Design, synthesis, biological evaluation, and docking studies of novel (imidazol-5-yl)pyrimidine-based derivatives as dual BRAFV600E/p38α inhibitors

European Journal of Medicinal Chemistry 215 (2021) 1 13277

Contents lists available at ScienceDirect

European Journal of Medicinal Chemistry

journal homepage: h ttp://www.elsevier.com/locate/ejmech

Design, synthesis, biological evaluation, and docking studies of novel

(imidazol-5-yl)pyrimidine-based derivatives as dual BRAF^V600E/p38

inhibitors

a

, b,

Eslam M.H. Ali ^{a c}, Rania Farag A. El-Telbany ^d, Mohammed S. Abdel-Maksoud ^e,

,

, b

Usama M. Ammar ^f, Karim I. Mersal ^{a b}, Seyed-Omar Zaraei ^a

,

Mohammed I. El-Gamal ^gⁱ, Se-In Choi ^j, Kyung-Tae Lee ^j, Hee-Kwon Kim ^k

,

h

,

, l

, b, *

Kwan Hyi Lee ^am, Chang-Hyun Oh ^a

^aCenter of Biomaterials, Korea Institute of Science & Technology (KIST School), Seoul, Seongbuk-gu, 02792, Republic of Korea

^bUniversity of Science & Technology (UST), Daejeon, Yuseong-gu, 34113, Republic of Korea

^cPharmaceutical Chemistry Department, Faculty of Pharmacy, Modern University for Technology and Information (MTI), Cairo, 12055, Egypt

^dBiochemistry Department, Faculty of Pharmacy, Modern University for Technology and Information (MTI), Cairo, 12055, Egypt

^eMedicinal & Pharmaceutical Chemistry Department, Pharmaceutical and Drug Industries Research Division, National Research Centre NRC (ID:

60014618)), Dokki, Giza, 12622, Egypt

^fStrathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 0NR, Scotland, United Kingdom

^gDepartment of Medicinal Chemistry, College of Pharmacy, University of Sharjah, Sharjah, 27272, United Arab Emirates

^hSharjah Institute for Medical Research, University of Sharjah, Sharjah, 27272, United Arab Emirates

ⁱDepartment of Medicinal Chemistry, Faculty of Pharmacy, University of Mansoura, Mansoura, 35516, Egypt

^jDepartment of Pharmaceutical Biochemistry, College of Pharmacy, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul, 130-701, Republic of Korea

^kDepartment of Nuclear Medicine, Molecular Imaging & Therapeutic Medicine Research Center, Jeonbuk National University Medical School and Hospital,

20 Geonji-ro, Deokjin-gu, Jeonju, 54907, Republic of Korea

^lResearch Institute of Clinical Medicine of Jeonbuk National University-Biomedical Research Institute of Jeonbuk National University Hospital, 20 Geonji-ro,

Deokjin-gu, Jeonju, 54907, Republic of Korea

^mKU-KIST Graduate School of Converging Science and Technology, Korea University, Seoul, 02841, Republic of Korea

a r t i c l e i n f o

a b s t r a c t

Article history:

The synergistic effect of dual inhibition of serine/threonine protein kinases that are involved in the same

signalling pathway of the diseases can exert superior biological beneﬁts for treatment of these diseases.

In the present work, a new series of (imidazol-5-yl)pyrimidine was designed and synthesized as dual

Received 29 October 2020

Received in revised form

21 January 2021

Accepted 1 February 2021

Available online 7 February 2021

inhibitors of BRAF^V600Eand p38

a kinases which are considered as key regulators in mitogen-activated

protein kinase (MAPK) signalling pathway. The target compounds were evaluated for dual kinase

inhibitory activity. The tested compounds exhibited nanomolar scale IC₅₀values against BRAF^V600Eand

low to sub-micromolar IC₅₀range against p38

BRAF^V600E/p38

inhibitor with IC₅₀values of 2.49 and 85 nM, respectively. Further deep investigation

revealed that compound 20h possesses inhibitory activity of TNF- production in lipopolysaccharide-

induced RAW 264.7 macrophages with IC₅₀value of 96.3 nM. Additionally, the target compounds efﬁ-

ciently frustrated the proliferation of LOX-IMVI melanoma cell line. Compound 20h showed a satisfactory

a. Compound 20h was identiﬁed as the most potent dual

Keywords:

a

BRAF^V600E

a

Imidazol-5-ylpyrimidine

MAPK14

Melanoma

Molecular docking

antiproliferative activity with IC₅₀value of 13

mM, while, compound 18f exhibited the highest cytotox-

TNF-

a

icity potency with IC₅₀value of 0.9 M. Compound 18f is 11.11-fold more selective toward LOX-IMVI

m

melanoma cells than IOSE-80PC normal cells. The newly reported compounds represent therapeuti-

cally promising candidates for further development of BRAF^V600E/p38

overcome the acquired resistance of BRAF mutant melanoma.

a inhibitors in an attempt to

1. Introduction

* Corresponding author. Center of Biomaterials, Korea Institute of Science &

Technology (KIST School), Seoul, Seongbuk-gu, 02792, Republic of Korea.

E-mail address: choh@kist.re.kr (C.-H. Oh).

Mitogen-activated protein kinases (MAPKs) are crucial signal-

ling components in transduction of extracellular stimuli to cellular

https://doi.org/10.1016/j.ejmech.2021.113277

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

responses [1]. In mammalian cells, MAPKs are classiﬁed into three

main families: ERKs (extracellular-signal-regulated kinases), JNKs

(c-Jun N-terminal kinases), and p38/SAPKs (stress-activated protein

kinases) [2].

compounds. Guided by SB203580 molecular interaction in p38a

kinase active site (Fig. 2) [34] (PDB: 3GCP), the target compounds

were designed to conserve the essential binding interactions.

Accordingly, a series of new imidazole-5-yl pyrimidine derivatives

(18a-i, 19a-i, 20a-i, and 21a-i) were designed and synthesized

conserving the fundamental binding interactions of both Dabra-

The MAP/ERK signalling pathway plays a crucial role in cancer

progression via promotion of cell proliferation and migration [3].

RAF kinases (ARAF, BRAF and CRAF) comprise a key component in

ERK signaling branch of MAPK cascade which starts upon binding

of RAF kinases with RAS as the upstream activator, followed by

mediating the MAPK signaling transduction to the MEK resulted to

the activation of downstream MEK1/2 and ERK1/2 [4,5]. Among the

RAF family, BRAF has higher basal activity and is a major activator of

MEK1/2 compared to ARAF and CRAF [6]. Notably, BRAF mutations

are associated with many human cancers and considered the most

frequent mutation in melanoma [7]. The most common BRAF mu-

tation is BRAF^V600Ewhen valine residue at site 600 is replaced by

glutamate residue [8]. BRAF^V600Emutation occurs in approximately

75% of patients with BRAF-mutant metastatic melanoma [9,10].

Consequently, promoted by tremendous efforts invested in devel-

oping small molecules BRAF^V600Einhibitors, vemurafenib (Zel-

boraf®, Roche) was the ﬁrst approved BRAF^V600Einhibitor in 2011

for the treatment of metastatic melanoma, followed by dabrafenib

(Taﬁnlar®, GlaxoSmithKline) approval in 2013 [1 1,12]. Sorafenib is a

multiple kinase inhibitor has also been shown to inhibit RAF ki-

nases including C-RAF and B-RAF (Fig. 1) [13]. Despite the prom-

ising clinical response, most BRAF^V600Emetastatic melanoma

patients eventually relapse and progress acquired resistance.

Cancer cells can develop resistance to BRAF^V600Einhibitors by

adopting several mechanisms; (1) genetic alteration leading to

constitutive activation of the MAPK pathway [14,15], (2) epigenetic

or transcriptome-based changes such as overexpression of MAP3K8,

cMet, and IL8 overexpression, and expression of BRAF^V600E-splicing

variants [16e18], (3) immunity enhancement as a response to BRAF

inhibitors followed by boosting the efﬁcacy of checkpoint inhibitors

[19,20]. Therefore, developing novel BRAF^V600Einhibitors became an

ultimate need to overcome the acquired resistance adopted against

the currently used inhibitors.

fenib and SB203580 in BRAF^V600Eand p38

a kinase domain,

respectively. The 2-aminopyrimidinyl moiety of the target com-

pounds is proposed to bind in the adenine hinge region of both

kinases through the interaction with the BRAF^V600ECys532 residue

and the p38a Met109 residue. The central imidazole ring is to bury

the inhibitors in the ribose pocket of the two kinases by its inter-

action with Val471 or Val38 residues. The back hydrophobic pocket

of both kinases was supposed to be occupied by the terminal 3-

methoxy (hydroxyl)-4-chloro (ﬂuoro) phenyl moiety (Fig. 2). Be-

sides the main molecular interactions, the target compounds

possess additional structural features so as to reinforce the binding

tendency in active sites of both kinases; the propylene linker in the

target compounds is proposed to extend the compound in-

teractions in the allosteric pocket of BRAF kinase and the front

hydrophobic pocket of p38

The dual enzyme inhibitory activity of the target compounds

was determined against BRAF^V600Eand p38

kinases, and the IC₅₀

a kinase (Fig. 2).

a

values were determined for the most potent inhibitors. The most

potent compound, 20h, was subjected to deep investigation in or-

der to test its inhibitory effect on the production of TNF-

a as a

downstream target of p38 kinase in lipopolysaccharide-

a

stimulated RAW 264.7 macrophages [35]. Furthermore, the target

compounds were evaluated for antiproliferative activity against

LOX-IMVI melanoma cell line, and the IC₅₀values were calculated.

2. Results and discussion

2.1. Chemistry

The preparation of N-3-aminopropyl unsubstituted and

substituted phenyl sulfonamide (3a-i) was achieved by coupling

propylene-1,3-diamine 1 with appropriate arylsulfonyl chloride

(2a-i) in dichloromethane in presence of triethylamine (TEA) at 0 ^ꢀC

(Scheme 1) [36].

The p38/SAPKs branch of the MAPK pathway is activated by the

upstream MKK3/6 kinases, and sometimes by autophosphorylation

[21,22]. By downstream regulation, p38 MAPK mediates the acti-

vation of a group of protein kinases including MAPK-activated ki-

nase 2 (MAPK2), mitogen- and stress-activated protein kinase 1

(MsK1), and MAP kinase-interacting serine/threonine kinase 1/2

(MNK1/2) [21,22]. MNK1/2 kinases are downstream targets of the

ERK and p38 MAPK pathways. MNK kinases mediate oncogenic

progression through its role in regulating mRNA translation [23].

The key intermediates 16 and 17 were synthesized through the

esteriﬁcation of the 4-ﬂuoro (chloro)-3-methoxybenzoic acid 4 and

5 with methanol in the presence of sulfuric acid to produce the

corresponding methyl esters 6 and 7. In a basic medium of lithium

bis(trimethylsilyl)amide (LiHMDS), the methyl group of 4-methyl-

2-(methylthio)pyrimidine 8 was activated by stirring for 30 min,

followed by dropwise addition of the methyl ester 6 or 7 to afford

the 1-(4-ﬂuoro (choro)-3-methoxyphenyl)-2-(2-(methylthio)pyr-

imidin-4-yl)ethan-1-one 9 and 10. Cyclization to the corresponding

imidazole derivatives 14 and 15 was carried out by oxidation of

compound 9 or compound 10 by HBr in DMSO to produce 1-(4-

ﬂuoro (choro)-3-methoxyphenyl)-2-(2-(methylthio)pyrimidin-4-

yl)ethane-1,2-dione (11 and 12), followed by condensation reac-

tion with benzaldehyde 13 in presence of ammonium acetate and

acetic acid to afford 4-(4-(4-ﬂuoro (chloro)-3-methoxyphenyl)-2-

phenyl-1H-imidazol-5-yl)-2-(methylthio)pyrimidine (14 and 15).

The methylthio group was oxidized using oxone to give the corre-

sponding methylsulfonyl analogues 4-(4-(4-ﬂuoro (chloro)-3-

methoxyphenyl)-2-phenyl-1 H-imidazol-5-yl)-2-(methylsulfonyl)

pyrimidine (16 and 17), respectively (Scheme 2).

Based on the crosstalk between p38

target), we proposed that developing dual inhibitors of BRAF^V600E

and p38 kinases could participate in overcoming the BRAF in-

a and ERK (BRAF downstream

a

hibitors resistance development.

Several ﬁve-membered ring based derivatives were reported in

literatures as MAP kinase inhibitors, in particular BRAF^V600Eand

p38a kinases [24e32] (Fig. 1). In a previous work, we designed a

series of pyridinyl imidazole derivatives based on the structure

features of dabrafenib. The reported compounds exhibited high

potency against BRAF^V600Ekinase, compound I showed the highest

activity (IC₅₀¼ 32 nM) [33]. In the recent work, guided by the

binding mode of dabrafenib in the BRAF kinase domain (PDB:

4XV2), a group of structural optimizations was conducted to

compound I in order to improve the binding afﬁnity of the new

designed compounds in the BRAF kinase binding site aiming to

boost their inhibitory activity.

The ﬁnal target compounds were obtained by coupling of

compounds 16 or 17 with appropriate amino reagents 3a-i using

N,N-diisopropylethylamine as a base. The mesyl group of com-

pounds 16 and 17 was displaced by the amino group to afford the

ﬁnal target compounds 18a-i and 19a-i. Demethylation was applied

Considering our purpose to develop dual BRAF^V600Eand p38

kinases inhibitors, the imidazole-based p38 inhibitor SB203580

(Fig. 1) was used a lead compound to design the new target

a

2

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

Fig. 1. Structures of reported 5-membered ring-based BRAF^V600Eand p38

a

kinase inhibitors.

by dropwise addition of boron tribromide at ꢁ70 ^ꢀC under nitrogen

in presence of tetrabutylammonium iodide in anhydrous

dichloromethane to obtain the hydroxyl derivatives 20a-i and 21a-i

(Scheme 3). Structures of the target compounds are illustrated in

Table 1.

substitution with electron-withdrawing groups such as Cl, Br, F in

compounds 18b-d and 19b-d slightly enhanced the BRAF^V600Eac-

tivity but faintly reduced the p38a activity, while meta-F substation

in compounds 18f and 19f had no signiﬁcant effect on the dual

inhibitory activity. Interestingly, substitution with bulky group like

CF₃in compounds 18e and 19e dramatically reduced the dual

inhibitory activity to about its half, however, replacement of phenyl

sulfonamide by naphthyl sulfonamide in compounds 18i and 19i

2.2. Biological evaluation

only declined the p38

BRAF^V600Ekinase. Introduction of electron-donating group CH₃at

the para position in compounds 18g and 19g had no effect in p38

inhibitory activity and slight improved the inhibitory effect against

BRAF^V600E. The highest improvement of methoxy derivatives dual

inhibitory activity was obtained by para substitution with methoxy

group in compounds 18h and 19h.

a activity but conserved the activity against

2.2.1. In vitro BRAF^V600E/p38

a kinase inhibitory activity at 10 mM

2.2.1.1. Screening of the methoxy derivatives. In order to identify the

methoxy derivatives that can afford dual inhibitory activity, both

ﬂuoro derivatives 18a-i and chloro analogues 19a-i bearing variable

aromatic sulfonamides were preliminarily screened for their dual

inhibitory activity through determining their inhibition percent-

a

ages at 10

m a kinase. The tested

M against BRAF^V600Eand p38

compounds showed a wide potency range. The results obtained in

Table 2, revealed that ﬂuoro derivatives 18a-i showed good inhib-

2.2.1.2. Screening of the hydroxyl derivatives. The methoxy de-

itory activity against both BRAF^V600Eand p38

a

kinase. While, the

corresponding chloro derivatives 19a-i emerged to be more potent

inhibitors of BRAF^V600Ethan p38

kinase. The diversity in activity

rivatives were subjected to further optimization. Aiming to improve

the BRAF^V600Eand p38

a kinase dual inhibitory activity, demethyla-

a

tion of methoxy group at the hydrophobic area into the corre-

sponding hydroxyl derivatives was required. The higher polarity of

the hydroxyl group would have proposed to facilitate the extension

into the accessible pocket, which would offer better

alongside the substitution on the aromatic sulfonamide reﬂect

certain structure-activity relationships (SAR) for the Ar moiety.

Compared to the unsubstituted compounds 18a and 19a, para

3

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

Fig. 2. Rational design of the new imidazole-5-yl pyrimidine derivatives 18a-i, 19a-i, 20a-i, and 21a-i as dual BRAF^V600Eand p38

teractions into BRAF^V600Ekinase domain; b) Proposed binding interaction of the target compounds into BRAF^V600Ekinase domain, c) SB203580 molecular interactions into p38

kinase domain; d) Proposed binding interaction of the target compounds into p38 kinase domain.

a kinase inhibitors. a) Dabrafenib molecular in-

a

Cl, Br, F, the p38

a

inhibition decreased slightly in case of compounds

20b-d, 21b, and 21c, in addition to a minute improvement in com-

pound 21c. Substitution either by para-(triﬂuoromethyl)phenyl or

naphthyl moieties in compounds 20e, 20i, 21e, and 21i, conserved

the activity of BRAF^V600Ewhile, dramatically dropped p38

a

activity.

Meta substitution with F group in compounds 20f and 21f, exhibited

no effect on the activity of both BRAF^V600Eand p38

a kinase. The

highest BRAF^V600Eactivity was observed upon sulfonamide substi-

tutionwith para OH group in compounds 20h and 21h, even though,

the OH substitution slightly decreased the p38

compounds.

a activity for the same

Scheme 1. Reagents and conditions: i) TEA, DCM, 0 ^ꢀC; rt, 18 h.

physicochemical properties. The hydroxyl-bearing derivatives 20a-i

and 21a-i were evaluated for their inhibitory activities against

2.2.2. Potential BRAF^V600E/p38

(IC₅₀) determination

a kinase dual inhibitory activity

BRAF^V600Eand p38

a kinase.

Based on the previous obtained inhibition percentage results,

ten compounds (20a, 20c, 20f-h, 21a, 21c, and 21f-h) that showed

potent dual inhibitory activities were subjected to kinase IC₅₀

As illustrated in Table 2, the structural optimization resulted in

remarkable improvement in the inhibitory activity against

BRAF^V600Eand p38

a

kinase, the inhibitory activity jumped from 42-

90% to 94e100% and from 19-88% to 85e92% for BRAF^V600Eand

p38 , respectively. The hydroxyl derivatives responded in a similar

determination. The dual BRAF^V600E/p38

a

inhibition of selected

compounds were evaluated at 5-dose concentrations (1 nM, 10 nM,

100 nM, 1 M and 10 M) in order to determine their IC₅₀values.

The ten compounds exhibited BRAF^V600EIC₅₀values in the nano-

molar range, and p38 IC₅₀in low to sub-micromolar range

(Table 3). Compound 20h emerged to be the most potent dual

a

m

way like their methoxy counterparts upon substitution on the sul-

fonamide moiety. However, the BRAF^V600Eactivity did not exhibit a

notable change by substitution with electron-withdrawing groups

a

4

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

Scheme 2. Reagents and conditions: i) MeOH, conc. H₂SO₄, 80 ^ꢀC, 12 h; ii) LiHMDS, THF, ꢁ70 ^ꢀC, 18 h; iii) HBr, DMSO, 55 ^ꢀC, 2 h; iv) NH₄OAc, CH₃COOH, 100 ^ꢀC, 4 h; v) oxone, MeOH/

H₂O, rt, 18 h.

Scheme 3. Reagents and conditions: i) DIPEA, DMSO, 80 ^ꢀC, 18 h; ii) BBr₃, DCM, TBAI, ꢁ70 ^ꢀC, 6 h.

inhibitor with IC₅₀values of 2.49 and 85 nM against BRAF^V600Eand

p38 , respectively. Compound 20h is 12.7-fold more potent than

staurosporine against BRAF^V600E. Similarly, it is 49.8-fold more

potent than PP2 against p38

concentration, respectively. In addition, its IC₅₀value equals

96.3 nM (Table 4). In addition, it is 53.8-fold more potent than BAY

11e7082 as inhibitor of TNF-a production.

a

.

2.2.4. In vitro cytotoxicity activity against LOX-IMVI melanoma cell

line and IOSE-80PC normal cells

2.2.3. TNF-

A further study was carried out for compound 20h in order to

determine its inhibitory effect on the production of TNF- in

lipopolysaccharide-stimulated RAW 264.7 macrophages. It was

compared with BAY 11e7082 as a reference standard TNF- pro-

duction inhibitor [37]. Compound 20h exerted 85.24% and 96.42%

inhibition of TNF- production when tested at 1 and 10

a production assay

The anticancer activity of the synthesized derivatives were

further determined using the MTT cytotoxicity assay on LOX-IMVI

melanoma cell line which shows overexpression of BRAF^V600Eki-

nase [38,39]. The compounds were tested at 5-dose concentrations

to calculate their IC₅₀values, and staurosporine was utilized as a

reference compound. According to the IC₅₀data presented in

a

mM

5

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

Table 1

Structures of the ﬁnal target compounds 18a-i, 19a-i, 20a-i, and 21a-i.

Compound No.

X

F

R

Ar

Compound No.

X

F

R

Ar

18a

OCH₃

20a

OH

18b

18c

18d

18e

18f

F

OCH₃

20b

20c

20d

20e

20f

F

OH

18g

18h

18i

F

OCH₃

20g

20h

20i

F

OH

19a

19b

19c

19d

19e

19f

Cl

OCH₃

21a

21b

21c

21d

21e

21f

Cl

OH

19g

19h

Cl

OCH₃

21g

21h

Cl

OH

6

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

Table 1 (continued)

Compound No.

X

R

Ar

Compound No.

X

R

Ar

19i

Cl

OCH₃

21i

Cl

OH

Table 2

Enzyme % inhibition activity of the ﬁnal target compounds 18a-i, 19a-i, 20a-i, and 21a-i at 10

m

M over BRAF^V600Eand p38 kinase.^a

a

Cpd.

kinase % inhibition at 10

BRAF^V600E

mM

Cpd.

kinase % inhibition at 10

BRAF^V600E

mM

p38a

/MAPK14

p38a/MAPK14

18a

18b

18c

18d

18e

18f

18g

18h

18i

19a

19b

19c

19d

19e

19f

19g

19h

19i

77.35% 2.25%

79.20% 1.97%

81.50% 3.07%

80.11% 0.75%

42.36% 2.68%

72.50% 5.10%

82.41% 0.89%

89.12% 0.77%

81.02% 1.26%

80.05% 3.85%

85.12% 2.87%

88.35% 0.98%

81.04% 1.09%

55.91% 1.82%

84.87% 0.69%

88.20% 1.69%

91.10% 0.23%

90.05% 3.45%

83.51% 2.18%

72.79% 3.76%

73.37% 2.66%

84.13% 1.13%

48.60% 0.06%

82.67% 2.77%

79.36% 2.86%

88.54% 0.97%

44.40% 9.59%

58.62% 1.22%

52.13% 3.13%

30.00% 6.11%

62.07% 0.64%

18.87% 0.15%

58.28% 0.91%

50.43% 0.29%

54.56% 1.05%

27.11% 1.71%

20a

20b

20c

20d

20e

20f

20g

20h

20i

21a

21b

21c

21d

21e

21f

21g

21h

21i

98.44% 0.89%

93.86% 0.69%

99.98% 0.60%

97.04% 1.80%

98.44% 0.55%

100.27% 1.56%

100.64% 0.36%

101.35% 1.19%

99.80% 0.86%

99.43% 1.27%

99.66% 0.89%

97.26% 0.58%

97.11% 0.79%

99.24% 0.54%

100.51% 0.86%

101.17% 1.79%

100.93% 1.63%

96.24% 0.97%

87.01% 3.32%

84.89% 5.82%

84.79% 5.95%

91.43% 3.09%

65.42% 0.07%

90.45% 2.53%

87.85% 0.39%

92.80% 3.48%

79.77% 3.10%

82.22% 4.00%

70.76% 7.00%

73.74% 2.89%

85.53% 3.88%

54.31% 4.04%

78.56% 3.98%

70.85% 0.25%

77.62% 1.53%

51.09% 0.80%

a

The results are expressed as means of duplicate experiments standard deviation (S.D.).

Table 3

activity with IC₅₀value of 0.9

(triﬂuoromethyl)benzenesulfonamido) and 19h (4-methoxyben

zenesulfonamido) with IC₅₀values of 1.78 M and 1.80 M, respec-

tively. Compounds 18a,18d,18g,18h,19a,19d, and 19f exhibited good

cytotoxicity activity with IC₅₀values ranging from 2.59 M to 7.47

compared to staurosporine (IC₅₀¼ 7.15 M). The rest of the methoxy

M).

Among the tested hydroxyl derivatives, compounds 20e, 21a

and 21d with 4-CF₃phenyl, phenyl, and 4-ﬂuorophenyl sulfon-

amide, respectively were exhibited the highest potency with IC₅₀

mM, followed by compounds 19e (4-

IC₅₀values of the ﬁnal target compounds 20a, 20c, 20f-h, 21a, 21c, and 21f-h in

addition to the reference standard compounds over BRAF^V600Eand p38

a

kinases.

m

Compound No.

BRAF^V600EIC₅₀(nM)

p38a IC₅₀(nM)

m

mM

20a

20c

20f

20g

20h

21a

21c

21f

21g

21h

Staurosporine

PP2

5.62

210

330

140

200

85

ND

m

26.30

30.30

8.68

2.49

4.25

21.80

14.80

8.04

7.81

31.60

ND

derivatives showed moderate activity (IC₅₀¼ 13.5e83.1

m

values of 2.45, 2.63, and 2.30

and 21g showed appreciated cytotoxicity effect with IC₅₀values of

5.66 M, 3.71 M, and 6.02 M, respectively in comparison with the

mM, respectively. Compounds 20c, 20i,

m

reference compound staurosporine. The rest of the hydroxyl com-

pounds revealed good ability to inhibit LOX-IMVI cell growth with

IC₅₀values ranging from 7.72 mM to 61.3 mM.

4,230

ND: not determined.

After that, the most potent compounds 18f, 19e, 20e, 21a, and

21d were tested against IOSE-80PC human ovarian epithelial cells

in 5-dose testing mode to investigate their selectivity indexes. The

dose-response curves are illustrated in Fig. 3. All the ﬁve com-

Table 4

Effect of compound 20h as inhibitor of TNF-

a production in lipopolysaccharide-

stimulated RAW 264.7 macrophages in comparison with BAY 11e7082.^a

pounds exerted less than 20% cell viability inhibition up to 10

mM

Compound

Result (% inhibition or IC₅₀

)

concentration (i.e. IC₅₀values are >10 M). Therefore, the selec-

m

20h

85.24% 1.88% inhibition at 1 mM

tivity indexes of compounds 18f, 19e, 20e, 21a, and 21d towards

LOX-IMVI melanoma cells than IOSE-80PC normal cells are more

than 11.11, 5.62, 4.08, 3.80, and 4.35 fold, respectively.

96.42% 0.95% inhibition at 10

IC₅₀¼ 96.3 1.2 nM

mM

BAY 11-7082

IC₅₀¼ 5,180 10 nM

a

The results are expressed as means S.D.

2.3. Docking study

Table 5, generally the hydroxyl derivatives 20 and 21 showed

higher ability to reduce LOX-IMVI cell line viability than their cor-

responding methoxy bearing derivatives 18 and 19.

In regard with the methoxy derivatives, compound 18f with the

terminal 3-ﬂuorophenyl sulfonamide showed incredible cytotoxicity

In attempt to study the binding characteristics of the target com-

pounds in the binding site of both BRAF^V600Eand p38

a kinases, mo-

lecular docking studies were conducted using Molecular Operating

Environment (MOE, 2014.0901) software. The two X-ray crystallo-

graphic structures of BRAF^V600Ein complex with Dabrafenib (PDB ID:

7

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

Table 5

IC₅₀cytotoxicity of target compounds 18a-i, 19a-i, 20a-i, and 21a-i against LOX-IMVI melanoma cell line.^a

Cpd.

Melanoma (LOX-IMVI) Cytotoxicity IC₅₀

(

m

M)

Cpd.

Melanoma (LOX-IMVI) Cytotoxicity IC₅₀(mM)

18a

18b

18c

18d

18e

18f

18g

18h

18i

19a

19b

19c

19d

19e

19f

19g

19h

19i

4.52 0.25

83.10 4.69

33.90 1.91

3.96 0.22

75.20 4.24

0.90 0.05

2.59 0.15

6.13 0.35

14.90 0.84

7.47 0.42

60.90 3.44

23.70 1.34

4.53 0.26

1.78 0.1

20a

20b

20c

20d

20e

20f

20g

20h

20i

21a

21b

21c

21d

21e

21f

21g

21h

21i

9.58 0.54

14.20 0.80

5.66 0.32

19.20 1.08

2.45 0.14

7.89 0.44

26.30 1.49

13.90 0.79

3.71 0.21

2.63 0.15

61.30 3.46

15.50 0.87

2.30 0.13

10.10 0.57

33.60 1.9

6.02 0.34

36.80 2.08

7.72 0.44

5.11 0.29

18.60 1.05

1.80 0.10

13.50 0.76

7.15 0.4

Staurosporine

a

The results are expressed as means of duplicate experiments S.D.

Fig. 3. Dose-response curves of compounds 18f, 19e, 20e, 21a, and 21d over IOSE-80PC human ovarian epithelial cell line.

4XV2) and p38

a

kinase in complex with SB203580 (PDB ID: 3GCP)

Molecular docking protocol was ﬁrstly validated by re-docking

were downloaded from the Protein Data Bank (PDB) [40,41].

of the co-crystalized ligands, Dabrafenib and SB203580, in the

8

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

binding sites of BRAF^V600Eand p38

simulation revealed the binding pattern of the two kinases (BRAF

and p38 ) with their corresponding co-crystalized ligands with

a, respectively. The docking

a

energy scores of ꢁ11.13 and ꢁ12.16 kcal/mol, respectively, and with

an RMSD of 0.162 and 0.460 Å, respectively, between the co-

crystalized native ligand and the docked poses. All the resulted

docking poses imitated all the essential molecular interactions

possessed by the native ligand in the binding active site of

BRAF^V600Eand p38

a (Table S1, Supplementary File).

As illustrated BRAF^V600Enative ligand (Dabrafenib) bound in the

adenine pocket via two H-bonds between N³, and 2-NH₂pyrimi-

dine and Cys532 in the hinge binding area, the hydrophobic pocket

was occupied by the 2-ﬂuorophenyl group, while the central thia-

zole ring was embedded in the ribose pocket through Arene-H

interaction with Val471. Additionally, dabrafenib interactions

extended the RAS selective pocket where, the sulfonamide moiety

formed H-bonds with Phe595 and Asp594, and the 2,6-

diﬂuorphenyl ring formed Arene-H interaction with Leu505

(Fig. 4a). However, p38a native ligand (SB203580) formed H-bond

with Met109 in the adenine hinge region, the hydrophobic back

pocket was occupied by the 4-ﬂuoro phenyl moiety, the central

imidazole ring embeds in the ribose pocket via arene-H-bond

interaction with Val38, the oxygen of the terminal methylsulﬁnyl

phenyl at position 2 of the imidazole ring binds in the phosphate

area by forming H-bond with Tyr35 residue (Fig. 4b).

Accordingly, the target compounds were designed to conserve

the essential molecular interactions of Dabrafenib and SB203580

into their corresponding kinase pockets. Analysis of the docking

results revealed that the target compounds 18e21 showed quite

similar binding interaction in both kinases with predicted docking

energy scores in the binding site of both BRAF^V600Eand p38

a kinase

ranging from ꢁ8.52 to ꢁ10.13 kcal/mol and ꢁ11.42 to ꢁ14.81 kcal/

mol, respectively (Table S1).

Most of the target compounds followed the molecular binding

pattern of both Dabrafenib and SB203580 (Table S1). As observed,

the central imidazole ring of the tested compounds 19d, 19g, 20a,

20h, and 20c buried into the ribose pocket of kinase active site via

Arene-H interaction with Val471 and Val38 of BRAF^V600Eand p38

a

kinase, respectively. In addition, the 2-aminopyrimidinyl arm was

found to be the essential motif responsible for the H-bond inter-

action with Cys532 and Met109 in the adenine hinge region of

BRAF^V600Eand p38

a kinase, respectively exempliﬁed by the binding

modes of compound 19b, 20h, 21f, and 21g. In both kinases, the

terminal 3-methoxy(hydroxy)-4-chloro(ﬂuoro) phenyl branch

embedded in the back hydrophobic pocket; in the BRAF^V600Eki-

nase, compounds 18d, 18f, 20c, 20e, 20h, and 20i exhibited Arene-H

interaction with Phe583, while the corresponding moiety in com-

pounds 19g and 21a interacted by Arene-H with Leu171. Further-

more, the propylene linker allowed the extension of the

sulfonamido moiety of compounds 18g,19d,19h, 20g, 20h, 21b, and

21d into the RAS selective pocket of BRAF^V600Ekinase through H-

bonding with Asp594. While, the extension of the sulfonamide in

Fig. 4. 3D molecular interaction diagram of: a) Dabrafenib in BRAF^V600Eactive site

(PDB ID: 4XV2), b) SB203580 in p38a kinase active site in (PDB ID: 3GCP).

3. Conclusion

In the current study, a series of imidazole-5-yl pyrimidine-based

dual BRAF^V600E/p38

inhibitors was designed and synthesized in

a

attempt to overwhelm the resistance acquired against BRAF in-

hibitors in BRAF^V600Emetastatic melanoma patients. Considering

the binding mode of dabrafenib in BRAF^V600Ekinase active site and

the front hydrophobic pocket of p38

a kinase binding sites was

achieved by H-bond formation with Try35 in compounds 18i and

19c. Obviously, the most potent inhibitor compound 20h

SB203580 with p38

a kinase, imidazolyl pyrimidin-2-amine was

(BRAF^V600EIC₅₀¼ 2.4 nM & p38

IC₅₀¼ 85 nM), exhibits typical

a

used as a core scaffold and a hinge binder in the active site of both

enzymes. Substitution on the 2-amino group with propylene linker

ended by different aromatic sulfonamides resulted in a wide range

of inhibitory activity against both kinases. Firstly, the kinase inhi-

bition assay was conducted to the methoxy derivatives 18a-i and

19a-i in order to identify the derivatives that can achieve optimal

molecular interactions of both Dabrafenib and SB203580 in the

active site of BRAF^V600Eand p38

a kinases. The 2-aminopyrimidine

binds in the adenine hinge region, the central imidazole is

embedded into the ribose pocket, the back hydrophobic pocket of

both kinases are occupied by the 4-ﬂuoro-3-hydroxy phenyl moi-

ety, and ﬁnally the terminal sulfonamide extended to the selective

RAS pocket in BRAF kinase via H-bond formation with Asp594

(Fig. 5 & Tables S1 and S2).

dual BRAF^V600E/p38

a inhibition. Compounds 18h and 19h having

the terminal p-methoxybenzenesulfonamido were the highest

active derivatives with % inhibition of 89% and 91% against

9

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

cytotoxic agent with IC₅₀value of 0.9 mM. Compound 18f is 11.11-

fold more selective against LOX-IMVI melanoma cells than IOSE-

80PC normal cell line. Molecular docking protocol was applied on

the target compounds to determine their molecular interaction

features in the active site of both kinases. Compound 20h

conserved the fundamental interactions of the two native ligands in

corresponding kinase active site.

Compound 20h is considered as a promising dual BRAF^V600E

p38 inhibitor that could be subjected for further optimization and

/

a

development aiming at improving its cytotoxicity activity so as to

overcome the resistance against currently used BRAF^V600Ein-

hibitors in BRAF-mutant metastatic melanoma.

4. Experimental

4.1. General

The melting points were measured on a Walden Precision

Apparatus Electrothermal 9300 and are uncorrected. Bruker Avance

(400 MHz spectrometer) was used to record ¹H NMR and ¹³C NMR

spectra. LC-MS analysis was conducted using LC-MS analyzer

(Waters Corporation, MA, USA). All the solvents and reagents were

obtained from commercial companies and used as such. The ﬁnal

and intermediate compounds were puriﬁed by ﬂash column

chromatography (silica gel, pore size 0.040e0.063 mm, 230e400

mesh) using laboratory reagent grade solvents.

4.1.1. Synthesis of N-(3-aminopropyl) unsubstituted and substituted

phenyl sulfonamide 3a-i

They were synthesized as per the previously reported method

[36].

4.1.2. Synthesis of the key intermediates 4-(4-(4-ﬂuoro (chloro)-3-

methoxyphenyl)-2-phenyl-1H-imidazol-5-yl)-2-(methylsulfonyl)

pyrimidine (16e17)

4.1.2.1. Methyl

4-ﬂuoro/chloro-3-methoxybenzoate

(6e7). A

mixture of 4-ﬂuoro(chloro)-3-methoxybenzoic acid (4(5),

0.03 mol) and methanol (50 mL) was heated under reﬂux until the

benzoic acid was dissolved in methanol then few drops of

concentrated sulfuric acid was added to the mixture and reﬂuxed

for 8 h. The resulting mixture was allowed to cool to rt, diluted with

water, and a saturated solution of NaHCO₃was added to the

mixture to neutralize the benzoic acid, extracted with ethyl acetate

(3 ꢂ 20 mL), dried over anhydrous Na₂SO₄, and evaporated in vacuo

to get the required ester compounds 6 and 7.

Fig. 5. 3D representation of compound 20h molecular interactions in the active site of

BRAF^V600Ekinase (a) and p38

a kinase (b).

BRAF^V600Eand 88% and 55% against p38

a. Demethylation of the

methoxy derivatives to the hydroxyl derivatives 20a-i and 21a-i

resulted in dramatically improvement of the activity against the

two enzymes. Notably, the hydroxyl derivatives 20h and 21h

exhibited the highest potency against BRAF^V600E(%

inhibition ¼ 101% and 100%) and good but not remarkable activity

4.1.2.2. 1-(4-Fluoro or chloro)-3-methoxyphenyl)-2-(2-(methylthio)

pyrimidin-4-yl)ethan-1-one (9-10). To a solution of and 4-methyl-

2-(methylthio)pyrimidine (8, 771 mg, 5.5 mmol) in THF (20 mL)

at ꢁ70 ^ꢀC, LiHMDS (11 mL, 1.0 M solution in THF, 10.8 mmol) was

slowly added under N₂to the reaction mixture maintaining the

temperature at ꢁ70 ^ꢀC. After 30 min, a solution of compound 6 or 7

(5.0 mmol) in THF (10 mL) was slowly added to the reaction

mixture under N₂at ꢁ70 ^ꢀC. The resulting mixture was stirred at

room temperature for 6 h. The mixture was quenched with satu-

rated aq. NH₄Cl (20 mL), and ethyl acetate (30 mL) was added for

extraction. The organic layer was separated, and the aqueous layer

was extracted with ethyl acetate (3 ꢂ 10 mL). The combined ethyl

acetate extract was washed with saline and dried over anhydrous

sodium sulfate, evaporated in vacuo to yield the title compounds 9

and 10, which were subjected to the next step without further

puriﬁcation.

against p38

a

(% inhibition ¼ 92% and 78%). Further 5-dose assay

was conducted for the highest active derivatives to determine their

IC₅₀, the tested compounds showed potent dual inhibitory activity

with IC₅₀in nanomolar range for BRAF^V600Eand IC₅₀in the nano-

molar range, and sub-micromolar for p38

emerged to be the most potent dual inhibitor with IC₅₀values of

2.49 and 85 nM against BRAF^V600Eand p38

, respectively. Com-

pound 20h was subjected to further investigation to evaluate its

inhibitory effect on TNF- production in lipopolysaccharide-

a. Compound 20h

a

induced RAW 264.7 macrophages, and exhibited high inhibitory

activity with IC₅₀value of 96.3 nM. The antiproliferative activity of

the target compounds was determined using the MTT cytotoxicity

assay on LOX-IMVI melanoma cell line. Compound 20h showed

moderate antiproliferative activity with IC₅₀value of 13 mM, while

the methoxy derivative 18f possessing terminal m-ﬂuo-

4.1.2.3. 1-(4-(Fluoro (chloro))-3-methoxyphenyl)-2-(2-(methylthio)

robenzenesulfonamido moiety emerged to be the most potent

pyrimidin-4-yl)ethane-1,2-dione (11e12). A solution of compound 9

10

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

or 10 (6.8 mmol) in DMSO (10 mL) was heated to 55 ^ꢀC. Hydro-

bromic acid (20.5 mmol) was added dropwise to the reaction

mixture. The reaction was stirred at 55 ^ꢀC for 2 h. The reaction

mixture was poured carefully to a saturated solution of NaHCO₃

(20 mL), extracted with ethyl acetate (3 ꢂ 10 mL), dried over

anhydrous sodium sulfate, and evaporated under vacuum to give

the required compounds 11 and 12, which were subjected to the

next step without further puriﬁcation.

4.1.3. Synthesis of the target compounds 18-21

4.1.3.1. Synthesis of N-(3-((4-(4-(4-(ﬂuoro and chloro)-3-

methoxyphenyl)-2-phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)

propyl) unsubstituted and substituted phenyl sulfonamide (18a-i and

19a-i). A mixture of compound 16 or 17 (0.7 mmol), appropriate

amine (3a-i, 1.4 mmol) and N,N-diisopropylethylamine (814 mg,

6.3 mmol) in anhydrous DMSO (2 mL). The reaction mixture was

allowed to stir at 90 ^ꢀC for 9 h. After that, it was cooled and

extracted between ethyl acetate (10 mL) and water (30 mL). The

organic layer was washed with water (3 ꢂ 30 ml). The organic layer

was dried over anhydrous Na₂SO₄and evaporated under vacuum.

The crude residue was subjected to column chromatography

(hexane, EtOAc; 1, 1) to give the titled compounds 18a-i and 19 a-i.

4.1.3.1.1. N-(3-((4-(4-(4-Fluoro-3-methoxyphenyl)-2-phenyl-1H-

imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzenesulfonamide

(18a). Yield: 70%; mp: 125e127 ^ꢀC; ¹H NMR (400 MHz, CDCl₃)

4.1.2.4. 4-(4-(4-(Fluoro and chloro)-3-methoxyphenyl)-2-phenyl-1H-

imidazol-5-yl)-2-(methylthio)pyrimidine (14e15). To a solution of

compound 11 or 12 (3.8 mmol) and benzaldehyde (13, 403 mg,

3.8 mmol) in acetic acid (9 mL), ammonium acetate (2.93 g,

38 mmol) was added. The reaction mixture was heated to 100 ^ꢀC for

4 h. The reaction mixture was poured in ammonia solution with

crushed ice. The resulted precipitate was ﬁltered, washed with

water (3 ꢂ 15 mL), and dried under vacuum to get the titled

compounds 14 and 15. They were puriﬁed by column chromatog-

raphy using silica gel and ethyl acetate/hexane eluting system.

4.1.2.4.1. 4-(4-(4-Fluoro-3-methoxyphenyl)-2-phenyl-1H-imida-

zol-5-yl)-2-(methylthio)pyrimidine (14). Yield: 85%; ¹H NMR

d

8.09 (d, J ¼ 6.6 Hz, 2H), 8.01 (d, J ¼ 5.0 Hz, 1H), 7.79 (d, J ¼ 7.2 Hz,

2H), 7.55 (t, J ¼ 7.2 Hz, 2H), 7.48 (s, 1H), 7.46 (d, J ¼ 3.1 Hz, 2H), 7.44

(s, 1H), 7.31 (s, 1H), 7.29 (s, 2H), 7.14 (d, J ¼ 8.0 Hz, 2H), 6.04 (s, 1H)

[AreH], 3.91 (s, 3H, OCH₃), 3.57 (brs, 2H, CH₂CH₂CH₂NHSO₂), 3.03

(brs, 2H, CH₂CH₂CH₂NHSO₂), 1.78 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³

C

NMR (100 MHz, CDCl₃) d 153.8,151.4,147.6,140.0,132.5,129.7,129.1,

129.0,128.9,126.8,126.1,122.0,116.2,116.0,114.6,106.4 [AreC], 56.3

(OCH₃), 40.2 (CH₂CH₂CH₂NHSO₂), 37.91 (CH₂CH₂CH₂NHSO₂), 29.80

(CH₂CH₂CH₂NHSO₂); LC/MS 599.1 (M^þþ 1).

(400 MHz, CDCl₃)

d

8.35 (d, J ¼ 5.2 Hz, 1H), 8.07 (d, J ¼ 3.8 Hz, 2H),

7.48e7.46 (m, 3H), 7.39 (d, J ¼ 8.0 Hz, 1H), 7.29 (s, 1H), 7.26 (d,

J ¼ 1.8 Hz, 1H), 7.11 (d, J ¼ 3.2 Hz, 1H) [AreH], 3.91 (s, 3H, OCH₃),

4.1.3.1.2. 4-Chloro-N-(3-((4-(4-(4-ﬂuoro-3-methoxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (18b). Yield: 65%; mp: 124e127 ^ꢀC; ¹H NMR (400 MHz,

2.52 (s, 3H, SCH₃); ¹³C NMR (100 MHz, CDCl₃)

d 172.6, 157.1, 155.1,

134.5, 130.3, 130.2, 129.8, 129.0, 126.4, 123.2, 122.0, 113.2 [AreC],

56.3 (OCH₃), 14.0 (SCH₃).

CDCl₃)

d

8.06 (t, J ¼ 3.2 Hz, 2H), 8.04 (d, J ¼ 3.2 Hz, 1H), 7.69 (d,

4.1.2.4.2. 4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-1H-imida-

J ¼ 3.8 Hz, 2H), 7.46e7.40 (m, 5H), 7.28 (d, J ¼ 8.2 Hz, 2H), 7.12 (d,

J ¼ 8.4 Hz, 2H) [AreH], 6.74 (brs, 1H, NH), 6.29 (brs, 1H, NH), 5.78

(brs, 1H, NH), 3.89 (s, 3H, OCH₃), 3.52 (brs, 2H, CH₂CH₂CH₂NHSO₂),

zol-5-yl)-2-(methylthio)pyrimidine (15). Yield: 87%; ¹H NMR

(400 MHz, CDCl₃)

(m, 4H), 7.35 (s, 2H), 7.02 (d, J ¼ 8.0 Hz, 1H) [AreH], 3.86 (s, 3H,

OCH₃), 2.43 (s, 3H, SCH₃); ¹³C NMR (100 MHz, CDCl₃)

173.0, 158.2,

d

8.36 (d, J ¼ 3.2 Hz, 1H), 8.27 (s, 2H), 7.50e7.43

3.01

(brs,

2H,

CH₂CH₂CH₂NHSO₂),

1.75

d

(brs,

2H,

d

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

153.9,147.6,138.9,

156.1, 135.1, 133.0, 131.3, 130.0, 128.8, 128.1, 126.4, 124.0, 122.0, 112.9

[AreC], 56.5 (OCH₃), 13.9 (SCH₃).

138.6, 129.7, 129.3, 128.9, 128.3, 126.0, 122.0, 116.2, 116.0, 114.6,

106.4 [AreC], 56.3 (OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 37.8

(CH₂CH₂CH₂NHSO₂), 29.9 (CH₂CH₂CH₂NHSO₂); LC/MS 594.8

(M^þþ 1).

4.1.2.5. 4-(4-(4-(Fluoro and chloro)-3-methoxyphenyl)-2-phenyl-1H-

imidazol-5-yl)-2-(methylsulfonyl)pyrimidine (16e17). A solution of

potassium peroxymonosulfate (852 mg, 5.6 mmol) in water (20 mL)

4.1.3.1.3. 4-Bromo-N-(3-((4-(4-(4-ﬂuoro-3-methoxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (18c). Yield: 65%; mp: 121e124 ^ꢀC; ¹H NMR (400 MHz,

was added dropwise to

a

solution of compound 14 or 15

CDCl₃)

d

8.04 (d, J ¼ 5.2 Hz, 3H), 7.60e7.55 (m, 4H), 7.47e7.39 (m,

(1.86 mmol) in MeOH (50 mL). The reaction mixture was allowed to

stir at rt for 9 h. The organic solvent was evaporated under vacuum.

The crude residue was extracted between dichloromethane (70 mL)

and water (30 mL). The organic layer was washed with brine so-

lution (3 ꢂ 30 mL). The organic layer was dried over anhydrous

Na₂SO₄and evaporated under vacuum. The crude solid residue was

puriﬁed through column chromatography (hexane/EtOAc: 2:1 v/v)

to give the titled products 16 and 17.

3H), 7.27 (t, J ¼ 8.4 Hz, 2H), 7.11 (d, J ¼ 8.5 Hz, 2H) [AreH], 6.73 (brs,

1H, NH), 6.35 (brs, 1H, NH), 5.72 (brs, 1H, NH), 3.87 (s, 3H, OCH₃),

3.49

CH₂CH₂CH₂NHSO₂), 1.73 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(100 MHz, CDCl₃) 151.3, 147.6, 147.5, 139.1, 132.3, 129.7, 129.0,

(brs,

2H,

CH₂CH₂CH₂NHSO₂),

2.99

(brs,

2H,

d

128.9,128.4,127.4, 126.0,121.9,116.2,116.0,114.6,106.5 [AreC], 56.3

(OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 37.8 (CH₂CH₂CH₂NHSO₂), 29.9

(CH₂CH₂CH₂NHSO₂); LC/MS 638.5 (M^þþ 1).

4.1.2.5.1. 4-(4-(4-Fluoro-3-methoxyphenyl)-2-phenyl-1H-imida-

4.1.3.1.4. 4-Fluoro-N-(3-((4-(4-(4-ﬂuoro-3-methoxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (18d). Yield: 70%; mp: 122e125 ^ꢀC; ¹H NMR (400 MHz,

zol-5-yl)-2-(methylsulfonyl)pyrimidine (16). Yield: 80%; ¹H NMR

(400 MHz, CDCl₃)

d

8.64 (d, J ¼ 5.4 Hz, 1H), 8.11e8.08 (m, 3H),

7.46e7.44 (m, 2H), 7.31e7.29 (m, 2H), 7.12 (s, 1H), 7.10e7.09 (m, 1H)

CDCl₃)

d

8.08 (d, J ¼ 7.8 Hz, 1H), 8.06 (s, 1H), 8.02 (d, J ¼ 4.8 Hz, 1H),

[AreH], 3.89 (s, 3H, OCH₃), 3.24 (s, 3H, SCH₃); ¹³C NMR (100 MHz,

7.79 (s, 2H), 7.47e7.42 (m, 3H), 7.28 (d, J ¼ 7.2 Hz, 2H), 7.14e7.10 (m,

CDCl₃)

d 165.6, 157.9, 147.9, 133.6, 130.7, 130.1, 129.1, 128.5, 126.7,

4H) [AreH], 6.78 (brs,1H, NH), 6.21 (brs,1H, NH), 3.91 (s, 3H, OCH₃),

121.9, 118.8, 114.8 [AreC], 56.4 (OCH₃), 39.0 (SCH₃).

3.53

CH₂CH₂CH₂NHSO₂), 1.77 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(100 MHz, CDCl₃) 165.9, 163.7, 152.9, 147.8, 144.0, 138.6, 136.1,

(brs,

2H,

CH₂CH₂CH₂NHSO₂),

3.01

(brs,

2H,

4.1.2.5.2. 4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-1H-imida-

zol-5-yl)-2-(methylsulfonyl)pyrimidine (17). Yield: 81%; ¹H NMR

d

(400 MHz, CDCl₃)

d

8.63 (d, J ¼ 5.4 Hz, 1H), 8.08e8.06 (m, 2H),

129.8,128.7,126.2,122.0,116.4,114.7,110.6,106.3,105.5 [AreC], 56.4

(OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 38.0 (CH₂CH₂CH₂NHSO₂), 29.7

(CH₂CH₂CH₂NHSO₂); LC/MS 577.1 (M^þþ 1).

7.45e7.43 (m, 3H), 7.36 (s, 1H), 7.29 (s, 1H), 7.24 (d, J ¼ 8.4 Hz, 1H),

7.09 (dd, J ¼ 1.8, 8.0 Hz, 1H) [AreH], 3.89 (s, 3H, OCH₃), 3.22 (s, 3H,

SCH₃); ¹³C NMR (100 MHz, CDCl₃)

d

157.9, 155.2, 130.6, 130.3, 130.1,

4.1.3.1.5. N-(3-((4-(4-(4-Fluoro-3-methoxyphenyl)-2-phenyl-1H-

imidazol-5-yl)pyrimidin-2-yl)amino)propyl)-4-(triﬂuoromethyl)ben-

zenesulfonamide (18e). Yield: 74%; mp: 121e123 ^ꢀC; ¹H NMR

129.0, 128.4, 126.6, 123.8, 121.9, 113.2 [AreC], 56.3 (OCH₃), 38.9

(SCH₃).

11

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

(400 MHz, CDCl₃)

d

8.06 (d, J ¼ 1.4 Hz, 1H), 8.04 (d, J ¼ 5.2 Hz, 2H),

1H), 7.25 (s, 1H), 7.13 (d, J ¼ 8.2 Hz, 1H) [AreH], 6.80 (brs, 1H, NH),

7.89 (d, J ¼ 7.1 Hz, 2H), 7.72 (s, 1H), 7.70 (s, 1H), 7.48e7.43 (m, 3H),

7.28 (d, J ¼ 7.2 Hz, 2H), 7.14e7.12 (m, 1H), 7.11 (s, 1H) [AreH], 6.76

(brs, 1H, NH), 6.51 (brs, 1H, NH), 5.93 (brs, 1H, NH), 3.91 (s, 3H,

OCH₃), 3.54 (brs, 2H, CH₂CH₂CH₂NHSO₂), 3.04 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.77 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

6.20 (brs, 2H, 2 NH), 3.91 (s, 3H, OCH₃), 3.53 (s, 2H,

CH₂CH₂CH₂NHSO₂), 3.01 (s, 2H, CH₂CH₂CH₂NHSO₂), 1.75 (s, 2H,

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

d

155.0,132.5,130.1,

129.7, 129.1, 128.9, 126.8, 126.2, 122.8, 122.2, 113.1, 106.5, 56.3

(OCH₃), 40.2 (CH₂CH₂CH₂NHSO₂), 38.0 (CH₂CH₂CH₂NHSO₂), 29.7

(CH₂CH₂CH₂NHSO₂); LC/MS 575.6 (M^þþ 1).

(100 MHz, CDCl₃)

120.1, 116.1, 114.8, 106.0, 101.2 [AreC], 56.4 (OCH₃), 40.2

d 143.7, 130.0, 128.9, 127.4, 126.3, 126.2, 126.2,

4.1.3.1.11. 4-Chloro-N-(3-((4-(4-(4-chloro-3-methoxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (19b). Yield: 65%; mp: 121e124 ^ꢀC; ¹H NMR (400 MHz,

(CH₂CH₂CH₂NHSO₂),

38.1

(CH₂CH₂CH₂NHSO₂),

29.6

(CH₂CH₂CH₂NHSO₂); LC/MS 626.9 (M^þþ 1).

4.1.3.1.6. 3-Fluoro-N-(3-((4-(4-(4-ﬂuoro-3-methoxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (18f). Yield: 68%; mp: 103e106 ^ꢀC. ¹H NMR (400 MHz,

CDCl₃)

d

8.07e8.02 (m, 3H), 7.67 (d, J ¼ 7.2 Hz, 2H), 7.43 (t, J ¼ 7.2 Hz,

4H), 7.38 (d, J ¼ 7.8 Hz, 2H), 7.23 (s, 1H), 7.11 (d, J ¼ 8.0 Hz, 1H)

[AreH], 6.74 (brs, 1H, NH), 6.31 (brs, 1H, NH), 5.59 (brs, 1H, NH),

3.88 (s, 3H, OCH₃), 3.49 (brs, 2H, CH₂CH₂CH₂NHSO₂), 3.00 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.73 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

CDCl₃)

d

8.07e8.03 (m, 3H), 7.57 (d, J ¼ 7.2 Hz, 1H), 7.49e7.40 (m,

5H), 7.29e7.25 (m, 2H), 7.24e7.22 (m, 1H), 7.13 (d, J ¼ 8 Hz, 2H)

[AreH], 6.75 (brs, 1H, NH), 6.40 (brs, 1H, NH), 5.90 (brs, 1H, NH),

3.90 (s, 3H, OCH₃), 3.55 (brs, 2H, CH₂CH₂CH₂NHSO₂), 3.04 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.77 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(100 MHz, CDCl₃)

d 155.0, 147.5, 138.9, 138.6, 130.1, 129.6, 129.3,

129.0, 128.9, 128.3, 126.0, 122.7, 122.2, 113.1, 106.7 [AreC], 56.2

(OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 37.8 (CH₂CH₂CH₂NHSO₂), 29.9

(CH₂CH₂CH₂NHSO₂); LC/MS 610.7 (M^þþ 1).

(100 MHz, CDCl₃)

d 163.6, 161.1, 158.6, 153.9, 147.7, 130.9, 129.8,

128.9, 126.1, 122.6, 122.0, 119.8, 116.2, 114.1, 106.4 [AreC], 56.3

(OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 37.9 (CH₂CH₂CH₂NHSO₂), 29.8

(CH₂CH₂CH₂NHSO₂); LC/MS 577 (M^þþ 1).

4.1.3.1.12. 4-Bromo-N-(3-((4-(4-(4-chloro-3-methoxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (19c). Yield: 70%; mp: 135e138 ^ꢀC; ¹H NMR (400 MHz,

4.1.3.1.7. N-(3-((4-(4-(4-Fluoro-3-methoxyphenyl)-2-phenyl-1H-

i m i d a z o l - 5 - y l ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p y l ) - 4 -

methylbenzenesulfonamide (18g). Yield: 60%; mp: 109e111 ^ꢀC; ¹H

CDCl₃) d 8.06e8.01 (m, 3H), 7.57e7.54 (m, 4H), 7.45e7.40 (m, 3H),

7.37 (d, J ¼ 8.4 Hz, 1H), 7.23 (d, J ¼ 1.6 Hz, 1H), 7.10 (d, J ¼ 7.2 Hz, 1H)

[AreH], 6.72 (brs, 1H, NH), 6.33 (brs, 1H, NH), 5.54 (brs, 1H, NH),

3.88 (s, 3H, OCH₃), 3.49 (brs, 2H, CH₂CH₂CH₂NHSO₂), 3.00 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.73 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

NMR (400 MHz, CDCl₃)

d

8.08 (d, J ¼ 6.8 Hz, 2H), 8.01 (d, J ¼ 5.2 Hz,

1H), 7.65 (d, J ¼ 7.6 Hz, 2H), 7.45e7.37 (m, 3H), 7.29e7.27 (m, 2H),

7.25e7.23 (m, 2H), 7.11 (d, J ¼ 8.8 Hz, 2H) [AreH], 6.72 (brs, 1H, NH),

(100 MHz, CDCl₃)

d 157.2, 155.0, 147.5, 139.1, 132.3, 130.1, 129.6,

6.02 (brs, 2H,

2

NH), 3.89 (s, 3H, OCH₃), 3.52 (brs, 2H,

129.1, 128.9,128.4,127.4, 126.0,122.7, 122.2,113.1, 106.8 [AreC], 56.3

(OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 37.7 (CH₂CH₂CH₂NHSO₂), 29.9

(CH₂CH₂CH₂NHSO₂); LC/MS 654.7 (M^þþ 1).

CH₂CH₂CH₂NHSO₂), 2.99 (brs, 2H, CH₂CH₂CH₂NHSO₂), 1.75 (brs, 2H,

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

d

143.3, 136.9,

129.7, 128.9, 126.9, 126.2, 116.2, 116.0, 114.6, 106.3 [AreC], 56.3

(OCH₃), 40.2 (CH₂CH₂CH₂NHSO₂), 38.0 (CH₂CH₂CH₂NHSO₂), 29.7

(CH₂CH₂CH₂NHSO₂); LC/MS 573.1 (M^þþ 1).

4.1.3.1.13. N-(3-((4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - yl ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p yl ) - 4 -

ﬂuorobenzenesulfonamide (19d). Yield: 62%; m.p.: 137e140 ^ꢀC; ¹H

4.1.3.1.8. N-(3-((4-(4-(4-Fluoro-3-methoxyphenyl)-2-phenyl-1H-

i m i d a z o l - 5 - y l ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p y l ) - 4 -

methoxybenzenesulfonamide (18h). Yield: 68%; mp: 113e115 ^ꢀC; ¹H

NMR (400 MHz, CDCl₃)

d

8.03 (d, J ¼ 5.6 Hz, 1H), 8.01e7.99 (m, 2H),

7.71 (s, 2H), 7.41e7.37 (m, 3H), 7.32 (d, J ¼ 8.0 Hz, 1H), 7.19 (d,

J ¼ 1.2 Hz, 1H), 7.09e7.05 (m, 3H) [AreH], 6.70 (brs, 1H, NH), 6.33

(brs, 1H, NH), 3.82 (s, 3H, OCH₃), 3.41 (brs, 2H, CH₂CH₂CH₂NHSO₂),

NMR (400 MHz, CDCl₃)

d

8.07 (d, J ¼ 6.8 Hz, 2H), 8.01 (d, J ¼ 5.2 Hz,

1H), 7.69 (d, J ¼ 8.4 Hz, 2H), 7.45e7.40 (m, 3H), 7.27 (d, J ¼ 8.0 Hz,

2H), 7.11 (d, J ¼ 8.8 Hz, 2H), 6.89 (d, J ¼ 8.8 Hz, 2H) [AreH], 6.71 (brs,

1H, NH), 5.98 (brs, 2H, 2 NH), 3.88 (s, 3H, OCH₃), 3.82 (s, 3H, OCH₃),

3.51 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.98 (t,

CH₂CH₂CH₂NHSO₂), 1.74 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(100 MHz, CDCl₃) 162.8, 153.8, 147.7, 131.5, 129.7, 129.0, 128.9,

122.0, 116.2, 116.0, 114.6, 114.2, 106.3, 56.3 (OCH₃), 55.6 (OCH₃), 40.1

2.94

(brs,

2H,

CH₂CH₂CH₂NHSO₂),

1.67

(brs,

2H,

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

d

166.2,162.0,157.4,

154.9,147.5,136.1,130.1,129.6,129.1,128.9,126.0,122.6,122.2,116.4,

113.1, 106.8 [AreC], 56.2 (OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 37.7

(CH₂CH₂CH₂NHSO₂), 29.8 (CH₂CH₂CH₂NHSO₂); LC/MS 594

(M^þþ 1).

J

¼

4.4 Hz, 2H,

d

4.1.3.1.14. N-(3-((4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-

1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)-4-(triﬂuoromethyl)

benzenesulfonamide (19e). Yield: 64%; m.p.: 129e132 ^ꢀC; ¹H NMR

(CH₂CH₂CH₂NHSO₂),

38.0

(CH₂CH₂CH₂NHSO₂),

29.7

(CH₂CH₂CH₂NHSO₂); LC/MS 588.8 (M^þþ 1).

4.1.3.1.9. N-(3-((4-(4-(4-Fluoro-3-methoxyphenyl)-2-phenyl-1H-

imidazol-5-yl)pyrimidin-2-yl)amino)propyl)naphthalene-1-

sulfonamide (18i). Yield: 65%; mp: 135e137 ^ꢀC; ¹H NMR (400 MHz,

(400 MHz, CDCl₃)

d

8.00 (d, J ¼ 4.8 Hz, 1H), 7.95 (s, 2H), 7.78 (s, 2H),

7.62 (d, J ¼ 8.0 Hz, 2H), 7.30 (s, 3H), 7.23 (d, J ¼ 6.8 Hz, 1H), 7.13 (s,

1H), 6.98 (d, J ¼ 7.2 Hz, 1H) [AreH], 6.65 (brs, 1H, NH), 3.72 (s, 3H,

OCH₃), 3.34 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.90 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.60 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

CDCl₃)

d

8.63 (d, J ¼ 3.8 Hz,1H), 8.2 (d, J ¼ 4.8 Hz,1H), 8.07e8.03 (m,

4H), 7.92 (d, J ¼ 7.4 Hz, 1H), 7.59e7.55 (m, 2H), 7.50 (d, J ¼ 8.2 Hz,

1H), 7.46e7.42 (m, 3H), 7.29e7.28 (m, 2H), 7.13 (d, J ¼ 8.2 Hz, 2H)

[AreH], 6.71 (brs, 1H, NH), 6.35 (brs, 1H, NH), 5.65 (brs, 1H, NH),

3.90 (s, 3H, OCH₃), 3.52 (brs, 2H, CH₂CH₂CH₂NHSO₂), 3.01 (t,

(100 MHz, CDCl₃)

d 162.2, 157.6, 154.8, 147.6, 143.7, 134.1, 129.9,

128.8,127.3, 126.2, 124.6,122.4,121.9, 119.2,113.1, 106.9 [AreC], 56.0

(OCH₃), 40.2 (CH₂CH₂CH₂NHSO₂), 29.9 (CH₂CH₂CH₂NHSO₂), 21.0

(CH₂CH₂CH₂NHSO₂); LC/MS 644.9 (M^þþ 1).

J

¼

3.8 Hz, 2H, CH₂CH₂CH₂NHSO₂), 1.70 (brs, 2H,

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

d

151.4, 147.7, 134.2,

4.1.3.1.15. N-(3-((4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - yl ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p yl ) - 3 -

ﬂuorobenzenesulfonamide (19f). Yield: 71%; m.p.: 136.5e139.5 ^ꢀC;

129.7, 129.1, 128.9, 128.3, 126.8, 126.0, 124.3, 122.0, 116.2, 114.6,

106.3, 56.3, 40.1 (CH₂CH₂CH₂NHSO₂), 37.9 (CH₂CH₂CH₂NHSO₂),

30.1 (CH₂CH₂CH₂NHSO₂); LC/MS 609.8 (M^þþ 1).

¹H NMR (400 MHz, CDCl₃)

d

8.04 (d, J ¼ 5.6 Hz, 1H), 8.04e7.99 (m,

4.1.3.1.10. N-(3-((4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-

1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzenesulfonamide

(19a). Yield: 69%; mp: 122e125 ^ꢀC; ¹H NMR (400 MHz, CDCl₃)

2H), 7.50 (s, 1H), 7.42e7.37 (m, 4H), 7.32 (d, J ¼ 8.4 Hz, 1H),

7.22e7.19 (m, 2H), 7.06 (d, J ¼ 8.0 Hz, 1H) [AreH], 6.69 (brs, 1H, NH),

6.49 (brs, 1H, NH), 3.82 (s, 3H, OCH₃), 3.44 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 2.98 (brs, 2H, CH₂CH₂CH₂NHSO₂), 1.69 (brs, 2H,

d

8.12 (d, J ¼ 3.8 Hz, 2H), 7.96 (s, 1H), 7.79 (d, J ¼ 7.2 Hz, 2H), 7.54 (t,

J ¼ 6.8 Hz, 2H), 7.46 (d, J ¼ 7.2 Hz, 2H), 7.46e7.41 (m, 3H), 7.29 (s,

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

d 163.6, 161.1, 157.4,

12

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

154.9,147.5,142.2,142.1,131.0, 130.1,129.6,128.9,126.0,122.6,119.8,

114.3, 106.8 [AreC], 56.2 (OCH₃), 40.2 (CH₂CH₂CH₂NHSO₂), 37.7

(CH₂CH₂CH₂NHSO₂), 29.9 (CH₂CH₂CH₂NHSO₂); LC/MS 593

(M^þþ 1).

7.19 (dd, J ¼ 1.2, 8.4 Hz, 1H), 7.18e7.10 (m, 1H), 7.08e6.99 (m, 1H)

[AreH], 6.82 (brs, 1H, NH), 3.28 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.86

(brs, 2H, CH₂CH₂CH₂NHSO₂), 1.61 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³

C

NMR (100 MHz, CD₃OD)

d 162.0, 157.2, 152.9, 150.5, 147.5, 144.7,

4.1.3.1.16. N-(3-((4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - y l ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p y l ) - 4 -

methylbenzenesulfonamide (19g). Yield: 68%; m.p.: 143e146 ^ꢀC; ¹H

140.2, 132.2, 129.4, 128.8, 128.6, 126.6, 125.9, 120.9, 119.0, 115.7,

106.6, 40.2 (CH₂CH₂CH₂NHSO₂), 37.6 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 545 (M^þþ 1).

NMR (400 MHz, CDCl₃)

d

8.03 (d, J ¼ 7.2 Hz, 3H), 7.61 (d, J ¼ 6.8 Hz,

4.1.3.2.2. 4-Chloro-N-(3-((4-(4-(4-ﬂuoro-3-hydroxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (20b). Yield: 65%; m.p.: 139e142 ^ꢀC; ¹H NMR

2H), 7.38 (d, J ¼ 6.8 Hz, 3H), 7.33 (d, J ¼ 8.0 Hz, 1H), 7.21 (s, 2H), 7.19

(s, 1H), 7.08 (d, J ¼ 8.0 Hz, 1H) [AreH], 6.68 (brs, 1H, NH), 6.68 (brs,

1H, NH), 6.14 (brs, 1H, NH), 3.83 (s, 3H, OCH₃), 3.44 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 2.95 (brs, 2H, CH₂CH₂CH₂NHSO₂),1.68 (brs, 2H,

(400 MHz, CD₃OD)

d

8.12 (d, J ¼ 6.8 Hz, 1H), 8.03 (d, J ¼ 6.8 Hz, 2H),

7.79 (dd, J ¼ 1.6, 8.4 Hz, 1H), 7.73 (d, J ¼ 6.4 Hz, 1H), 7.62 (d,

J ¼ 8.8 Hz, 1H), 7.53e7.43 (m, 6H), 7.19e7.00 (m, 3H) [AreH],

3.34e3.30 (m, 2H, CH₂CH₂CH₂NHSO₂), 2.88 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.62 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

d 162.0,157.3,154.9,

147.5, 143.3, 136.9, 130.0, 129.7, 129.2, 128.8, 126.8, 126.1, 122.5,

122.2, 113.1, 106.7 [AreC], 56.2 (OCH₃), 40.2 (CH₂CH₂CH₂NHSO₂),

37.8 (CH₂CH₂CH₂NHSO₂), 29.8 (CH₂CH₂CH₂NHSO₂); LC/MS 590.8

(M^þþ 1).

(100 MHz, CD₃OD)

d 162.0, 152.9, 150.5, 147.5, 144.7, 139.1, 138.3,

129.4, 128.3, 126.0, 120.9, 119.0, 117.7, 115.4, 114.5, 106.5, 105.5

[AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.5 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 579.1 (M^þþ 1).

4.1.3.1.17. N-(3-((4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - y l ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p y l ) - 4 -

methoxybenzenesulfonamide (19h). Yield: 68%; m.p.: 124e127 ^ꢀC;

4.1.3.2.3. 4-Bromo-N-(3-((4-(4-(4-ﬂuoro-3-hydroxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (20c). Yield: 75%; m.p.: 132.5e135.5 ^ꢀC; ¹H NMR

¹H NMR (400 MHz, CDCl₃)

d

8.05e8.02 (m, 3H), 7.66 (d, J ¼ 7.8 Hz,

2H), 7.42e7.37 (m, 3H), 7.34 (d, J ¼ 8.4 Hz, 1H), 7.22 (d, J ¼ 1.6 Hz,

1H), 7.09 (dd, J ¼ 1.6, 8.1 Hz,1H), 6.87 (d, J ¼ 8.4 Hz, 2H) [AreH], 6.68

(brs, 1H, NH), 6.04 (brs, 1H, NH), 5.59 (brs, 1H, NH), 3.84 (s, 3H,

OCH₃), 3.80 (s, 3H, OCH₃), 3.45 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.95

(400 MHz, CD₃OD)

d

8.09 (d, J ¼ 4.6 Hz, 1H), 8.01 (d, J ¼ 7.2 Hz, 2H),

7.70e7.62 (m, 4H), 7.47 (t, J ¼ 3.8 Hz, 2H), 7.42 (t, J ¼ 7.2 Hz, 1H), 7.17

(dd, J ¼ 1.6, 8.4 Hz,1H), 7.13e7.10 (m,1H), 7.08e6.87 (m,1H), 6.86 (s,

1H) [AreH], 3.31 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.87 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.60 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(brs, 2H, CH₂CH₂CH₂NHSO₂), 1.69 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³

C

NMR (100 MHz, CDCl₃)

129.1, 129.0, 128.9, 126.1, 122.5, 114.2, 113.1, 106.7 [AreC], 56.2

(OCH₃), 55.6 (OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂), 37.9

d 162.8, 161.9, 157.2, 154.9, 147.5, 131.5, 130.1,

(100 MHz, CD₃OD)

d 162.0, 157.2, 152.9, 150.5, 147.5, 144.6, 139.6,

132.0, 129.4, 129.1, 128.6, 128.3, 126.7, 126.0, 120.8, 115.0, 106.6

[AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.5 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 624.8 (M^þþ 1).

(CH₂CH₂CH₂NHSO₂), 29.8 (CH₂CH₂CH₂NHSO₂); LC/MS 605.8

(M^þþ 1).

4.1.3.1.18. N-(3-((4-(4-(4-Chloro-3-methoxyphenyl)-2-phenyl-

1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)naphthalene-1-

sulfonamide (19i). Yield: 65%; m.p.: 125e128 ^ꢀC; ¹H NMR

4.1.3.2.4. 4-Fluoro-N-(3-((4-(4-(4-ﬂuoro-3-hydroxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (20d). Yield: 70%; m.p.: 129.2e132.2 ^ꢀC; ¹H NMR

(400 MHz, CDCl₃)

d

8.62 (s, 1H), 8.15 (d, J ¼ 6.8 Hz, 1H), 8.04e7.99

(400 MHz, CD₃OD)

d

8.10 (d, J ¼ 5.2 Hz, 1H), 8.01 (d, J ¼ 7.2 Hz, 2H),

(m, 4H), 7.90e7.88 (m, 1H), 7.56e7.51 (m, 2H), 7.44 (t, J ¼ 7.2 Hz,

1H), 7.46e7.32 (m, 4H), 7.21 (s, 1H), 7.08 (d, J ¼ 7.8 Hz, 1H) [AreH],

6.66 (brs, 1H, NH), 6.50 (brs, 1H, NH), 3.83 (s, 3H, OCH₃), 3.41 (brs,

2H, CH₂CH₂CH₂NHSO₂), 2.95 (brs, 2H, CH₂CH₂CH₂NHSO₂), 1.61 (brs,

7.88e7.81 (m, 2H), 7.48 (t, J ¼ 7.2 Hz, 2H), 7.43 (t, J ¼ 7.2 Hz, 1H),

7.23e7.15 (m, 3H), 7.18e7.12 (m, 1H), 7.10e7.09 (m, 1H), 6.86 (s, 1H)

[AreH], 3.32 (d, J ¼ 6.8 Hz, 2H, CH₂CH₂CH₂NHSO₂), 2.87 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.62 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CDCl₃)

d

162.1, 157.4,

(100 MHz, CD₃OD) d 166.1, 163.6, 161.9, 157.1, 152.9, 150.5, 147.5,

154.9, 147.5, 135.0, 130.1, 129.5, 129.1, 128.8, 128.1, 126.0, 124.3,

122.5, 113.1, 106.7 [AreC], 56.2 (OCH₃), 40.1 (CH₂CH₂CH₂NHSO₂),

30.0 (CH₂CH₂CH₂NHSO₂), 21.1 (CH₂CH₂CH₂NHSO₂); LC/MS 626.7

(M^þþ 1).

144.5, 136.6, 129.4, 128.6, 126.0, 120.9, 119.0, 115.9, 115.6, 106.5

[AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.6 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 563.1 (M^þþ 1).

4.1.3.2.5. N-(3-((4-(4-(4-Fluoro-3-hydroxyphenyl)-2-phenyl-1H-

imidazol-5-yl)pyrimidin-2-yl)amino)propyl)-4-(triﬂuoromethyl)ben-

zenesulfonamide (20e). Yield: 72%; m.p.: 138.5e141.5 ^ꢀC; ¹H NMR

4.1.3.2. General procedure for demethylation of compounds 18a-i and

19a-i to form the hydroxyl analogues 20a-i and 21a-i. To a solution

of compound 18a-i or 19a-i (0.35 mmol) and TBAB (58 mg,

0.18 mmol) in dry methylene chloride (3 mL), BBr₃(3.5 mmol) was

added dropwise at ꢁ70 ^ꢀC under N₂. The reaction mixture was

allowed to stir at the same temperature for 1 h, and then allowed to

warm to room temperature and stirred for another 3 h. The mixture

was quenched with saturated aqueous Na₂CO₃until alkalinization.

Ethyl acetate (5 mL) was added for extraction and the organic layer

was separated. The aqueous layer was extracted with ethyl acetate

(2 ꢂ 3 mL). The combined organic extract was washed with saline,

and dried over anhydrous Na₂SO₄. After evaporation of ethyl ace-

tate, the residue was puriﬁed by short column chromatography

(silica gel, ethyl acetate then switching to ethyl acetate-methanol

4:1 v/v) to yield the title compounds 20a-i and 21a-i.

(400 MHz, CD₃OD)

d

8.12 (d, J ¼ 5.4 Hz, 1H), 8.02 (d, J ¼ 7.1 Hz, 3H),

7.96 (d, J ¼ 7.2 Hz, 1H), 7.87e7.81 (m, 2H), 7.52e7.43 (m, 3H), 7.18

(dd, J ¼ 1.84, 8.4 Hz, 1H), 7.15e7.10 (m, 1H), 7.09e7.00 (m, 1H), 6.87

(s, 1H) [AreH], 3.34e3.30 (m, 2H, CH₂CH₂CH₂NHSO₂), 2.91 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.64 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(100 MHz, CD₃OD)

d 162.0, 157.2, 150.5, 147.5, 144.7, 144.4, 133.3,

129.0, 128.5, 127.3, 125.9, 124.9, 122.2, 120.7, 119.0, 115.4, 106.5

[AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.5 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 612.9 (M^þþ 1).

4.1.3.2.6. 3-Fluoro-N-(3-((4-(4-(4-ﬂuoro-3-hydroxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (20f). Yield: 74%; m.p.: 133e136 ^ꢀC; ¹H NMR

(400 MHz, CD₃OD)

d

8.11 (d, J ¼ 4.6 Hz, 1H), 8.02 (d, J ¼ 7.2 Hz, 2H),

7.6 (d, J ¼ 7.8 Hz, 1H), 7.53e7.41 (m, 5H), 7.34e7.29 (m, 1H), 7.18 (dd,

J ¼ 2.2, 8.4 Hz, 1H), 7.15e7.10 (m, 1H), 7.07e6.99 (m, 1H), 6.84 (s, 1H)

[AreH], 3.34e3.30 (m, 2H, CH₂CH₂CH₂NHSO₂), 2.91 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.63 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

4.1.3.2.1. N-(3-((4-(4-(4-Fluoro-3-hydroxyphenyl)-2-phenyl-1H-

imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzenesulfonamide

(20a). Yield: 72%; m.p.: 144e147 ^ꢀC; ¹H NMR (400 MHz, CD₃OD)

d

8.06 (d, J ¼ 4.6 Hz, 1H), 8.01 (d, J ¼ 7.2 Hz, 2H), 7.82e7.76 (m, 2H),

(100 MHz, CD₃OD)

129.1, 128.5, 125.9, 122.5, 120.8, 119.2, 119.0, 115.4, 113.5, 106.5

d 162.0, 157.2, 150.5, 147.5, 144.7, 142.6, 131.0,

7.51 (t, J ¼ 6.8 Hz, 1H), 7.45 (t, J ¼ 8.0 Hz, 4H), 7.38 (d, J ¼ 6.8 Hz, 1H),

13

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

[AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.6 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 563.1 (M^þþ1).

4.1.3.2.12. 4-Bromo-N-(3-((4-(4-(4-chloro-3-hydroxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (21c). Yield: 72%; m.p.: 141e144 ^ꢀC; ¹H NMR

4.1.3.2.7. N-(3-((4-(4-(4-Fluoro-3-hydroxyphenyl)-2-phenyl-1H-

i m i d a z o l - 5 - y l ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p y l ) - 4 -

methylbenzenesulfonamide (20g). Yield: 70%; m.p.: 136.5e139.5 ^ꢀC;

(400 MHz, CD₃OD)

d

8.08 (d, J ¼ 5.2 Hz, 1H), 7.99 (d, J ¼ 7.1 Hz, 2H),

7.66e7.58 (m, 4H), 7.46e43 (m, 2H), 7.41e7.37 (m, 1H), 7.32 (d,

J ¼ 5.2 Hz, 1H), 7.16 (d, J ¼ 1.8 Hz, 1H), 7 (dd, J ¼ 1.8, 5.2 Hz, 1H), 6.85

(s, 1H) [AreH], 3.24 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.85 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.58 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

¹H NMR (400 MHz, CD₃OD)

d

8.09 (d, J ¼ 5.2 Hz, 1H), 8.01 (d,

J ¼ 7.4 Hz, 2H), 7.69e7.59 (m, 2H), 7.49e7.40 (m, 3H), 7.30e7.25 (m,

2H), 7.17 (dd, J ¼ 1.9, 8.4 Hz, 1H), 7.14e7.07 (m, 1H), 7.01e6.98 (m,

1H), 6.83 (s, 1H) [AreH], 3.33e3.29 (m, 2H, CH₂CH₂CH₂NHSO₂),

(100 MHz, CD₃OD)

d 162.0, 157.3, 152.7, 147.7, 139.6, 132.1, 129.6,

2.85

(brs,

2H,

CH₂CH₂CH₂NHSO₂),

1.61

(brs,

162.0, 157.2,

2H,

129.1, 128.3, 126.7, 126.0, 121.3, 120.5, 117.6, 106.7 [AreC], 40.3

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CD₃OD)

d

(CH₂CH₂CH₂NHSO₂),

37.6

(CH₂CH₂CH₂NHSO₂),

29.4

152.9, 150.5, 147.5, 144.5, 143.1, 137.3, 129.3, 128.6, 126.6, 126.0,

120.9, 119.0, 115.6, 106.5 [AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.6

(CH₂CH₂CH₂NHSO₂), 29.4 (CH₂CH₂CH₂NHSO₂); LC/MS 559

(M^þþ 1).

(CH₂CH₂CH₂NHSO₂); LC/MS 641.6 (M^þþ 1).

4.1.3.2.13. N-(3-((4-(4-(4-Chloro-3-hydroxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - yl ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p yl ) - 4 -

ﬂuorobenzenesulfonamide (21d). Yield: 70%; m.p.: 145.5e148.5 ^ꢀC;

4.1.3.2.8. N-(3-((4-(4-(4-Fluoro-3-hydroxyphenyl)-2-phenyl-1H-

i m i d a z o l - 5 - y l ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p y l ) - 4 -

hydroxybenzenesulfonamide (20h). Yield: 74%; m.p.: 155e158 ^ꢀC;

¹H NMR (400 MHz, CD₃OD)

d

8.13 (d, J ¼ 5.2 Hz, 1H), 8.02 (d,

J ¼ 6.8 Hz, 2H), 7.89e7.83 (m, 2H), 7.52e7.43 (m, 3H), 7.36 (d,

J ¼ 8.4 Hz, 1H), 7.28e7.20 (m, 2H), 7.16 (d, J ¼ 1.6 Hz, 1H), 7.06e6.95

(m, 1H), 6.93 (s, 1H) [AreH], 3.22 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.85

¹H NMR (400 MHz, CD₃OD)

d

8.10 (d, J ¼ 5.2 Hz, 1H), 8.01 (d,

J ¼ 7.2 Hz, 2H), 7.61 (d, J ¼ 8.2 Hz, 2H), 7.50e7.41 (m, 3H), 7.19 (dd,

J ¼ 1.9, 8.2 Hz, 1H), 7.15e7.10 (m, 1H), 7.04e7.01 (m, 1H), 6.85 (d,

J ¼ 8.8 Hz, 2H) [AreH], 3.34e3.30 (m, 2H, CH₂CH₂CH₂NHSO₂), 2.84

(brs, 2H, CH₂CH₂CH₂NHSO₂), 1.60 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³

C

NMR (100 MHz, CD₃OD)

d 156.9, 152.7, 147.7, 136.6, 129.6, 129.5,

129.4, 128.6, 128.6, 126.0, 125.8, 121.2, 120.6, 117.5, 115.9, 115.7, 106.5

[AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.6 (CH₂CH₂CH₂NHSO₂), 29.3

(CH₂CH₂CH₂NHSO₂); LC/MS 580.9 (M^þþ 1).

(brs, 2H, CH₂CH₂CH₂NHSO₂), 1.63 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³

C

NMR (100 MHz, CD₃OD)

d 161.3, 157.2, 152.9, 150.5, 147.5, 144.7,

130.2, 129.4, 128.8, 128.5, 126.0, 120.9, 119.0, 115.6, 113.9, 106.5

[AreC], 40.1 (CH₂CH₂CH₂NHSO₂), 37.6 (CH₂CH₂CH₂NHSO₂), 29.3

(CH₂CH₂CH₂NHSO₂); LC/MS 561.1 (M^þþ 1).

4.1.3.2.14. N-(3-((4-(4-(4-Chloro-3-hydroxyphenyl)-2-phenyl-

1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)-4-(triﬂuoromethyl)

benzenesulfonamide (21e). Yield: 72%; m.p.: 160e163 ^ꢀC; ¹H NMR

4.1.3.2.9. N-(3-((4-(4-(4-Fluoro-3-hydroxyphenyl)-2-phenyl-1H-

imidazol-5-yl)pyrimidin-2-yl)amino)propyl)naphthalene-1-

sulfonamide (20i). Yield: 72%; m.p.: 140e143 ^ꢀC; ¹H NMR

(400 MHz, CD₃OD)

d

8.15 (d, J ¼ 5.2 Hz, 1H), 8.03 (d, J ¼ 7.6 Hz, 3H),

7.97 (d, J ¼ 7.6 Hz,1H), 7.89e7.83 (m, 2H), 7.54e7.43 (m, 3H), 7.36 (d,

J ¼ 8.4 Hz, 1H), 7.17 (s, 1H), 7.04 (d, J ¼ 8.8 Hz, 1H), 6.95 (s, 1H)

[AreH], 3.28 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.90 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.63 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(400 MHz, CD₃OD)

d

8.64 (d, J ¼ 8.8 Hz, 1H), 8.12 (d, J ¼ 6.8 Hz, 1H),

8.02 (d, J ¼ 4.8 Hz,1H), 7.99 (t, J ¼ 7.2 Hz, 3H), 7.86 (d, J ¼ 8.0 Hz,1H),

7.6 (t, J ¼ 1.2 Hz, 1H), 7.58e7.50 (m, 1H), 7.48e7.44 (m, 3H),

7.42e7.38 (m, 1H), 7.14 (d, J ¼ 7.2 Hz, 1H), 7.05e7.01 (m, 1H), 6.89 (s,

1H) [AreH], 3.17 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.83 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 1.49 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(100 MHz, CD₃OD)

d 157.0, 129.3, 129.1, 128.6, 128.6, 127.4, 127.3,

126.0, 125.8, 124.7, 121.2, 118.8 [AreC], 40.3 (CH₂CH₂CH₂NHSO₂),

38.2 (CH₂CH₂CH₂NHSO₂), 29.5 (CH₂CH₂CH₂NHSO₂); LC/MS 630.7

(M^þþ 1).

(100 MHz, CD₃OD)

d

161.9, 157.1, 152.9, 150.4, 147.4, 144.6, 135.1,

4.1.3.2.15. N-(3-((4-(4-(4-Chloro-3-hydroxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - yl ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p yl ) - 3 -

ﬂuorobenzenesulfonamide (21f). Yield: 70%; m.p.: 155e158 ^ꢀC; ¹H

133.7, 129.4, 128.2, 127.6, 126.5, 124.4, 123.9, 120.8, 115.6, 106.5

[AreC], 40.1 (CH₂CH₂CH₂NHSO₂), 37.6 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 595.1 (M^þþ 1).

NMR (400 MHz, CD₃OD)

d

8.09 (d, J ¼ 5.2 Hz, 1H), 7.99 (d, J ¼ 7.2 Hz,

4.1.3.2.10. N-(3-((4-(4-(4-Chloro-3-hydroxyphenyl)-2-phenyl-

1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzenesulfonamide

(21a). Yield: 70%; m.p.: 133.5e136.5 ^ꢀC; ¹H NMR (400 MHz,

2H), 7.65e7.58 (m, 1H), 7.53e7.49 (m, 2H), 7.48e7.73 (m, 3H),

7.42e7.38 (m, 1H), 7.34e7.31 (m, 1H), 7.29e7.27 (m, 1H), 7.16 (d,

J ¼ 1.6 Hz, 1H), 7.02 (dd, J ¼ 1.6, 8.4 Hz, 1H), 6.86 (s, 1H) [AreH], 3.26

(brs, 2H, CH₂CH₂CH₂NHSO₂), 2.87 (brs, 2H, CH₂CH₂CH₂NHSO₂), 1.61

CD₃OD)

d

8.10 (d, J ¼ 5.4 Hz, 1H), 8.01 (d, J ¼ 6.4 Hz, 2H), 7.77 (d,

J ¼ 7.2 Hz, 2H), 7.53 (d, J ¼ 6.8 Hz, 1H), 7.50e7.46 (m, 4H), 7.46e7.41

(brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CD₃OD)

d 163.7,

(m, 1H), 7.34 (d, J ¼ 8.0 Hz, 1H), 7.16 (d, J ¼ 1.8 Hz, 1H), 7.01 (dd,

161.2, 157.2, 152.7, 147.6, 142.5, 132.4, 130.9, 129.3, 128.6, 126.0,

122.6, 121.2, 120.5, 119.2, 113.5, 106.6 [AreC], 40.3

J

¼

1.8, 8.4 Hz, 1H), 6.86 (s, 1H) [AreH], 3.26 (brs, 2H,

CH₂CH₂CH₂NHSO₂), 2.85 (brs, 2H, CH₂CH₂CH₂NHSO₂), 1.59 (brs, 2H,

(CH₂CH₂CH₂NHSO₂),

37.6

(CH₂CH₂CH₂NHSO₂),

29.4

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CD₃OD)

d

161.9, 157.2,

(CH₂CH₂CH₂NHSO₂); LC/MS 580.8 (M^þþ 1).

152.7, 147.6, 140.3, 132.1, 129.5, 129.1, 128.8, 128.6, 128.6, 126.5,

126.0, 121.2, 120.5, 117.5, 106.6 [AreC], 40.2 (CH₂CH₂CH₂NHSO₂),

37.6 (CH₂CH₂CH₂NHSO₂), 29.4 (CH₂CH₂CH₂NHSO₂); LC/MS 562

(M^þþ 1).

4.1.3.2.16. N-(3-((4-(4-(4-Chloro-3-hydroxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - yl ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p yl ) - 4 -

methylbenzenesulfonamide (21g). Yield: 69%; m.p.: 138e141 ^ꢀC; ¹H

NMR (400 MHz, DMSO‑d₆)

d 10.21 (s, 1H), 8.23 (s, 1H), 8.11 (d,

4.1.3.2.11. 4-Chloro-N-(3-((4-(4-(4-chloro-3-hydroxyphenyl)-2-

phenyl-1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)benzene-

sulfonamide (21b). Yield: 72%; m.p.: 127e130 ^ꢀC; ¹H NMR

J ¼ 6.4 Hz, 2H), 7.64 (d, J ¼ 7.2 Hz), 7.5 (t, J ¼ 7.8 Hz, 2H), 7.42 (d,

J ¼ 6.0 Hz, 2H), 7.34 (d, J ¼ 7.8 Hz, 4H), 7.07 (s, 1H), 6.76 (s, 1H) [Ar

and exchangeable H], 3.36 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.63 (brs,

2H, CH₂CH₂CH₂NHSO₂), 1.36 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR

(400 MHz, CD₃OD)

d

8.11 (d, J ¼ 5.2 Hz, 1H), 8.01 (d, J ¼ 7.2 Hz, 2H),

7.79 (t, J ¼ 8.4 Hz,1H), 7.72 (d, J ¼ 8.4 Hz,1H), 7.51e7.40 (m, 6H), 7.33

(d, J ¼ 8.2 Hz, 1H), 7.16 (d, J ¼ 1.6 Hz, 1H), 7.01 (dd, J ¼ 1.8, 8.2 Hz,

1H), 6.87 (s, 1H) [AreH], 3.25 (brs, 2H, CH₂CH₂CH₂NHSO₂), 2.85

(100 MHz, DMSO‑d₆) d 162.3, 142.9, 139.7, 138.1, 130.4, 130.0, 129.2,

128.5, 127.0, 125.8, 125.4 [AreC], 30.9 (CH₂CH₂CH₂NHSO₂), 29.6

(CH₂CH₂CH₂NHSO₂), 21.4 (CH₂CH₂CH₂NHSO₂); LC/MS 576.9

(M^þþ 1).

(brs, 2H, CH₂CH₂CH₂NHSO₂), 1.59 (brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³

C

NMR (100 MHz, CD₃OD)

d

161.9, 157.3, 152.7, 147.7, 139.1, 138.3,

4.1.3.2.17. N-(3-((4-(4-(4-Chloro-3-hydroxyphenyl)-2-phenyl-

1 H - i m i d a z o l - 5 - yl ) p y r i m i d i n - 2 - y l ) a m i n o ) p r o p yl ) - 4 -

hydroxybenzenesulfonamide (21h). Yield: 70%; m.p.: 159e162 ^ꢀC;

129.5, 129.1, 129.1, 128.6, 126.0, 121.2, 120.5, 118.9, 117.6, 106.6, 105.5

[AreC], 40.2 (CH₂CH₂CH₂NHSO₂), 37.6 (CH₂CH₂CH₂NHSO₂), 29.4

(CH₂CH₂CH₂NHSO₂); LC/MS 596.8 (M^þþ 1).

¹H NMR (400 MHz, CD₃OD)

d

8.11 (d, J ¼ 5.2 Hz, 1H), 8.01 (d,

14

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

J ¼ 7.2 Hz, 2H), 7.61 (d, J ¼ 8.4 Hz, 2H), 7.51e7.42 (m, 3H), 7.35 (d,

J ¼ 8.0 Hz, 1H), 7.17 (d, J ¼ 6.0 Hz, 1H), 7.03 (dd, J ¼ 2.0, 8.0 Hz, 1H),

6.85 (d, J ¼ 8.8 Hz, 3H) [AreH], 3.27 (brs, 2H, CH₂CH₂CH₂NHSO₂),

4.3. Docking study

The X-ray crystal structures of BRAF V600E oncogenic mutant

kinase in complex with dabrafenib (PDB ID: 4XV2) and p38 in

2.82

(brs,

2H,

CH₂CH₂CH₂NHSO₂),

1.61

(brs,

2H,

a

CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CD₃OD)

d

161.9, 161.3,

complex with SB203580 (PDB ID: 3GCP) were downloaded from

the protein data bank (www.rcsb.org) in PDB format. The 2D

structure of the target compounds were drawn using ChemDraw

software. Molecular Operating Environment (MOE, 2014.0901)

software was used for the molecular docking operation of the target

compounds 18a-i, 19a-i, 20a-i, and 21a-i with BRAF^V600Ekinase

157.2,152.7,147.7,130.2,129.5,129.1,128.6,126.0,121.2,120.5,120.5,

117.5, 115.2, 113.9, 106.5 [AreC], 40.1 (CH₂CH₂CH₂NHSO₂), 37.6

(CH₂CH₂CH₂NHSO₂), 29.3 (CH₂CH₂CH₂NHSO₂); LC/MS 577.1

(M^þþ 1).

4.1.3.2.18. N-(3-((4-(4-(4-Chloro-3-hydroxyphenyl)-2-phenyl-

1H-imidazol-5-yl)pyrimidin-2-yl)amino)propyl)naphthalene-1-

sulfonamide (21i). Yield: 70%; m.p.: 129.5e132.5 ^ꢀC; ¹H NMR

enzyme domain (PDB ID: 4XV2) and p38a kinase enzyme domain

(PDB ID: 3GCP). Both kinases were prepared for the molecular

docking procedure by applying 3D protonation of both enzyme

amino acids and the native ligands (dabrafenib and SB203580). In

addition, water of crystallization was removed from both BRAF^V600E

(400 MHz, CD₃OD)

d

8.65 (t, J ¼ 8.4 Hz, 1H), 8.19e8.12 (m, 1H), 8.05

(d, J ¼ 5.2 Hz, 1H), 7.99 (d, J ¼ 6.4 Hz, 3H), 7.89 (d, J ¼ 3.8 Hz, 1H),

7.61 (t, J ¼ 7.2 Hz, 1H), 7.53 (t, J ¼ 7.8 Hz, 1H), 7.46e7.41 (m, 4H), 7.25

(d, J ¼ 7.2 Hz, 1H), 7.12 (s, 1H), 6.89 (d, J ¼ 6.4 Hz, 1H) [AreH], 3.15

(brs, 2H, CH₂CH₂CH₂NHSO₂), 2.82 (brs, 2H, CH₂CH₂CH₂NHSO₂),1.48

kinase and p38a kinase domains. The active site of both enzymes

was isolated. The docking simulation of native ligands (dabrafenib

(brs, 2H, CH₂CH₂CH₂NHSO₂); ¹³C NMR (100 MHz, CD₃OD)

d 161.7,

and SB203580) with the active site of BRAF^V600Ekinase (PDB ID:

157.0,152.7,147.6, 135.2,134.3,133.7, 132.4,129.1, 128.6,127.6,126.5,

125.8, 124.4, 121.2, 117.5, 106.5 [AreC], 40.1 (CH₂CH₂CH₂NHSO₂),

37.6 (CH₂CH₂CH₂NHSO₂), 29.4 (CH₂CH₂CH₂NHSO₂); LC/MS 612.6

(M^þþ 1).

4XV2) and p38a kinase (PDB ID: 3GCP) was investigated in order to

validate the docking protocol. Both 3D protonation and energy

minimization were performed for the target compounds using

MOE, 2014.0901 software.

Declaration of competing interest

4.2. Biological assays

The authors declare that they have no known competing

ﬁnancial interests or personal relationships that could have

appeared to inﬂuence the work reported in this paper.

4.2.1. In vitro BRAF^V600E/p38

a kinase inhibitory activity

Thermo Fisher Scientiﬁc, SelectScreen Kinase Proﬁling Services

was used for screening of the target compounds in single point

concentrations mode, and 5-points titration mode for IC₅₀deter-

mination. Staurosporine and PP2 were tested as reference standard

Acknowledgments

compounds in BRAF^V600Eand p38

a

assays, respectively.

Assay protocol: as reported on ThermoFisher Scientiﬁc website

using 100 M concentration of ATP (https://www.thermo ﬁ sher.

The authors are thankful to Korea Institute of Science and

Technology (KIST) for ﬁnancially supporting this work (project

2E30341 & 2E31130).

m

com/ae/en/home/industrial/pharma-biopharma/drug-discovery-

development/target-and-lead-identi ﬁ cation-and-validation/

kinasebiology/kinase-activity-assays.html).

Appendix A. Supplementary data

Supplementary data to this article can be found online at

https://doi.org/10.1016/j.ejmech.2021.113277.

4.2.2. TNF-a production assay

RAW 264.7 macrophages were obtained from the Korean Cell

Line Bank (Seoul, Korea). Cells were plated at 2 ꢂ 10⁵cells per well

in 24-well plates and incubated overnight. Following treatment

with various compound 20h concentrations (0.05, 0.1, 0.2, 0.4, 0.8,

References

[1] M. Aouadi, B. Binetruy, L. Caron, Y. Le Marchand-Brustel, F. Bost, Role of

MAPKs in development and differentiation: lessons from knockout mice,

Biochimie 88 (2006) 1091e1098.

[2] Y. Zhang, C. Dong, Regulatory mechanisms of mitogen-activated kinase

signaling, Cell. Mol. Life Sci. 64 (2007) 2771e2789.

[3] E.K. Kim, E.-J. Choi, Compromised MAPK signaling in human diseases: an

update, Arch. Toxicol. 89 (2015) 867e882.

[4] A.M. Kidgera, L.K. Rushwortha, J. Stellziga, J. Davidsona, C.J. Bryantb,

C. Bayleyb, E. Caddyeb, T. Rogersc, S.M. Keysea, C.J. Cauntb, Dual-speci ﬁ city

Phosphatase 5 Controls the Localized Inhibition, Propagation, and Trans-

forming Potential of ERK Signaling, 2017.

or 1.6

10.0 M) for 1 h, cells were treated with lipopolysaccharide (1

ml) for 24 h. Productions of TNF- , in the cell cultured supernatants

mM) or BAY 11e7082 concentrations (0.5, 1.0, 5.0, and

m

g/

a

were measured using EIA kits (R&D Systems, MN, USA) according to

the manufacturer’s instructions [42].

[5] R. Roskoski Jr., Targeting ERK1/2 protein-serine/threonine kinases in human

cancers, Pharmacol. Res. 142 (2019) 151e168.

4.2.3. In vitro cytotoxicity assay

[6] P.I. Poulikakos, C. Zhang, G. Bollag, K.M. Shokat, N. Rosen, RAF inhibitors

transactivate RAF dimers and ERK signalling in cells with wild-type BRAF,

Nature 464 (2010) 427e430.

[7] H. Arkenau, R. Kefford, G. Long, Targeting BRAF for patients with melanoma,

Br. J. Canc. 104 (2011) 392e398.

[8] X. Liu, S. Qu, R. Liu, C. Sheng, X. Shi, G. Zhu, A.K. Murugan, H. Guan, H. Yu,

Y. Wang, TERT promoter mutations and their association with BRAF V600E

mutation and aggressive clinicopathological characteristics of thyroid cancer,

J. Clin. Endocrinol. Metab. 99 (2014) E1130eE1136.

[9] J.A. Jakob, R.L. Bassett Jr., C.S. Ng, J.L. Curry, R.W. Joseph, G.C. Alvarado,

M.L. Rohlfs, J. Richard, J.E. Gershenwald, K.B. Kim, NRAS mutation status is an

independent prognostic factor in metastatic melanoma, Cancer 118 (2012)

4014e4023.

LOX-IMVI human melanoma cell line was obtained from

American Type Culture Collection (ATCC). The cells were cultured

using DMEM (Invitrogen/Life Technologies) supplemented with

10% FBS (Hyclone), 10 mg/mL of insulin (Sigma), and 1% penicillin-

streptomycin. All of the other chemicals and reagents were pur-

chased from Sigma or Invitrogen. Plate cells (cells density

1.2e1.8 ꢂ 10,000 cells/well) in a volume of 100

mL complete growth

L of the tested compound per well in a 96-well

medium þ 100

m

plate for 24 h before the MTT assay [43].

IOSE-80PC (human ovarian epithelial cell) were cultured in

RPMI 1640, which also added FBS and penicillin-streptomycin. Cells

were seeded (1 ꢂ 10⁵/mL) and incubated with various concentra-

[10] A.M. Menzies, T. Lum, J.S. Wilmott, J. Hyman, R.F. Kefford, J.F. Thompson,

S. O ’Toole, G.V. Long, R.A. Scolyer, Intrapatient homogeneity of BRAFV600E

expression in melanoma, Am. J. Surg. Pathol. 38 (2014) 377e382.

[11] G. Kim, A.E. McKee, Y.-M. Ning, M. Hazarika, M. Theoret, J.R. Johnson, Q.C. Xu,

tions (0, 0.001, 0.01, 0.1, 1, or 10 mM) of tested compounds for 24 h.

15

E.M.H. Ali, R.F.A. El-Telbany, M.S. Abdel-Maksoud et al.

European Journal of Medicinal Chemistry 215 (2021) 113277

X-ray structure of p38 bound to TAK-715: comparison with three classic

inhibitors, Acta Crystallogr. Sect. D Biol. Crystallogr. 68 (2012) 1041e1050.

[29] Y. Jiang, H. Gram, M. Zhao, L. New, J. Gu, L. Feng, F. Di Padova, R.J. Ulevitch,

J. Han, Characterization of the structure and function of the fourth member of

S. Tang, R. Sridhara, X. Jiang, FDA approval summary: vemurafenib for treat-

ment of unresectable or metastatic melanoma with the BRAFV600E mutation,

Clin. Canc. Res. 20 (2014) 4994e5000.

a

[12] L. Odogwu, L. Mathieu, G. Blumenthal, E. Larkins, K.B. Goldberg, N. Grif ﬁ n,

K. Bijwaard, E.Y. Lee, R. Philip, X. Jiang, FDA approval summary: dabrafenib

and trametinib for the treatment of metastatic non-small cell lung cancers

harboring BRAF V600E mutations, Oncol. 23 (2018) 740.

p38 group mitogen-activated protein kinases, p38 d , J. Biol. Chem. 272 (1997)

30122e30128.

[30] Z. Zhang, B. Wang, S. Wu, Y. Wen, X. Wang, X. Song, J. Zhang, L. Hou, W. Chen,

PD169316, a speci ﬁ c p38 inhibitor, shows antiviral activity against Entero-

virus71, Virology 508 (2017) 150e158.

[31] K. Ward, J. Proksch, K. Salyers, L. Azzarano, J. Morgan, T. Roethke, J. McSurdy-

Freed, M. Levy, B. Smith, SB-242235, a selective inhibitor of p38 mitoge-

nactivated protein kinase. I: preclinical pharmacokinetics, Xenobiotica 32

(2002) 221e233.

[13] S. Wilhelm, C. Carter, M. Lynch, T. Lowinger, J. Dumas, R.A. Smith, B. Schwartz,

R. Simantov, S. Kelley, Discovery and development of sorafenib: a multikinase

inhibitor for treating cancer, Nat. Rev. Drug Discov. 5 (2006) 835e844.

[14] W. Hugo, H. Shi, L. Sun, M. Piva, C. Song, X. Kong, G. Moriceau, A. Hong,

K.B. Dahlman, D.B. Johnson, Non-genomic and immune evolution of mela-

noma acquiring MAPKi resistance, Cell 162 (2015) 1271e1285.

[15] T. Amaral, T. Sinnberg, F. Meier, C. Krepler, M. Levesque, H. Niessner, C. Garbe,

The mitogen-activated protein kinase pathway in melanoma part Ieactivation

and primary resistance mechanisms to BRAF inhibition, Eur. J. Canc. 73 (2017)

85e92.

[32] M.M. Madkour, H.S. Anbar, M.I. El-Gamal, Current Status and Future Prospects

of p38 a /MAPK14 kinase and its inhibitors, Eur. J. Med. Chem. 213 (2021)

113216.

[33] E.M. Ali, M.S. Abdel-Maksoud, U.M. Ammar, K.I. Mersal, K.H. Yoo, P. Jooryeong,

C.-H. Oh, Design, synthesis, and biological evaluation of novel imidazole de-

rivatives possessing terminal sulphonamides as potential BRAFV600E in-

hibitors, Bioorg. Chem. 106 (2020) 104508.

[34] G. Wagner, S. Laufer, Small molecular anti-cytokine agents, Med. Res. Rev. 26

(2006) 1e62.

[35] A.D. Bachstetter, L.J. Van Eldik, The p38 MAP kinase family as regulators of

proin ﬂ ammatory cytokine production in degenerative diseases of the CNS,,

Aging Dis. 1 (2010) 199.

[36] M.S. Abdel-Maksoud, M.-R. Kim, M.I. El-Gamal, M.M.G. El-Din, J. Tae, H.S. Choi,

K.-T. Lee, K.H. Yoo, C.-H. Oh, Design, synthesis, in vitro antiproliferative

evaluation, and kinase inhibitory effects of a new series of imidazo [2, 1-b]

thiazole derivatives, Eur. J. Med. Chem. 95 (2015) 453e463.

[16] C.M. Johannessen, J.S. Boehm, S.Y. Kim, S.R. Thomas, L. Wardwell, L.A. Johnson,

C.M. Emery, N. Stransky, A.P. Cogdill, J. Barretina, COT drives resistance to RAF

inhibition through MAP kinase pathway reactivation, Nature 468 (2010)

968e972.

[17] B. Sanchez-Laorden, A. Viros, M.R. Girotti, M. Pedersen, G. Saturno, A. Zambon,

D. Niculescu-Duvaz, S. Turajlic, A. Hayes, M. Gore, BRAF inhibitors induce

metastasis in RAS mutant or inhibitor-resistant melanoma cells by reac-

tivating MEK and ERK signaling, Sci. Signal. 7 (2014) ra30-ra30.

[18] P.I. Poulikakos, Y. Persaud, M. Janakiraman, X. Kong, C. Ng, G. Moriceau, H. Shi,

M. Ate ﬁ , B. Titz, M.T. Gabay, RAF inhibitor resistance is mediated by dimer-

ization of aberrantly spliced BRAF (V600E), Nature 480 (2011) 387e390.

[19] M. Kuske, D. Westphal, R. Wehner, M. Schmitz, S. Beissert, C. Praetorius,

F. Meier, Immunomodulatory effects of BRAF and MEK inhibitors: implications

for melanoma therapy, Pharmacol. Res. 136 (2018) 151e159.

[37] S.D.S. Mendes, A. Candi, M. Vansteenbrugge, M.-R. Pignon, H. Bult,

K.Z. Boudjeltia, C. Munaut, M. Raes, Microarray analyses of the effects of NF-k B

[20] K.M. Ilieva, I. Correa, D.H. Josephs, P. Karagiannis, I.U. Egbuniwe,

M.J. Cafferkey, J.F. Spicer, M. Harries, F.O. Nestle, K.E. Lacy, Effects of BRAF

mutations and BRAF inhibition on immune responses to melanoma, Mol.

Canc. Therapeut. 13 (2014) 2769e2783.

[21] K. Ono, J. Han, The p38 signal transduction pathway activation and function,

Cell. Signal. 12 (2000) 1e13.

[22] A. Cuenda, S. Rousseau, p38 MAP-kinases pathway regulation, function and

role in human diseases, Biochim. Biophys. Acta Mol. Cell Res. 1773 (2007)

1358e1375.

or PI3K pathway inhibitors on the LPS-induced gene expression pro ﬁ le in

RAW264. 7 cells: synergistic effects of rapamycin on LPS-induced MMP9-

overexpression, Cell. Signal. 21 (2009) 1109e1122.

[38] K.P. Hoe ﬂ ich, D.C. Gray, M.T. Eby, J.Y. Tien, L. Wong, J. Bower, A. Gogineni,

J. Zha, M.J. Cole, H.M. Stern, Oncogenic BRAF is required for tumor growth and

maintenance in melanoma models, Canc. Res. 66 (2006) 999e1006.

[39] H. Davies, G.R. Bignell, C. Cox, P. Stephens, S. Edkins, S. Clegg, J. Teague,

H. Woffendin, M.J. Garnett, W. Bottomley, Mutations of the BRAF gene in

human cancer, Nature 417 (2002) 949e954.

[23] P.P. Roux, J. Blenis, ERK and p38 MAPK-activated protein kinases: a family of

protein kinases with diverse biological functions, Microbiol. Mol. Biol. Rev. 68

(2004) 320e344.

[40] C. Zhang, W. Spevak, Y. Zhang, E.A. Burton, Y. Ma, G. Habets, J. Zhang, J. Lin,

T. Ewing, B. Matusow, RAF inhibitors that evade paradoxical MAPK pathway

activation, Nature 526 (2015) 583e586.

[24] A. Hauschild, J.-J. Grob, L.V. Demidov, T. Jouary, R. Gutzmer, M. Millward,

P. Rutkowski, C.U. Blank, W.H. Miller Jr., E. Kaempgen, Dabrafenib in BRAF-

mutated metastatic melanoma: a multicentre, open-label, phase 3 rando-

mised controlled trial, Lancet 380 (2012) 358e365.

[41] J.R. Simard, M.u Getlik, C. Grütter, V. Pawar, S. Wulfert, M. Rabiller, D. Rauh,

Development of a ﬂ uorescent-tagged kinase assay system for the detection

and characterization of allosteric kinase inhibitors, J. Am. Chem. Soc. 131

(2009) 13286e13296.

[25] P. Koelblinger, O. Thuerigen, R. Dummer, Development of encorafenib for

BRAF-mutated advanced melanoma, Curr. Opin. Oncol. 30 (2018) 125.

[26] M. Barancık, V. Bohacova, J. Kvackajova, S. Hudecova, O.g Krizanova, A. Breier,

SB203580, a speci ﬁ c inhibitor of p38-MAPK pathway, is a new reversal agent

of P-glycoprotein-mediated multidrug resistance, Eur. J. Pharmaceut. Sci. 14

(2001) 29e36.

[42] H.-S. Han, J.-S. Shin, S.-B. Lee, J.C. Park, K.-T. Lee, Cirsimarin, a ﬂ avone gluco-

side from the aerial part of Cirsium japonicum var. ussuriense (Regel) Kitam.

ex Ohwi, suppresses the JAK/STAT and IRF-3 signaling pathway in LPS-

stimulated RAW 264.7 macrophages, Chem. Biol. Interact. 293 (2018) 38e47.

ꢀ

ꢁꢀ

ꢁ

ꢀ

ꢁ

ꢀ

ꢁ

[43] J.C. Stockert, R.W. Horobin, L.L. Colombo, A. Blazquez-Castro, Tetrazolium salts

and formazan products in Cell Biology: viability assessment, ﬂ uorescence

imaging, and labeling perspectives, Acta Histochem. 120 (2018) 159e167.

[27] P.B.R. Acrylamide, C.C. Kit, S.S.D.G. Stain, Pexmetinib (ARRY-614).

[28] R. Azevedo, M. van Zeeland, H. Raaijmakers, B. Kazemier, J. de Vlieg, A. Oubrie,

́

16

Article Doi

Design, synthesis, biological evaluation, and docking studies of novel (imidazol-5-yl)pyrimidine-based derivatives as dual BRAFV600E/p38α inhibitors

DOI: 10.1016/j.ejmech.2021.113277

Source and publish data:

Authors:

Article abstract of DOI:10.1016/j.ejmech.2021.113277

Full text of DOI:10.1016/j.ejmech.2021.113277

Products guided by the article

R&D Labs maybe for 616196-43-1

Relevant to this article

Hot Product