4 mmol) kali bromat vo bnh cu

c cha 3ml acid acetic bng, khuy k trong 30 pht. Sau , nh t t 3ml (12,4

mmol) acid hydrobromic 40%, mu vng xut hin. Tip tc khuy hn hp phn

ng nhit phng trong 1 gi 30 pht.

23

Sau khi kt thc phn ng, chuyn hn hp phn ng sang cc c m cha

50ml nc xut hin kt ta, tip tc khuy trong 15 pht. Sau lc ta, ra ta

bng nc ct nhiu ln. Kt tinh li trong ethanol 60o. Sy ta 60-70oC.

Kt qu:

- Cm quan: cht bt mu trng.

- Rf = 0,79 (h dung mi khai trin ethylacetat:n-hexan = 7:3).

- tonc=162-165oC.

- Khi lng: m = 1,18 g.


b. Tng hp 4-(1H-benzimidazol-2-yl)-2-bromo-6-methoxyphenol (II)

S phn ng:

S 3.4: Tng hp 4-(1H-benzimidazol-2-yl)-2-bromo-6-methoxyphenol

(II)

Tin hnh:

Thm 0,54 g (5 mmol) o-phenylendiamin v 1,18 g (5 mmol) bromovanillin

vo bnh cu 50 ml cha 10 ml DMF, khuy cho ti khi tan hon ton. Sau , thm

t t 0,95 g (5 mmol) natri metabisulfit vo hn hp phn ng. Duy tr phn ng

90oC trong 3 gi.

Sau khi kt thc phn ng, t t hn hp phn ng vo cc c m c cha

50ml nc , xut hin kt ta. Lc kt ta, kt tinh li trong ethanol 60o, sy kh

60-70oC.

Kt qu:

- Cm quan: cht bt mu vng xanh.

- Rf = 0,64 (h dung mi khai trin cloroform: methanol = 9:1).

- tonc = 164-166oC.

24

- Khi lng: m = 1,27 g.


3.1.1.3. Tng hp 2-(4-benzyloxy)-3-bromo-5-methoxyphenyl)-1H-benzimidazol

(III)

a. Tng hp 4-benzyloxy-3-bromo-5-methoxybenzaldehyd

S phn ng:

S 3.5: Tng hp 4-benzyloxy-3-bromo-5-methoxybenzaldehyd

Tin hnh:

Cho 1,16 g (5 mmol) bromovanillin vo bnh cu 50 ml cha 10ml methanol,

khuy cho n khi tan hon ton. Tip tc cho 0,69 g (5 mmol) kali carbonat v

0,83 g (5 mmol) kali iodid vo hn hp, khuy trong 30 pht. Nh t t 0,6 ml (5

mmol) benzyl clorid vo hn hp phn ng. un hi lu 70oC trong 4 gi.

Sau khi kt thc phn ng t t hn hp phn ng vo cc c m cha

30ml nc lnh, khuy k thy xut hin cc git du khng tan. Dng 30 ml

diclomethan (CHCl2) chit (chit 3 ln mi ln 10 ml). Lm khan dch chit

diclomethan bng natri sulfat khan, ct loi dung mi thu c sn phm.

Kt qu:

- Cm quan: cht lng mu vng nu.

- Khi lng: m = 1,08 g.


b. Tng hp 2-(4-benzyloxy)-3-bromo-5-methoxyphenyl)-1H-benzimidazol (III).

S phn ng:

25

S 3.6: Tng hp 2-(4-benzyloxy)-3-bromo-5-methoxyphenyl)-1H-

benzimidazol (III).

Tin hnh:

Thm 0,64 g (2 mmol) 4-benzyloxy-3-bromo-5-methoxybenzaldehyd vo

bnh cu 50 ml cha 10 ml DMF, cho 0,38 g (2 mmol) natri metabisulfit vo, khuy

k trong 30 pht. Sau , cho 0,22 g (2 mmol) o-phenylendiamin vo hn hp phn

ng. un phn ng 90oC trong 3 gi.

Sau khi kt thc phn ng, chuyn t t hn hp phn ng sang cc c m

cha 50ml nc lnh, xut hin kt ta, khuy mnh. Lc ta, kt tinh li trong

ethanol 70o, sy kh 60-70oC.

Kt qu:

- Cm quan: cht bt mu vng nht.

- Rf = 0,82 (h dung mi khai trin cloroform: methanol = 9: 1).

- tonc = 211-213oC.

- Khi lng: m = 0,68 g.


3.1.1.4. Tng hp ethyl 2-[2-(4-benzyloxy-3-bromo-5-methoxyphenyl)-1H-

benzimidazol-1-yl] acetat (IV)

S phn ng:

S 3.7: Tng hp ethyl 2-[2-(4-benzyloxy-3-bromo-5-methoxyphenyl)-

1H-benzimidazol-1-yl] acetat (IV)

26

Tin hnh:

Thm 0,41 g (1 mmol) cht III vo bnh cu 50 ml c cha 10 ml DMF

khuy cho ti khi tan ht. Tip tc cho 0,14 g (1 mmol) kali carbonat vo bnh phn

ng, khuy trong 1 gi. Sau , nh t t 0,14 ml (1,3 mmol) ethyl cloroacetat

(d=1,1450 g/ml) vo hn hp phn ng. Khuy hn hp phn ng nhit phng

trong 24 gi.

Sau khi phn ng kt thc, lc hn hp phn ng loi mui v c.

dch lc vo cc c m cha 30 ml nc lnh, ri chit bng ethyl acetat (chit 3

ln, mi ln 20 ml). Lm khan dch chit ethyl acetat bng natri sulfat khan. Ct

quay chn khng n ht dung mi thu c sn phm.

Kt qu:

- Cm quan: cht rn mu nu .

- Rf = 0,87 (h dung mi khai trin cloroform: methanol = 9: 1).

- tonc = 78-80oC.

- Khi lng: m = 0,29 g.


Nh vy, 4 cht I-IV c tng hp. Sau y l bng tm tt kt qu tng

hp ha hc:

Bng 3.1: Kt qu tng hp ha hc

STT Cht CTPT Cm quan Hiu sut

(%)

1 I C14H12N2O2 Tinh th mu trng xm 90,2

2 II C14H11BrN2O2 Cht bt mu vng xanh 79,6

3 III C21H17BrN2O2 Cht bt nu vng nht 83,1

4 IV C25H23BrN2O4 Cht rn mu nu 58,6

3.1.2. Kim tra tinh khit

Kim tra tinh khit ca 4 cht tng hp c bng SKLM v o nhit

nng chy nh sau:

27

Tin hnh chy SKLM bn cht I-IV vi 2 h dung mi khai trin:

ethylacetat : n-hexan (7: 3) v cloroform: methanol (9: 1).

Kt qu trn sc k i vi 2 h dung mi khai trin thy 1 vt r, gn,

khng c vt ph khi soi di nh sng t ngoi bc sng 254 nm.

Bn cht t I-IV sau khi tinh ch u dng rn v c o nhit nng

chy.

Gi tr Rf v nhit nng chy (tonc) ca cc cht c tm tt trong bng

sau:

Bng 3.2: Gi tr Rf v nhit nng chy (to

nc) ca cc cht

Cht H dung mi khai trin Rf Tonc (

oC)

I Ethylacetat: n-hexan = 7: 3 0,51 219-221

II Cloroform: methanol = 9: 1 0,64 164-166

III Cloroform: methanol = 9: 1 0,82 211-213

IV Cloroform: methanol = 9: 1 0,87 78-80

Nhn xt: Vi cc h dung mi khai trin, cc cht tng hp c cho vt

gn r trn bn sc k v nhit nng chy ca cc cht c khong dao ng hp

20C. Nh vy, c th kt lun s b cc cht tng hp c l tinh khit.

28

3.1.3. Xc nh cu trc

xc nh cu trc ha hc, cc cht I-IV c tin hnh o ph hng

ngoi (IR), ph khi lng (MS), ph cng hng t ht nhn proton (1H-NMR).

3.1.3.1. Kt qu ph hng ngoi (IR)

Thng qua vic phn tch ph IR ca cc cht tng hp, nhn bit c cc

di hp ph c trng ca dao ng ha tr (), dao ng bin dng () ca cc

nhm chc v cc lin kt in hnh c trong cu trc phn t ca chng nh C=C

thm, O-H, N-H, C=N, C-N, C=O ester[2,5] Ngoi ra vi cc nhm th khc

nhau, mi cht li c nhng pic c trng ring. S liu phn tch ph hng ngoi

ca 5 cht c trnh by trong bng sau:

Bng 3.3: Kt qu ph hng ngoi ca cc cht I-IV

Cht O-H C-O alcol N-H C=C C-O-C C=N C=O

I 3341,11 1217,47 3511,15 1504,25 1136,26 1656,52

II 3159,83 1463,61 1226,76 1628,60

III 3057,72 1464,20 1232,98 1630,57

IV 1459,22 1212,17 1562,63 1746,49

Nhn xt: Qua bng nhn thy s liu phn tch ph IR hon ton ph hp

vi cc ti liu tham kho [2,5] cng nh cu trc d kin ca 4 cht I-IV.

3.1.3.2. Kt qu ph khi lng (MS)

Tin hnh ghi ph MS ca cc cht I-IV kt qu thu c nh sau:

Bng 3.4: Kt qu ph khi lng ca cc cht I-IV

Cht Cng thc cu to CTPT Khi

lng m/z

I

C14H12N2O2 240 238,4

[M-H]-

29

Cht Cng thc cu to CTPT Khi

lng m/z

II

C14H11BrN2O2 318 319/321

[M+H]+

III

C21H17BrN2O2 408 409/411

[M+H]+

IV

C25H23BrN2O4 494 495/497

[M+H]+

Nhn xt: T bng trn cho thy cc cht c gi tr ph khi lng ph hp

vi khi lng phn t ca cc hp cht d kin [2,5]. Cc cht II, III, IV do c

nguyn t brom trong phn t nn ph khi lng xut hin pic ng v ca brom

ng vi t l t nhin (79Br: 81Br = 0,981)

3.1.3.3. Kt qu ph cng hng t ht nhn proton (1H-NMR)

khng nh chc chn cu trc ca cc cht tng hp c, chng ti tin

hnh ghi ph cng hng t ht nhn proton (1H-NMR) ca cc cht I-IV kt qu

c trnh by trong bng sau:

Bng 3.5: Kt qu ph cng hng t ht nhn proton ca cc cht I-IV

Cht Cng thc cu to 1H-NMR

I

1H-NMR (500MHz, DMSO,

ppm): 3,88 (3H, s, O-CH3),

6,9 (1H, d, J = 4,1 Hz, C5-H),

7,15 (2H, dd, C5-H + C6-H),

7,47 (1H, d, J = 3,5 Hz, C6-H),

30


7,60 (2H, m, C4-H +C7-H),

7,74 (1H, d, C2-H),

9,53 (1H, s, O-H),

12,65 (1H, s, N-H)

II


ppm): 3,95 (3H, s, O-CH3),

7,18 (2H, dd, C5-H + C6-H),

7,56 (2H, s, C2-H + C6-H),

7,77 (1H, d, C4-H),

7,93 (1H, d, C7-H),

10,02 (1H, s, O-H)

III


ppm): 3,98 (3H, s, O-CH3),

5,10 (2H, s, O-CH2),

7,29 (2H, dd, C5-H + C6-H),

7,32 (1H, d, CHAr),

7,38 (2H, d, 2CHAr),

7,51 (2H, d, C4-H + C7-H),

7,66 (2H, dd, 2CHAr),

7,88 (1H, s, C6-H),

8,00 (1H, s, C2-H)

IV


ppm): 1,15 (3H, t, CH2-CH3),

3,92 (3H, s, O-CH3),

4,11-4,15 (2H, m, CH2-CH3),

5,08 (2H, s, N-CH2),

5,27 (2H, s, O-CH2),

7,28-7,32 (2H, m, CHAr),

31


7,35-7,42 (4H, m, CHAr),

7,47 (1H, d, C2-H),

7,53 (2H, d, C5-H + C6-H),

7,61 (1H, d, C4-H),

7,70 (1H, d, C7-H)

Ghi ch: dch chuyn ha hc (ppm); s: singlet; d: double; t: triplet; m:

muntiplet.

Nhn xt: Qua vic phn tch ph , thy s c mt ca cc proton ph hp

vi cng thc cu to ca cc cht I-IV [2,5].

3.2. TH TC DNG SINH HC

Da vo mt s nghin cu v tc dng sinh hc ca mt s dn cht 2-

arylbenzimidazol, chng ti tin hnh th tc dng khng nm, khng khun ca

cc cht I, II, III, IV v th hot tnh gy c t bo vi cht III, IV.

3.2.1. Th hot tnh khng khun

3.2.1.1. Ging vi sinh vt kim nh

Th tc dng khng khun c thc hin i vi 5 vi khun Gram(+) v 5

vi khun Gram() sau:

Vi khun Gram(-) gm:

Escherichia coli ATCC 25992 (EC)

Proteus mirabilis BV 108 (Pro)

Shigella flexneri DT 112 (Shi)

Salmonella typhi DT 220 (Sal)

Pseudomonas aeruginosa VM 201 (Pseu)

Vi khun Gram(+) gm:

Staphylococus aureus ATCC 1128 (Sta)

Bacillus pumilus ATCC 10241 (Bp)

Bacillus subtilis ATCC 6633 (Bs)

32

Bacillus cereus ATCC 9946 (Bc)

Sarvina lutea ATCC 9341 (SL)

Khng sinh i chng: Streptomycin (Strep) 14 g/ml

Penicillin (Peni) 27 UI/ml

3.2.1.2. Mi trng th nghim

Thnh

. phn

MT

NaCl

(%)

Cao tht

(%)

Pepton

(%)

Thch

(%) Nc pH

Canh thang

nui cy VSV 0,5 0,3 0,5 0,0

Va

100 ml

7,0-7,4

Thch thng 0,5 0,3 0,5 1,6

3.2.1.3. 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.

3.2.1.4. Tin hnh

Mu th c pha vo dung mi thch hp 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 I, II, III, IV, sau mi ln tm cc khoanh giy lc c cha mu th

u c sy nhit thp hn 60oC n kh ht dung mi.

Chun b mi trng v cy vi sinh vt kim nh: vi sinh vt kim nh

c cy vo mi trng canh thang, ri nui cy cho pht trin trong t m 37oC

trong thi gian 18 - 24 gi n nng 107 t bo/ml (kim tra bng pha long v

dy dch chun). Mi trng thch thng v trng (tit trng 118oC/30 pht) c

lm lnh v 45 - 500C v c cy ging vi sinh vt kim nh vo vi t l 2,5

33

ml/100 ml. Lc trn vi sinh vt 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 vi sinh vt kim nh theo s

nh sn.

cc a petri c mu th c t nh trn trong t m to = 37oC trong

18 24 gi, 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).

3.2.1.5. Kt qu th nghim

Bng 3.6: 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

I 9,31

0,69 - - - - - -

12,03

1,05 -

9,00

1,22

II - - 8,53

1,12 - - - - - -

8,47

0,83

III 8,24

1,20

9,45

0,72 - - -

12,37

0,82 -

10,06

0,47 - -

IV - - - - - 11,94 - - - -

34

Mu ng knh vng v khun [mm] ( D , s)

Bc Bs Bp SL Sal EC Shi Pro Pseu Sta

0,75

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

Kt lun:

- Mu I c tc dng c ch 3 chng vi khun th (Proteus mirabilis,

Staphylococus aureus, Bacillus cereus).

- Mu II c tc dng c ch 2 chng vi khun th (Staphylococus aureus,

Bacillus pumilus).

- Mu III c tc dng c ch 4 chng vi khun th (Escherichia coli, Proteus

mirabilis, Bacillus subtilis, Bacillus cereus).

- Mu IV c tc dng c ch 1 chng vi khun th (Escherichia coli).

3.2.2. Th hot tnh khng nm

3.2.2.1. Ging vi sinh vt kim nh

S dng 4 chng nm:

Candida albicans (Ca)

Aspergillus niger (As)

Saccharomyces cerevisiae (Sa)

Penicilium sp. (Pe)

Cht i chng: Intraconazol (Intra) 50 g/ml

3.2.2.2. Mi trng th nghim

Mi trng th nghim trong nghin cu c thnh phn nh sau:

35

Thnh .

. phn

MT

Glucose

(%)

Pepton

(%)

Thch

(%) Nc pH

Sabouraud nui cy 2,0 1,0 0,0 Va

100ml 7,0-7,4 Sabouraud th

nghim 2,0 1,0 1,6

3.2.2.3. Nguyn tc

Th tc dng khng nm 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 nm.

Php th c thc hin ti B mn Vi sinh - Sinh hc, Trng i hc

Dc H Ni.

3.2.2.4. Tin hnh

Mu th c pha vo dung mi thch hp 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 I, II, III, IV, sau mi ln tm cc khoanh giy lc c cha mu th

u c sy nhit thp hn 60oC n kh ht dung mi.

Chun b mi trng v cy vi sinh vt kim nh: nm kim nh (C.

albicans) c cy vo mi trng Sabouraud lng, ri nui cy cho pht trin

trong t m 28 - 30oC trong thi gian 24 - 48 gi n nng 107 t bo/ml (kim

tra bng pha long v dy dch chun). i vi Aspergillus niger th s dng huyn

conidi trong dung dch Tween 80 0,2% v trng. Mi trng Sabouraud v trng

(tit trng 118oC/30 pht) c lm lnh v 45 - 50oC v c cy ging vi sinh vt

kim nh vo vi t l 2,5 ml/100 ml. Lc trn vi nm 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.

36

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 (4 th nghim song song).

cc a petri c mu th c t nh trn trong t m to = 28 - 30oC

trong 24 -48h, ri sau ly ra c kt qu, o ng knh vng v nm nu c (s

dng thc kp Panmer chnh xc 0,02 mm).

nh gi kt qu: da trn ng knh vng v nm 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 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 = 3).

3.2.2.5. Kt qu th nghim

Tc dng i vi vi nm th hin bng sau:

Bng 3.7: Kt qu th tc dng khng nm

Mu ng knh vng v nm [mm] ( D , s)

Ca As Sa Pe

I - - 8,16

0,65 -

II - - 8,20

0,17 -

III - - 7,72

0,82 -

IV 8,78

0,72 -

8,30

0,83 -

37

Mu ng knh vng v nm [mm] ( D , s)

Ca As Sa Pe

Intra 16,33

2,01

15,26

0,41

9,73

0,43

12,39

0,75

Kt lun:

- Mu I, II, III c tc dng vi 1 chng nm (Saccharomyces cerevisiae).

- Mu IV c tc dng vi 2 chng nm (Saccharomyces cerevisiae, Candida

albicans) .

3.2.3. Th hot tnh gy c t bo

3.2.3.1. Nguyn tc

Th hot tnh gy c t bo theo phng php ca Skehan cng cng s

(1990) v Likiwitayawuid cng cng s (1993) hin ang c p dng ti Vin

nghin cu ung th Quc gia ca M (NCI) v Trng i hc Dc, i hc tng

hp Illinois, Chicago, M.

Php th c thc hin ti phng Sinh hc thc nghim (Vin ha hc cc

hp cht thin nhin Vin Hn lm Khoa hc v Cng ngh Vit Nam).

3.2.3.2. Cch tin hnh

Cc dng t bo ung th ngi c cung cp bi ATCC gm: Hep-G2

(Hepatocellular carcinoma t bo ung th gan), Lu (Lung cancer Ung th phi).

Cc dng t bo c nui cy ph hp: DMEM (Dulbeccos Modified

Eagle Medium) hoc MEME (Minimum Essential Medium with Eagles salt) c b

sung thm L-gluthamin, natri pyruvat, 10% huyt thanh b v cc thnh phn cn

thit khc iu kin tiu chun (5% CO2, 37oC, m 98%, v trng tuyt i).

Ty thuc vo c tnh tng dng t bo khc nhau, thi gian cy cng khc nhau.

T bo pht trin pha logarit s c s dng th c tnh.

Th c t bo: 200 M dung dch t bo pha logarit nng 3x104 t

bo/ml vo mi ging (a 96 ging) trong mi trng thch hp cho mi dng t

bo. Mu th c x l vi t bo cc nng pha long khc nhau sao cho t

38

n nng thch hp. 37oC, 5% CO2, 3 ngy. Ging iu khin gm 200 M

dung dch t bo nng 3x104 t bo/ml trong iu kin nh trn, thm 50 M

MTT (1 mg/ml pha trong mi trng nui cy khng huyt thanh), 37oC trong 4

gi, loi b mi trng, thm 100 M DMSO lc u, c kt qu bc sng

495-515nm trn my ELISA.

Cht chun i chng c kh nng dit t bo: Ellipithin, Vinblastin hoc

Taxol pha trong DMSO.

3.2.3.3. Kt qu

a. Cch tnh kt qu

Gi tr CS: l kh nng sng st ca t bo nng no ca cht th

tnh theo % so vi i chng. Da trn kt qu o c ca chng OD (ngy 0),

DMSO 10% v so snh vi gi tr OD khi trn mu tm gi tr CS (%) theo cng

thc:

( ) ( )

( ) ( )

Gi tr CS % sau khi tnh theo cng thc trn c a vo tnh ton excel

tm ra % trung bnh lch tiu chun ca php th c lp li 3 ln theo cng

thc ca Ducan nh sau: lch tiu chun

=( )

Cc mu c biu hin hot tnh (CS < 50%) s c chn ra th nghim

tip tm gi tr IC50.

b.aKt qu thc nghim

39

Bng 3.8: Kt qu th hot tnh gy c t bo ca 2 cht III, IV

STT K hiu mu Nng % t bo cn sng st

Kt lun Hep-G2 Lu

DMSO 100,000,0 100,00,0 m tnh

Chng(+)

Ellipithin 0,20,0 1,20,2

Dng

tnh

1 Cht III 5 g/ml 87,70,9 89,60,8 m tnh

2 Cht IV 20 g/ml 60,10,2 92,31,5 m tnh

Nhn xt: cc cht III, IV khng c hot tnh gy c t bo ng k trn c

2 dng t bo ung th Hep-G2 v Lu.

3.3. BN LUN

3.3.1. V tng hp ha hc

3.3.1.1. Phn ng ng vng to nhn benzimidazol v phn ng th trn nhn

thm

Phn ng ng vng ca o-phenylendiamin vi cc dn cht khc nhau ca

vanillin s dng cht xc tc Na2S2O5 din ra trong thi gian ngn vi hiu sut kh

cao.

Vic tng hp cc nhm th mi trn nhn phenyl nu i t cht I din ra rt

kh khn, sn phm thu c khng nh mong mun v nhim nhiu tp. Nu i t

vanillin, tin hnh phn ng bromo ha hoc alkyl ho trc sau ng vng vi

o-phenylendiamin cho sn phm nh mong mun vi hiu sut cao.

40

3.3.1.2. Phn ng th v tr N-1 trn nhn benzimidazol

Phn ng th v tr N-1 vi cc tc nhn ethyl cloroacetat c hiu sut phn

ng thp, c th l do cc tc nhn ny l tc nhn alkyl ha yu.

iu kin phn ng: phn ng cn thc hin trong mi trng khan nc, do

dung mi, ha cht, thit b cn c lm kh trc khi tin hnh phn ng.

iu ny c th gii thch nh sau: tc nhn alkyl ha y l ester, nu trong mi

trng c mt ca nc di s tc dng ca kim xc tc (K2CO3), chng s b

thy phn to thnh acid monocloroacetic, s lm gim hiu sut phn ng.

Cht xc tc: phn ng th v tr N-1 trn nhn benzimidazol s dng

K2CO3 lm xc tc va c vai tr hot ha, va c vai tr phn ng vi HCl sinh ra

to thnh KHCO3, do khng to ra nc trong qu trnh phn ng.

T l mol gia tc nhn th v dn cht benzimidazol: Khi tng hoc gim t

l ny u lm nh hng ti hiu sut phn ng. nh hng ny c th gii thch

nh sau: khi t l ny < 1, phn ng xy ra khng hon ton v lm gim hiu sut.

Mt khc nu t l ny > 1,5, c th xy ra qu trnh to mui amoni bc 4 ti nit

v tr s 3 trn nhn benzimidazol, hiu sut phn ng cng gim.

- Phn ng gia ethyl cloroacetat v cht III vi t l s mol 1,3 : 1 th sn

phm IV thu c l dn cht th vo v tr N-1.

Dung mi: s dng dung mi l DMF cho hiu sut cao hn dung mi

aceton. Tuy nhin, vic tinh ch ly sn phm t DMF tn nhiu thi gian hn

rt nhiu so vi aceton.

3.3.2. V tc dng sinh hc

3.3.2.1. V tc dng khng khun

C 4 cht I-IV u c tc dng khng khun, kt qu cho thy nhm dn cht

c tc dng trn VK Gram(-) tt hn Gram(+), c th nh sau:

- Cht I c tc dng c ch 3 VK th (1 chng Gram(-): Proteus mirabilis; 2

chng Gram(+): Staphylococus aureus, Bacillus cereus), tc dng c ch

trn VK Gram(-) tt hn VK Gram(+) (ng knh vng v khun trn VK

Gram(-) ln hn VK Gram(+)).

41

- Cht II c tc dng c ch 2 VK Gram(+): Staphylococus aureus, Bacillus

pumilus).

- Cht III c tc dng c ch 4 VK th (2 chng Gram(-): Escherichia coli,

Proteus mirabilis; 2 chng Gram(+): Bacillus subtilis, Bacillus cereus), tc

dng c ch trn VK Gram(-) tt hn VK Gram(+).

- Cht IV c tc dng c ch 1 VK th (chng Gram(-): Escherichia coli).

3.3.2.2. V tc dng khng nm

C 4 cht I-IV u c tc dng khng nm C. albicans, ring cht IV c tc

dng trn c Saccharomyces cerevisiae.

3.3.2.3. V tc dng gy c t bo

Cc cht III, IV khng c hot tnh gy c t bo ng k trn c 2 dng t

bo ung th Hep-G2 v Lu.

42

KT LUN V XUT

KT LUN

T nhng kt qu nghin cu trnh by trn y, chng ti rt ra mt s kt

lun sau:

1. tng hp c 4 dn cht ca 2-arylbenzimidazol vi cc nhm th khc

nhau trn nhn phenyl v v tr N-1. Cu trc cc cht trung gian v sn phm

c xc nh bng phng php phn tch cc ph IR, MS, 1H-NMR.

2. th c tc dng khng nm, khng khun ca cc cht I-IV v th c

tc dng khng t bo ung th ca cc cht III, IV trn 2 dng t bo ung th

Hep-G2 v Lu. Kt qu cho thy c 2 cht u khng c tc dng

XUT

Vi cc kt qu t c, hy vng nghin cu ny c th ng gp mt phn

vo cng trnh nghin cu rng hn v tng hp v th tc dng sinh hc ca dn

cht 2-arylbenzimidazol. tip tc pht trin kt qu, chng ti a ra mt s

xut sau:

1. Tip tc nghin cu phn ng tng hp dn cht 2-arylbenzimidazol vi cc

nhm th khc nhau trn nhn phenyl v v tr N-1.

2. Tip tc th nghim cc hot tnh sinh hc ca dn cht 2-arylbenzimidazol nh

tc dng khng t bo ung th, tc dng khng nm, khng khungp phn

vo vic tm ra nhm dn cht mi c tc dng sinh hc trin vng.

TI LIU THAM KHO

Ting Vit

1. Phan nh Chu (2005), Cc qu trnh c bn tng hp hu c, NXB khoa

hc & k thut, H Ni, tr. 30-44, 64-85.

2. Nguyn Hu nh, Trn Th (1999), ng dng mt s phng php ph

nghin cu cu trc phn t, Nh xut bn gio dc, H Ni, tr. 27-88, 199-

275, 296-333.

3. T Minh Kong v cng s (2006), K thut sn xut dc phm, Tp I,

NXB Y hc, H Ni, tr. 24-31, 42-48, 121-127.

4. Nguyn nh Luyn (2009), K thut ha dc, Tp I, Th vin trng i

hc Dc H Ni, H Ni, tr. 48-53.

5. Nguyn nh Triu, Cc phng php phn tch vt l v ha l 2000, Tp I,

NXB Khoa hc & K thut, H Ni, tr. 29-145, 290-400.

Ting Anh

6. Ahn Chan Mug, Tak Jung Ae, Choi Sun Ju (2000), "Synthesis andin vitro

antitumor activity of 2-alkyl, 2-aryl, and 2-piperazinyl benzimidazole-4, 7-

dione derivatives", Archives of Pharmacal Research, 23(4), pp. 288-301.

7. Alper Sabiha, Temiz Arpaci zlem, ener Aki Esin, et al. (2003), "Some

new bi-and ter-benzimidazole derivatives as topoisomerase I inhibitors", Il

Farmaco, 58(7), pp. 497-507.

8. Balram Soni, Mahendra Singh Ranawat, Anil Bhandari, et al. (2012),

"Synthesis and in vitro antitumor activity of benzimidazole derivatives",

Int.J.Drug Res.Tech, 2 (7), pp. 479-485.

9. Chen Allan Y., Yu Chiang, Gatto Barbara, et al. (1993), "DNA minor

groove-binding ligands: a different class of mammalian DNA topoisomerase

I inhibitors", Proceedings of the National Academy of Sciences, 90(17), pp.

8131-8135.

10. Chen Guo-Feng, Dong Xiao-Yun (2012), "Facile and selective synthesis of

2-substituted benzimidazoles catalyzed by FeCl 3/Al 2 O 3", Journal of

Chemistry, 9(1), pp. 289-293.

11. Enders D., Ley S. V., Mehta G., et al. (2008), "Topics in Heterocyclic

Chemistry", Synthesis, 14, pp.

12. Estrada-Soto Samuel, Villalobos-Molina Rafael, Aguirre-Crespo Francisco,

et al. (2006), "Relaxant activity of 2-(substituted phenyl)-1 H-

benzimidazoles on isolated rat aortic rings: Design and synthesis of 5-nitro

derivatives", Life sciences, 79(5), pp. 430-435.

13. Gker Hakan, Alp Mehmet, Yldz Sulhiye (2005), "Synthesis and potent

antimicrobial activity of some novel N-(alkyl)-2-phenyl-1H-benzimidazole-

5-carboxamidines", Molecules, 10(11), pp. 1377-1386.

14. Kilcigil Glgn Ayhan, Altanlar Nurten (2006), "Synthesis and antifungal

properties of some benzimidazole derivatives", Turkish Journal of

Chemistry, 30(2), pp. 223.

15. Kim Jung Sun, Sun Qun, Gatto Barbara, et al. (1996), "Structure-activity

relationships of benzimidazoles and related heterocycles as topoisomerase I

poisons", Bioorganic & medicinal chemistry, 4(4), pp. 621-630.

16. Liu Ju-Fang, Huang Yuan-Li, Yang Wei-Hung, et al. (2012), "1-Benzyl-2-

Phenylbenzimidazole (BPB), a Benzimidazole Derivative, Induces Cell

Apoptosis in Human Chondrosarcoma through Intrinsic and Extrinsic

Pathways", International journal of molecular sciences, 13(12), pp. 16472-

16488.

17. Nagawade Rahul R., Shinde Devanand B. (2006), "BF3 OEt 2 promoted

solvent-free synthesis of benzimidazole derivatives", Chinese Chemical

Letters, 17(4), pp. 453.

18. Oksuzoglu Emine, Tekiner-Gulbas Betul, Alper Sabiha, et al. (2008), "Some

benzoxazoles and benzimidazoles as DNA topoisomerase I and II inhibitors",

Journal of Enzyme Inhibition and Medicinal Chemistry, 23(1), pp. 37-42.

19. Romero-Castro Aurelio, Len-Rivera Ismael, vila-Rojas Laura Citlalli, et

al. (2011), "Synthesis and preliminary evaluation of selected 2-aryl-5 (6)-

nitro-1H-benzimidazole derivatives as potential anticancer agents", Archives

of Pharmacal Research, 34(2), pp. 181-189.

20. Savall Brad M., Fontimayor Jill R., Edwards James P. (2009), "Selective

phenol alkylation for an improved synthesis of 2-arylbenzimidazole H< sub>

4 receptor ligands", Tetrahedron Letters, 50(21), pp. 2490-2492.

21. Sener E., Yalcin Ismail, Temiz O., et al. (1997), "Synthesis and structure-

activity relationships of some 2, 5-dissubtituted benzoxazoles and

benzimidazoles as antimicrobial agents", Farmaco, 52, pp. 99-103.

22. Sharma Mukesh C., Kohli Dharm V, Sharma Smita (2011), "Benzimidazoles

derivatives with (2-{6-Chloro-5-nitro-1-[2-(1H-tetrazol-5-yl) biphenyl-4-

ylmethyl] 1H-benzoimidazol-2-yl}-phenyl)-(substituted-benzylidene)-amine

with potential angiotensin II receptor antagonists as antihypertensive

activity", International Journal of Drug Delivery, 2(3), pp.

23. Tewey Kathleen M., Chen G. L., Nelson E. M., et al. (1984), "Intercalative

antitumor drugs interfere with the breakage-reunion reaction of mammalian

DNA topoisomerase II", Journal of Biological Chemistry, 259(14), pp. 9182-

9187.

24. Vsquez David, Lagos Carlos F., Mella-Raipan, Jaime, Gonzlez Luis, et al.

(2007), "1-benzoyl-2-(2-nitrophenyl)-1H-benzimidazole derivatives: A novel

approach to the development of new HIV-1 reverse transcriptase inhibitors",

Journal of the Chilean Chemical Society, 52(4), pp. 1281-1287.

25. Wright John B. (1951), "The Chemistry of the Benzimidazoles", Chemical

Reviews, 48(3), pp. 397-541.

26. Yurtta Leyla, Demirayak eref, ifti Glen A, et al. (2013), "Synthesis

and Biological Evaluation of Some 1, 2-Disubstituted Benzimidazole

Derivatives as New Potential Anticancer Agents", Archiv der Pharmazie, pp.

27. Chen Allan Y., Yu Chiang, Bodley Annette, et al. (1993), "A new

mammalian DNA topoisomerase I poison Hoechst 33342: cytotoxicity and

drug resistance in human cell cultures", Cancer research, 53(6), pp. 1332-

1337.

28. Khholzer Birgit, Prather Randall S. (2001), "Synchronization of porcine

fetal fibroblast cells with topoisomerase-inhibitor Hoechst 33342", Animal

reproduction science, 66(1), pp. 109-116.

29. Tonelli Michele, Simone Matteo, Tasso Bruno, et al. (2010), "Antiviral

activity of benzimidazole derivatives. II. Antiviral activity of 2-

phenylbenzimidazole derivatives", Bioorganic & medicinal chemistry, 18(8),

pp. 2937-2953.

PH LC

Ph lc 1: Ph hng ngoi (IR) ca cht I

Ph lc 2: Ph khi lng (MS) ca cht I

Ph lc 3.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht I

Ph lc 3.2: Ph gin cng hng t ht nhn proton (1H-NMR) ca cht I

Ph lc 4: Ph hng ngoi (IR) ca cht II

Ph lc 5.1: Ph khi lng (MS) ca cht II


Ph lc 6.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht II

Ph lc 6.2: Ph gin cng hng t ht nhn proton (1H-NMR) ca cht II

Ph lc 7: Ph hng ngoi (IR) ca cht III

Ph lc 8.1: Ph khi lng (MS) ca cht III


Ph lc 9.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht III

Ph lc 9.2: Ph gin cng hng t ht nhn proton (1H-NMR) ca cht III


Ph lc 10: Ph hng ngoi (IR) ca cht IV

Ph lc 11.1: Ph khi lng (MS) ca cht IV


Ph lc 12.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht IV

Ph lc 12.2: Ph gin cng hng t ht nhn proton (1H-NMR) ca cht IV

Ph lc13.1: Kt qu th tc dng gy c t bo ca cht III, IV



Ph lc 1: Ph hng ngoi (IR) ca cht I

Ph lc 2: Ph khi lng (MS) ca cht I

Ph lc 3.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht I

Ph lc 3.2: Ph gin cng hng t ht nhn proton (1H-NMR) ca cht I

Ph lc 4: Ph hng ngoi (IR) ca cht II

Ph lc 6.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht II

Ph lc 6.2: Ph gin cng hng t ht nhn proton (1H-NMR) ca cht II

Ph lc 7: Ph hng ngoi (IR) ca cht III

Ph lc 9.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht III

Ph lc 10: Ph hng ngoi (IR) ca cht IV

Ph lc 12.1: Ph cng hng t ht nhn proton (1H-NMR) ca cht IV

Ph lc 12.3: Ph gin cng hng t ht nhn proton (1H-NMR) ca cht IV

Ph lc 13.1: Kt qu th tc dng gy c t bo ca cht III, IV

Documents

Nghiên cứu tổng hợp và thử tác dụng sinh học một số dẫn chất 2-Arylbenzimidazol.pdf