This document discusses the role of aminopyrimidinylisoindolines as AXL kinase inhibitors in cancer therapy, highlighting their potential in targeting overexpressed kinases linked to various cancers. It details the synthesis, clinical evaluation, and results of novel compounds, identifying one particularly effective compound (comp. xxi) with low IC50 values against AXL family kinases. The conclusion suggests that comp. xxi can serve as a lead molecule for further kinase inhibition studies.

1. Role of Aminopyrimidinylisoindolines as AXL
Kinase Inhibitors in Cancer Therapy
Journal Club- 1
Presented By
Vanktesh Kumar
(11912659)
Pharmaceutical Chemistry
LOVELY INSTITUTE OF TECHNOLOGY
SCHOOL OF PHARMACEUTICAL SCIENCES, LPU

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Contents
• Introduction
• Lead Molecule
• Scheme for synthesis
• Clinical evaluation
• Results
• Conclusion

3. Introduction
• Tyrosine Kinases are class of important mediators of the signaling cascade, determining key roles in
diverse biological processes like growth, differentiation, apoptosis in response to external and internal
stimuli.
• Tyrosine Kinases is an enzyme that can transfer a phosphate group from ATP to a protein in a cell and
acts as an “On” or “Off” switch in many cellular functions.
• AXL Kinase is a type of transmembrane-receptor tyrosine kinase which belongs to TAM class having
sub-families:- TYRO3, AXL, and MER.
• Responsible for immune enhancement and drug resistance overcome using intercellular antibodies
elimination
• Overexpression of TAM is associated with the various types of cancer such as leukemia, myeloma,
breast cancer and other oncological conditions.
1
References
• Perkins, L.A.; Johnson, M.R.; Melnick, M.B.; Perrimon, N.; The non-receptor protein tyrosine phosphatase corkscrew functions in multiple receptor tyrosine kinase pathways in Drosophila; Developmental
biology, 1996, 180(1), 63-81.
• National cancer institute. Introduction to cancer; 2019; website https://www.cancer.gov/about-cancer/understanding/types (Accessed on Aug 16, 2020).

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• Due to all these reasons are emerged as potential oncological
target, because overexpression of AKL Kinase is a benchmark in
various types of cancer.
• Many Pyrimidine derivatives or nitrogen containing heterocyclic
scaffold were found to have promising kinase inhibiting activity
so aminopyrimidinylisoindolines were employed as AKL kinase
inhibitor.
Fig: Represents the structure of AKL kinase and MER kinase.
References
Choi MJ, Roh EJ, Hur W, Lee SH, Sim T, Oh CH, Lee SH, Kim JS, Yoo KH. Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors. Bioorganic &
Medicinal Chemistry Letters. 2018., 15;28(23-24):3761-5.

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Lead Molecule
• In literature survey it was found that pyrimidine containing molecules are effective against the signal cascade
mechanism inhibition.
• Aminopyrimidinylisoindolines (C) derivatives are evaluated against the Fedratinib, taking it as reference for activity
evaluation.
• Aminopyridine was present on the hinge region of the molecule.
References
Choi MJ, Roh EJ, Hur W, Lee SH, Sim T, Oh CH, Lee SH, Kim JS, Yoo KH. Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors. Bioorganic &
Medicinal Chemistry Letters. 2018., 15;28(23-24):3761-5.

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Scheme for synthesis
• A total 23 molecules were prepared using aminopyrimidine derivative of the lead molecule (C) using following
scheme.
• The lead molecule was prepared using three step reaction and both were connected using N-methylation reaction.
Reagents and condition:
(i) tetrabutylammoniumbromide, dimethylsulfuric acid,
toluene,
15% aqueous NaOH solution, rt, 24 h, 51%;
(ii) methanesulfonyl chloride, pyridine, rt, 30 min,98%;
(iii) N-bromosuccinimide, AIBN, CH2Cl2, reflux,
4 h, 73%.
References
Choi MJ, Roh EJ, Hur W, Lee SH, Sim T, Oh CH, Lee SH, Kim JS, Yoo KH. Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors. Bioorganic &
Medicinal Chemistry Letters. 2018., 15;28(23-24):3761-5.

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Clinical Evaluation
• After synthesis and preparation compounds were tested using an in vitro method on different cell lines.
• A proliferation inhibition activity was checked on cell lines having cancerous cell.
• These cell lines were:-
 U87-MG
 MDA-MB-231
 A549
 MV4-11
 Hep G2
 IMR-32
 HCC-78
 NOMO-1
 OVCAR-3
• All the cell lines were firstly tested for their proliferative activity and then deployed in the evaluation process.
References
Choi MJ, Roh EJ, Hur W, Lee SH, Sim T, Oh CH, Lee SH, Kim JS, Yoo KH. Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors. Bioorganic &
Medicinal Chemistry Letters. 2018., 15;28(23-24):3761-5.

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Sr. no. Compound no. U87-MG (µM) OVCAR-3 (µM) Hep G2 (µM) A549 (µM) HeLa (µM)
1 Comp. I >30 0.63 1.8 24.8 7.0
2 Comp. II 6.2 >30 25.6 7.3 16.5
3 Comp. III >30 >30 >30 >30 17.2
4 Comp. IV >30 >30 22.1 >30 5.1
5 Comp. V 29.4 >30 >30 >30 14.0
6 Comp. VI >30 >30 19.1 20.7 10.6
7 Comp. VII >30 >30 >30 >30 >30
8 Comp. VIII >30 >30 >30 >30 >30
9 Comp. IX 28.3 >30 19.6 25.5 17.4
10 Comp. X >30 >30 16.8 >30 6.5
11 Comp. XI >30 >30 >30 >30 10.9
12 Comp. XII >30 >30 >30 >30 5.3
13 Comp. XIII >30 >30 >30 >30 >30
14 Comp. XIV 16.8 >30 10.3 >30 9.2
References
Choi MJ, Roh EJ, Hur W, Lee SH, Sim T, Oh CH, Lee SH, Kim JS, Yoo KH. Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors. Bioorganic &
Medicinal Chemistry Letters. 2018., 15;28(23-24):3761-5.
Results

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Sr.
no.
Compound no. U87-MG (µM) OVCAR-3 (µM) Hep G2 (µM) A549 (µM) HeLa (µM)
15 Comp. XV 5.5 21.5 9.9 6.7 4.3
16 Comp. XVI >30 >50 16.0 15.2 11.1
17 Comp. XVII 29.8 >50 8.6 6.2 7.3
18 Comp. XVIII 19.5 24.9 23.0 19.0 4.3
19 Comp. XIX >30 >50 >30 >30 23.9
20 Comp. XX 6.8 2.4 1.9 2.5 0.79
21 Comp. XXI 22.1 0.97 1.4 1.3 0.73
22 Comp. XXII 4.7 1.6 3.8 3.3 1.3
23 Comp. XXIII >30 >30 9.8 12.1 6.3
References
Choi MJ, Roh EJ, Hur W, Lee SH, Sim T, Oh CH, Lee SH, Kim JS, Yoo KH. Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors. Bioorganic &
Medicinal Chemistry Letters. 2018., 15;28(23-24):3761-5.

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• The most effective compound was found to be Comp. XXI which has IC50 of 0.00005 µM against AXL family.
• There after best enzymatic inhibition was also produced by the comp. XXI having IC50= <0.00050, 0.025, and
0.050 μM for AXL, MER, and TYRO3, respectively.
• The comp. XXI can be further used as lead molecule for other kinase enzyme inhibition because it showed
satisfactory inhibition on various other kinase enzyme also.
• But out of all kinases such as Aurora A, AKL, FAK, FLT3 etc. AKL family kinase inhibition was very much
effective.
Conclusion
References
Choi MJ, Roh EJ, Hur W, Lee SH, Sim T, Oh CH, Lee SH, Kim JS, Yoo KH. Design, synthesis, and biological evaluation of novel aminopyrimidinylisoindolines as AXL kinase inhibitors. Bioorganic &
Medicinal Chemistry Letters. 2018., 15;28(23-24):3761-5.

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