B Y T TRNG I HC DC H NI

NGUYN NGC VNG

TNG HP V TH TC DNG SINH

HC MT S DN CHT CA

ACID 2-PHENYLTHIAZOLIDIN-4-

CARBOXYLIC

KHA LUN TT NGHIP DC S

H NI 2013

B Y T

TRNG I HC DC H NI

NGUYN NGC VNG

TNG HP V TH TC DNG SINH

HC MT S DN CHT CA

ACID 2-PHENYLTHIAZOLIDIN-4-

CARBOXYLIC

KHA LUN TT NGHIP DC S

Ngi hng dn:

1. PGS. TS. Nguyn nh Luyn

2. DS. Nguyn Th Lan Hng

Ni thc hin:

B mn Cng nghip dc

Trng i hc Dc H Ni

H NI 2013

LI CM N

Trc ht ti xin by t lng knh trng v bit n su sc n

thy gio PGS. TS. Nguyn nh Luyn, 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. Nguyn

Th Lan Hng, DS. Phm Th Hin 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 Hn Lm Khoa hc v Cng ngh Vit Nam,

Phng phn tch Hu c - Vin Hn lm Khoa hc v Cng ngh

Vit Nam, 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

Nguyn Ngc Vng

MC LC

MC LC

DANH MC CH, K HIU VIT TT

DANH MC CC BNG

DANH MC CC HNH V

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

Chng 1. TNG QUAN ................................................................................. 2

1.1. TC DNG SINH HC CA AICD 2- PHENYLTHIAZOLIDIN-4-

CARBOXYLIC V DN CHT ............................................................. 2

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

1.1.2. Tc dng gin mch, h huyt p ........................................................ 6

1.1.3. Tc dng c ch virus cm A .............................................................. 7

1.1.4. Tc dng chng ung th ...................................................................... 8

1.2. PHNG PHP TNG HP CC DN CHT CA ACID 2-

PHENYLTHIAZOLIDIN-4-CARBOXYLIC ......................................... 12

1.2.1. Phn ng ng vng to acid 2-phenylthiazolidin-4-carboxylic ...... 12

1.2.1.1. Tng hp TCA t L-cystein v dn cht benzaldehyd ................ 12

1.2.1.2. Tng hp TCA t L-cystein hydroclorid monohydrat v dn cht

benzaldehyd ................................................................................. 12

1.2.2. Phn ng v tr N-3 ca acid 2-phenylthiazolidin-4-carboxylic .... 13

1.2.3. Phn ng trn nhm acid COOH ca acid 2-phenylthiazolidin-4

carboxylic. ........................................................................................ 15

Chng 2. I TNG V PHNG PHP NGHIN CU ................... 17

2.1. NGUYN LIU ....................................................................................... 17

2.2. DNG C, THIT B ............................................................................. 18

2.3. NI DUNG NGHIN CU .................................................................... 19

2.4. PHNG PHP NGHIN CU ............................................................ 19

2.4.1. Tng hp ha hc .............................................................................. 19

2.4.2. Kim tra tinh khit ....................................................................... 19

2.4.3. Xc nh cu trc .............................................................................. 19

2.4.4. Th hot tnh sinh hc ...................................................................... 20

Chng 3. THC NGHIM, KT QU V BN LUN .......................... 21

3.1. TNG HP HA HC ........................................................................... 21

3.1.1. Tng hp acid 2-phenylthiazolidin-4-carboxylic v dn cht .......... 21

3.1.1.1. Tng hp acid 2-phenylthiazolidin-4-carboxylic (2a) ................. 21

3.1.1.2. Tng hp acid 2-(4-hydroxy-3-methoxyphenyl)thiazolidin-4-

carboxylic (2b) ............................................................................ 22

3.1.1.3. Tng hp acid 2-(4-clorophenyl)thiazolidin-4-carboxylic (2c) .. 23

3.1.1.4. Tng hp acid 2-(4-methoxyphenyl)thiazolidin-4-carboxylic (2d)

..................................................................................................... 23

3.1.2. Tng hp acid 3-tert-butoxycarbonyl-2-phenylthiazolidin-4-

carboxylic v dn cht ...................................................................... 24

3.1.2.1. Tng hp acid 3-tert-butoxycarbonyl-2-phenylthiazolidin-4-

carboxylic (3a) ............................................................................ 24

3.1.2.2. Tng hp acid 3-tert-butoxycarbonyl-2-(4-hydroxy-3-methoxy

phenyl)thiazolidin-4-carboxylic (3b). ......................................... 25

3.1.2.3. Tng hp acid 3-tert-butoxycarbonyl-2-(4-clorophenyl)

thiazolidin-4-carboxylic(3c) ........................................................ 25

3.1.2.4. Tng hp acid 3-tert-butoxycarbonyl-2-(4-methoxyphenyl)

thiazolidin-4-carboxylic (3d) ...................................................... 26

3.1.3. Tm tt kt qu tng hp c ......................................................... 27

3.2. KIM TRA TINH KHIT ................................................................ 28

3.3. XC NH CU TRC ......................................................................... 29

3.3.1. Ph hng ngoi (IR) .......................................................................... 29

3.3.2. Ph khi lng (MS) ........................................................................ 31

3.3.3. Ph cng hng t ht nhn proton (1H-NMR) ............................... 31

3.4. HOT TNH SINH HC ........................................................................ 33

3.4.1. Hot tnh khng khun ...................................................................... 33

3.4.2. Hot tnh khng nm ......................................................................... 36

3.5. BN LUN ............................................................................................. 39

3.5.1. V tng hp ha hc ......................................................................... 39

3.5.1.1. Tng hp acid 2-phenylthiazolidin-4-carboxylic v dn cht ..... 39

3.5.1.2. Tng hp acid N-tert-butoxycarbonylthiazolidin-4-carboxylic v

dn cht ....................................................................................... 39

3.5.2. V hot tnh sinh hc ........................................................................ 40

KT LUN V KIN NGH ......................................................................... 41

1. KT LUN ................................................................................................. 41

2. KIN NGH ................................................................................................ 42

TI LIU THAM KHO

PH LC

DANH MC CH, K HIU VIT TT

AngI Angiotensin I

ACE Enzym chuyn ha Angiotensin

Boc2O Di-tert-butyl dicarbonat

DCM Dicloromethan

DMF N,N-dimethylformamid

DTIC Dimethylthiazen - imidazol - carboxamid

EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimid

HOBt Hydroxybenzotriazol

1H-NMR Ph cng hng t ht nhn proton

(1H - Nuclear Magnetic resonance spectroscopy)

IC50 Nng c ch 50% i tng th

(Inhibition concentration at 50%)

IR Ph hng ngoi (Infrared spectroscopy)

MIC Nng c ch ti thiu

(Minimum inhibitory concentration)

MS Ph khi lng (Mass spectrometry)

Rf H s lu gi (Retension factor)

SKLM Sc k lp mng

T0nc Nhit nng chy

TEA Triethylamin

THF Tetrahydrofuran

TCA Acid 2-phenylthiazolidin-4-carboxylic

VSV Vi sinh vt

VK Vi khun

DANH MC CC BNG

Bng 1.1: Kt qu th tc dng sinh hc khng nm ca mt s dn cht ...... 6

Bng 1.2: Kt qu th tc dng c ch AngI v ACE ca mt s cht. ........... 6

Bng 1.3: Kt qu tc dng c ch virus thng qua c ch NA. ..................... 8

Bng 1.4: Kt qu tc dng chng ung th caca (4R)-2-phenyl-

N-tetradecylthiazolidin-4-carboxamid hydroclorid (17a) ................ 9

Bng 1.5: Kt qu khng ung th ca cc cht 2-(4-acetamidophenyl)-N-

hexadecylthiazolidin-4-carboxamid (18a) ...................................... 10

Bng 1.6: Kt qu tc dng chng ung th ca 2-phenyl-3-propionyl

thiazolidin-4-carboxylat ethyl ester (19) ........................................ 11

Bng 2.1: Danh mc cc nguyn liu ............................................................. 17

Bng 2.2: Danh mc cc thit b, dng c...................................................... 18

Bng 3.1: Kt qu tng hp ha hc .............................................................. 27

Bng 3.3: Kt qu kim tra tinh khit bng SKLM v o nhit nng

chy ................................................................................................. 29

Bng 3.2: S liu ph IR (cm-1) ca cc cht 3a-d ........................................ 30

Bng 3.3: S liu phn tch ph khi lng ca cc cht 3a-d ...................... 31

Bng 3.4: S liu phn tch ph 1H-NMR ca cc cht 3a-d ........................ 31

Bng 3.5: ng knh vng v khun ca 3a-3d (D mm). ........................... 35

Bng 3.6: ng knh vng v nm ca 3a-3d (D mm). .............................. 38

DANH MC CC HNH V

Hnh 1.1: CTCT ca acid 2-phenylthiazolidin-4-carboxylic ........................... 2

Hnh 1.2: CTCT ca acid 2-(5-fluoro-2-hydroxyphenyl)thiazolidin-4-

carboxylic (2a) ................................................................................. 2

Hnh 1.3: CTCT ca acid 3-(tert-butoxycarbonyl)-2-(5-fluoro-2-hydroxy

phenyl)-1,3-thiazolidin-4-carboxylic (2b) ....................................... 3

Hnh 1.4: CTCT ca (4-benzylpiperazin-1-yl)-(2-(4-fluorophenyl)thiazolidin

-4-yl)methanon (9a) ......................................................................... 5

Hnh 1.5: CTCT ca (4R)-2-phenyl-N-tetradecylthiazolidin-4-carboxamid

hydroclorid (17a) ............................................................................. 9

Hnh 1.6: CTCT ca 2-(4-acetamidophenyl)-N-hexadecylthiazolidin-4-

carboxamid (18a) ........................................................................... 10

Hnh 1.7: CTCT ca 2-phenyl-3-propionylthiazolidin-4-carboxylat ethyl

ester (19) ......................................................................................... 10

Hnh 3.1: S tng hp acid 2-phenylthiazolidin-4-carboxylic v dn cht.

........................................................................................................ 21

Hnh 3.2: SKLM ca cht 3a ......................................................................... 24

Hnh 3.3: SKLM ca cht 3b ......................................................................... 25

Hnh 3.4: SKLM ca cht 3c .......................................................................... 26

Hnh 3.5: SKLM ca cht 3d ......................................................................... 27

1

T VN

Thuc s dng trong cng tc phng v cha bnh c nhiu ngun gc

khc nhau, trong cc thuc hin nay ch yu c ngun gc t tng hpv

bn tng hp ha hc. Trong lnh vc nghin cu v pht trin thuc mi, cc

nh nghin cu thng da trn cu trc ca cc cht ang c dng lm

thuc hoc cc cht c tc dng dc l, c trin vng to ra cc cht mi

d on c tc dng tt hn, t c tnh hn v c hiu qu hn trong iu tr.

Acid 2-phenylthiazolin-4-carboxylic v dn cht l mt dy cht d

vng c nghin cu mt cch h thng v mt ha hc cng nh tc

dng sinh hc. Nhiu cng trnh nghin cu cng b cho thy cc dn cht

ca acid 2-phenylthiazolidin-4-carboxylic c hot tnh sinh hc a dng nh:

khng nm, khng khun [6,12,20], chng ung th [18], khng virus [9],

khng lao [14] v nhiu tc dng dc l khc [8,15].

Vi mong mun gp phn nghin cu lm a dng v cc cc dn cht

ca acid 2-phenylthiazolidin-4-carboxylic, chng ti thc hin ti Tng

hp v th tc dng sinh hc mt s dn cht ca acid 2-

phenylthiazolidin-4-carboxylic gm hai mc tiu chnh sau y:

1. Tng hp c mt s dn cht ca acid 2-phenylthiazolidin-4-

carboxylic.

2. Th hot khng nm, khng khun vi cc cht tng hp c.

2

Chng 1. TNG QUAN

1.1. TC DNG SINH HC CA AICD 2- PHENYLTHIAZOLIDIN-4-

CARBOXYLICV DN CHT

Acid 2-phenylthiazolidin-4-carboxylic c cng thc cu to nh sau:

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

Nm 2009, Z. C. Song v cng s tng hp cc dn cht ca TCA v

th tc dng tc dng khng khun in vitro vi Bacillus subtilis ATCC 6633,

Staphylococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 13525,

Escherichia coli ATCC 35218 v c so snh vi kanamycin B, penicillin G.

Hu ht cc cht u c tc dng khng khun, c bit l cht acid 2-

(5-fluoro-2-hydroxyphenyl)-1,3-thiazolidin-4-carboxylic (2a) c ch P.

aeruginosa (IC50 = 1,5625 g/ml).`

Hnh 1.1: CTCT ca acid 2-phenylthiazolidin-4-carboxylic

Hnh 1.2: CTCT ca acid 2-(5-fluoro-2-hydroxyphenyl)thiazolidin-4-

carboxylic (2a)

3

Trong nghin cu ny, Z. C. Song v cng s tng hp thm 9 dn cht

acid 3-tert-butoxycarbonyl-2-phenylthiazolidin-4-carboxylic (TBCA) t TCA

tng ng v tert-butyloxycarbonyl anhydrid (Boc2O). Kt qu cho thy, cc

cht TBCA c hot tnh khng khun tng rt nhiu so vi TCA tng ng.

ng ch nht l acid 3-(tert-butoxycarbonyl)-2-(5-fluoro-2-hydroxy

phenyl)-1,3-thiazolidin-4-carboxylic (2b) c hot tnh khng P. aeruginosa

(IC50 = 0,195 g/ml) mnh hn kanamycin B (IC50 = 3,125 g/ml) v

penicillin G (IC50 = 6,25 g/ml) [20].

Nm 2011, K. A. El-Sharkawy v cng stng hp dn cht acid N-

acetyl-2-phenylthiazolidin-4-carboxylic v th tc dng khng khun, khng

nm vi: Bacillius subtilis, Bacillus pumilus, Pseudomonas aeruginosa,

Escherichia coli v nm Candida utilis. Kt qu th c so snh vi hai cht

i chiu l: sulfadimidin, amoxycillin trihydrat.

Cc cht (3a-c) c hot tnh khng khun B. subtilis, B. pumilis, P.

aeruginosa, E. coli (MIC = 75-250 g/ml). Trong hp cht (3b): 3-acetyl-

2-phenylthiazolidin-4-carboxylic-L-alanin methyl ester ch tc dng vi B.

subtilis v B. biomilus (MIC = 125 g/ml). Cc cht (4-8) u c hot tnh

Hnh 1.3: CTCT ca acid 3-(tert-butoxycarbonyl)-2-(5-fluoro-2-

hydroxyphenyl)-1,3-thiazolidin-4-carboxylic (2b)

3a, R1 = H

3b, R2 = CH3

3c, R3 = CH2CH2SCH3

4

khng khun (MIC = 75-250 g/ml) vi B. subtilis, P. aeruginosa, E.coli, B.

biomil nhng hu ht u khng cho thy kh nng c ch nm Candida

utilis [6].

Nm 2011, D. Sriram v cng s tng hp 64 dn cht TCA v th

hot tnh khng vi khun lao: Mycobacterium tuberculosis H37Rv (MTB),

Mycobacterium smegmatis (MC2). Hot tnh khng lao c so snh vi cc

khng sinh isoniazid (INH), rifampicin (RIF), ciprofloxacin.

R1 = H, OCH3

R2 = H, F, NO2, OH

Ar= , , ,

i vi Mycobacterium tuberculosis, c 64 cht u cho tc dng c

ch rt tt (MIC = 0,12-20,94 M), trong 17 cht c MIC nh hn 1 M.

Khi so snh vi isoniazid (MIC = 0,66 M) th c 13 cht c tc dng tt hn.

Ngoi ra c 3 hp cht c hot tnh mnh hn rifampicin (MIC = 0,23 M)

v 42 hp cht khng khun tt hn ciprofloxacin (MIC = 4,71 M). ng

4, R1 = H

5, R1 = CH3

6, R1 = CH2CH2SCH3

7, R2 = H

8, R2 = Cl

5

ch nht l cht (4-benzylpiperazin-1-yl)-(2-(4-fluorophenyl)thiazolidin-4-

yl)methanon (9a) c tc dng c ch vi khun lao rt tt (MIC = 0,12 M)

gp 5,5 ln isoniazid v 1,9 ln so vi rifampicin.

i vi Mycobacterium smegmatis, c 64 cht u th hin kh nng

c ch (MIC = 1,23-39,5M) tt hn isoniazid (MIC >123 M) v rifampicin

(MIC = 45,57 M). Trong c 8 hp cht c hot tnh hn ciprofloxacin

(MIC = 2,35 M) [14].

Nm 2003, Y. S. Prabhakar cng cng s tng hp dn cht v tr

2,3,4 trn nhn thm ca TCA v ng thi th tc dng khng nm i vi

Candida albicans (CA), Cryptococus neoformans (CN), Tricophyton

mentagrophytes (TM) v Aspergillus fumigatus (AF).

Kt qu cho thy tt c cc cht c hot tnh khng nm yu, tuy nhin

i vi Candida albicans (CA) th nh hng ca cc nhm th ti kh nng

khng CA l khng ng k [12].

Hnh 1.4: CTCT ca (4-benzylpiperazin-1-yl)-(2-(4-fluorophenyl)

thiazolidin-4-yl)methanon (9a)

6

Bng 1.1:Kt qu th tc dng sinh hc khng nm ca mt s dn cht

Cht R R1 Y -log MIC (M/ml)

CA CN TM AF

10a 4-Cl CO2tBu OH 0,54 1,44 0,84 0,84

10b 4-F CO2tBu OH 0,51 0,82 0,82 -

10c 4-OH CO2tBu OH 0,51 0,81 0,81 -

Ch thch: CA: Candida albicans, CN: Cryptococus neoformans, TM:

Tricophyton mentagrophytes, AF: Aspergillus fumigatus

1.1.2. Tc dng gin mch, h huyt p

Nm 1984, Junichi Iwao v cng s tng hp cc dn cht acid 2-

phenylthiazolidin-4-carboxylic v th tc dng gin mch in vivo trn chut.

Cc dn cht u c tc dng gin mch thng qua c ch angiotensin I

(AngI) v angiotensin I-converting enzym (ACE) nn c tc dng lm h

huyt p [17].

Bng 1.2: Kt qu th tc dng c ch AngI v ACE ca mt s cht.

Angiotensin I

IC50 (M)

ACE

IC50 (M)

11

0,21

1,3

7

Angiotensin I

IC50 (M)

ACE

IC50 (M)

12

0,9

13,0

13

0,5

0,42

14

0,7 3,6

1.1.3. Tc dng c ch virus cm A

Nm 2011, Yu Liu v cng s tng hp dn cht TCA, th tc dng

c ch virus cm A thng qua c ch enzym neuraminidase (NA) ca virus.

Tt c cc hp cht u c tc dng c ch NA vi IC50 = 30,4 - 0,14

M, trong cht acid 2-(2-hydroxy-3-methoxyphenyl)thiazolidin-4-

carboxylic (16) c hot tnh mnh nht (IC50 = 0,14 M) nhng yu hn 7 ln

so vi cht i chiu oseltamivir carboxylat [9].

8

Bng 1.3: Kt qu tc dng c ch virus thng qua c ch NA.

1.1.4. Tc dng chng ung th

Nm 2007, W. Li v cng s tng hp dn cht amid ca TCA, th tc

dng c ch s pht trin ca cc dng t bo ung th trong khi u c tnh

SKMEL-188, WM-164 v t bo to si fibroblast.

Hu ht cc cht u th hin c tc dng c ch, c bit l hp cht

(4R)-2-phenyl-N-tetradecylthiazolidin-4-carboxamid (17a) c tc dng tt

trn c 3 dng t bo hn sorafenib v DTIC nhng yu hn taxol trn 2 dng

t bo l SKMEL-188 v WM-164. Tuy nhin cht 17a c tc dng chn lc

trn fibroblast (IC50 = 6,1 M) tt hn taxol (IC50> 10 M) [18].

Hp cht IC50 (M)

15

23,4

16

0,14

Oseltamivir carboxylat 0,02

9

Bng 1.4: Kt qu tc dng chng ung th caca (4R)-2-phenyl-N-

tetradecylthiazolidin-4-carboxamid hydroclorid (17a)

T bo

Cht

SKMEL-188

(IC50 M)

WM-164

(IC50 M)

Fibroblast

(IC50 M)

17a 0,57 0,46 6,1

Sorafenib 4,3 4,7 >100

Taxol 3,8 x 10-3 6,1 x 10-3 > 10

DTIC > 100 > 100 Khng

Ch thch: SKMEL-188, WM-164: khi u c tnh.

Fibroblast: T bo to si.

Nm 2009, Yan Lu v cng s tng hp cc dn cht amid ca TCA

(18) v th tc dng chng ung th invitro trn cc dng t bo trong khi u

c tnh v ung th tuyt tin lit.

Hu ht cc cht u th hin tc dng c ch cc t bo ung th, c

bit l hp cht 2-(4-acetamidophenyl)-N-hexadecylthiazolidin-4-carboxamid

Hnh 1.5: CTCT ca (4R)-2-phenyl-N-tetradecylthiazolidin-4-

carboxamid hydroclorid (17a)

10

(18a) c tc dng rt tt trn tt c cc t bo th. Tuy nhin kh nng chng

ung th ca 18a cn yu hn rt nhiu so vi colchicin [10].

Bng 1.5: Kt qu khng ung th ca cc cht2-(4-acetamidophenyl)-N-

hexadecylthiazolidin-4-carboxamid (18a)

Cht IC50 SEM (M)

B16-F1 A 375 DU 145 PC-3 LNCaP PPC-1

18a 2,2 0,3 2,1 0,2 1,7 0,1 1,2 0,1 1,0 0,1 0,4 0,1

Colchicin 0,029

0,003

0,020

0,003

0,010

0,002

0,011

0,001

0,016

0,004

0,020

0,001

Ch thch: A 375, B16-F1: Khi u c tnh

DU 145, PC-3, LNCaP, PPC-1: T bo ung th tuyn tin lit

Nm 2012, F. E. Onen-Bayram v cng s tng hp v th tc dng

sinh hc chng ung th ca (2RS,4R)-2-phenyl-3-propionylthiazolidin-4-

carboxylat ethyl ester (19) trn cc dng t bo ung th gan, v, trc trng

v ni mc t cung. Kt qu c so snh vi 2 cht chng ung th c

bit n l camptothecin (CPT) v 5-fluorouracil (5-FU) [7].

Hnh 1.6: CTCT ca 2-(4-acetamidophenyl)-N-hexadecylthiazolidin-

4-carboxamid (18a)

Hnh 1.7: CTCT ca 2-phenyl-3-propionylthiazolidin-4-carboxylat

ethyl ester (19)

11

Bng 1.6: Kt qu tc dng chng ung th ca 2-phenyl-3-

propionylthiazolidin-4-carboxylat ethyl ester (19)

M Dng t bo Cht 19

IC50 (M)

CPT

IC50 (M)

5FU

IC50 (M)

Gan

HepG2 10,01,5 0,01 5,7

Huh7 5,30,93 0,15 30,7

MV 0,410,5

12

1.2. PHNG PHP TNG HP CC DN CHT CA ACID 2-

PHENYLTHIAZOLIDIN-4-CARBOXYLIC

1.2.1. Phn ng ng vng to acid 2-phenylthiazolidin-4-carboxylic

Phn ng ng vng to TCA ch yu c tng hp t nguyn liu

L-cystetin (hoc L-cystein hydroclorid monohydrat) v cc dn cht

benzaldehyd.

1.2.1.1. Tng hp TCA t L-cystein v dn cht benzaldehyd

Khuy hn hp L-cystein v benzaldehyd trong ethanol (methanol)

nhit phng. Sau khi phn ng kt thc, ta sinh ra c lc, ra bng

ethanol (methanol), sy n kh thu c cht 20 [5,10,15].

1.2.1.2. Tng hp TCA t L-cystein hydroclorid monohydrat v dn cht

benzaldehyd

Ha tan L-cystein hydroclorid monohydrat v CH3COOK (NaHCO3

hoc CH3COONa) trong nc nhit phng. Sau thm t t

benzaldehyd trong ethanol (methanol) vo dung dch trn. Sau khi phn ng

kt thc, ta sinh ra c lc, ra, sy thu c sn phm 21 [4,8,9,13,19].

Nghin cu gn y nht, F. Esra Onen-Bayram, Irem Durmaz v cng

s tng hp TCA t L-cystein hydroclorid monohydrat vi hiu sut

100%.

13

Cho cc ht pellets NaOHvo L-cystein hydrochlorid monohydrate

trong nc. Sau khi tan hon ton, cho thm ethanol, benzaldehyd v khuy

trong 3h nhit phng. Kt ta c lc, ra bng nc v sy thu c

cht 22. Hiu sut phn ng 100% [7].

1.2.2. Phn ng v tr N-3 ca acid 2-phenylthiazolidin-4-carboxylic

Nm 1986, L. A. Pavlova tng hp thnh cng hp cht acid N-tert-

butyloxycarbonyl-2-phenylthiazolidin-4-carboxylic (23) trong Et3N/DMF.

Thm t t Et3N vo hn hp TCA trong DMF n tan hon ton. Nh

git t t Boc2O vo dung dch trn, khuy nhit phng trong 2h. Tin

hnh chit bng ethylacetat. Dch chit c ra bng KHSO4 5% v dung

dch mui NaCl bo ha, ct quay chn khng thu c sn phm 23 [11].

Nm 1991, Howard-Lock tng hpcht acid 3-benzoyl-2-

phenylthiazolidin-4-carboxylic (24) trong pyridin.

00C, ha tan TCA trong pyridin. Khuy v nh dn dn benzoyl

clorid cho ti khi hn hp phn ng chuyn sang dng du mu vng. Phn

14

ng din ra trong 24h trong t lnh. Ct quay chn khng thu c cn th,

sau ha tan trong DCM v ra bng nc. Dch DCM c lm khan, lc,

v ct quay chn khng n kit. Tip tc ha tan trong methanol v acid ha

ti pH 2-3 bng HCl, thm nc t t cho n khi dung dch c mu hi c.

yn hn hp ny nhit phng cho ti khi tinh th hnh thnh v kt

tinh li trong methanol nng, thu c cht rn 24 mu vng nht (hiu sut

60%) [16].

Nm 2009, Z. C. Song v cng s tng hp 9 dn cht acid 3-tert-

butoxycarbonyl-2-phenylthiazolidin-4-carboxylic (TBCA) t TCA tng ng

v tert-butoxycarbonyl anhydrid (Boc2O).

Quy trnh tng hp nh sau:

Ha tan TCA v NaOH trong 1,4-dioxan 00C trong 2h. Thm t t

Boc2O v khuy 00C trong 1h v 5h tip theo nhit phng. Hn hp

phn ng c ra bng ethyl acetat loi ht dung mi, sau chit 3 ln

bng ethyl acetat. Dch chit c ra bng dung dch mui NaCl bo ha,

lm khan bng MgSO4 khan, ct quay ti kh thu c sn phm rn mu

trng 25. Hiu sut 87-96% [20].

Nm 2011, Yu Liu v cng s nghin cu, tng hp dn cht acid 3-(2-

cloro-acetyl)-2-phenylthiazolidin-4-carboxylic (26).

15

Quy trnh phn ng nh sau: TCA, NaHCO3 c ha tan trong nc,

thm nh git ClCH2COCl v khuy nhit phng cho n khi khng cn

nguyn liu. iu chnh n pH 8-9 bng NaHCO3, ra bng DCM. Acid ha

pha nc bng acid citric n pH 2-3, sau chit bng DCM. Dch chit

c lm khan v ct quay chn khng thu c sn phm th. Tinh ch bng

sc k ct thu c 26. Hiu sut 70,2% [9].

1.2.3. Phn ng trn nhm acid COOH ca acid 2-phenylthiazolidin-

4carboxylic.

Nm 2009, Yan Lu v cng s tng hp dn cht amid (2RS, 4R)-2-

aryl-N-(3,4,5-trimethoxyphenyl)thiazolidin-4-carboxamid.

(a) Boc2O; NaOH 1N; 1,4-dioxan, H2O

(b) EDCI; HOBt; TEA; 3,4,5-trimethoxyanilin

(c) TFA, CH2Cl2

Ha tan TCA trong NaOH v 1,4-dioxan, sau lm lnh 00C. Tin

hnh nh dn dn di-tert-butyldicarbonat v khuy nhit phng trong 1h,

ct quay chn khng v ra bng ethyl acetat. Pha nc cn li c acid ha

ti pH = 4 bng HCl 1N hoc KHSO4 5%, sau tin hnh chit bng ethyl

acetat. Dch chit c lm khan bng MgSO4 khan, lc v ct quay chn

16

khng thu c cht rn mu trng l acid TCA c bo v nhm chc

amin bng Boc.

Ha tan cht rn, EDCI v HOBT trong CH2Cl2 nhit phng trong

10 pht. Sau cho thm 3,4,5-trimethoxyanilin v Et3N, khuy nhit

phng t 6-8h. Pha long phn ng bng CH2Cl2 v ra ln lt vi nc,

mui NaCl bo ha, NaHCO3 v lm khan bng MgSO4 khan. Ct quay chn

khng loi b dung mi thu c sn phm du. Loi b nhm bo v Boc

bng cch khuy vi TEA trong CH2Cl2 t 1-8h nhit phng. Hn hp

phn ng c c m c bng ct quay di p sut gim, sau ra vi

NaCl bo ha, NaHCO3 v lm khan. Ct quay thu c sn phm27. Hiu

sut 69,5% [10].

Nm 2012, F. E. Onen-Bayram v cng s tng hp dn cht ester ca

TCA l 2-phenylthiazolidin-4-carboxylat ethyl ester (28).

Ha tan thionyl clorid trong ethanol 00C. Khuy v thm t t TCA

vo dung dch trn. Phn ng din ra nhit phng v khuy trong 24h.

Ct quay chn khng loi dung mi. Cn th c ha tan trong CH2Cl2,

ra vi dung dch NaHCO3 bo ha v nc, lm khan bng NaSO4. Ct quay

chn khng thu c cht 28. Hiu sut 73% [7].

17

Chng 2. I TNG V PHNG PHP NGHIN CU

2.1. NGUYN LIU

Kha lun s dng nguyn vt liu, thit b, dng c ca phng th

nghim Tng hp Ha dc B mn Cng nghip Dc, bao gm

Bng 2.1: Danh mc cc nguyn liu

STT Nguyn liu Ngun gc

1 Acid acetic Merck

2 Acid hydrocloric Trung Quc

3 Aceton Trung Quc

4 4-methoxybenzaldehyd Trung Quc

5 Benzaldehyd Trung Quc

6 Boc2O Merck

7 n-Butanol Trung Quc

8 4-clorobenzaldehyd Trung Quc

9 L-cystein hydroclorid monohydrat Trung Quc

10 Ether Trung Quc

11 Ethanol Trung Quc

12 n-Hexan Trung Quc

13 Methanol Trung Quc

14 Natri hydrocarbonat Trung Quc

15 NaOH Trung Quc

16 THF Trung Quc

17 Vanilin Trung Quc

18

2.2. DNG C, THIT B

Bng 2.2: Danh mc cc thit b, dng c

STT Thit b, dng c Xut x

1 Bnh chit 125ml, 50ml c

2 Bnh cu 1 c, 2 c, 3 c 50ml, 100ml c

3 Bn mng silicagel GF254 c

4 Cn k thut Trung Quc

5 My cy quay Buchi B480 Thy S

6 My o nhit ng nng chy EZ-Melt M

7 My o ph hng ngoi Impact-410 (Vin Ha hc -

Vin Hn lm Khoa hc v Cng ngh Vit Nam) M

8

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

Bruker AV-500 (Vin Ha hc - Vin Hn lm Khoa hc

v Cng ngh Vit Nam)

M

9

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

Varian 910MS (Vin Ha hc - Vin Hn lm Khoa hc

v Cng ngh Vit Nam)

M

10

My o ph khi lng 1100 Series LC/MSD Trap

(Vin Ha hc Vin Hn lm Khoa hc v Cng ngh

Vit Nam)

M

11 My khuy t c b phn gia nhit IKA c

12 My sy n hng ngoi c

13 Phu lc Buchner Trung Quc

14 T sy Memmert c

19

2.3. NI DUNG NGHIN CU

Tng hp cc dn cht acid 2-phenylthiazolidin-4-carboxylic (2a-d) v dn

cht acid 3-tert-butoxycarbonyl-2-phenylthiazolidin-4-carboxylic (3a-d) c

cu trc ha hc nh sau:

Xc nh cu trc ca cc cht 3a-d tng hp bng cch o ph hng

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

Th hot tnh khng khun, khng nm in vitro ca cc cht 3a-d.

2.4. PHNG PHP NGHIN CU

2.4.1. Tng hp ha hc

Da trn nhng nguyn tc, phng php c bn ca ha hc hu

tng hp cc cht d kin.

2.4.2. Kim tra tinh khit

Chy SKLM trn bn mng silicagel F254, soi kim tra di n t

ngoi 254 nm.

o nhit nng chy trn my o nhit nng chy EZ-Melt.

2.4.3. Xc nh cu trc

Da trn cc kt qu phn tch ph IR, MS, 1H-NMR.

a: R1 = R2 = H

b: R1 = OCH3, R2 = OH

c: R1 = H, R2= Cl

d: R1 = H, R2 = OCH3

20

Ph IR: ti phng phn tch cu trc phn t Vin Ha hc Vin Hn lm

Khoa hc v Cng ngh Vit Nam, cc cht c ghi ph hng ngoi

trn my vi k thut vin nn KBr ghi 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 [1,2].

Ph MS: My o ph khi lng 1100 Series LC/MSD Trap Vin Ha hc

- Vin Hn lm Khoa hc v Cng ngh Vit Nam.

Nguyn tc chung ca phng php ph khi lng l dng cc electron

mang nng lng cao bn ph phn t hp cht hu c trung ha thnh cc

ion phn t mang in tch dng hoc ph v thnh cc mnh ion, cc

gc. Cc ion c phn tch ri c ghi li tn hiu theo t s khi

lng/in tch (m/ze) ca chng. V xc sut to thnh ion c z>1 l rt

nh v v in tch ca 1 electron khng i nn thng thng m/ze chnh

l khi lng ca ion [3].

Ph cng hng t ht nhn proton (1H-NMR) c ghi trn my Bruker

AV-500 ti Vin Ha hc - Vin Hn lm Khoa hc v Cng ngh Vit

Nam.

2.4.4. Th hot tnh sinh hc

Th tc dng khng khun, khng nm tin hnh theo phng php

khuch tn trn thch, thc hin ti b mn Vi sinh- Sinh hc ca Trng i

hc Dc H Ni.

21

Chng 3. THC NGHIM, KT QU V BN LUN

3.1. TNG HP HA HC

3.1.1. Tng hp acid 2-phenylthiazolidin-4-carboxylic v dn cht

3.1.1.1. Tng hp acid 2-phenylthiazolidin-4-carboxylic (2a)

a. S phn ng

b. Cch tin hnh

Ha tan 3,51g (0,02mol) L-cystein hydroclorid monohydrat trong 16ml

nc ct vo bnh cu 3 c 100ml sch, kh c sc kh N2. Kh N2 c

sc lin tc trong qu trnh phn ng. Cho dn dn 1,7g (0,02mol) NaHCO3

Hnh 3.1: S tng hp acid 2-phenylthiazolidin-4-carboxylic v

dn cht.

22

cho n khi ht bt kh sinh ra. Sau thm t t 2,2ml (0,02mol)

benzaldehyd trong 20ml ethanol tuyt i vo dung dch trn.Tin hnh phn

ng nhit phng trong 24h. Theo di phn ng bng SKLM vi h dung

mi n-butanol: acid acetic: nc = 9:2:2,5. Kt thc phn ng lc ly ta, ra

bng nc v ethanol, sy kh. Kt tinh li trong ethanol thu c cht 2a.

c. Kt qu

Cm quan: tinh th hnh kim, mu trng.

Lng sn phm th: 4,0g.

Sau kt tinh: m2a = 3,1g.

Hiu sut phn ng: H2a= 71,8%.

Rf = 0,74.

T0nc = 160-1620C.

3.1.1.2. Tng hp acid 2-(4-hydroxy-3-methoxyphenyl)thiazolidin-4-

carboxylic(2b)

a. S phn ng

b. Cch tin hnh

Tin hnh tng t nh phn ng tng hp cht 2a nhng thay 2,2ml

cht 1a bng 3,1g (0,02mol) cht vanillin (1b).

c. Kt qu

Cm quan: bt kt tinh mu trng.

Khi lng sn phm: m2b = 3,5g.

Hiu sut phn ng: H2b= 68,6%.

Rf = 0,73. T0nc = 168-1700C.

23

3.1.1.3. Tng hp acid 2-(4-clorophenyl)thiazolidin-4-carboxylic(2c)

a. S phn ng

b. Cch tin hnh

Tin hnh tng t nh phn ng tng hp cht 2a nhng thay 2,2ml

cht 1a bng 2,81g (0,02mol) 4-clorobenzaldehyd (1c).Sn phm th c kt

tinh li trong hn hp aceton/nc (1/1).

c. Kt qu

Cm quan: tinh th hnh kim, mu trng.

Khi lng sn phm: m2c = 2,28g.

Hiu sut: H2c = 54,7%.

Rf = 0,71.

T0nc = 162-1640C.

3.1.1.4. Tng hp acid 2-(4-methoxyphenyl)thiazolidin-4-carboxylic (2d)

a. S phn ng

b. Cch tin hnh

Tin hnh tng t nh phn ng tng hp cht 2a nhng thay 2,2ml

cht 1a bng 2,43ml (0,02mol) 4-methoxybenzaldehyd (1d).

c. Kt qu

Cm quan: bt kt tinh trng.

Khi lng sn phm: m2d = 3,8g. Hiu sut: H2d = 79,5%.

Rf = 0,75. T0nc = 166-1700C.

24

3.1.2. Tng hp acid 3-tert-butoxycarbonyl-2-phenylthiazolidin-4-

carboxylic v dn cht

3.1.2.1. Tng hp acid 3-tert-butoxycarbonyl-2-phenylthiazolidin-4-

carboxylic (3a)

a. S phn ng

b. Cch tin hnh

Cho 4,5ml NaOH 1N vo hn hp gm 1,0g (4,78mmol) cht 2a v

8ml THF nhit phng. Khuy n tan hon ton trong khong 10 pht,

sau thm t t dung dch gm 1,1ml (4,78mmol) Boc2O trong 4ml THF.

Sau khi khuy nhit phng trong 18h, hn hp phn ng c ra bng

ether, sau acid ha bng HCl 18% ti pH 1 v chit bng ether. Dch chit

ether c ra bng nc, lm khan bng Na2SO4 khan. Ct quay loi

dung mi thu c cht rn mu trng v tin hnh kt tinh li trong n-

hexan/ether thu c 3a.

Kim tra tinh khit ca sn phm bng SKLM vi h methanol: n-

Hexan = 7:3.

c. Kt qu

Cm quan: tinh th hnh lng tr, mu trng.

Khi lng sn phm: m3a = 0,81g.

Hiu sut: H3a = 54,7%.

Rf = 0,60.

T0nc = 170-1740C.

Hnh 3.2: SKLM ca cht 3a

25

3.1.2.2. Tng hp acid 3-tert-butoxycarbonyl-2-(4-hydroxy-3-methoxyphenyl)

thiazolidin-4-carboxylic (3b).

a. S phn ng

b. Cch tin hnh

Tin hnh tng t nh phn ng tng hp cht 3a vi cc nguyn liu:

1,0g (3,92mmol) cht acid 2-(4-hydroxy-3-methoxyphenyl)thiazolidin-4-

carboxylic (2b) trong 8ml THF.

4,0ml NaOH 1N.

1ml Boc2O/4ml THF.

c. Kt qu

Cm quan: bt tinh th mu trng.

Khi lng sn phm: m3b = 0,7g.

Hiu sut: H3b = 50,3%.

Rf = 0,47.

T0nc = 167-1700C.

3.1.2.3. Tng hp acid 3-tert-butoxycarbonyl-2-(4-clorophenyl)thiazolidin-4-

carboxylic (3c)

a. S phn ng

b. Cch tin hnh

Hnh 3.3: SKLM ca cht 3b

26

Tin hnh tng t nh phn ng tng hp cht 3a vi cc nguyn liu

nh sau:

1,0g (4,1mmol) acid 2-(4-clorophenyl)thiazolidin-4-carboxylic (2c) trong

8ml THF.

4,2ml NaOH 1N.

1ml Boc2O trong 4ml THF.

c. Kt qu

Cm quan: tinh th hnh lng tr mu trng, trong sut.

Khi lng sn phm: m3c = 0,9g.

Hiu sut: H3c = 63,8 %.

Rf = 0,54.

T0nc = 176-1780C.

3.1.2.4. Tng hp acid 3-tert-butoxycarbonyl-2-(4-methoxyphenyl)thiazolidin-

4-carboxylic (3d)

a. S phn ng

b. Cch tin hnh

Tin hnh tng t nh phn ng tng hp cht 3a vi cc nguyn liu:

1,0g (4,18mmol) acid 2-(4-methoxyphenyl)thiazolidin-4-carboxylic (3d)

trong 8ml THF.

4ml NaOH 1N.

1ml Boc2O trong 4ml THF.

c. Kt qu

Cm quan: tinh th hnh lng tr trong sut mu trng.

Hnh 3.4: SKLM ca cht 3c

27

Khi lng sn phm: m3d= 0,82 g.

Hiu sut: H3d = 58,1 %.

Rf = 0,63.

T0nc = 180-1850C.

3.1.3. Tm tt kt qu tng hp c

Bng 3.1: Kt qu tng hp ha hc

STT Cht R1 R2 R3 CTCT

KLPT Cm quan

Hiu

sut

(%)

1 2a H H H C11H13NO4S

255,29

Tinh th hnh

kim, mu trng 71,8

2 2b OCH3 OH H C11H13NO4S

225,29

B kt tinh

trng 68,6

3 2c H Cl H C10H10ClNO2S

243,71

Tinh th hnh

kim, mu trng 54,7

4 2d H OCH3 H C11H13NO3S

239,29

Bt kt tinh

trng 79,5

Hnh 3.5: SKLM ca cht 3d

28

STT Cht R1 R2 R3 CTCT

KLPT Cm quan

Hiu

sut

(%)

5 3a H H Boc C15H19NO4S

309,41

Tinh th hnh

lng tr, mu

trng.

54,7

6 3b OCH3 OH Boc C17H23NO6S

355,41

Bt kt tinh

trng 50,3

7 3c H Cl Boc C15H18ClNO4S

343,82

Tinh th hnh

lng tr, mu

trng

63,8

8 3d H OCH3 Boc C16H21NO5S

339,41

Tinh th hnh

lng tr, mu

trng

58,1

3.2. KIM TRA TINH KHIT

Sau khi tng hp cc cht, chng ti tin hnh kim tra tinh khit

ca cc sn phm bng SKLM trn bn mng silicagel F254:

Cc cht 2a-2d c ha tan trong ethanol chm sc k. Pha ng

chy sc k l h dung mi n-butanol: acid acetic: nc = 9:2:2,5.

Cc cht 3a-3d c ho tan trong methanol. Pha ng chy sc k

l h dung mi methanol: n-hexan = 7:3.

Sau khi chy sc k, soi bn mng di n t ngoi bc sng 254

nm, cc cht th u cho mt vt gn, r, khng c vt ph.

ng thi, chng ti tin hnh o nhit nng chy ca cc cht tng

hp c. Kt qu o nhit nng chy cho thy cc cht u c im chy

r rng.

29

Bng 3.3: Kt qu kim tra tinh khit bng SKLM v o nhit nng

chy

STT Cht

Rf

T0nc (0C) n-butanol: acid acetic: nc

(9:2:2,5)

methanol: n-hexan

(7:3)

1 2a 0,74 - 160-162

2 2b 0,73 - 168-170

3 2c 0,71 - 162-164

4 2d 0,75 - 166-170

5 3a - 0,6 170-174

6 3b - 0,47 167-170

7 3c - 0,54 176-178

8 3d - 0,63 180-185

3.3. XC NH CU TRC

Chng ti tin hnh o ph hng ngoi (IR), ph khi lng (MS),

ph cng hng t proton (1H-NMR) xc nh cu trc cc cht 3a-d.

3.3.1. Ph hng ngoi (IR)

Qua nghin cu ph ca cc cht, chng ti nhn bit c cc di hp

th c trng ca cc nhm chc v lin kit in hnh ca cc cht 3a-d

Ph IR c ghi ph lc 1.1 1.4. Kt qu phn tch s liu ph

hng ngoi c ghi trong bng 3.2.

30

Bng 3.2: S liu ph IR (cm-1) ca cc cht 3a-d

Tn

cht

max ca cc cht (cm-1)

O-H

phenol CH C=O amid C=O acid C-S

3a - 2930 1722 1633 618

3b 3464 2929 1714 1635 614

3c - 2976 1732 1638 605

3d - 2935 1731 1637 598

Cc cht 3a - 3d u c:

Di hp th nm trong khong t 2929 2976 cm-1 c trng cho nhm

CH ca cc cht.

Di hp th nm trong khong 1714 1732 cm-1 c trng cho nhm C=O

ca amid.

Di hp th nm trong khong 1633 - 1638 cm-1 c trng cho nhm C=O

ca acid.

Di hp th nm trong khong 594 618 cm-1 c trng cho nhm C-S

Cht 3a c di hp ph 3464 cm-1 c trng cho nhm OH trong cng

thc phn t.

a: R1 = R2 = H

b: R1 = OCH3, R2 = OH

c: R1 = H, R2 = Cl

d: R1 = H, R2 = OCH3

31

3.3.2. Ph khi lng (MS)

Bng 3.3: S liu phn tch ph khi lng ca cc cht 3a-d

Cht Cng thc

phn t

Khi lng

phn t m/z Ch thch

3a C15H19NO4S 309,38 332,0 [M+Na]+

3b C17H23NO6S 355,41 354,0 [M-H]+

3c C15H18ClNO4S 343,82 342,0 [M-H]+

3d C16H21NO5S 339,41 338,0 [M-H]+

Trn ph ca 4 cht u xut hin pic phn t [M-H]+ hoc [M+Na]+

c s khi ng bng khi lng phn t M hoc [M+Na] nh d kin.

3.3.3. Ph cng hng t ht nhn proton (1H-NMR)

Cc ph c ghi cc ph lc 3.1 3.4. S liu phn tch ph 1H-

NMR ca cc cht 3a-d c trnh by trong bng 3.4.

Bng 3.4: S liu phn tch ph 1H-NMR ca cc cht 3a-d

Cht Cng thc cu to 1H-NMR (DMSO-d6) , (ppm)

3a

1,07-1,36 (9H, s, 3CH3); 3,16 (1H, d,

C5-Ha); 3,44 (1H, d, C5-Hb); 4,56 (1H, d,

C4-H); 5,94 (1H, s, C2-H); 7,25-7,32 (3H,

m, C3-H + C4

-H + C5-H); 7,62 (2H, s,

C2-H + C6

-H ) ppm

3b

1,05-1,35 (3H, s, 3CH3); 3,09-3,17 (1H,

dd, C5-Ha); 3,38-3,44 (1H, dd, C5-Hb);

3,72 (3H, s, O-CH3); 4,53-4,69 (1H, d,

C4-H); 5,86-6,03 (1H, s, C2-H); 6,66 (1H,

d, C5-H); 6,9 (1H, s, C2-H);7,38-7,46

(1H, d, C6-H); 8,88 (1H,s, OH) ppm

32

Cht Cng thc cu to 1H-NMR (DMSO-d6) , (ppm)

3c

1,07-1,35 (9H, s, 3CH3); 3,07-3,17 (2H,

m, C5-2H); 4,57-4,71 (1H, d, C4-H); 5,96-

6,11 (1H, s, C2-H); 7,35 (1H, d, C2-H);

7,39 (1H, d, C6-H); 7,65 (2H, s, C3-H +

C5-H) ppm

3d

1,08-1,35 (9H, s, 3CH3); 3,06-3,17 (2H,

d, C5-H2); 3,45 (3H, s, OCH3); 4,56 (1H,

d, C4-H); 5,96 (1H,s, C2-H); 7,38 (2H, d,

C3-H + C5-H); 7,66 (2H, s, C2-H + C5-

H) ppm

Ghi ch: : dch chuyn ha hc c ly theo gi tr trung bnh (ppm); s:

singlet; d: doublet; dd: doublet doublet, m: multiplet.

Nhn xt:

Ph ca 4 cht u c cc pic cho php nhn dng proton v s lng

proton thng qua cc s liu v dch chuyn ha hc v cng ca cc

pic, c th nh sau:

= 1,05 - 1,36 ppm c trng cho 9 proton ca -C(CH3)3 ca 4 cht 3a-3d.

= 3,06 - 3,44 ppm c trng cho 2 proton v tr C5 ca 4 cht 3a-3d.

= 3,45 - 3,75 ppm c trng cho 3 proton ca nhm O-CH3 ca cc cht

3b, 3d.

= 4,53 - 4,71 ppm c trng cho 1 proton v tr C4 ca 4 cht 3a-3d.

= 5,86 - 6,11 ppm c trng cho 1 proton v tr C2 ca 4 cht 3a-3d.

= 6,66 7,66 ppm c trng cho cc proton vng benzen ca cc cht

3a-3d.

= 8,88 ppm c trng cho 1 proton -OH phenol ca cht 3b.

33

Nh vy, vi cc kt qu phn tch ph IR, MS v 1H-NMR nh trnh

by trn, c th kt lun l cc cht 3a-d tng hp c c cu trc ph hp

vi cu trc d kin.

3.4. HOT TNH SINH HC

3.4.1. Hot tnh khng khun

a. Cc chng vi sinh vt kim nh do b mn Vi sinh - Sinh hc, trng i

hc Dc H Ni cung cp.

Vi khun Gram(-)

Escherichia coli ATCC 25922 (EC)

Proteus mirabilis BV 108 (Pro)

Shigella flexneri DT 102 (Shi)

Salmonella typhi DT 220 (Sal)

Pseudomonas aeruginosa VM 201 (P.A)

Vi khun Gram(+)

Staphylococus aureus ATCC 1128 (Sta)

Bacillus pumilus ATCC 10242 (Bp)

Bacillus subtilisATCC 6633 (Bs)

Bacillus cereus ATCC 9946 (Bc)

Sarcina lutea ATCC 9341 (SL)

Khng sinh i chng: Streptomycin: 14g/ml

Penicillin: 27 UI/ml

b. Mi trng th nghim

Mi trng canh thang nui cy VK kim nh:

NaCl 0,5% ; pepton 0,5% ; cao tht 0,3 % ; nc v 100ml.

Mi trng thch thng :

NaCl 0,5% ; pepton 0,5% ; cao tht 0,3 % ; thch 1,6% ; nc v 100ml.

34

c. Phng php th (nh gi hot tnh khng sinh bng phng php

khuch tn).

Nguyn tc: mu th (c cha hot cht th) c t ln lp thch dinh

dng cy VSV kim nh, hot cht t mu khuch tn vo mi trng

thch s c ch s pht trin ca VSV kim nh to thnh vng v khun.

Tin hnh:

Cc khoanh giy lc v trng v c sy kh c tm vi 3 ln mi

dung dch mu th, sau mi ln tm cc khoanh giy lc c cha mu th

u c sy < 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 18-24h n nng 107 t bo/ml (kim tra bng pha long v

dy dch chun). Mi trng thch 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 MT

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 t0C = 370C

trong 18-24h, ri sau ly ra c kt qu, o ng knh vng v khun

nu c (thc kp Panmer chnh xc 0,02mm).

nh gi 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

35

D : ng knh trung bnh vng v khun (mm)

iD : ng knh vng v khun th i (mm)

s : lch thc nghim chun c hiu chnh

n : s th nghim lm song song (n = 3)

d. Kt qu thc nghim

Bng 3.5:ng knh vng v khun ca 3a-3d (D mm).

Cht

VSV 3a 3b 3c 3d Peni Strep

Bc - - - - 19,38

0,37

21,50

0,66

Bs - - 8,53

0,61 -

20,47

0,7

20,16

1,21

Bp 8,40

0,67 -

8,63

0,43 - - -

SL - - - - 12,91

0,47

12,73

0,41

Sal - - - - - 13,73

1,5

EC - - - - - 13,2

1,05

Shi - - - - - 12,07

0,89

Pro 8,45

0,39 -

9,42

0,41 - -

15,53

0,81

P.A - - - - - 17,37

0,67

Sta 9,00

1,22 -

8,47

0,83 -

11,42

0,67

17,37

0,12

Ch thch: Peni: Penicillin, Strep: Streptomycin

e. Kt lun

Cht 3b, 3d khng c tc dng vi vi khun.

36

Cht 3a tc dng trn 3 vi khun th gm: Bacillus pumilis (D = 8,40

0,67 mm), Proteus mirabilis (D = 8,45 0,39 mm), Staphylococus aureus

(9,00 1,22 mm).

Cht 3c tc dng trn 4 vi khun th gm: Bacillus subtilis (8,53 0,61

mm), Bacillus pumilis (8,63 0,43 mm), Proteus mirabilis (9,42 0,41

mm), Staphylococus aureus (8,47 0,83).

3.4.2. Hot tnh khng nm

a. Cc chng VSV kim nh do b mn Vi sinh Sinh hc, trng i hc

Dc H Ni cung cp

Vi Nm

Candida albicans (Ca)

Saccharomyces cerevisiae (Sa)

Aspergillus niger (As)

Penicilium sp. (Pe)

Cht i chiu: Itraconazol 50 g/ml

b. Mi trng kim nghim

Mi trng Sabouraud lng nui cy C. albicans:

Pepton 1,0% ; glucose 2,0 %; nc v 100ml

Mi trng th nghim Sabouraud:

Pepton 1,0% ; glucose 2,0 %; thch 1,6%; nc v 100ml

c. Phng php th (nh gi hot tnh khng sinh bng phng php

khuch tn)

Nguyn tc: mu th (khoanh giy lc c tm hot cht th) c t

ln lp thch dinh dng cy VSV kim nh, hot cht t mu th

khuch tn vo mi trng thch s c ch s pht trin ca VSV kim

nh to thnh vng v nm.

Tin hnh:

37

Mu th c pha vo dung mi thch hp (ethanol) vi nng 500

g/ml, 5,0 g/ml, v 10,0 g/ml. Cc khoanh giy lc v trng v c

sy kh c tm 3 ln vi dung dch mu th, sau mi ln tm cc khoanh

giy lc c cha mu th u c sy < 600C n kh ht dung mi.

Chun b mi trng v cy VSV kim nh:

Nm kim nh (C. albicans) c cy vo mi trng Sabouraud lng, ri

nui cy cho pht trin trong t m 28-300C trong thi gian 18-24h 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 c v trng (tit trng 1180C/30

pht) c lm lnh v 45-500C v c cy ging vi nm kim nh vo

vi t l 2,5 ml/ 100ml. Lc trn vi nm kim nh phn tn trong mi

trng thch, ri a petri v trng vi th thch 20ml/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 (4 thc nghim song song).

cc a petri c mu th c t trn trong t m t0C = 28-300C trong

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

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

nh gi kt qu: Da vo 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 khun (mm)

iD : ng knh vng v khun th i (mm)

38

s : lch thc nghim chun c hiu chnh

n : s th nghim lm song song (n = 4)

d. Kt qu thc nghim

Bng 3.6: ng knh vng v nm ca 3a-3d(D mm).

Cht

VSV 3a 3b 3c 3d Intra

Ca 9,35

1,18 -

9,17

0,53

9,23

0,31

16,33

2,01

As - - - 9,62

1,03

15,26

0,41

Sa 9,42

0,97

8,16

1,16 -

9,08

0,59

9,73

0,43

Pe - 8,07

1,14 - -

12,39

0,75

Ch thch: Intra : Itraconazol

e. Kt lun: cc mu th c kh nng khng nm

Cht 3a tc dng trn 2 vi nm: Candida albicans (D = 9,35 1,18 mm),

Saccharomyces cerevisiae (D = 9,42 0,97 mm).

Cht 3b tc dng trn Saccharomyces cerevisiae (D = 8,16 1,16 mm) v

Penicilium sp. (D = 8,07 1,14 mm).

Cht 3c ch tc dng trn Candida albicans (D = 9,17 0,53 mm).

Cht 3d c tc dng trn 3 vi nm: Candida albicans (D = 9,23 0,31

mm), Aspergillus niger (D = 9,62 1,03 mm), Saccharomyces cerevisiae

(D = 9,08 0,59 mm).

39

3.5. BN LUN

3.5.1. V tng hp ha hc

3.5.1.1. Tng hp acid 2-phenylthiazolidin-4-carboxylic v dn cht

Phn ng tng hp 2a-d c hiu sut tng i thp l do trong phn

ng ng vng to thiazolidin t L-cystein hydroclorid monohydrat th phi

thm xc tc base NaHCO3 chuyn dng mui L-cystetin hydroclorid v

dng L-cystein base (dng t do). Khi L-cystein rt d b oxy ha to

thnh cystin v vy nh hng rt nhiu ti hiu sut phn ng. Do trong

qu trnh phn ng phi sc kh N2 loi ht kh O2 trong mi trng phn

ng tng hiu sut phn ng.

Vic kt tinh li cc cht 2a-d trong ethanol hoc hn hp aceton/nc

(1:1) cng lm mt mt phn sn phm.

3.5.1.2. Tng hp acid N-tert-butoxycarbonylthiazolidin-4-carboxylic v dn

cht

Hiu sut tng hp 3a-d thp, iu ny c th gii thch nh sau:

Th nht l cc cht 2a-d rt khng bn trong mi trng kim, khi nh

NaOH 1N vo hn hp cha 2a-d v THF c th lm cho phn hy 2a-d.

40

gim s phn hy cc cht 2a-d th nn nh t t NaOH n khi tan ht

2a-d th dng li.

Th 2 l trong qu trnh tinh ch sn phm bng cch kt tinh li trong hn

hp dung mi diethyl ether/n-hexan (1/1) cng lm mt mt phn sn

phm.

3.5.2. V hot tnh sinh hc

Kt qu cho thy cc cht 3b v 3d khng c hot tnh khng khun i

vi cc VSV kim nh, tuy nhin 3b v 3d li cho c tc dng khng nm rt

tt. Cht 3b khng c c Saccharomyces cerevisiae v Penicilium sp.. iu

c bit l ch duy nht 3b khng c vi vi nm Penicilium sp.. Cht 3d c

tc dng chng nm i vi 3 vi nm l C. albicans, S. ceravisiae, A. niger.

Cht 3a v 3c c tc dng khng khun kh tt, u c ch B. pumilus,

P. mirabilis, S. aureus, v cht 3c cn c ch vi khun B. subtilis. Tuy nhin

kh nng khng nm ca 3a v 3c tng i yu . Cht 3a c tc dng c ch

A. albicans, S. ceravitiae. Cht 3c ch c tc dng trn A. albicans.

Kt qu cho thy khi gn thm nhm th -Cl vo nhn thm th hot

tnh khng khun tng ln rt nhiu. Tuy nhin khi gn thm OCH3 th

khng cn hot tnh khng khun nhng nh hng rt nhiu ti hot tnh

khng nm. c bit hp cht 3d khng li 3/4 vi nm l C. albicans, S.

ceravitiae, A. niger v ch c duy nht cht 3b khng li Penicillium sp..

41

KT LUN V KIN NGH

1. KT LUN

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

kt lun sau:

1) tng hp c cc dn cht:

Acid 3-tert-butoxycarbonyl-2-phenylthiazolidin-4-carboxylic (3a)

Acid 3-tert-butoxycarbonyl-2-(4-hydroxy-3-methoxyphenyl)thiazolidin-4-

carboxylic (3b)

Acid 3-tert-butoxycarbonyl-2-(4-clorophenyl)thiazolidin-4carboxylic (3c)

Acid 3-tert-butoxycarbonyl-2-(4-methoxyphenyl)thiazolidin-4-carboxylic

(3d)

2) th hot tnh khng khun, khng nm ca cc cht 3a-3d.

Hot tnh khng khun: ch c 3a v 3c c tc dng khng khun, trong

cht 3c c hot tnh tt nht, khng li 4/10 chng VSV kim nh l

Bacillus subtilis, Bacills pumilus, Proteus mirabili, Staphylococcus aureus.

Hot tnh khng nm: c 4 cht u c kh nng khng nm vi VSV kim

nh, hp cht 3d c tc dng vi 3/4 VSV kim nh gm Candida

albicans, Aspergillus niger, Saccharomyces cerevisiae. iu c bit l ch

c duy nht cht 3b khng li Penicilium sp..

42

2. KIN NGH

tip tc pht trin cc nghin cu t c, chng ti c mt s

kin ngh sau:

Nghin cu thay i cc iu kin, cng nh cc tc nhn trong cc phn

ng nng cao hiu sut ca cc dn cht acid 3-tert-butoxycarbonyl-2-

phenylthiazolidin-4-carboxylic.

Tng hp cc dn cht khc ca acid 2-phenylthiazolidin-4-carboxylic v

th hot tnh sinh hc, nhm nh gi su rng hn v nh hng ca cu

trc ti hot tnh ca cc dn cht TCA v c nhng nh hng cho vic

xy dng cu trc ca cc cht tng hp tip theo.

Th cc tc dng sinh hc khc ca cc dn cht 2-phenylthiazolidin-4-

carboxylic: tc dng chng ung th, tc dng h huyt p

TI LIU THAM KHO

TING VIT

1. Trn Mnh Bnh (2003), Phn tch cu trc cc hp cht hu c, B

mn Ha Hu c, Trng i hc Dc H Ni.

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.

3. Nguyn nh Triu (2005), Cc phng php phn tch vt l v ha l

tp II, Nh xut bn Khoa hc v K thut, H Ni, tr 9.

TING ANH

4. Chavan S. P., Chittiboyina Amar G, Ramakrishna Guduru, et al. (2005),

"An unusual stereochemical outcome of radical cyclization: synthesis

of (+)-biotin", Tetrahedron, 61(39), pp. 9273-9280.

5. Ding S. M. (2010),"3-(tert-Butoxycarbonyl)-2-(4-chlorophenyl)-1,3-

thiazolidine-4-carboxylic acid", Acta Crystallographica Section E:

Structure Reports Online, 66(10), pp. 2633-2633.

6. ElSharkawy K. A. (2011), "Synthesis and Antimicrobial Activity of 2-

Substituted-3-Acetylthiazolidine-4-Carbonyl-Amino acid Derivatives",

Journal of Pharmaceutical Sciences and Research, 3(1), pp. 1005-

1014.

7. Esra Onen-Bayram F, Durmaz Irem, Scherman Daniel, et al. (2012), "A

novel thiazolidine compound induces caspase-9 dependent apoptosis in

cancer cells", Bioorganic & medicinal chemistry, 20, pp. 5094-5012.

8. Khan Khalid Mohammed, Ullah Zia, Lodhi Muhammad Arif, et al.

(2006), "Successful computer guided planned synthesis of (4R)-

thiazolidine carboxylic acid and its 2-substituted analogues as urease

inhibitors", Molecular diversity, 10(2), pp. 223-231.

9. Liu Yu, Jing Fanbo, Xu Yingying, et al. (2011), "Design, synthesis and

biological activity of thiazolidine-4-carboxylic acid derivatives as novel

influenza neuraminidase inhibitors", Bioorganic & medicinal

chemistry, 19(7), pp. 2342-2348.

10. Lu Yan, Li Chien-Ming, Wang Zhao, et al. (2009), "Discovery of 4-

substituted methoxybenzoyl-aryl-thiazole as Novel Anticancer Agents:

Synthesis, biological evaluation, and structure activity relationships",

Journal of medicinal chemistry, 52(6), pp. 1701-1711.

11. Pavlova L. A., Komarova T. V., Davidovich Yu A., et al. (1986),

"Synthesis of derivatives of 2-phenylthiazolidine-4-carboxylic acid and

a study of their radioprotective properties", Pharmaceutical Chemistry

Journal, 20(9), pp. 653-658.

12. Prabhakar Yenamandra S., Jain P., Khan Z.K., et al. (2003), "Synthesis

and QSAR Studies on the Antifungal Activity of 2, 3, 4-Substituted

Thiazolidines", QSAR & Combinatorial Science, 22(4), pp. 456-465.

13. Schubert Maxwell P. (1936), "Compounds of thiol acids with

aldehydes", Journal of Biological Chemistry, 114(1), pp. 341-350.

14. Sriram D., Yogeeswari P., Patrisha J. T., et al. (2011), "Exploring aryl

thiazolidine carboxamides as a new class of antimycobacterials",

Newsletter, pp. 181-195.

15. Ha Young Mi, Park Yun Jung, Lee Ji Yeon, et al. (2012), "Design,

synthesis and biological evaluation of 2-(substituted phenyl)

thiazolidine-4-carboxylic acid derivatives as novel tyrosinase

inhibitors", Biochimie, 94(2), pp. 533-540.

16. Howard-Lock H. E., Lock C. J. L., Martins M. L. (1991), "Amino

acid/zwitterion equilibria II: vibrational and NMR studies of substituted

thiazolidine-4-carboxylic acids", Canadian journal of chemistry,

69(11), pp. 1721-1727.

17. Iwao Junichi, Oya Masayuki, Baba Toshio, et al., Antihypertensive 4-

thiazolidinecarboxylic acids (substituted phenyl derivatives). 1984,

Google Patents.

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activity relationship studies of arylthiazolidine amides as selective

cytotoxic agents for melanoma", Anticancer Research, 27(2), pp. 883-

888.

19. Paul B., Korytnyk W.(1976), "Cysteine derivatives with reactive groups

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20. Song Zhong-Cheng, Ma Gao-Yuan, Lv Peng-Cheng, et al. (2009),

"Synthesis, structure and structureactivity relationship analysis of 3-

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potential antibacterial agents", European journal of medicinal

chemistry, 44(10), pp. 3903-3908.

PH LC

Ph lc 1.1: Ph IR ca cht 3a

Ph lc 1.2: Ph IR ca cht 3b

Ph lc 1.3: Ph IR ca cht 3c

Ph lc 1.4: Ph IR ca cht 3d

Ph lc 2.1: Ph MS ca cht 3a

Ph lc 2.2: Ph MS ca cht 3b

Ph lc 2.3: Ph MS ca cht 3c

Ph lc 2.4: Ph MS ca cht 3d

Ph lc 3.1: Ph 1H-NMR ca cht 3a

Ph lc 3.2: Ph 1H-NMR ca cht 3b

Ph lc 3.3: Ph 1H-NMR ca cht 3c

Ph lc 3.4: Ph 1H-NMR ca cht 3d

Ph lc 1.1: Ph IR ca cht 3a

Ph lc 1.2: Ph IR cht 3b

Ph lc 1.3:Ph IR ca cht 3c

Ph lc 1.4: Ph IR ca cht 3d

Ph lc 2.1: Ph MS ca cht 3a

Ph lc 2.2: Ph MS ca cht 3b

Ph lc 2.3: Ph MS ca cht3c

Ph lc 2.4: Ph MS ca cht 3d

Ph lc 3.1: Ph1H-NMR ca cht 3a

Ph lc 3.2: Ph1H-NMR ca cht 3b

Ph lc 3.3: Ph 1H-NMR ca cht 3c

Ph lc 3.4: Ph 1H-NMR ca cht 3d