2-Thiopyrimidine/chalcone hybrids: design, synthesis, ADMET prediction, and anticancer evaluation as STAT3/STAT5a inhibitors

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY

https://doi.org/10.1080/14756366.2020.1740922

RESEARCH PAPER

2

-Thiopyrimidine/chalcone hybrids: design, synthesis, ADMET prediction, and

anticancer evaluation as STAT3/STAT5a inhibitors

Phoebe F. Lamie and John N. Philoppes

Department of Pharmaceutical Organic Chemistry, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt

ABSTRACT

ARTICLE HISTORY

A novel 2-thiopyrimidine/chalcone hybrid was designed, synthesised, and evaluated for their cytotoxic

activities against three different cell lines, K-562, MCF-7, and HT-29. The most active cytotoxic derivatives

were 9d, 9f, 9n, and 9p (IC50¼0.77–1.74 mM, against K-562 cell line), 9a and 9r (IC50¼1.37–3.56 mM

against MCF-7 cell line), and 9a, 9l, and 9n (IC ¼2.10 and 2.37 mM against HT-29 cell line). Compounds

Received 11 January 2020

Revised 2 March 2020

Accepted 2 March 2020

5

0

KEYWORDS

9

a, 9d, 9f, 9n, and 9r were further evaluated for their cytotoxicity against normal fibroblast cell line WI38.

2

-Thiopyrimidine; chalcone;

Moreover, STAT3 and STAT5a inhibitory activities were determined for the most active derivatives 9a, 9d,

STAT; cytotoxicity;

computational analysis

9

f, 9n, and 9r. Dual inhibitory activity was observed in compound 9n (IC50¼113.31 and 50.75 mM, against

STAT3 and STAT5a, respectively). Prediction of physicochemical properties, drug likeness score, pharmaco-

kinetic and toxic properties was detected.

GRAPHICAL ABSTRACT

Introduction

A critical signalling intermediate in cancer cells, specially leu-

kaemia, breast and colon cancer cells is called signal transducer

1

,2

One of the main causes of mortality all over the world is cancer

The highest prevalence for cancer death is being for stomach,

breast, prostatic, lung, and colon .

The most common female cancer around the world is breast

cancer. It represents for 16% of all female cancers and 18.2% of all

cancer death causes including both males and females .

On the other hand, about two million new cases are diagnosed

every year for colorectal cancer. Thus, making it as one of the

.

1

3–17

and activator of transcription (STAT) protein family

.

They are cytoplasmic transcription factors. STAT family consists

of seven members, STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b,

and STAT6. STAT2, STAT4, and STAT6 are responsible for regula-

tion of immune response. While, STAT1, STAT3, and STAT5 can

control cell cycle (cyclin D1, D2, c-Myc), cell survival (Bcl-xl, Bcl2,

3

4

Mcl-1), and angiogenesis (HIF1a, VEGFR) through regulation of

18,19

gene expression

.

5

,6

most common causes of cancer-related death

.

STAT can be activated either by receptor tyrosin kinases like

JAKs, PDGFR, EGFR, and FLT3, or through non-receptor tyrosin kin-

ases, Src, Brk, and Bcr-Abl. Also, activation of STAT may be from

activation of cytokines (IL-6), growth factors or negative feedback

Another common cause of cancer death is leukaemia, cancer in

blood-forming cells of the bone marrow, which is chemoresist-

7–11

ant

. Although, treatment of cancer using chemotherapeutic

2

0–24

agents is still used for several cancer types including breast, colon mechanisms

.

and leukaemia cancers, high toxicity level of chemotherapeutic

drugs limit their use .

Phosphorylation of STATs transforms them to active form caus-

ing their homo- or heterodimerisation then migration to the

1

2

CONTACT Phoebe F. Lamie

feebi.lamey@pharm.bsu.edu.eg;

feby.farag@yahoo.com

Department of Pharmaceutical Organic Chemistry, Faculty of

Pharmacy, Beni-Suef University, Beni-Suef, 62514, Egypt

Supplemental data for this article can be accessed here .

ß 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,

distribution, and reproduction in any medium, provided the original work is properly cited.

2

P. F. LAMIE AND J. N. PHILOPPES

nucleus to control gene expression. Over activation of STAT level synthesised derivatives were evaluated against leukaemia (K-562),

20

can lead to tumorigenesis .

breast (MCF-7), and colon (HT-29) cancer cell lines. Inhibitory activ-

Several studies have demonstrated that blocking STAT3 or ities of the most potent bioactive molecules against STAT3 and

STAT5 signalling pathway led to apoptosis in tumour cells. While, STAT5a were measured, aiming at finding more effective anti-

normal cells were able to survive even under a very low concen- cancer therapeutics.

tration of STAT3 or STAT5 and also capable of growing using

2

1,25

other mechanisms

.

Therefore, development of new anti-cancer agents with less

toxicity and overcoming chemotherapeutic drug resistance can be

achieved by: (1) using drugs that target two or three activators of

Experimental

Chemistry

2

4

8

STAT3 or (2) combined targeting of STAT3 and STAT5 .

Melting points were measured on the Griffin apparatus and were

It was found that a potent STAT3 inhibitor, S31-201 (I, uncorrected. Determination of IR spectra was achieved using

Figure 1), could inhibit proliferation of hepatocellular and breast

carcinoma in mice .

Shimadzu IR-435 spectrophotometer with KBr discs and values

1

6

ꢀ

1

13

were obtained in cm . H NMR and C NMR were recorded on

Moreover, compound S3I-201.1066 (II, Figure 1), containing sul-

phonamide group could inhibit STAT3 function in both breast and

1

13

Bruker instrument at 400 MHz for H NMR and 100 MHz for

NMR spectrophotometer (Faculty of Pharmacy, Mansoura

University, Mansoura, Egypt), in DMSO-d (as a solvent), D O using

C

2

6

myeloma cancer cells (EC ¼10 and 16 mM, respectively) .

50

6

2

Another compound, curcumine analogue, FLLL32 (III, Figure 1),

showed potent inhibitory activity in many human cancer cell lines

such as breast, colorectal, melanoma, and myeloma by preventing

TMS as an internal standard and chemical shifts (d) were

expressed in parts per million (ppm) compared to internal stand-

ard, TMS (d ¼ 0 ppm). Coupling constant (J) values were expressed

in Hertz (Hz). Signal splitting patterns were designated as follows:

s, singlet; d, doublet, t, triplet; q, quartette; m, multiplet. The elec-

tron impact (EI) mass spectra were carried out using Hewlett

Packard 5988 spectrometer (Palo Alto, CA) at Faculty of Science,

Cairo University, Giza, Egypt. Microanalysis was calculated for C, H,

N on Perkin-Elmer 2400 at the Microanalytical centre, Faculty of

Science, Cairo University, Egypt and was within ±0.4% of theoret-

ical values. The progress of the reaction and purity of products

were monitored by thin layer chromatography (TLC), pre-coated

plastic sheets, 0.2 mm silica gel with UV indicator (Macherey-

Nagel, D u€ ren, Germany). All used reagents and solvents were pur-

chased from the Aldrich Chemical Company (Milwaukee, WI).

20,27

STAT3 dimerisation and downstream functioning

.

Moreover, treatment with chalcone IV (Figure 1) caused signifi-

2

8

cant decrease in STAT3 level in leukaemia HL-60 cell line .

Pyrazolo[1,5-a]pyrimidine/chalcone hybrid V (Figure 1) showed

promising anti-proliferative activity by down regulation of STAT3

2

9

in MDA-MB-231 cells .

Additionally, NCI library identified two compounds (BP-1108

and BP-1075) as the most potent STAT5 in K562 leukaemia cell

3

0

lines through down regulation of STATs-defendant genes .

In medicinal chemistry, a very well-known heterocycle is pyr-

imidine. It takes its importance from its presence in thymidine,

cytosine and uracil bases, the building blocks of DNA and RNA

3

1,32

nucleic acids

-Thiopyrimidines (2-TPs), also named as 2-mercaptopyrimi-

dines, are one of the most important class of pyrimidines.

.

2

They attract the biochemists attention due to their wide range General method for preparation of compounds 4a–f

of applications in preparation of cardiotonic drugs, antitubercular A mixture of the appropriate chalcone derivative 3a–f (0.01 mol),

3

3,34

and anti-inflammatory agents

.

thiourea (0.76 g, 0.01 mol), and KOH (0.11 g, 0.02 mol) in absolute

3

Moreover, 2-TPs were evaluated for their anticancer activity . ethanol (20 ml) was heated under reflux temperature for 12 h. The

They were reported to have potent antitumor activity against leu- resulting solution was evaporated to dryness or (a precipitate in

kaemia, colon and breast cell lines such as compounds VI–IX case of 3c and 3f). The obtained residue was solubilised in water,

3

5–38

(Figure 1)

.

filtered and dried. The crude product was crystallised from etha-

Synthesis of 2-TPs derivatives could be achieved from reaction nol/DMF (8:2) to get compounds 4a–f.

39

of chalcone derivatives with thiourea . As chalcones constitute an

important group of natural products, their biological activities

were arisen from their chemical structure, a,b-unsaturated car-

4

-(4-Methoxyphenyl)-6-p-tolylpyrimidine-2-thiol (4a). Yield 82%;

ꢁ

ꢀ1

yellow powder; (ethanol 95%); mp 149–151 C; IR (cm ): 3413

4

0

bonyl group .

1

(NH), 3192 (CH aromatic), 2958 (CH aliphatic); H NMR (400 MHz,

Many synthesised chalcones were reported to have potent

in vitro anticancer activity against human colon carcinoma, non-

DMSO-d ) d 2.50 (s, 3H, CH ), 4.43 (s, 3H, OCH ), 7.24–7.59 (m, 4H,

6

3

p-methoxyphenyl H-3, H-5, p-tolyl H-3, H-5), 7.79–7.91 (m, 4H, pyr-

imidine H-5, p-methoxyphenyl H-2, H-6, NH, D O exchangeable),

.21 (d, J¼ 8.4 Hz, 2H, p-tolyl H-2, H-6); C NMR (100 MHz, DMSO-

) d 21.2 (CH ), 55.6 (OCH ), 101.0 (pyrimidine C-5), 114.7 (p-

41–44

small cell lung carcinoma, and breast cancer

.

2

Thus, both chloro- and dimethylamino-derivatives of com-

1

3

8

d

pound X (Figure 1), showed cytotoxic activity against human leu-

4

0

6

3

kaemia cells with CC50¼2.17 and 2.06 lM, respectively . While,

4

5

methoxyphenyl C-3, C-5), 126.1 (p-tolyl C-2, C-6), 128.1 (p-tolyl C-3,

C-5), 129.4 (p-methoxyphenyl C-2, C-6), 130.9 (p-methoxyphenyl C-

1), 134.5 (p-tolyl C-1), 136.7 (p-tolyl C-4), 159.2 (p-methoxyphenyl

C-4), 164.6 (pyrimidine C-4) 172.9 (pyrimidine C-6), 175.2 (pyrimi-

compound XI (Figure 1) was apoptosis inducer in A549 cells .

In light of the above facts, and as a part of our previously pub-

46,47

lished anticancer research articles

, our scope in this research

was to design and synthesised a new series of 2-TP/chalcone

hybrids (Figure 1), through molecular hybridisation, by merging:

þ

dine C-2); EIMS (m/z): 309.00 (M þ 1, 31.75%), 308.00 (M , 74.33%),

3

5

16 2

07.00 (100%); Anal. Calcd. for C18H N OS (308.40): C, 70.10; H,

.23; N, 9.08. Found: C, 70.31; H, 5.07; N, 8.88.

(i) 2-Thiopyrimidine scaffold, such as in compounds (VI–IX), (ii)

chalcone part from compounds (III–V, X, XI), (iii) choosing sub-

stituents on phenyl rings of pyrimidine C-4, pyrimidine C-6, and

chalcone as in compounds (I–V), and (iv) amide linkage to mimic 4-(4-Methoxyphenyl)-6-(4-nitrophenyl)pyrimidine-2-thiol (4b). Yield

ꢁ

39

that in compounds (I, II, VI, X, XI). The cytotoxic activities of the 75%; yellow crystals; mp 63–65 C .

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY

3

Figure 1. The designed strategy for 2-TP/chalcone hybrids as new anticancer STAT inhibitors.

4

-[4-(2-Chloroethoxy)phenyl]-6-(4-methoxyphenyl)pyrimidine-2-

2

5, H-6, p-methoxyphenyl H-3, H-5, NH, D O exchangeable),

1

3

thiol (4c). Yield 69%; yellow powder; (ethanol 95%); mp 8.22–8.30 (m, 3H, p-methoxyphenyl H-2, H-6, pyrimidine H-5);

C

ꢁ

ꢀ1

2

82–284 C; IR (cm ): 3436 (NH), 3192 (CH aromatic), 2927 (CH NMR (100 MHz, DMSO-d

6

) d 42.9 (CH

2 3 2

Cl), 55.8 (OCH ), 68.1 (OCH ),

1

aliphatic); H NMR (400 MHz, DMSO-d ) d 3.83–4.37 (m, 7H, OCH , 105.0 (pyrimidine C-5), 114.8 (p-methoxyphenyl C-3, C-5), 115.4 (p-

6

3

2 2

OCH , CH Cl), 6.96–7.10 (m, 7H, p-chloroethoxyphenyl H-2, H-3, H- chloroethoxyphenyl C-3, C-5), 121.8 (p-chlorophenyl C-2, C-6),

4

P. F. LAMIE AND J. N. PHILOPPES

1

2

27.9 (p-methoxyphenyl C-1), 128.5 (p-methoxyphenyl C-2, C-6), 7.91–7.94 (m, 4H, phenyl H-2, H-3, H-5, H-6), 10.84 (s, 1H, NH, D O

1

3

29.4 (p-chloroethoxyphenyl C-2, C-6), 130.0 (p-chloroethoxy- exchangeable); C NMR (100 MHz, DMSO-d

), 56.4 (2OCH ), 113.7 (p-methoxyphenylacroyl C-3, C-5), 118.8

phenyl C-4), 162.5 (pyrimidine C-4) 176.1 (pyrimidine C-6), 179.3 (p-methoxyphenyl C-3, C-5), 120.1 (COCH¼CH), 121.7 (phenyl C-2,

pyrimidine C-2); Anal. Calcd. for C H ClN O S (372.87): C, 61.20; C-6), 127.5 (p-methoxyphenylacroyl C-1), 127.6 (p-tolyl C-2, C-6),

) d 22.3 (CH ), 36.2

6

3

phenyl C-1), 160.6 (p-methoxyphenyl C-4), 161.7 (p-chloroethoxy- (CH

2

3

(

19

17

2 2

H, 4.60; N, 7.51. Found: C, 61.41; H, 4.57; N, 7.75.

129.0 (p-methoxyphenyl C-2, C-6), 129.1 (p-methoxyphenyl C-1),

1

30.2 (p-tolyl C-3, C-5), 130.8 (p-methoxyphenylacroyl C-2, C-6),

31.4 (phenyl C-3, C-5), 132.0 (p-tolyl C-4), 133.5 (phenyl C-4),

4

-(4-Chlorophenyl)-6-p-tolylpyrimidine-2-thiol (4d). Yield 91%;

ꢁ

47

136.9 (p-tolyl C-1), 144.3 (phenyl C-1), 154.4 (COCH¼CH), 155.1 (p-

yellow crystals; mp 180–182 C .

methoxyphenylacroyl C-4), 155.4 (p-methoxyphenyl C-4), 164.1

(

1

7

pyrimidine C-6), 167.8 (pyrimidine C-4), 172.3 (pyrimidine C-2),

73.0 (CONH), 190.0 (CO); Anal. Calcd. for C H N O S (601.20): C,

36 31 3 4

1.86; H, 5.19; N, 6.98. Found: C, 71.67; H, 5.07; N, 6.93.

4

6

3

7

-(4-Chlorophenyl)-6-(4-nitrophenyl)pyrimidine-2-thiol (4e). Yield

ꢁ

ꢀ1

5%; yellow powder; (ethanol 95%); mp 263–265 C; IR (cm ):

1

434 (NH), 3064 (CH aromatic); H NMR (400 MHz, DMSO-d

.60 (d, J¼ 8.4 Hz, 2H, p-chlorophenyl H-3, H-5), 7.96 (s, 1H, NH,

O exchangeable), 8.33–8.34 (m, 4H, p-chlorophenyl H-2, H-6, p-

6

) d

(

ZE)-N-f4-[3-(3,4-Dimethoxyphenyl)acryloyl]phenylg-2-[4-(4-

D

2

methoxyphenyl)-6-p-tolylpyrimidin-2-ylthio]acetamide (9b). Yield

65%; yellow powder; mp 165–167 C; IR (cm ): 3324 (NH), 3061

(

3

2

nitrophenyl H-2, H-6), 8.54 (d, J¼ 8.4 Hz, 2H, p-nitrophenyl H-3, H-

ꢁ

ꢀ1

13

5

), 8.66 (s, 1H, pyrimidine H-5); C NMR (100 MHz, DMSO-d

6

) d

1

CH aromatic), 2925 (CH aliphatic), 1663 (broad, 2C¼O); H NMR

1

01.3 (pyrimidine C-5), 123.6 (p-chlorophenyl C-2, C-6), 128.0 (p-

6 3 3

400 MHz, DMSO-d ) d 2.36 (s, 3H, CH ), 3.81 (s, 3H, OCH ), 3.82 (s,

nitrophenyl C-3, C-5), 128.9 (p-chlorophenyl C-1), 129.3 (p-chloro-

phenyl C-3, C-5), 129.5 (p-nitrophenyl C-2, C-6), 133.5 (p-chloro-

phenyl C-4), 139.2 (p-nitrophenyl C-1), 150.2 (p-nitrophenyl C-4),

H, OCH ), 3.86 (s, 3H, OCH ), 4.23 (s, 2H, CH ), 6.97 (d, J¼ 8.4 Hz,

3

2

H, p-methoxyphenyl H-3, H-5), 7.03 (d, J¼ 8.0 Hz, 1H, dimethoxy-

phenyl H-5), 7.28 (d, J¼ 8.0 Hz, 1H, dimethoxyphenyl H-6), 7.37 (d,

1

2

1

64.6 (pyrimidine C-4) 176.3 (pyrimidine C-6), 180.4 (pyrimidine C-

); Anal. Calcd. for C16 10ClN S (343.79): C, 55.90; H, 2.93; N,

2.22. Found: C, 60.12; H, 2.87; N, 12.46.

J¼ 12.0 Hz, 1H, COCH¼CH), 7.54 (s, 1H, dimethoxyphenyl H-2),

H

3 2

O

7

.69 (d, J¼ 12.0 Hz, 1H, COCH¼CH), 7.83–7.87 (m, 3H, p-tolyl H-3,

H-5, pyridine H-5), 8.18–8.23 (m, 6H, phenyl H-2, H-6, p-methoxy-

phenyl H-2, H-6, p-tolyl H-2, H-6), 8.31 (d, J¼ 8.4 Hz, 2H, phenyl H-

4

-[4-(2-Chloroethoxy)phenyl]-6-(4-chlorophenyl)pyrimidine-2-thiol

13

3

, H-5), 10.84 (s, 1H, NH, D O exchangeable); C NMR (100 MHz,

2

ꢁ

(4f). Yield 62%; yellow powder; (ethanol 95%); mp 242–244 C; IR

DMSO-d

6 3 2 3 3

) d 21.4 (CH ), 36.4 (CH ), 55.8 (OCH ), 56.0 (OCH ), 56.1

ꢀ

1

(cm ): 3417 (NH), 3066 (CH aromatic), 2927 (CH aliphatic);

H

(

OCH ), 107.4 (pyrimidine C-5), 111.0 (dimethoxyphenyl C-2), 111.9

3

NMR (400 MHz, DMSO-d ) d 3.98 (t, J¼ 8.4 Hz, 2H, CH Cl), 4.36 (t,

6

2

(dimethoxyphenyl C-5), 114.6 (dimethoxyphenyl C-6), 118.8 (p-

methoxyphenyl C-3, C-5), 119.8 (COCH¼CH), 124.4 (phenyl C-2, C-

6), 127.7 (p-tolyl C-2, C-6), 128.0 (p-methoxyphenyl C-2, C-6), 129.6

J¼ 8.4 Hz, 2H, OCH ), 7.07–7.12 (m, 4H, p-chloroethoxyphenyl H-3,

2

H-5, p-chlorophenyl H-2, H-6), 7.31 (s, 1H, NH, D O exchangeable),

2

7

.60 (d, J¼ 8.4 Hz, 2H, p-chlorophenyl H-3, H-5), 7.96 (s, 1H, pyrimi-

(

p-methoxyphenyl C-1), 130.3 (p-tolyl C-3, C-5), 130.4 (p-tolyl C-1),

33.0 (dimethoxyphenyl C-1), 133.6 (phenyl C-3, C-5), 141.7 (p-tolyl

C-4), 143.9 (phenyl C-4), 144.4 (phenyl C-1), 149.4 (dimethoxy-

dine H-5), 8.31 (d, J¼ 8.4 Hz, 2H, p-chloroethoxyphenyl H-2, H-6);

1

3

6 2 2

C NMR (100 MHz, DMSO-d ) d 43.4 (CH Cl), 68.5 (OCH ), 103.9

(

pyrimidine C-5), 115.3 (p-chloroethoxyphenyl C-3, C-5), 120.2 (p- phenyl C-3), 150.0 (dimethoxyphenyl C-4), 151.6 (COCH¼CH),

chlorophenyl C-2, C-6), 128.5 (p-chlorophenyl C-1), 129.4 (p-chlor- 162.3 (p-methoxyphenyl C-4), 164.2 (pyrimidine C-6), 164.3 (pyrimi-

oethoxyphenyl C-2, C-6), 129.5 (p-chlorophenyl C-3, C-5), 130.7 (p- dine C-4), 167.7 (pyrimidine C-2), 170.8 (CONH), 187.8 (CO); Anal.

chloroethoxyphenyl C-1), 136.6 (p-chlorophenyl C-4), 161.7 (p- Calcd. for C H N O S (631.21): C, 70.34; H, 5.27; N, 6.65. Found:

3

7 33 3 5

chloroethoxyphenyl C-4), 162.8 (pyrimidine C-4) 176.3 (pyrimidine C, 70.54; H, 5.07; N, 6.71.

C-6), 179.0 (pyrimidine C-2); EIMS (m/z): 376.95 (M þ 1, 16.83%),

þ

3

75.90 (M , 19.31%), 55.10 (100%); Anal. Calcd. for C18

H

14ClN

2

OS

(

5

ZE)-2-[4-(4-Methoxyphenyl)-6-p-tolylpyrimidin-2-ylthio]-N-f4-[3-

3,4,5-trimethoxyphenyl)acryloyl]phenylgacetamide (9c). Yield

(377.29): C, 57.30; H, 3.74; N, 7.42. Found: C, 57.41; H, 3.57; N, 7.68.

ꢁ

ꢀ1

6%; yellow powder; mp 195–197 C; IR (cm ): 3290 (NH), 2998

1

(

CH aromatic), 2927 (CH aliphatic), 1665 (broad, 2C¼O); H NMR

400 MHz, DMSO-d ) d 2.36 (s, 3H, CH ), 3.72 (s, 3H, OCH ), 3.81 (s,

H, OCH ), 3.87 (s, 6H, 2OCH ), 4.24 (s, 2H, CH ), 6.95 (s, 2H, trime-

thoxyphenyl H-2, H-6), 6.98–7.28 (m, 4H, p-methoxyphenyl H-3, H-

, p-tolyl H-3, H-5), 7.69 (d, J¼ 12.0 Hz, 1H, COCH¼CH), 7.87–8.19

m, 3H, COCH¼CH, p-tolyl H-2, H-6), 8.22–8.31 (m, 7H, p-methoxy-

phenyl H-2, H-6, pyrimidine H-5, phenyl H-2, H-3, H-5, H-6), 10.86

General method for preparation of compounds 9a–r

(

3

6

3

A mixture of pyrimidine derivatives 4a–f (0.01 mol), acetyl chloride

derivatives 8a–c (0.01 mol), and catalytic amount of TEA is stirred

in acetonitrile (20 ml) for 24 h. The solution was evaporated to dry-

ness. The obtained residue was solubilised in ice cold water and

neutralised with conc. HCl. The obtained solid was filtered, dried

and crystallised from ethanol/DMF (8:2).

3

2

5

(

13

(s, 1H, NH, D O exchangeable); C NMR (100 MHz, DMSO-d ) d

2

6

2

1

1.4 (CH

3

), 36.4 (CH

2

), 55.8 (OCH

3

), 56.5 (2OCH

3

), 60.6 (OCH ),

(

ZE)-2-[4-(4-Methoxyphenyl)-6-p-tolylpyrimidin-2-ylthio]-N-f4-[3-

06.8 (trimethoxyphenyl C-2, C-6), 107.4 (pyrimidine C-5), 114.5 (p-

4 methoxyphenyl)acryloyl]phenylgacetamide (9a). Yield 82%; methoxyphenyl C-3, C-5), 118.8 (phenyl C-2, C-6), 121.5

ꢁ

ꢀ1

yellow powder; mp 226–228 C; IR (cm ): 3257 (NH), 3039 (CH (COCH¼CH), 126.9 (p-tolyl C-2, C-6), 127.7 (trimethoxyphenyl C-1),

1

aromatic), 2925 (CH aliphatic), 1795, 1663 (2C¼O); H NMR 128.5 (p-methoxyphenyl C-1), 129.6 (p-methoxyphenyl C-2, C-6),

(

400 MHz, DMSO-d ) d 1.86 (s, 3H, CH ), 4.41 (s, 3H, OCH ), 4.43 (s, 130.4 (p-tolyl C-3, C-5), 131.0 (p-tolyl C-4), 132.8 (phenyl C-3, C-5),

6 3 3

3

H, OCH ), 4.84 (s, 2H, CH ), 6.67 (d, J¼ 8.4 Hz, 2H, p-methoxyphe- 133.5 (p-tolyl C-1), 140.0 (phenyl C-4), 141.7 (trimethoxyphenyl C-

3

2

nylacroyl H-3, H-5), 7.10 (d, J¼ 7.6 Hz, 2H, p-methoxyphenyl H-3, 4), 144.0 (phenyl C-1), 144.4 (COCH¼CH), 153.5 (trimethoxyphenyl

H-5), 7.24 (d, J¼ 8.4 Hz, 2H, p-methoxyphenylacroyl H-2, H-6), 7.33 C-3, C-5), 162.3 (p-methoxyphenyl C-4), 164.2 (pyrimidine C-6),

(

d, J¼ 6.0 Hz, 2H, p-tolyl H-3, H-5), 7.44–7.74 (m, 7H, COCH¼CH, 164.3 (pyrimidine C-4), 167.7 (pyrimidine C-2), 170.7 (CONH), 187.9

þ

pyrimidine H-5, p-methoxyphenyl H-2, H-6, p-tolyl H-2, H-6), (CO); EIMS (m/z): 662.05 (M þ 1, 5.50%), 661.05 (M , 12.94%),

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY

5

3

22.05 (100%); Anal. Calcd. for C38

H

35

N

3

O

6

S (661.22): C, 68.97; H, 3.87 (s, 6H, 2OCH

methoxyphenyl H-3, H-5), 7.23 (s, 2H, trimethoxyphenyl H-2, H-6),

.69 (d, J¼ 12.0 Hz, 1H, COCH¼CH), 7.86–7.92 (m, 3H, COCH¼CH,

p-methoxyphenyl H-2, H-6), 8.21 (d, J¼ 8.0 Hz, 2H, phenyl H-2, H-

3

), 4.27 (s, 2H, CH

2

), 6.98 (d, J¼ 8.0 Hz, 2H, p-

5

.33; N, 6.35. Found: C, 68.78; H, 5.17; N, 6.24.

7

(

ZE)-2-[4-(4-Methoxyphenyl)-6-(4-nitrophenyl)pyrimidin-2-ylthio]-

6

), 8.27 (d, J¼ 8.0 Hz, 2H, phenyl H-3, H-5), 8.33 (d, J¼ 8.0 Hz, 2H,

N-f4-[3-(4-methoxyphenyl)acryloyl]phenylgacetamide (9d). Yield

ꢁ

ꢀ1

p-nitrophenyl H-2, H-6), 8.40 (s, 1H, pyrimidin H-5), 8.56 (d,

5

7%; yellow powder; mp 183–185 C; IR (cm ): 3407 (NH), 3063

1

J¼ 8.0 Hz, 2H, p-nitrophenyl H-3, H-5), 10.89 (s, 1H, NH, D O

2

(

CH aromatic), 2930 (CH aliphatic), 1664 (broad, 2C¼O); H NMR

1

3

6 2

exchangeable); C NMR (100 MHz, DMSO-d ) d 36.4 (CH ), 55.8

400 MHz, DMSO-d ) d 3.83 (s, 3H, OCH ), 3.84 (s, 3H, OCH ), 4.24

6

3

(

6

(

1

OCH

), 109.0 (pyrimidine C-5), 114.6 (p-methoxyphenyl C-3, C-5), 118.8

phenyl C-2, C-6), 121.4 (COCH¼CH), 124.2 (p-nitrophenyl C-3, C-5),

28.1 (trimethoxyphenyl C-1), 129.1 (p-nitrophenyl C-2, C-6), 129.8

p-methoxyphenyl C-2, C-6), 130.4 (phenyl C-3, C-5), 130.7 (p-

3 3 3

), 56.5 (2OCH ), 60.6 (OCH ), 106.8 (trimethoxyphenyl C-2, C-

s, 2H, CH ), 6.98 (d, J¼ 8.8 Hz, 2H, p-methoxyphenylacryloyl H-3,

2

H-5), 7.03 (d, J¼ 8.8 Hz, 2H, p-methoxyphenyl H-3, H-5), 7.69 (d,

J¼ 15.6 Hz, 1H, COCH¼CH), 7.80–7.86 (m, 5H, COCH¼CH, p-

methoxyphenylacryloyl H-2, H-6, p-methoxyphenyl H-2, H-6), 8.18

(

d, J¼ 8.8 Hz, 2H, phenyl H-2, H-6), 8.27 (d, J¼ 8.8 Hz, 2H, phenyl

methoxyphenyl C-1), 132.9 (phenyl C-4), 140.0 (trimethoxyphenyl

C-4), 142.3 (p-nitrophenyl C-1), 144.3 (phenyl C-1), 144.4

(COCH¼CH), 149.3 (p-nitrophenyl C-4), 153.5 (trimethoxyphenyl C-

H-3, H-5), 8.33 (d, J¼ 8.8 Hz, 2H, p-nitrophenyl H-2, H-6), 8.40 (s,

1

H, pyrimidine H-5), 8.56 (d, J¼ 8.8 Hz, 2H, p-nitrophenyl H-3, H-5),

13

1

0.86 (s, 1H, NH, D O exchangeable); C NMR (100 MHz, DMSO-

2

3

, C-5), 162.2 (p-methoxyphenyl C-4), 162.6 (pyrimidine C-6), 164.9

d

6

) d 36.4 (CH

14.7 (p-methoxyphenylacryloyl C-3, C-5), 114.8 (p-methoxyphenyl

C-3, C-5), 118.8 (phenyl C-2, C-6), 119.8 (p-nitrophenyl C-3, C-5),

2 3 3

), 55.8 (OCH ), 55.9 (OCH ), 109.1 (pyrimidine C-5),

(

pyrimidine C-4), 167.6 (pyrimidine C-2), 171.3 (CONH), 187.9 (CO);

1

þ

EIMS (m/z): 693.00 (M þ 1, 0.93%), 692.00 (M , 1.33%), 55.10

(

32 4 8

100%); Anal. Calcd. for C37H N O S (692.19): C, 64.15; H, 4.66; N,

1

24.2 (COCH¼CH), 127.8 (p-nitrophenyl C-2, C-6), 128.2 (p-methox-

8

.09. Found: C, 63.98; H, 4.57; N, 7.89.

yphenyl C-2, C-6), 129.1 (p-methoxyphenylacryloyl C-1), 129.8 (p-

methoxyphenyl C-1), 130.0 (p-methoxyphenylacryloyl C-2, C-6),

1

4

1

2

30.3 (phenyl C-3, C-5), 131.1 (p-nitrophenyl C-1), 133.1 (phenyl C- (ZE)-2-f4-[4-(2-Chloroethoxy)phenyl]-6-(4-methoxyphenyl)pyrimi-

), 142.4 (phenyl C-1), 143.8 (COCH¼CH), 149.3 (p-nitrophenyl C-4), din-2-ylthiog-N-f4-[3-(4-methoxyphenyl)acryloyl]phe-

61.7 (p-methoxyphenylacryloyl C-4), 162.2 (p-methoxyphenyl C-4),

62.6 (pyrimidine C-6), 164.9 (pyrimidine C-4), 167.6 (pyrimidine C-

), 171.3 (CONH), 187.8 (CO); Anal. Calcd. for C35H N O S (632.17):

28 4 6

ꢁ

nylgacetamide (9g). Yield 52%; yellow powder; mp 280–282 C; IR

ꢀ

1

(

cm ): 3431 (NH), 3039 (CH aromatic), 2935 (CH aliphatic), 1598

1

(

broad, 2C¼O); H NMR (400 MHz, DMSO-d

6

) d 3.70 (s, 3H, OCH

3

),

C, 66.44; H, 4.46; N, 8.86. Found: C, 66.42; H, 4.39; N, 8.74.

3

.83–3.85 (m, 5H, OCH and CH Cl), 4.21–4.23 (m, 4H, OCH

3

2

and

CH ), 7.02–7.16 (m, 6H, p-methoxyphenyl H-3, H-5, p-methoxyphe-

2

(

ZE)-N-f4-[3-(3,4-Dimethoxyphenyl)acryloyl]phenylg-2-[4-(4-

nylacryloyl H-3, H-5 and p-chloroethoxyphenyl H-3, H-5), 7.71–7.96

(m, 9H, p-methoxyphenyl H-2, H-6, p-methoxyphenylacryloyl H-2,

H-6, p-chloroethoxyphenyl H-2, H-6, phenyl H-2, H-6 and

COCH¼CH), 8.16 (d, J¼ 8.4 Hz, 2H, phenyl H-3, H-5), 8.18–8.20 (m,

methoxyphenyl)-6-(4-nitrophenyl)pyrimidin-2-ylthio]-acetamide

ꢁ

ꢀ1

(9e). Yield 54%; yellow powder; mp 225–227 C; IR (cm ): 3431

(

NH), 3039 (CH aromatic), 2924 (CH aliphatic), 1656 (broad, 2C¼O);

1

2

H, pyrimidin H-5 and COCH¼CH), 10.89 (s, 1H, NH, D

2

O

H NMR (400 MHz, DMSO-d ) d 3.83 (s, 3H, OCH ), 3.87 (s, 6H,

6

3

1

3

exchangeable); C NMR (100 MHz, DMSO-d

CH Cl), 55.4 (OCH ), 55.8 (OCH ), 68.9 (OCH ), 107.8 (pyrimidine C-

), 114.1 (p-methoxyphenylacroyl C-3, C-5), 114.7 (p-methoxy-

phenyl C-3, C-5), 114.8 (p-chloroethoxyphenyl C-3, C-5), 121.3

6 2

) d 36.2 (CH ), 40.6

2

3

OCH ), 4.27 (s, 2H, CH

2

), 6.99 (d, J¼ 8.4 Hz, 2H, p-methoxyphenyl

(

5

2

3

2

H-3, H-5), 7.03 (d, J¼ 8.8 Hz, 1H, dimethoxyphenyl H-5), 7.39 (d,

J¼ 8.8 Hz, 1H, dimethoxyphenyl H-6), 7.55 (s, 1H, dimethoxyphenyl

H-2), 7.68 (d, J¼ 15.2 Hz, 1H, COCH¼CH), 7.82–7.88 (m, 3H,

COCH¼CH, p-methoxyphenyl H-2, H-6), 8.20 (d, J¼ 8.8 Hz, 2H, phe-

nyl H-2, H-6), 8.28 (d, J¼ 8.8 Hz, 2H, phenyl H-3, H-5), 8.34 (d,

J¼ 8.8 Hz, 2H, p-nitrophenyl H-2, H-6), 8.41 (s, 1H, pyrimidine H-5),

(

COCH¼CH), 122.1 (phenyl C-2, C-6), 127.4 (p-chloroethoxyphenyl

C-1), 127.5 (p-methoxyphenylacroyl C-1), 128.3 (p-chloroethoxy-

phenyl C-2, C-6), 129.5 (p-methoxyphenyl C-2, C-6), 130.3 (p-

methoxyphenylacroyl C-2, C-6), 131.1 (phenyl C-3, C-5), 133.5 (phe-

nyl C-4), 144.0 (phenyl C-1), 144.3 (p-methoxyphenyl C-1), 145.4

8

.57 (d, J¼ 8.8 Hz, 2H, p-nitrophenyl H-3, H-5), 10.87 (s, 1H, NH,

13

D O exchangeable); C NMR (100 MHz, DMSO-d ) d 36.4 (CH ),

5

1

2

6

2

(

COCH¼CH), 159.3 (p-chloroethoxyphenyl C-4), 159.8 (p-methoxy-

5.9 (OCH ), 56.0 (OCH ), 56.2 (OCH ), 109.1 (pyrimidine C-5),

3 3 3

11.0 (dimethoxyphenyl C-2), 112.0 (dimethoxyphenyl C-5), 114.7

phenylacroyl C-4), 160.6 (p-methoxyphenyl C-4), 162.8 (pyrimidine

C-6), 164.9 (pyrimidine C-4), 168.6 (pyrimidine C-2), 172.3 (CONH),

(

1

p-methoxyphenyl C-3, C-5), 118.8 (dimethoxyphenyl C-6), 119.8

phenyl C-2, C-6), 124.2 (p-nitrophenyl C-3, C-5), 124.4 (COCH¼CH),

1

3 5

89.9 (CO); Anal. Calcd. for C37H32ClN O S (665.18): C, 66.71; H,

4

.84; N, 6.31. Found: C, 66.98; H, 4.57; N, 6.28.

28.0 (p-nitrophenyl C-2, C-6), 128.2 (p-methoxyphenyl C-2, C-6),

29.1 (dimethoxyphenyl C-1), 129.8 (p-methoxyphenyl C-1), 130.3

(

phenyl C-3, C-5), 132.0 (phenyl C-4), 142.4 (phenyl C-1), 143.8 (p- (ZE)-2-f4-[4-(2-Chloroethoxy)phenyl]-6-(4-methoxyphenyl)pyrimi-

nitrophenyl C-1), 144.4 (COCH¼CH), 149.4 (dimethoxyphenyl C-4),

49.4 (dimethoxyphenyl C-3), 153.5 (p-nitrophenyl C-4), 162.2 (p-

methoxyphenyl C-4), 162.6 (pyrimidine C-6), 165.0 (pyrimidine C-

), 167.6 (pyrimidine C-2), 171.3 (CONH), 187.8 (CO); Anal. Calcd.

for C36 S (662.18): C, 65.24; H, 4.56; N, 8.45. Found: C,

5.35; H, 4.71; N, 8.24.

din-2-ylthiog-N-f4-[3-(3,4-dimethoxyphenyl)acryloyl]phe-

1

ꢁ

nylgacetamide (9h). Yield 52%; yellow powder; mp 135–137 C; IR

ꢀ1

(

cm ): 3426 (NH), 3067 (CH aromatic), 2928 (CH aliphatic), 1657

4

1

(

3

4

broad, 2C¼O); H NMR (400 MHz, DMSO-d

6

) d 3.70 (s, 3H, OCH

3

),

30 4 7

H N O

.82 (s, 3H, OCH ), 3.86 (s, 3H, OCH ), 3.96 (t, J¼ 7.2 Hz, 2H, CH Cl),

3

2

6

.23 (s, 2H, CH ), 4.34 (t, J¼ 7.2 Hz, 2H, OCH ), 6.82–7.12 (m, 7H, p-

2

methoxyphenyl H-3, H-5, p-chloroethoxyphenyl H-3, H-5, dime-

thoxyphenyl H-2, H-5, H-6), 7.38 (d, J¼ 15.2 Hz, 1H, COCH¼CH),

(

ZE)-2-[4-(4-Methoxyphenyl)-6-(4-nitrophenyl)pyrimidin-2-ylthio]-

N-f4-[3-(3,4,5-trimethoxyphenyl)acryloyl]phenylgacetamide (9f). 7.67–7.86 (m, 5H, COCH¼CH, p-methoxyphenyl H-2, H-6, p-chlor-

ꢁ

ꢀ1

Yield 69%; yellow powder; mp 254–256 C; IR (cm ): 3265 (NH), oethoxyphenyl H-2, H-6), 8.18–8.29 (m, 5H, phenyl H-2, H-6, phe-

1

3

103 (CH aromatic), 2933 (CH aliphatic), 1663 (broad, 2C¼O);

H

),

nyl H-3, H-5, pyrimidin H-5), 10.89 (s, 1H, NH, D

2

O exchangeable);

1

3

NMR (400 MHz, DMSO-d ) d 3.71 (s, 3H, OCH ), 3.82 (s, 3H, OCH

6

3

6

C NMR (100 MHz, DMSO-d ) d 42.2 (CH

2

), 43.4 (CH Cl), 55.8

2

6

P. F. LAMIE AND J. N. PHILOPPES

(

OCH

3

), 56.1 (OCH

3

), 56.2 (OCH

3

), 75.1 (OCH

2

), 109.0 (pyrimidine C- (COCH¼CH), 161.7 (p-methoxyphenyl C-4), 163.4 (pyrimidine C-6),

5

1

3

), 111.5 (dimethoxyphenyl C-2), 111.9 (dimethoxyphenyl C-5), 164.9 (pyrimidine C-4), 167.6 (pyrimidine C-2), 171.1 (CONH), 187.8

14.5 (p-methoxyphenyl C-3, C-5), 115.1 (p-chloroethoxyphenyl C- (CO); EIMS (m/z): 607.20 (M þ 2, 1.54%), 606.15 (M þ 1, 1.28%),

þ

, C-5), 118.8 (phenyl C-2, C-6), 119.8 (COCH¼CH), 124.4 (dime- 605.15 (M , 2.65%), 57.10 (100%); Anal. Calcd. for C35

3 3

H28ClN O S

thoxyphenyl C-6), 127.3 (dimethoxyphenyl C-1), 127.4 (p-chloroe- (605.15): C, 69.35; H, 4.66; N, 6.93. Found: C, 69.41; H, 4.87; N, 7.13.

thoxyphenyl C-1), 128.0 (p-chloroethoxyphenyl C-2, C-6), 128.7 (p-

methoxyphenyl C-1), 129.6 (p-methoxyphenyl C-2, C-6), 130.3

(

ZE)-2-[4-(4-Chlorophenyl)-6-p-tolylpyrimidin-2-ylthio]-N-f4-[3-

3,4-dimethoxyphenyl)acryloyl]phenylgacetamide (9k). Yield 57%;

(

phenyl C-3, C-5), 131.3 (phenyl C-4), 143.9 (phenyl C-1), 144.4

COCH¼CH), 149.4 (dimethoxyphenyl C-4), 151.6 (dimethoxyphenyl

ꢁ

ꢀ1

yellow powder; mp 144–146 C; IR (cm ): 3273 (NH), 3039 (CH

C-3), 157.2 (p-chloroethoxyphenyl C-4), 161.2 (p-methoxyphenyl C-

1

aromatic), 2924 (CH aliphatic), 1665 (broad, 2C¼O); H NMR

4

), 162.2 (pyrimidine C-6), 164.3 (pyrimidine C-4), 167.6 (pyrimidine

(400 MHz, DMSO-d ) d 2.35 (s, 3H, CH ), 3.85 (s, 3H, OCH ), 3.86 (s,

6 3 3

C-2), 170.7 (CONH), 187.6 (CO); Anal. Calcd. for C H ClN O S

38

34

3

6

3H, OCH ), 4.24 (s, 2H, CH ), 7.01 (d, J¼ 8.4 Hz, 1H, dimethoxy-

3

2

(695.19): C, 65.56; H, 4.92; N, 6.04. Found: C, 65.74; H, 5.07; N, 5.99.

phenyl H-5), 7.27 (d, J¼ 8.0 Hz, 2H, p-tolyl H-3, H-5), 7.37 (d,

J¼ 8.4 Hz, 1H, dimethoxyphenyl H-6), 7.50 (d, J¼ 8.8 Hz, 2H, p-

(

ZE)-2-f4-[4-(2-Chloroethoxy)phenyl]-6-(4-methoxyphenyl)pyrimi- chlorophenyl H-3, H-5), 7.53 (s, 1H, dimethoxyphenyl H-2), 7.69 (d,

din-2-ylthiog-N-f4-[3-(3,4,5-trimethoxyphenyl)acryloyl]phe-

J¼ 11.6 Hz, 1H, COCH¼CH), 7.81 (d, J¼ 11.6 Hz, 1H, COCH¼CH),

ꢁ

7

.85 (d, J¼ 8.0 Hz, 2H, p-tolyl H-2, H-6), 8.18 (d, J¼ 8.0 Hz, 2H, phe-

nylgacetamide (9i). Yield 54%; yellow powder; mp 116–118 C; IR

ꢀ

1

nyl H-2, H-6), 8.22 (d, J¼ 8.0 Hz, 2H, phenyl H-3, H-5), 8.28 (s, 1H,

pyrimidine H-5), 8.34 (d, J¼ 8.8 Hz, 2H, p-chlorophenyl H-2, H-6),

(cm ): 3417 (NH), 3039 (CH aromatic), 2934 (CH aliphatic), 1658

1

(

3

broad, 2C¼O); H NMR (400 MHz, DMSO-d ) d 3.71 (s, 3H, OCH ),

6

3

1

3

1

d

0.91 (s, 1H, NH, D

2

O exchangeable); C NMR (100 MHz, DMSO-

.72 (s, 3H, OCH

3

), 3.82 (t, J¼ 3.6 Hz, 2H, CH

2

Cl), 3.88 (s, 6H,

) d 21.4 (CH ), 36.2 (CH ), 56.1 (OCH ), 56.8 (OCH ), 108.3 (pyr-

6 3 2 3 3

2

3

OCH ), 4.23 (s, 2H, CH

2

), 4.34 (t, J¼ 3.6 Hz, 2H, OCH

2

,), 7.25 (s, 2H,

imidine C-5), 111.0 (dimethoxyphenyl C-2), 111.9 (dimethoxy-

phenyl C-5), 118.8 (phenyl C-2, C-6), 119.6 (dimethoxyphenyl C-6),

1

C-2, C-6), 128.0 (dimethoxyphenyl C-1), 129.3 (p-chlorophenyl C-3,

C-5), 129.6 (p-tolyl C-3, C-5), 129.9 (phenyl C-3, C-5), 130.3 (p-tolyl

C-4), 133.0 (p-tolyl C-1), 133.3 (p-chlorophenyl C-1), 134.0 (p-chlor-

ophenyl C-4), 135.1 (phenyl C-4), 136.6 (phenyl C-1), 144.7

trimethoxyphenyl H-2, H-6), 7.68–7.72 (m, 3H, p-chloroethoxy-

phenyl H-3, H-5 and COCH¼CH), 7.81 (d, J¼ 8.8 Hz, 2H, p-methox-

yphenyl H-3, H-5), 7.90 (d, J¼ 8.8 Hz, 2H, phenyl H-3, H-5), 7.95 (d,

J¼ 8.2 Hz, p-chloroethoxyphenyl H-2, H-6) , 8.20–8.22 (m, 4H, phe-

nyl H-2, H-6 and p-methoxyphenyl H-2, H-6), 8.29–8.31 (m, 2H,

19.8 (p-tolyl C-2, C-6), 124.3 (COCH¼CH), 127.8 (p-chlorophenyl

COCH¼CH and pyrimidin H-5), 10.71 (s, 1H, NH, D O exchange-

2

1

3

6 2 2

able); C NMR (100 MHz, DMSO-d ) d 43.4 (CH ), 55.4 (CH Cl), 55.8

(

COCH¼CH), 149.4 (dimethoxyphenyl C-4), 151.6 (dimethoxyphenyl

C-3), 163.4 (pyrimidine C-6), 164.9 (pyrimidine C-4), 167.6 (pyrimi-

dine C-2), 171.1 (CONH), 187.9 (CO); Anal. Calcd. for C H ClN O S

(

OCH ), 56.5 (2OCH ), 60.6 (OCH ), 68.6 (OCH ), 106.5 (trimethoxy-

3 3 3 2

phenyl C-2, C-6), 106.9 (pyrimidine C-5), 114.1 (p-methoxyphenyl

C-3, C-5), 114.9 (p-chloroethoxyphenyl C-3, C-5), 118.9 (phenyl C-2,

C-6), 119.8 (COCH¼CH), 121.5 (trimethoxyphenyl C-1), 129.6 (p-

chloroethoxyphenyl C-1), 129.7 (p-chloroethoxyphenyl C-2, C-6),

36 30

3 4

(635.16): C, 67.97; H, 4.75; N, 6.61. Found: C, 67.85; H, 4.58; N, 6.42.

1

30.4 (phenyl C-3, C-5), 130.7 (p-methoxyphenyl C-1), 131.0 (p- (ZE)-2-[4-(4-Chlorophenyl)-6-p-tolylpyrimidin-2-ylthio]-N-f4-[3-

methoxyphenyl C-2, C-6), 131.6 (phenyl C-4), 140.0 (trimethoxy- (3,4,5-trimethoxyphenyl)acryloyl]phenylgacetamide (9l). Yield

ꢁ

ꢀ1

phenyl C-4), 143.7 (phenyl C-1), 144.4 (COCH¼CH), 153.5 (trime- 62%; yellow powder; mp 234–236 C; IR (cm ): 3280 (NH), 3088

1

thoxyphenyl C-3, C-5), 160.6 (p-methoxyphenyl C-4), 161.7 (p- (CH aromatic), 2927 (CH aliphatic), 1663 (broad, 2C¼O); H NMR

chloroethoxyphenyl C-4), 164.1 (pyrimidine C-6), 164.0 (pyrimidine (400 MHz, DMSO-d ) d 2.36 (s, 3H, CH ), 3.71 (s, 3H, OCH ), 3.86 (s,

6

3

C-4), 168.6 (pyrimidine C-2), 172.3 (CONH), 187.9 (CO); Anal. Calcd. 6H, 2OCH ), 4.25 (s, 2H, CH ), 7.23 (s, 2H, trimethoxyphenyl H-2, H-

3

2

for C39

H

36ClN

3

O

7

S (725.20): C, 64.50; H, 5.00; N, 5.79. Found: C, 6), 7.28 (d, J¼ 8.0 Hz, 2H, p-tolyl H-3, H-5), 7.52 (d, J¼ 8.4 Hz, 2H,

p-chlorophenyl H-3, H-5), 7.69 (d, J¼ 12.8 Hz, 1H, COCH¼CH), 7.87

d, J¼ 8.0 Hz, 2H, p-tolyl H-2, H-6), 7.93 (d, J¼ 12.8 Hz, 1H,

COCH¼CH), 8.19–8.24 (m, 4H, phenyl H-2, H-3, H-5, H-6), 8.31 (s,

6

4.38; H, 4.98; N, 5.53.

(

ZE)-2-[4-(4-Chlorophenyl)-6-p-tolylpyrimidin-2-ylthio]-N-f4-[3-(4-

1

H, pyrimidine H-5), 8.35 (d, J¼ 8.4 Hz, 2H, p-chlorophenyl H-2, H-

methoxyphenyl)acryloyl]phenylgacetamide (9j). Yield 59%; yellow

1

3

ꢁ

ꢀ1

6), 10.86 (s, 1H, NH, D O exchangeable); C NMR (100 MHz,

powder; mp 242–244 C; IR (cm ): 3256 (NH), 3038 (CH aromatic),

2

1

DMSO-d ) d 21.4 (CH ), 36.4 (CH ), 56.0 (2OCH ), 60.6 (OCH ),

2

917 (CH aliphatic), 1663 (broad, 2C¼O); H NMR (400 MHz,

6

3

2

3

1

06.8 (trimethoxyphenyl C-2, C-6), 108.3 (pyrimidine C-5), 118.8

DMSO-d ) d 2.36 (s, 3H, CH ), 3.82 (s, 3H, OCH ), 4.24 (s, 2H, CH ),

6

3

2

(

phenyl C-2, C-6), 121.5 (COCH¼CH), 127.9 (p-tolyl C-2, C-6), 129.3

7

2

5

.02 (d, J¼ 8.8 Hz, 2H, methoxyphenyl H-3, H-5), 7.27 (d, J¼ 8.0 Hz,

trimethoxyphenyl C-1), 129.9 (p-chlorophenyl C-2, C-6), 130.4 (p-

H, p-tolyl H-3, H-5), 7.52 (d, J¼ 8.8 Hz, 2H, p-chlorophenyl H-3, H-

), 7.69 (d, J¼ 11.6 Hz, 1H, COCH¼CH), 7.82 (d, J¼ 11.6 Hz, 1H,

chlorophenyl C-3, C-5), 130.4 (p-tolyl C-3, C-5), 130.7 (phenyl C-3,

C-5), 132.8 (p-tolyl C-4), 133.3 (p-tolyl C-1), 135.1 (phenyl C-4),

COCH¼CH), 7.83–7.85 (m, 4H, p-tolyl H-2, H-6, p-methoxyphenyl

H-2, H-6), 8.16 (d, J¼ 8.8 Hz, 2H, phenyl H-2, H-6), 8.22 (d,

J¼ 8.8 Hz, 2H, phenyl H-3, H-5), 8.30 (s, 1H, pyrimidine H-5), 8.34

1

36.6 (p-chlorophenyl C-1), 140.0 (p-chlorophenyl C-4), 142.0 (tri-

methoxyphenyl C-4), 144.0 (phenyl C-1), 144.4 (COCH¼CH), 153.5

trimethoxyphenyl C-3, C-5), 163.4 (pyrimidine C-6), 164.9 (pyrimi-

dine C-4), 167.6 (pyrimidine C-2), 171.1 (CONH), 187.8 (CO); Anal.

Calcd. for C37 32ClN S (665.18): C, 66.71; H, 4.84; N, 6.31. Found:

C, 66.57; H, 4.76; N, 6.37.

(

d, J¼ 8.8 Hz, 2H, p-chlorophenyl H-2, H-6), 10.84 (s, 1H, NH, D

2

O

1

3

exchangeable); C NMR (100 MHz, DMSO-d

6 3

) d 21.4 (CH ), 36.4

H

3 5

O

(

CH ), 55.8 (OCH ), 108.3 (pyrimidine C-5), 114.8 (p-methoxyphenyl

2

3

C-3, C-5), 118.8 (phenyl C-2, C-6), 119.8 (COCH¼CH), 127.8 (p-tolyl

C-2, C-6), 129.3 (p-chlorophenyl C-2, C-6), 129.6 (p-chlorophenyl C-

3

1

, C-5), 129.9 (p-tolyl C-3, C-5), 130.3 (p-methoxyphenyl C-2, C-6), (ZE)-2-[4-(4-Chlorophenyl)-6-(4-nitrophenyl)pyrimidin-2-ylthio]-N-

31.1 (phenyl C-3, C-5), 133.0 (p-methoxyphenyl C-1), 133.3 (p-tolyl f4-[3-(4-methoxyphenyl)acryloyl]phenylgacetamide (9m). Yield

ꢁ

ꢀ1

C-4), 135.1 (p-tolyl C-1), 136.6 (phenyl C-4), 142.0 (p-chlorophenyl 55%; yellow powder; mp 190–192 C; IR (cm ): 3403 (NH), 3066

1

C-1), 143.8 (phenyl C-1), 143.8 (p-chlorophenyl C-4), 148.0 (CH aromatic), 2927 (CH aliphatic), 1658 (broad, 2C¼O); H NMR

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY

7

6 3 2

(400 MHz, DMSO-d ) d 3.83 (s, 3H, OCH ), 4.29 (s, 2H, CH ), 7.03 (d, (p-nitrophenyl C-3, C-5), 129.3 (p-nitrophenyl C-2, C-6), 129.4

J¼ 8.8 Hz, 2H, p-methoxyphenyl H-3, H-5), 7.55 (d, J¼ 8.8 Hz, 2H, (p-chlorophenyl C-2, C-6), 129.8 (p-chlorophenyl C-3, C-5), 130.5

p-chlorophenyl H-3, H-5), 7.72 (d, J¼ 11.6 Hz, 1H, COCH¼CH), (trimethoxyphenyl C-1), 130.8 (phenyl C-3, C-5), 132.9 (p-chloro-

7

.77–7.93 (m, 5H, COCH¼CH, p-methoxyphenyl H-2, H-6, phenyl phenyl C-1), 134.8 (p-chlorophenyl C-4), 137.0 (phenyl C-4), 140.0

H-2, H-6), 8.17 (d, J¼ 8.8 Hz, 2H, phenyl H-3, H-5), 8.28 (d, (trimethoxyphenyl C-4), 142.1 (phenyl C-1), 143.9 (p-nitrophenyl C-

J¼ 8.8 Hz, 2H, p-nitrophenyl H-2, H-6), 8.40 (d, J¼ 8.8 Hz, 2H, p- 1), 144.4 (COCH¼CH), 149.5 (p-nitrophenyl C-4), 153.5 (trimethoxy-

chlorophenyl H-2, H-6), 8.50 (s, 1H, pyrimidine H-5), 8.58 (d, phenyl C-3, C-5), 162.8 (pyrimidine C-6), 164.2 (pyrimidine C-4),

J¼ 8.8 Hz, 2H, p-nitrophenyl H-3, H-5), 10.89 (s, 1H, NH, D O 167.5 (pyrimidine C-2), 171.6 (CONH), 187.8 (CO); EIMS (m/z):

2

1

3

þ

exchangeable); C NMR (100 MHz, DMSO-d ) d 38.8 (CH ), 55.8 696.20 (M , 0.11%), 86.15 (100%); Anal. Calcd. for C H ClN O S

6

2

36 29

4 7

(

1

3

OCH

3

), 109.7 (pyrimidine C-5), 114.8 (p-methoxyphenyl C-3, C-5), (696.14): C, 62.02; H, 4.19; N, 8.04. Found: C, 61.98; H, 4.35; N, 7.98.

18.8 (phenyl C-2, C-6), 120.1 (COCH¼CH), 123.2 (p-nitrophenyl C-

, C-5), 124.3 (p-nitrophenyl C-2, C-6), 125.8 (p-chlorophenyl C-2,

(

ZE)-2-f4-[4-(2-Chloroethoxyphenyl)phenyl]-6-(4-chlorophenyl)-

C-6), 127.9 (p-methoxyphenyl C-1), 129.4 (p-chlorophenyl C-3, C-5),

29.4 (p-methoxyphenyl C-2, C-6), 133.5 (phenyl C-3, C-5), 133.8

p-chlorophenyl C-1), 134.4 (p-chlorophenyl C-4), 134.5 (phenyl C-

pyrimidin-2-ylthiog-N-f4-[3-(4-methoxyphenyl)acryloyl]phe-

1

(

4

1

ꢁ

nylgacetamide (9p). Yield 52%; yellow powder; mp 136–138 C; IR

ꢀ

1

(

cm ): 3181 (NH), 3043 (CH aromatic), 2927 (CH aliphatic), 1663

), 141.9 (p-nitrophenyl C-1), 144.3 (phenyl C-1), 145.4 (COCH¼CH),

1

broad, 2C¼O); H NMR (400 MHz, DMSO-d ) d 3.83 (s, 3H, OCH

Cl), 4.25 (s, 2H, CH

), 4.32 (t, J¼ 6.9 Hz,

H, OCH ), 6.90–7.14 (m, 4H, methoxyphenyl H-3, H-5, p-chloroe-

6

3

),

49.2 (p-nitrophenyl C-4), 161.8 (p-methoxyphenyl C-4), 162.4 (pyr-

3

2

.97 (t, J¼ 6.9 Hz, 2H, CH

2

imidine C-6), 164.6 (pyrimidine C-4), 167.8 (pyrimidine C-2), 171.5

2

(CONH), 187.5 (CO); Anal. Calcd. for C H ClN O S (636.12): C,

34 25 4 5

thoxyphenyl H-3, H-5), 7.43–7.69 (m, 10H, COCH¼CH, COCH¼CH,

phenyl H-2, H-6, p-methoxyphenyl H-2, H-6, p-chloroethoxyphenyl

H-2, H-6, p-chlorophenyl H-3, H-5), 8.12–8.35 (m, 5H, phenyl H-3,

6

4.10; H, 3.96; N, 8.79. Found: C, 64.24; H, 4.06; N, 8.47.

(

ZE)-2-[4-(4-Chlorophenyl)-6-(4-nitrophenyl)pyrimidin-2-ylthio]-N-

f4-[3-(3,4-dimethoxyphenyl)acryloyl]phenylgacetamide (9n).

Yield 53%; yellow powder; mp 147–149 C; IR (cm ): 3256 (NH),

H-5, pyrimidine H-5, p-chlorophenyl H-2, H-6), 10.88 (s, 1H, NH,

13

D

2

O exchangeable); C NMR (100 MHz, DMSO-d

6

) d 36.2 (CH

2

),

ꢁ

ꢀ1

4

1

3

3.4 (CH Cl), 54.6 (OCH ), 55.8 (OCH ), 107.8 (pyrimidine C-5),

2 3 2

1

3

079 (CH aromatic), 2922 (CH aliphatic), 1660 (broad, 2C¼O);

H

14.8 (p-methoxyphenyl C-3, C-5), 115.3 (p-chloroethoxyphenyl C-

, C-5), 118.8 (phenyl C-2, C-6), 121.8 (COCH¼CH), 127.4 (p-chlor-

NMR (400 MHz, DMSO-d ) d 3.82 (s, 3H, OCH ), 3.87 (s, 3H, OCH ),

4

7

6

3

.29 (s, 2H, CH ), 7.03 (d, J¼ 8.4 Hz, 1H, dimethoxyphenyl H-5),

2

oethoxyphenyl C-1), 128.5 (p-methoxyphenyl C-1), 128.6 (p-chloro-

phenyl C-2, C-6), 129.4 (p-chlorophenyl C-3, C-5), 129.6 (p-

methoxyphenyl C-2, C-6), 129.6 (p-chloroethoxyphenyl C-2, C-6),

.39 (d, J¼ 8.4 Hz, 1H, dimethoxyphenyl H-6), 7.53–7.55 (m, 3H, p-

chlorophenyl H-3, H-5, dimethoxyphenyl H-2), 7.70 (d, J¼ 15.6 Hz,

1

H, COCH¼CH), 7.82–7.87 (m, 3H, p-chlorophenyl H-2, H-6,

1

30.7 (p-chlorophenyl C-1), 132.7 (phenyl C-3, C-5), 133.9 (p-chlor-

ophenyl C-4), 140.1 (phenyl C-4), 143.0 (phenyl C-1), 144.4

COCH¼CH), 159.4 (p-chloroethoxyphenyl C-4), 159.9 (p-methoxy-

phenyl C-4), 162.8 (pyrimidine C-6), 164.3 (pyrimidine C-4), 167.3

pyrimidine C-2), 171.4 (CONH), 187.8 (CO); Anal. Calcd. for

S (669.13): C, 64.48; H, 4.36; N, 6.27. Found: C, 64.36;

H, 4.19; N, 6.35.

COCH¼CH), 8.19 (d, J¼ 8.8 Hz, 2H phenyl H-2, H-6), 8.27 (d,

J¼ 8.8 Hz, 2H phenyl H-3, H-5), 8.38 (d, J¼ 8.4 Hz, 2H p-nitrophenyl

H-2, H-6), 8.49 (s, 1H, pyrimidine H-5), 8.57 (d, J¼ 8.4 Hz, 2H, p-

(

1

3

nitrophenyl H-3, H-5), 10.88 (s, 1H, NH, D O exchangeable);

C

2

(

NMR (100 MHz, DMSO-d

1

thoxyphenyl C-5), 118.8 (phenyl C-2, C-6), 119.8 (dimethoxyphenyl

C-6), 124.2 (p-nitrophenyl C-3, C-5), 124.3 (COCH¼CH), 128.0 (p-

nitrophenyl C-2, C-6), 129.2 (dimethoxyphenyl C-1), 129.4 (p-chlor-

6 2 3 3

) d 36.5 (CH ), 56.0 (OCH ), 56.1 (OCH ),

36 2 3 4

C H29Cl N O

09.9 (pyrimidine C-5), 111.0 (dimethoxyphenyl C-2), 111.9 (dime-

(

ZE)-2-f4-[4-(2-Chloroethoxyphenyl)phenyl]-6-(4-chlorophenyl)-

ophenyl C-2, C-6), 129.8 (p-chlorophenyl C-3, C-5), 130.3 (phenyl pyrimidin-2-ylthiog-N-f4-[3-(3,4-dimethoxyphenyl)acryloyl]phe-

ꢁ

C-3, C-5), 133.1 (p-chlorophenyl C-1), 134.7 (p-chlorophenyl C-4), nylgacetamide (9q). Yield 49%; yellow powder; mp 210–212 C; IR

ꢀ

1

37.0 (phenyl C-4), 142.1 (p-nitrophenyl C-1), 143.8 (phenyl C-1), (cm ): 3429 (NH), 3061 (CH aromatic), 2927 (CH aliphatic), 1663

44.4 (COCH¼CH), 149.4 (p-nitrophenyl C-4), 151.6 (dimethoxy- (broad, 2C¼O); H NMR (400 MHz, DMSO-d ) d 3.72–3.82 (m, 5H,

1

6

phenyl C-4), 152.3 (dimethoxyphenyl C-3), 162.7 (pyrimidine C-6), OCH and CH Cl), 3.83–3.88 (m, 5H, OCH and OCH ), 4.34 (s, 2H,

3

2

3

2

1

64.1 (pyrimidine C-4), 167.5 (pyrimidine C-2), 171.6 (CONH), 187.8 CH ), 6.98–7.15 (m, 4H, dimethoxyphenyl H-5, H-6, p-chloroethoxy-

2

(

CO); Anal. Calcd. for C35

H

27ClN

4 6

O S (666.13): C, 63.01; H, 4.08; N, phenyl H-3, H-5), 7.31–7.40 (m, 2H, dimethoxy H-2 and

8

.40. Found: C, 62.89; H, 4.15; N, 8.37.

COCH¼CH), 7.52–7.70 (m, 4H, p-chlorophenyl H-3, H-5 and p-chlor-

oethoxyphenyl H-2, H-6), 7.75–7.88 (m, 4H, phenyl H-2, H-6 and p-

chlorophenyl H-2, H-6), 7.95 (d, J¼ 8.8 Hz, 2H, phenyl H-3, H-5),

(

ZE)-2-[4-(4-Chlorophenyl)-6-(4-nitrophenyl)pyrimidin-2-ylthio]-N-

f4-[3-(3,4,5-trimethoxyphenyl)acryloyl]phenylgacetamide (9o).

Yield 47%; yellow powder; mp 268–270 C; IR (cm ): 3371 (NH),

8

.28 (d, J¼ 12.0 Hz, 1H, COCH¼CH), 8.46 (s, 1H, pyrimidine H-5),

13

ꢁ

ꢀ1

2

10.76 (s, 1H, NH, D O exchangeable); C NMR (100 MHz, DMSO-

1

d

6

) d 38.4 (CH

OCH ), 107.3 (pyrimidine C-5), 111.5 (dimethoxyphenyl C-2), 112.7

), 7.24 (s, 2H, trimethoxyphenyl H-2, H-6), 7.55 (d, (dimethoxyphenyl C-5), 114.9 (p-chloroethoxyphenyl C-3, C-5),

2

2 2 3 3

), 42.9 (CH Cl), 56.1 (OCH ), 56.6 (OCH ), 68.6

3

059 (CH aromatic), 2935 (CH aliphatic), 1656 (broad, 2C¼O);

NMR (400 MHz, DMSO-d ) d 3.72 (s, 3H, OCH ), 3.87 (s, 6H, 2OCH

.29 (s, 2H, CH

H

(

2

6

3

),

4

J¼ 8.8 Hz, 2H, p-chlorophenyl H-3, H-5), 7.70 (d, J¼ 15.6 Hz, 1H, 121.1 (phenyl C-2, C-6), 121.3 (COCH¼CH), 122.5 (dimethoxyphenyl

COCH¼CH), 7.87 (d, J¼ 8.8 Hz, 2H, p-chlorophenyl H-2, H-6), 7.92 C-6), 127.3 (dimethoxyphenyl C-1), 127.4 (p-chloroethoxyphenyl C-

(

d, J¼ 15.6 Hz, 1H, COCH¼CH), 8.21 (d, J¼ 8.8 Hz, 2H, phenyl H-2, 1), 128.1 (p-chloroethoxyphenyl C-2, C-6), 128.9 (p-chlorophenyl C-

H-6), 8.28 (d, J¼ 8.8 Hz, 2H, phenyl H-3, H-5), 8.39 (d, J¼ 8.8 Hz, 2, C-6), 129.3 (p-chlorophenyl C-3, C-5), 131.4 (phenyl C-3, C-5),

H, p-nitrophenyl H-2, H-6), 8.51 (s, 1H, pyrimidine H-5), 8.58 (d, 133.9 (p-chlorophenyl C-1), 134.3 (p-chlorophenyl C-4), 135.5 (phe-

J¼ 8.8 Hz, 2H, p-nitrophenyl H-3, H-5), 10.89 (s, 1H, NH, D

nyl C-4), 144.3 (phenyl C-1), 145.1 (COCH¼CH), 149.3 (dimethoxy-

exchangeable); C NMR (100 MHz, DMSO-d ) d 36.5 (CH ), 56.6 phenyl C-4), 149.7 (dimethoxyphenyl C-3), 151.1 (p-

2

O

1

3

6

2

(

2OCH ), 60.6 (OCH ), 106.9 (trimethoxyphenyl C-2, C-6), 110.0 chloroethoxyphenyl C-4), 162.5 (pyrimidine C-6), 164.4 (pyrimidine

3 3

(pyrimidine C-5), 118.8 (phenyl C-2, C-6), 121.5 (COCH¼CH), 124.3 C-4), 168.7 (pyrimidine C-2), 172.7 (CONH), 189.7 (CO); Anal. Calcd.

8

P. F. LAMIE AND J. N. PHILOPPES

for C37

H

31Cl

2 3

N O

5

S (699.14): C, 63.43; H, 4.46; N, 6.00. Found: C, Predicted pharmacokinetic and toxicity properties

6

3.33; H, 4.27; N, 6.11.

Pharmacokinetic properties (absorption, distribution, metabolism,

and excretion) through determination of human intestinal absorp-

tion (HIA), in vitro caco-2 cell permeability, in vitro Madin-Darby

Canine Kidney (MDCK) cell permeability, plasma protein binding

(

ZE)-2-f4-[4-(2-Chloroethoxyphenyl)phenyl]-6-(4-chlorophenyl)-

pyrimidin-2-ylthiog-N-f4-[3-(3,4,5-trimethoxyphenyl)acryloyl]phe-

(

PPB), blood–brain binding (BBB), skin permeability, p-glycoprotein

Pgp), and cytochrome p450 isoforms inhibition data, in addition

ꢁ

nylgacetamide (9r). Yield 53%; yellow powder; mp 221–223 C; IR

ꢀ

1

(cm ): 3421 (NH), 3061 (CH aromatic), 2931 (CH aliphatic), 1665

to toxicity (Ames test, rodent carcinogenicity assay and hERG-

1

(

3

broad, 2C¼O); H NMR (400 MHz, DMSO-d

6

) d 3.70 (s, 3H, OCH

3

),

49

inhibition) were evaluated through preADMET online server .

.85 (s, 6H, 2OCH

.45 (t, J¼ 7.2 Hz, 2H, OCH

3

), 3.96 (t, J¼ 7.2 Hz, 2H, CH Cl), 4.24 (s, 2H, CH

2

4

2

), 7.03–7.20 (m, 4H, trimethoxyphenyl

H-2, H-6, chloroethoxyphenyl H-3, H-5), 7.29 (d, J¼ 12.0 Hz, 1H, Statistical analysis

COCH¼CH), 7.51–7.60 (m, 4H, p-chlorophenyl H-3, H-5, chloroe-

Data obtained were expressed as means ± standard deviations

thoxyphenyl H-2, H-6), 7.87–7.96 (m, 3H, phenyl H-2, H-6,

COCH¼CH), 8.18–8.32 (m, 5H, phenyl H-3, H-5, p-chlorophenyl H-2,

ꢂ

SDs). The results were considered significant when p ˂ 0.05 or

(

ꢂꢂ

p ˂ 0.005 using Student’s t-test was compared to reference

1

3

H-6, pyrimidine H-5), 10.91 (s, 1H, NH, D

2

O exchangeable);

Cl), 55.5 (2OCH

), 106.7 (trimethoxyphenyl C-2, C-6), 109.4

pyrimidine C-5), 114.9 (p-chloroethoxyphenyl C-3, C-5), 118.8

C

3

),

drugs. The obtained values were representative of triplicate inde-

pendent experiments.

NMR (100 MHz, DMSO-d

7.8 (OCH ), 60.8 (OCH

3

(

6

) d 36.8 (CH

2

), 43.9 (CH

2

5

2

phenyl C-2, C-6), 121.5 (COCH¼CH), 126.4 (trimethoxyphenyl C-1), Results and discussion

1

2

27.4 (p-chloroethoxyphenyl C-1), 128.1 (p-chloroethoxyphenyl C-

, C-6), 129.5 (p-chlorophenyl C-2, C-6), 129.8 (p-chlorophenyl C-3,

Chemistry

C-5), 132.7 (phenyl C-3, C-5), 135.4 (p-chlorophenyl C-1), 136.8 (p- The target 2-TP/chalcone hybrids 9a–r were prepared from two

synthesised starting materials 4a–f and 8a–c, as depicted in

Schemes 1–3.

Heating under reflux condition chalcone derivatives 3a–f (syn-

thesised from condensation of p-methoxy/chlorobenzaldehyde

chlorophenyl C-4), 140.3 (phenyl C-4), 141.7 (trimethoxyphenyl C-

), 143.0 (phenyl C-1), 144.4 (COCH¼CH), 153.5 (trimethoxyphenyl

C-3, C-5), 159.4 (p-chloroethoxyphenyl C-4), 162.2 (pyrimidine C-6),

64.1 (pyrimidine C-4), 167.7 (pyrimidine C-2), 171.7 (CONH), 187.7

CO); EIMS (m/z): 728.10 (M-1, 0.28%), 58.10 (100%); Anal. Calcd.

for C38 S (729.15): C, 62.47; H, 4.55; N, 5.75. Found: C,

2.53; H, 4.76; N, 5.58.

4

1

(

1a&b with p-methyl/nitro/or ethoxychloroacetophenone 2a–c)

and thiourea in presence of KOH afforded 2-TP derivatives 4a–f.

H

2 3 6

33Cl N O

3

9,50

The method was reported for compounds 4b and 4d

6

(Scheme 1).

The other starting materials, chloroacetyl chalcone derivatives

8

a–c were obtained by stirring p-aminochalcone derivatives 7a–c

Biological evaluations

with chloroacetyl chloride, K CO in chloroform at room tempera-

ture (Scheme 2).

S-Alkylation of 2-TPs 4a–f with acetylated chalcones 8a–c was

achieved in acetonitrile using TEA as a base catalysis to obtain the

target compounds 9a–r in 47–82% yield.

2

3

2

4

Cytotoxic assay

To investigate cytotoxic activity of the final target compounds

9a–r, MTT assay was performed. Three different cell lines were

used, leukaemia (K-562), breast (MCF-7) and colon (HT-29) cell

lines. Cisplatin and erlotinib were the reference drugs used in this

study. Half maximal concentration at which 50% of cells were

viable was calculated as IC in lM, according to cytotoxic assay

1

13

H NMR and C NMR spectroscopic tools were used to confirm

1

formation of the target derivatives 9a–r. Thus, H NMR spectra of

compounds 9a–r displayed a singlet signal at d 3.82–4.84 ppm

attributed to (SCH

50

2

CO) protons. Additionally, protons of chalcone

4

7

reported protocol .

fragment appeared as two doublet signals at d 7.29–7.72 ppm and

7

.69–8.31 ppm with coupling constant J¼ 11.6–15.6 Hz.

Furthermore, amide NH proton appeared as a singlet signal at d

10.71–10.91 ppm.

STAT3/STAT5a assays

1

3

Both K-562 and MCF-7 cell lines were seeded overnight in plates,

then 10 mM of test compounds (9a and 9r for MCF-7 cells; 9d, 9f, peak at

and 9n for K-562 cells) or reference drug pacritinib was added for Moreover, two carbonyl carbons at d 170.71–173.07 ppm and

C NMR spectra of compounds 9a–r showed appearance of a

36.21–43.44 ppm characterised to SCH carbon.

d

2

1

87.58–190.01 ppm related to (NHCO) and (C¼O), respectively,

2

4 h. A nuclear extract kit was used to extract nuclear fractions

were also appeared (Scheme 3).

from treated cells using the manufacture’s procedure. STAT3 and

STAT5a activations were analysed using the collected nuclear

extracts (20 mg) through TransAM STAT3 and STAT5a activation

assay guided by the manufacture’s protocol. The obtained results

Biological activity

were expressed in the form of mean ± SD. Each experiment was Cytotoxic activity

done in triplicate.

All target compounds 9a–r were screened against three different

cancer cell lines, leukaemia (K-562), breast (MCF-7), and colon (HT-

9). MTT assay was used. Both cisplatin and erlotinib were used as

2

the reference drugs. Cytotoxicity results are recorded in Table 1.

Regarding cytotoxic activity of the test compounds against leu-

Biological properties

The target compounds 9a–r were drawn using ChemDraw Ultra kaemia (K-562) cell line, compounds 9d, 9f, 9n, and 9p were the

1

0.0. Biological properties and drug likeness were predicted using most potent compounds with IC₅₀ranged from 0.77 to 1.74 lM if

48

online computational tool Molinspiration .

compared to cisplatin, the reference drug (IC50¼2.31 lM). Their

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY

9

Scheme 1. Synthesis of 2-thiopyrimidine derivatives 4a–f.

Scheme 2. Synthesis of chloroacetyl aminochalcone derivatives 8a–c.

common feature was presence of one or more para substituted

Compounds 9b and 9k showed weak inhibitory activity with

phenyl ring(s) with electron withdrawing group (NO , Cl) at pyr- IC₅₀values equal to 22.45 and 28.65 lM, respectively. Both of

2

imidine core.

them have dimethoxyphenyl ring on chalcone part, and p-tolyl

Compounds 9b, 9e, 9g–j, 9m, and 9q exhibited potent inhibi- ring at pyrimidine core.

tory activity with IC50 values ranged from 3.17 to 9.71 lM, if com-

By inspecting cytotoxicity results of HT-29 cell line, compounds

pared to the second reference erlotinib (IC : 9.85 lM). P- 9a, 9l, and 9n showed IC₅₀(2.10–2.37 lM) near in potency to the

50

Methoxyacyloyl derivative 9k, with IC50 value ¼ 9.95 lM, was reference drug cisplatin (IC ¼1.12 lM). Compounds 9c, 9e, 9j, 9k,

5

0

nearly equal in potency to erlotinib. Compounds 9a, 9c, 9o, and 9m, 9o, 9q, and 9r showed significant activity with IC values

5

0

9

r showed moderate inhibitory activity (IC ¼10.67–18.40 lM). The between 3.62 and 8.70 lM, compared to erlotinib (IC ¼9.20 lM).

50

lowest inhibitory activity was observed in compound 9l While, rest of the compounds exhibited weak inhibitory activity

(

IC50¼42.60 lM), bearing p-chlorophenyl and p-tolyl rings at pyr- (IC ¼9.41–25.92 lM). Results showed that no effect was observed

5

0

imidine scaffold, beside, trimethoxyphenyl chalcone hybrid.

regarding substituents on the two hybrid structures pyrimidine

Concerning MCF-7 cell line, the most active derivative was 9r and chalcone.

(

IC ¼1.37 lM) compared to the reference drug cisplatin

Finally, dual cytotoxic activity was observed for compound 9f

IC50¼6.62 lM). It is characterised by presence of p-chlorophenyl (against K-562 and MCF-7 cell lines), and for compound 9a (on MCF-

5

0

ring and p-chloroethoxyphenyl ring at pyrimidine core together 7 and HT-29 cell lines) and 9n (against K-562 and HT-29 cell lines).

with trimethoxy chalcone part.

Other compounds exerted excellent activity were 9a, 9c, 9f, 9j,

9

m, and 9o (IC ¼3.56–6.26 lM). Additionally, compound 9q was Cytotoxicity against normal cell line (WI38)

5

0

nearly equipotent to the reference drug cisplatin (IC50¼6.81 lM). To know cytotoxicity of the most active compounds, they were

Derivatives 9g and 9h with IC50¼10.40 and 7.70 lM, respectively, tested against normal human fibroblast cell line (WI38) and IC50

were more potent than erlotinib (IC ¼10.64 lM).

values are represented as in Figure 2. Cisplatin was used as a ref-

50

Moderate activity was observed in compounds 9d, 9e, 9i, 9l, erence drug. All the test compounds showed higher IC50 values

n, and 9p (IC50¼11.47–18.74 lM). (29.19–40.13 mM) than the reference drug (18.86 mM) except

9

1

0

P. F. LAMIE AND J. N. PHILOPPES

Scheme 3. Synthesis of the target compounds 9a–r.

compound 9a which exerted cytotoxic activity (IC ¼17.09 mM) STAT3 and STAT5a was observed mainly in compound 9n. For this

50

slightly less than cisplatin.

compound, both phenyl rings on pyrimidine core were para-sub-

stituted with electron withdrawing groups (NO₂and Cl), beside

presence of disubstituted methoxyphenyl ring at chalcone hybrid.

STAT3 and STAT5a inhibitory activity determination

The most active compounds in cytotoxic assay against leukaemia

cell line K-562 and human breast adenocarcinoma cells MCF-7

Biological properties

were further tested as inhibitors for STAT3 and STAT5a enzymes. Molinspiration was used to predict bioactivity scores for all the

49

Pacritinib, an inhibitor for both STAT3 and STAT5a was used in target compounds 9a–r. The obtained results are recorded in

this study as a reference drug.

Table 3. It was found that most of the prepared compounds had

The results are listed in Table 2. They indicated that the test bioactivity values in the range ꢀ0.5 to 0.00. This revealed that the

compounds showed inhibitory activity against both STAT3 and designed pyrimidine/chalcone derivatives might be involved in

STAT5a. Compounds 9d, 9n, and 9r were the most active against moderate interactions with G-protein-coupled receptors (GPCRs)

STAT3. Additionally, compound 9n was the most effective as and protease inhibitors. However, the bioactivity prediction was

STAT5a inhibitor. Compounds 9a, 9f, and 9r had also strong not in the standard range against other receptors such as ion

inhibitory activity against STAT5a. Dual inhibitory activity against channel modulator, kinases and nuclear receptor ligand.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY

11

Table 1. Cytotoxicity results of pyrimidine/chalcone hybrids 9a–r against three different cancer cell lines.

R¹

N

R²

H

N

OCH₃

R³

N

S

O

R

O

9

a-r

(

IC50 lM)±SD

1

2

3

Compound

R

K-562

MCF-7

HT-29

9

a

b

c

d

e

f

g

h

i

j

k

l

m

n

o

p

q

r

OCH

Cl

3

CH

NO

OCH

3

H

CH

OCH

H

OCH

H

OCH

H

OCH

H

OCH

H

OCH

H

OCH

18.40 ± 0.76

3.62 ± 0.08

11.42 ± 0.04

0.77 ± 0.03

7.05 ± 0.28

1.37 ± 0.03

3.17 ± 0.11

7.07 ± 0.31

9.71 ± 0.41

4.26 ± 0.06

9.95 ± 0.29

42.60 ± 1.99

3.86 ± 0.14

1.05 ± 0.02

12.35 ± 0.72

1.74 ± 0.04

5.77 ± 0.16

10.67 ± 0.77

2.31 ± 0.09

9.85 ± 0.51

3.56 ± 0.14

2.20 ± 0.04

16.36 ± 0.76

8.70 ± 0.28

25.92 ± 1.67

7.31 ± 0.36

9.41 ± 0.43

10.55 ± 0.47

18.77 ± 0.88

11.47 ± 0.23

6.26 ± 0.34

5.61 ± 0.18

2.37 ± 0.07

7.90 ± 0.22

2.10 ± 0.06

3.62 ± 0.08

11.64 ± 0.63

6.06 ± 0.29

4.74 ± 0.27

1.12 ± 0.06

9.20 ± 0.41

3

22.45 ± 1.57

4.25 ± 0.13

14.16 ± 0.74

18.74 ± 0.92

5.77 ± 0.22

10.40 ± 0.64

7.70 ± 0.32

11.47 ± 0.81

6.26 ± 0.24

28.65 ± 1.39

13.46 ± 0.62

3.90 ± 0.09

17.36 ± 0.75

3.62 ± 0.084

11.64 ± 0.49

6.81 ± 0.25

1.37 ± 0.07

6.62 ± 0.29

10.64 ± 0.58

3

2

OCH

H

OCH

H

OCH

H

OCH

H

3

2

CH

2

Cl

3

CH

3

CH

3

CH

3

NO

OCH

2

3

2

CH

2

CH

2

CH

2

Cl

OCH

3

Cisplatin

Erlotinib

Figure 2. Cytotoxicity (IC50) of the most active derivatives and cisplatin against WI38 cell line.

Drug likeness is a complex balance between various molecular cardiac toxicity was estimated using Pharmacokinetics/PreADMET

5

2

properties like, molecule size, hydrogen bonding characters, elec- Toxicity Predictor (Table 4).

51

tronic distribution and hydrophobicity . The results in Table 3

Absorption refers to the process by which the drug can go to

showed that all the final target compounds had positive predict- the systemic circulation through the organs of the body. Several

able score values which stranded for good drug likeness behav- routes for absorption such as oral absorption (human intestinal

iour, especially compound 9r as represented in Figure 3.

absorption, HIA), skin permeability (SP, logKp), and permeability

through certain cells such as Caco2 (derived from human colon

adenocarcinoma cells) and MDCK cells were measured.

Predicted pharmacokinetic and toxicity properties

By inspecting results recorded in Table 4, it was found that

Prediction of the major pharmacokinetic parameters such as compounds 9a–r showed good intestinal absorption all above

absorption, distribution, metabolism, and excretion, in addition to 97.25% (permissible limit: 70–100%abs). Skin permeability was

toxicological properties, such as mutagenicity, carcinogenicity, and found to be slightly less than acceptable range (ꢀ2.5 logKp).

1

2

P. F. LAMIE AND J. N. PHILOPPES

Moreover, moderate permeability through in vitro Caco2 cells

ranged from 54.34 to 28.65 nm/sc were observed. While, low val- was obtained by measuring AMES test (to predict mutagenicity of

ues were detected for in vitro MDCK cells.

the compounds), carcino-Mouse/Rate (to test carcinogenicity

Prediction of toxicological behaviour of the test compounds

The second property is the distribution, through which the of the compounds), and hERG-inhibition (to check cardiac toxicity

transformation of the molecules from one tissue or organ to of the target synthesised molecules) (Table 5). Half of test com-

another can be predicted. Blood–brain barrier (BBB) and PPB were pounds showed non-mutagenic behaviour in AMES test. All com-

two distribution parameters used in this study. BBB permits the pounds had negative carcinogenic effect in mouse and rats, in

diffusion of hydrophobic and small molecules to the brain. It is an addition to medium risk as cardiotoxic agents. From the predicted

important predictor for central nervous system (CNS) drug discov- ADMET properties of the novel synthesised compounds, it was jus-

ery. Moreover, the measured of percentage of a molecule bound

to plasma protein (%PPB) was also helpful in prediction of distri-

bution for the novel target compounds.

tified that they may have good characters as lead compounds.

Results showed that all the test compounds displayed strong Structure–activity relationship of target compounds

PPB value (90.94–99.63%) indicating prolonged half-lives and lim-

ited brain penetration. Consequently, BBB (unbound brain-to-

plasma ratio) was low in most compounds except in nitrophenyl

chalcone. There was a relationship between presence of small

containing derivatives 9d–9f and 9m–9o (0.37–0.57) which was

medium and around the acceptable range to be CNS active com-

pounds (>0.4).

Structure–activity relationship (SAR) study for the target com-

pounds 9a–r focussed on two important scaffolds, pyrimidine and

electron donating group such as –CH

3

, (D

s

), large electron donat-

ing group such as –OCH CH Cl, (D ) or electron withdrawing

2 2 l

group, –NO , (W) in para position of phenyl ring at pyrimidine C-4

2

Metabolism, the biotransformation or chemical modification of

exogenous compounds to increase their water solubility by

increasing their hydrophobicity facilitating their excretion can be

predicted either in phase I or phase II. Cytochrome P450 isoforms,

calculate the ability of the test compounds to be inhibitor to drug

metabolising enzymes such as CYP2C19, CYP2C9, CYP2D6,

CYP3A4, and CYP1A2. Moreover, glycoprotein (P-gp) inhibition

measured to predict excretion property of the target compounds.

The test compounds showed good inhibitory behaviour for

CYP2C9 and CYP3A4 and did not show inhibition behaviour for

CYP2C19 and CYP3A4. All compounds had inhibitory effect on

P-gp.

and electron donating group, –OCH , (D) or electron withdrawing

3

group, –Cl, (W) in para position of phenyl ring at pyrimidine C-6

with that of (mono-, di-, or tri-)methoxyphenyl ring of chalcone

part, and between cytotoxic activities on the three different tested

cell lines K-562, MCF-7, and HT-29, as represented in Figure 4.

In compounds 9a–c, pyrimidine core carried p-tolyl group and

p-methoxyphenyl group at C-4 and C-6, respectively. Cytotoxic

activity against K-562 cell line was maximised in compound 9b

bearing dimethoxyphenyl chalcone moiety then decreased in 9c

and 9a (trimethoxyphenyl and methoxyphenyl chalcones,

respectively).

The order of reactivity was altered when evaluated against

MCF-7 or HT-29 cell lines, where 9a>9c>9b.

For compounds 9d–f, replacement of p-tolyl group at pyrimi-

dine C-4 with p-nitrophenyl group and keeping p-methoxyphenyl

group at pyrimidine C-6 constant, led to variation in cytotoxic

activity. Thus, compound 9d (with p-methoxyphenyl chalcone

part) was the most potent against K-562 cell line, than 9f (trime-

thoxyphenyl chalcone analogue) and finally 9e (dimethoxyphenyl

chalcone analogue). While against MCF-7, the order was

Table 2. STAT3 and STAT5a inhibitory activity of compounds 9a, 9d, 9f, 9n, 9r

and reference drug pacritinib.

Inhibition IC50 (mM)

Compound/no.

STAT3

STAT5a

9

a/MCF7

d/K562

f/K562

n/K562

r/MCF7

242.53 ± 9.24

160.01 ± 4.59

244.74 ± 11.07

113.31 ± 3.22

148.69 ± 3.81

79.47 ± 2.17

65.49 ± 2.55

83.78 ± 3.28

116.31 ± 4.13

77.65 ± 2.91

9f>9d>9e. For HT-29, 9e was the most potent than 9f and at

50.75 ± 1.26 last 9d.

63.24 ± 1.57

54.35 ± 1.09

69.81 ± 1.82

Regarding compounds 9g–i, they characterised by bearing

electron donating groups at para position of two phenyl rings at

pyrimidine C-4 and C-6; however, presence of large sized electron

Pacritinib/MCF7

Pacritinib/K562

Table 3. Biological properties, prediction, and drug likeness of the target compounds.

Compound

GPCR ligand

Ion channel modulator

Kinase inhibitor

Nuclear receptor ligand

Protease inhibitor

Drug likeness score

9

a

b

c

d

e

f

g

h

i

j

k

l

m

n

o

p

q

r

ꢀ0.43

ꢀ0.56

ꢀ0.73

ꢀ0.61

ꢀ0.78

ꢀ0.99

ꢀ0.69

ꢀ0.88

ꢀ1.11

ꢀ0.39

ꢀ0.50

ꢀ0.64

ꢀ0.55

ꢀ0.69

ꢀ0.88

ꢀ0.61

ꢀ0.79

ꢀ1.00

ꢀ1.03

ꢀ1.27

ꢀ1.55

ꢀ1.25

ꢀ1.53

ꢀ1.86

ꢀ1.44

ꢀ1.75

ꢀ2.09

ꢀ0.94

ꢀ1.16

ꢀ1.41

ꢀ1.13

ꢀ1.39

ꢀ1.69

ꢀ1.31

ꢀ1.60

ꢀ1.92

ꢀ0.58

ꢀ0.78

ꢀ1.01

ꢀ0.83

ꢀ1.07

ꢀ1.35

ꢀ0.81

ꢀ1.08

ꢀ1.39

ꢀ0.52

ꢀ0.68

ꢀ0.89

ꢀ0.75

ꢀ0.95

ꢀ1.21

ꢀ0.71

ꢀ0.96

ꢀ1.24

ꢀ0.69

ꢀ0.90

ꢀ1.17

ꢀ0.92

ꢀ1.17

ꢀ1.50

ꢀ1.01

ꢀ1.29

ꢀ1.64

ꢀ0.63

ꢀ0.81

ꢀ1.06

ꢀ0.83

ꢀ1.06

ꢀ1.36

ꢀ0.91

ꢀ1.17

ꢀ1.49

ꢀ0.43

ꢀ0.51

ꢀ0.62

ꢀ0.53

ꢀ0.65

ꢀ0.80

ꢀ0.56

ꢀ0.70

ꢀ0.87

ꢀ0.44

ꢀ0.50

ꢀ0.58

ꢀ0.52

ꢀ0.62

ꢀ0.74

ꢀ0.54

ꢀ0.66

ꢀ0.81

0.49

0.54

0.96

0.07

0.11

0.49

0.83

0.79

1.18

0.82

0.80

1.07

0.43

0.40

0.63

1.14

1.05

1.40

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY

13

2 2

donating group such as –OCH CH Cl at phenyl ring of pyrimidine against MCF-7. For HT-29, 9n was the most potent than 9o and

C-4 led to increase its lipophilic character.

finally 9m.

In compounds 9p–r, incorporation of p-chloroethoxyphenyl group

The order of cytotoxic activity against K-562 was found to be

9

(

9

g (methoxyphenyl chalcone)>9h (dimethoxyphenyl chalcone)>9i at pyrimidine C-4, while keeping p-chlorophenyl group at pyrimidine

trimethoxyphenyl chalcone), and for MCF-7 cell line was, C-6 constant, resulted in changing order of cytotoxic activity against

h>9g>9i. While converted to be 9i>9g>9h in case of HT-29 K-562 to be 9p (methoxyphenyl chalcone)>9q (dimethoxyphenyl

cell line.

chalcone), and still the least potent compound was 9r (trimethoxy-

In 9j–l derivatives, electron withdrawing group (–Cl) was intro- phenyl chalcone). However, for MCF-7 and HT-29, it was observed

duced to para position of phenyl ring at pyrimidine C-6, while pyr- that 9r was the most potent than 9q and finally 9p.

imidine C-4 carried small sized electron donating group (–CH ) at

3

para position of its phenyl ring.

Conclusions

The most active compound was 9j (methoxyphenyl chalcone)

in both K-562 and MCF-7 cell lines, while 9l (trimethoxyphenyl

chalcone) was the most potent against HT-29 cell line.

Compounds 9m–o, p-tolyl ring were replaced with p-nitro-

phenyl ring at pyrimidine C-4, while p-chlorophenyl ring at pyrimi-

dine C-6 was kept constant.

The most potent derivatives against K-562 were dimethoxy-

phenyl chalcone 9n, than methoxyphenyl chalcone derivative 9m.

While, trimethoxyphenyl chalcone derivative 9o and methoxy-

phenyl chalcone analogue 9m showed nearly equal potency

A novel series of 2-TP/chalcone hybrids 9a–r was designed to be

as anticancer agents. They were synthesised and identified using

different spectroscopic techniques. Their cytotoxic activities

against three different cancerous cell lines, K-562, MCF-7, and HT-

2

9 were evaluated. The synthesised compounds showed strong to

moderate cytotoxic activities especially against K-562 and MCF-7

cell lines. The highest cytotoxic activity against K-562 cell line was

observed in compounds 9d, 9f, 9n, and 9p with IC₅₀values in the

range of 0.77–1.74 mM, compared to the reference drug, cisplatin

(

IC ¼2.31 mM). For cytotoxic activity against MCF-7 cell line, com-

5

0

pounds 9a, 9c, 9f, 9j, 9m, 9o, and 9r exhibited the highest

Table 5. Toxicity assessment of the target synthesised compounds 9a–r.

Compound

AMES

Carcino-Mouse

Carcino-Rat

hERG-inhibition

9

a

b

c

d

e

f

g

h

i

j

k

l

m

n

o

p

q

r

Mutagen

Negative

Medium

Non-mutagen

Mutagen

Non-mutagen

Mutagen

Non-mutagen

Mutagen

Non-mutagen

Figure 3. Drug likeness score value (1.40) for compound 9r.

Table 4. Pharmacokinetic properties assessment of the target synthesised compounds 9a–r.

Absorption

Distribution

Metabolism (CYP) and excretion

Compound

HIA (%)

SP LogP (cm/h)

Caco2 (nm/sc)

MDCK (nm/sc)

BBB (c.brain/c.blood)

PPB %

2C19

2C9

2D6

3A4

Pgp Inh.

9

a

b

c

d

e

f

g

h

i

j

k

l

m

n

o

p

q

r

97.50

97.36

97.25

98.78

99.22

99.39

97.69

97.54

97.42

97.95

97.83

97.69

97.49

97.96

98.56

98.11

97.99

97.86

ꢀ1.72

ꢀ1.73

ꢀ1.74

ꢀ2.26

ꢀ2.25

ꢀ2.22

ꢀ2.09

ꢀ2.03

ꢀ1.96

ꢀ1.74

ꢀ1.73

ꢀ2.28

ꢀ2.27

ꢀ2.14

ꢀ2.10

ꢀ2.05

54.34

54.31

54.26

28.65

29.60

30.34

37.98

38.68

39.36

51.82

52.19

52.52

33.88

35.90

37.81

42.99

43.47

43.93

0.05

0.06

0.04

0.05

0.06

0.07

0.04

0.05

0.03

0.49

0.45

0.38

0.02

0.08

0.05

0.04

0.37

0.50

0.57

0.05

0.03

98.31

99.63

97.83

92.53

96.23

96.95

92.10

95.16

95.17

90.94

94.17

94.05

93.03

91.43

92.17

92.90

91.65

91.71

No

Yes

No

Yes

Inh.

1

4

P. F. LAMIE AND J. N. PHILOPPES

Figure 4. SAR study of the target compounds 9a–r.

activities with IC50 values of 1.37–6.26 mM (cisplatin Disclosure statement

IC ¼6.62 mM). While, moderate cytotoxic activity was noticed for

5

0

The authors declare that there is no any conflict of interest.

test compounds against colon HT-29 cell line. The most potent

derivatives

IC50¼2.10–2.37 mM), if compared with cisplatin (IC50¼1.12 mM).

The most active derivatives 9a, 9d, 9f, 9n, and 9r (either

between

them

were

9a,

9l,

and

9n

(

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Article Doi

2-Thiopyrimidine/chalcone hybrids: design, synthesis, ADMET prediction, and anticancer evaluation as STAT3/STAT5a inhibitors

DOI: 10.1080/14756366.2020.1740922

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