Recent achievements in targeting cancer cells with VEGFR-2 inhibitor nitrogenous rings

Shereen M. Maher & Hamada S. Abulkhair

Pharmaceutical Chemistry Department, Faculty of Pharmacy, Horus University - Egypt,

International Coastal Road, New Damietta 34518, Egypt

Abstract
Affecting individuals of all age intervals, cancer cuts through all societies, causing global distress.
As announced by the World Health Organization, cancer is the second most prevalent cause of
death worldwide. Now, we live in the era of thriving research that has seen the expansion in the
field of cancer research in a vibrant and multidisciplinary society that pursues modern and
innovative ways to tackle this disease. Vascular endothelial growth factor II (VEGFR-2) is a
member of the tyrosine kinase that plays a vital role in the regulation of angiogenesis and hence is
critical to supply oxygen and nutrition for cancer cells. Therefore, it has been verified as a rational
target for chemotherapeutic agents. many attempts have been conducted over the last five years to
identify VEGFR-2 inhibitor molecules. In the heart of these attempts. nitrogenous heterocycles
were the most common ring that are involved in the core structures of the identified VEGFR-2
inhibitor molecules. So. we will discuss in this review the recent achievements in targeting cancer
cells with VEGFR-2 inhibitor nitrogenous rings over the last five years (2017-2021). This report
will emphasize on the synthetic strategies adopted to develop the target molecules, their biological
activity and their suggested interaction with the specified biological target.

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1 Introduction
Cancer is a worldwide health problem. As stated by world health organization about 10 million
deaths have been recorded in 2020 as a consequence of malignant diseases. Angiogenesis is a
critical step in the growth and metastasis of cancer cells. Targeting one of angiogenic factors is
essential step in cancer limitation. VEGFR-2 is the most contributing factor in this process, so
suppression of VEGF/VEGFR signaling system disrupt angiogenesis effectively.

In medicinal chemistry, nitrogenous heterocycles, especially the five-membered rings, are a

valuable source of therapeutic agents. More than 80% of the FDA-approved drugs over the last
decade are nitrogen-containing heterocyclic molecules [1]. They were incorporated in many
clinically used anticancer drugs [2]. As well numerous candidates with potential anticancer activity
and VEGFR-2 inhibition have been recently developed [3,4]. The number of novel five-membered
nitrogenous heterocycles with promising applications in medicine is ever-growing [5]. In this
review, we consolidate the recent achievements in the development of novel nitrogen-containing
five-membered heterocycles and their diverse anticancer and VEGFR-2 inhibitory activities,
reported over the past five year (2017 to early 2021). This review highlights the synthetic pathways
and the use of prepared candidates in drug design and the development of VEGFR-2 inhibitors.

2 Literature Survey
At the end of 2020, Thanks to Egyptian-Saudi cooperation, a group of pyrazoline derivatives was
synthesized according to procedures presented in Scheme 1. The main idea in this procedure is the
Claisen – Schmidt condensation reaction type. They used different cancer cells such as prostate
(PC-3), hepatocellular (HepG2), and breast (MDA-MB-) to determine the cytotoxic effect of the
synthesized compounds. It was ranging between (1.30–7.18μM) As well, they evaluated their
inhibitory activity on VEGFR-2. Only two compounds (AZ-2a & AZ-3b; Figure 5) showed the
promising inhibitory effect against VEGFR-2. Their inhibitory effect against VEGFR-2 was (IC50
= 0.22 and 0.21μM respectively). They show a good docking bind to the receptor shown in (Table
1 & Figure 1) [6].

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 Scheme 1: Synthetic procedure (Claisen-Schmidt condensation)

In the middle of 2019, a group of Indian and Australian researchers have cooperated and
synthesized a number of pyrazole-benzothiazole derivatives according to Scheme 2. They
hypothesized that when 1-interacting unsubstituted acetophenones with phenyl hydrazine then
cyclocondensation and oxidation to produced compounds. 2-Coupling the produced compounds
with appropriate amine in presence of suitable catalysts for this chemical reaction. With these
pervious steps, we will get a group of compounds that have been proven to be effective as a
VEGFR-2 inhibitor comparing to FDA approved drug axitinib. The pervious compounds have
been tested on many cells and the result was amazing such as HT-29 (colon), PC-3 (prostate),
A549 (lung), and U87MG (brain). The IC50 was ranging from 3.17 to 10.7 µM. among them,
(Compound AZ-2c-3a; Figure 5) show the best inhibitory effect to these tested cells with 3.17
µM (PC-3), 3.32 µM (HT-29), 3.87 µM (A549) and 6.77 µM (U87MG). Moreover, its potent
inhibitory effect, it is highly selective 2-3 times toward cancer cells than normal cells and 9-15fold

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more selective to cancer cells than axitinib. Best effect comes from best binding positions with
receptors so docking results illustrated in Table 1) [7].

Figure 1:Docking of compound AZ-2a & AZ-3b with VEGFR-2

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 Scheme 2: General procedure for preparation pyrazole-benzothiazole derivatives

At the end of 2019, a group of Chinese scientists has synthesized a number of sulfonamide-
incorporating pyrazolylbenzamides with the objective of targeting VEGFR-2. The adopted
strategy for synthesis of the target molecules is shown in Scheme 3. This involves the condensation
of arene sulfonyl chlorides with two isomeric forms of aminobenzoic acid followed by subsequent
condensation of the produced sulfonamides with previously prepared N1-substituted-4-
aminopyrazoles. The cytotoxic activity of the eighteen synthesized compounds has been evaluated
against cervical, breast, adenocarcinoma, and hepatocarcinoma cancer cells. As well, their
VEGFR-2 inhibitory potential has been assessed. Results revealed that the p-tolylpyrazole
(compound AZ-3d-2b; Figure 5) has showed the best inhibitory activity both against the tested
cells and the biological target (IC50 = 1.08 μM, 2.44 μM, and 0.34 μM against MCF-7, HepG-2,
and VEGFR-2, respectively). These findings were at the same inhibitory levels of the FDA-
approved antitumor drug vandetanib. Additionally, compound 3 effectively induced the apoptotic
cell death of MCF-7 cells. Despite the excellent effect of this compound as cytotoxic against cancer
cells, it was found less toxic to the human renal epithelial cells (293T) (CC50 = 110.47
µM).Docking experiment result summarized in (Table 1&Figure 2) [8].

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 Figure 2: Binding mode of compound 1 with VEGFR-2

In the same year another group of Arab researchers have synthesized a number of
biphenylimidazol-diones treated with N-alkylacetamide fragments as illustrated in Scheme 4.
They followed this strategy in synthesis: i) heating 5,5-diphenylhydantoin and ethyl bro
monoacetate in acetone containing K2CO3. ii)condensation of the resulting ester with hydrazine
in refluxing ethanol. iii)Reacting the produced intermediate with adequate amount of
benzaldehyde derivatives to obtain their target compounds 2. They tested the cytotoxic activity of
all the synthesized compounds on some of cancer cells such as HeLa, human lung adenocarcinoma
(A549), and human breast adenocarcinoma (MDA-MB-231) cell in comparison to Docetaxil
(standard drug). Among all compounds, (compound AZ-13e; Figure 5) has shown the best effect
among them. Its average inhibitory effect reached 59 µM against tested cancer cells. VEGFR2 has
inhibited greatly by this compound up to (IC50= 0.09 µM). They attributed this effect to indole ring
added as (R) and fluorine atom on position 5. Docking results is shown in Table1 and Figure 3
[9].

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Scheme 3: Synthetic strategy of benzoylamide pyrazole

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 Scheme 4: Synthesis strategy of imidazole-diones

Figure 3: Compound 2 binding with VEGFR-2

In 2017, pyridine derivatives containing pyrazolone and triazole moieties have been prepared
according to the procedures provided in scheme 5. They started with substitution, reduction,
hydrazinolysis in Methanecarbonitrile (MeCN), then reaction with N, N-dimethylformamide
dimethyl acetal (DMF-DMA) and finally cyclization the resulting compound to obtain the desired
products. By analyzing and assessment of the resulted products against Human umbilical vein
endothelial cells (HUVEC ), (Compound AZ-4f; Figure 5) has shown the best inhibitory activity

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against (HUVEC IC50=0.30 μM ).Further investigation was done against VEGFR-2 compared
with the FDA approved drug cabozantinib. The inhibitory effect was(IC50= 0.19 μM) which is
amazing . The binding model illustrated in Table 1 & Figure 4 [10].

Figure 4: Docking of compound 3 with VEGFR2

Scheme 5:- Procedure steps of flurorophenoxy pyridine derivatives

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It is clear that this research point has attracted great interest from Chinese scholars. In the same
year, other scientists prepared some other compounds that aim to treat cancer by inhibiting
VEGFR2. They believed that these five membered rings (pyrazoline) has a great effect on its anti
tumor activity. So they prepared some chemical compounds according to the Scheme 6. By
analyzing these compounds on A549, HepG2, MCF-7, and PC-3 cancer cell lines. Comparing to
the FDA approved drug (Sorafenib),they found that compound AZ-2g (Figure 5) show the best
antiVEGFR2 (IC50=0.56 µM). This in turn helps reduce angiogenesis efficiently, which is mainly
involved in metastasis of cancer cells. Molecular docking analysis were performed shown in n
(Table 1).

Scheme 6: Synthetic route of target compounds

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 Figure 5: 5-membered nitrogenous compounds

Table 1: Results of docking interactions of VEGFR-2 targeting molecules

Score Bond length Interacting Residue

Comp. ID.
(kcal/mole) (Å) Residue

1 -12.52 2.79 CYS 919

- 3.05 Asp 1046
2 - - ASN 923
- - LEU 840
3 -57.56 - LYS868
- - Glu885
- - CYS 919
- - Asp1046
4 -9.8 - Glu 917
- - CYS 919
- - LEU 840
- - ASN 923

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 Score Bond length Interacting Residue
Comp. ID.
(kcal/mole) (Å) Residue

5 - - CYS 919
- - Val 898
- - LYS 868
6 - - ASN923
- - GLU885

3 References:
[1] M.M. Heravi, V. Zadsirjan, Prescribed drugs containing nitrogen heterocycles: an overview,
RSC Adv. 10 (2020) 44247–44311. https://doi.org/10.1039/D0RA09198G.

[2] T. Olivier, A. Haslam, V. Prasad, Anticancer Drugs Approved by the US Food and Drug
Administration From 2009 to 2020 According to Their Mechanism of Action, JAMA Netw.
Open. 4 (2021) e2138793. https://doi.org/10.1001/jamanetworkopen.2021.38793.

[3] K. El-Adl, H. Sakr, S.S.A. El-Hddad, A.G.A. El-Helby, M. Nasser, H.S. Abulkhair, Design,
synthesis, docking, ADMET profile, and anticancer evaluations of novel thiazolidine-2,4-
dione derivatives as VEGFR-2 inhibitors, Arch. Pharm. (Weinheim). 354 (2021).
https://doi.org/10.1002/ardp.202000491.

[4] A.-G.A. El-Helby, R.R.A. Ayyad, H. Sakr, K. El-Adl, M.M. Ali, F. Khedr, Design,
Synthesis, Molecular Docking, and Anticancer Activity of Phthalazine Derivatives as
VEGFR-2 Inhibitors, Arch. Pharm. (Weinheim). 350 (2017) 1700240.
https://doi.org/10.1002/ardp.201700240.

[5] incy Joseph, M.C. Sanu, D. Chacko, A Review on Five Membered Nitrogen Containing
Heterocyclic Compounds with Various Biological Activities, Int. J. Pharm. Sci. Rev. Res.
69 (2021). https://doi.org/10.47583/ijpsrr.2021.v69i01.002.

[6] O.M. Alkamaly, N. Altwaijry, R. Sabour, M.F. Harras, Dual EGFR/VEGFR2 inhibitors and
apoptosis inducers: Synthesis and antitumor activity of novel pyrazoline derivatives, Arch.
Pharm. (Weinheim). 354 (2021). https://doi.org/10.1002/ardp.202000351.

Page 12 of 13
[7] V.G. Reddy, T.S. Reddy, C. Jadala, M.S. Reddy, F. Sultana, R. Akunuri, S.K. Bhargava, D.
Wlodkowic, P. Srihari, A. Kamal, Pyrazolo-benzothiazole hybrids: Synthesis, anticancer
properties and evaluation of antiangiogenic activity using in vitro VEGFR-2 kinase and in
vivo transgenic zebrafish model, Eur. J. Med. Chem. 182 (2019) 111609.
https://doi.org/10.1016/j.ejmech.2019.111609.

[8] F.-Q. Shen, L. Shi, Z.-F. Wang, C.-R. Wang, J.-J. Chen, Y. Liu, H.-Y. Qiu, H.-L. Zhu,
Design, synthesis, biological evaluation of benzoyl amide derivatives containing nitrogen
heterocyclic ring as potential VEGFR-2 inhibitors, Bioorg. Med. Chem. 27 (2019) 3813–
3824. https://doi.org/10.1016/j.bmc.2019.07.007.

[9] H.M. Alkahtani, M.M. Alanazi, F.S. Aleanizy, F.Y. Alqahtani, A. Alhoshani, F.E. Alanazi,
A.A. Almehizia, A.N. Abdalla, M.G. Alanazi, A.S. El-Azab, A.A.M. Abdel-Aziz,
Synthesis, anticancer, apoptosis-inducing activities and EGFR and VEGFR2 assay
mechanistic studies of 5,5-diphenylimidazolidine-2,4-dione derivatives: Molecular docking
studies, Saudi Pharm. J. 27 (2019) 682–693. https://doi.org/10.1016/j.jsps.2019.04.003.

[10] W. Gu, Y. Dai, H. Qiang, W. Shi, C. Liao, F. Zhao, W. Huang, H. Qian, Discovery of novel
2-substituted-4-(2-fluorophenoxy) pyridine derivatives possessing pyrazolone and triazole
moieties as dual c-Met/VEGFR-2 receptor tyrosine kinase inhibitors, Bioorg. Chem. 72
(2017) 116–122. https://doi.org/10.1016/j.bioorg.2017.04.001.

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