Design and synthesis of novel imatinib analogs as antitumor multi-target kinase inhibitors / iten mamdouh fawzy abd el-moteleb ; supervision khaled abouzid mohamed, khairia mohamed youssef ,nasser saad mohamed ,deena samy lasheen.

Thesis (PH.D.)- Ain Shams University. Faculty of Pharmacy. Pharmaceutical Chemistry Department,2018.

Includes bibliographical references (p.195 - 212 ).

Cancer is a life-threatening disease and a leading cause of death. Although a range of therapies based on chemotherapy, surgery, and radiotherapy are available, yet they are of limited efficacy. Moreover, current anticancer regimens are associated with significant levels of toxicity and serious adverse side-effects. Hence, many research projects have been focused on developing new chemotherapies with minimal side-effects on mammalian cells. Targeted therapies became one of the eminent principles of drug design and development nowadays to combat cancer.
Imatinib (Gleevec, STI571) became one of the most popular targeted drugs which was approved by FDA for the treatment of chronic myeloid leukemia (CML) as well as various types of cancer. It was proved that imatinib had a potential role in the treatment of non-small cell lung cancer. Additionally, imatinib showed effective inhibition to all epithelial colon cancer cell lines. In fact, imatinib possess loads of proposed mechanisms to fight cancer, the foremost of which is that it acts as tyrosine kinase inhibitor especially against Bcr-Abl. Surprisingly, imatinib anti-tumor activity expressed another evolved mechanism; it had the ability to suppress CrKl phosphorylation level. Moreover, it arrested cell cycle at Go/G1 of K562 cells and dramatically activated caspase-3 together with downstream substrates PARP thus it induced apoptosis.
Unfortunately, imatinib faced resistance in CML patients, due to a point mutation in the kinase domain of Bcr-Abl which leads to disruption in the binding site of imatinib on the tyrosine kinase. Consequently, extensive research had led to the production of second generation of tyrosine kinase inhibitors (TKIs) as dasatnib, nilotinib, bosutinib and ponatinib which showed less resistance and less intolerance than imatinib. Although many new potent (TKIs) are in market, imatinib still represents the front-line therapy for cancer treatment.
The current study aimed to design “two series” of analogs to act as anti-tumor agents. Synthesis of the designed compounds was then accomplished and their structures were confirmed by various spectral and microanalytical data.
This study involved the synthesis of the following unavailable reported intermediates:
1) 4-(4-Nitrophenyl)morpholine (2a)
2) 1-(4-Nitrophenyl)-4-phenylpiperazine (2b)
3) 2-[4-(4-Nitrophenyl)piperazin-1-yl]ethanol (2c)
4) 4-Morpholinoaniline (3a)
5) 4-(4-Phenylpiperazin-1-yl)aniline (3b)
6) 2-[4-(4-Aminophenyl)piperazin-1-yl]ethanol (3c)
7) 4-(4-Nitrophenethyl)morpholine (4a)
8) 2-(4-(4-Nitrophenethyl)piperazin-1-yl)ethanol (4c)
9) 4-(2-Morpholinoethyl)aniline (5a)
10) 2-[4-(4-Aminophenethyl)piperazin-1-yl]ethanol (5c)
11) 4-[2-(4-Nitrophenoxy)ethyl]morpholine (7)
12) 4-(2-Morpholinoethoxy)aniline (8)
13) 4-Morpholinobenzaldehyde (9a)
14) 4-(4-(2-hydroxyethyl)piperazin-1-yl)benzaldehyde (9b)
15) 4-Morpholinobenzoic acid (10a)
16) 4-(4-(2-hydroxyethyl)piperazin-1-yl)benzoic acid (10b)
17) 4-(2-Morpholinoethoxy)benzaldehyde (11a)
18) 4-(2-Morpholinoethoxy)benzoic acid (12a)
19) 4-(Chloromethyl)benzoyl chloride (17)
20) 4-(Chloromethyl)-N-(4-methyl-3-nitrophenyl)benzamide (18)
21) Ethyl 2,4-diaminofuro[2,3-d]pyrimidine-6-carboxylate (21)
22) 2,4-Diaminofuro[2,3-d]pyrimidine-6-carboxylic acid (22)
Also, it comprised the following new intermediates:
1) 1-(4-Nitrophenethyl)-4-phenylpiperazine (4b)
2) 4-[2-(4-Phenylpiperazin-1-yl)ethyl]aniline (5b)
3) 4-[2-(4-Phenylpiperazin-1-yl)ethoxy]benzaldehyde (11b)
4) 4-(2-(4-Phenylpiperazin-1-yl)ethoxy)benzoic acid (12b)
5) N-(4-Methyl-3-nitrophenyl)-4-morpholinobenzamide (13a)
6) 4-[4-(2-Hydroxyethyl)piperazin-1-yl]-N-(4-methyl-3-nitrophenyl)benzamide (13b)
7) N-(3-amino-4-methylphenyl)-4-morpholinobenzamide (14a)
8) N-(3-amino-4-methylphenyl)-4-[4-(2-hydroxyethyl)piperazin-1-yl)]benzamide (14b)
9) N-(4-methyl-3-nitrophenyl)-4-(2-morpholinoethoxy)benzamide (15a)
10) N-(4-methyl-3-nitrophenyl)-4-[2-(4-phenylpiperazin-1-yl)ethoxy]benzamide (15b)
11) N-(3-amino-4-methylphenyl)-4-(2-morpholinoethoxy)benzamide (16a)
12) N-(3-amino-4-methylphenyl)-4-[2-(4-phenylpiperazin-1-yl)ethoxy]benzamide (16b)
13) N-(3-amino-4-methylphenyl)-4-[4-(2-hydroxyethyl)piperazin-1-yl)methyl]benzamide (20)
14) 3-(9H-purin-6-ylamino)-4-methylbenzoic acid (29)
15) 1-[Phenyl 2-(6-Chloro-9H-purin-9-yl)] ethan-1-one (36a)
16) 1-[(4-Methoxyphenyl)-2-(6-Chloro-9H-purin-9-yl)] ethan-1-one (36b)
17) 1-([1,1-Biphenyl]-4-yl)-2-(6-chloro-9H-purin-9-yl)ethan-1-one (36c)
18) 3-{9-[2-(Phenyl)-2-oxoethyl]-9H-purin-6-ylamino}-4-methylbenzoic acid (37a)
19) 3-{9-[2-(4-Methoxy-phenyl)-2-oxoethyl]-9H-purin-6-ylamino}-4-methylbenzoic acid (37 b)
20) 3-{9-[2-([1,1-Biphenyl]-4-yl)-2-oxoethyl)]-9H-purin-6-ylamino}-4-methylbenzoic acid (37 c)
And finally, the study involved the synthesis and the characterization of the following new targeted compounds:
1) 2,4-Diamino-N-[4-(4-phenylpiperazin-1-yl)phenyl]furo[2,3-d]pyrimidine-6-carboxamide (24a)
2) 2,4-Diamino-N-{4-[4-(2-hydroxyethyl)piperazin-1-yl]phenyl}furo[2,3-d]pyrimidine-6-carboxamide (24b)
3) 2,4-Diamino-N-{4-[2-(4-phenylpiperazin-1-yl)ethyl]phenyl}furo[2,3-d]pyrimidine-6-carboxamide (25a)
4) 2,4-Diamino-N-{4-[2-(4-(2-hydroxyethyl)piperazin-1-yl)ethyl]phenyl}furo[2,3-d]pyrimidine-6-carboxamide (25b)
5) 2,4-Diamino-N-[2-methyl-4-(4-morpholinobenzamido)phenyl]furo[2,3-d]pyrimidine-6-carboxamide (26a)
6) 2,4-Diamino-N-{4-[4-(4-(2-hydroxyethyl)piperazin-1-yl)benzamido]-2-methylphenyl}furo[2,3-d]pyrimidine-6-carboxamide (26b)
7) 2,4-Diamino-N-{2-methyl-4-[4-(2-morpholinoethoxy)benzamido]phenyl}furo[2,3-d]pyrimidine-6-carboxamide (27a)
8) 2,4-Diamino-N-{2-methyl-4-[4-(2-(4-phenylpiperazin-1-yl)ethoxy)benzamido]phenyl}furo[2,3-d]pyrimidine-6-carboxamide (27b)
9) 2,4-Diamino-N-{4-[4-((4-(2-hydroxyethyl)piperazin-1-yl)methyl)benzamido]-2-methylphenyl}furo[2,3-d]pyrimidine-6-carboxamide (28)
10) 3-(9H-purin-6-ylamino)-4-methyl-N-(4-morpholinophenyl)benzamide (31a)
11) 3-(9H-purin-6-ylamino)-N-{4-[4-(2-hydroxyethyl)piperazin-1-yl]phenyl}-4-methylbenzamide (31b)
12) 3-(9H-purin-6-ylamino)-4-methyl-N-[4-(2-morpholinoethyl)phenyl]benzamide (32a)
13) 3-(9H-purin-6-ylamino)-4-methyl-N-{4-[2-(4-phenylpiperazin-1-yl)ethyl]phenyl}benzamide (32b)
14) 3-(9H-purin-6-ylamino)-N-{4-[2-(4-(2-hydroxyethyl)piperazin-1-yl)ethyl]phenyl}-4-methylbenzamide (32c)
15) 3-(9H-purin-6-ylamino)-N-{4-[2-(4-(2-chloroethyl)piperazin-1-yl)ethyl]phenyl}-4-methylbenzamide (33)
16) 3-(9H-purin-6-ylamino)-4-methyl-N-[4-(2-morpholinoethoxy)phenyl]benzamide (34)
17) 3-{9-[2-(Phenyl)-2-oxoethyl]-9H-purin-6-ylamino}-N-(4-morpholinophenyl)-4-methyl-1-benzamide (39a)
18) 3-{9-[2-(Phenyl)-2-oxoethyl]-9H-purin-6-ylamino}-N-[4-(4-hydroxyethyl-piperazine)phenyl]-4-methyl-1-benzamide (39b)
19) 3-{9-[2-([1,1 Biphenyl]-4-yl)-2-oxoethyl)]-9H-purin-6-ylamino}-N-[4-(4-phenylpiperazin -1-yl)phenyl]-4-methyl-1-benzamide (40)
20) 3-{9-[2-(1-(4-Methoxy)phenyl)-2-oxoethyl)]-9H-purin-6-ylamino}-N-{4-[2-(4-phenylpiperazin-1-yl)ethyl]phenyl}-4-methyl-1-benzamide (41)
21) 3-{9-[2-(1-(4-methoxy)phenyl)-2-oxoethyl]-9H-purin-6-ylamino}-N-[4-(2-morpholinoethoxy)phenyl]-4-methyl-1-benzamide (42)
In search for the mechanism of action of the newly synthesized compounds various bioassay studies were performed. Anti-tumor activity of the newly synthesized compounds was achieved through in vitro kinase inhibitory assay against Bcr-Abl and Ck-1 performed at BPS Bioscience at 10 µM concentration. The compounds didn’t show potency against those enzymes and hence field alignment study was performed to unleash the reasons. In addition, anti-proliferative evaluation was carried out against both (A549) non-small lung and (Caco) colon cancer cell lines. The compounds showed moderate to high antitumor activity displaying IC50 = 0.14-5.07 µM on both cell lines. Furtherly, three of the most active agents; (24a, 31b and 39a) were subjected to flow cytometry cell cycle and caspase-3 colorimetric assays.
Finally, a comprehensive molecular modeling studies including molecular docking, ADMET and 3D-QSAR based pharmacophore studies were also performed to narrate the antitumor activity of the synthesized compounds with the essential chemical features within their structures.

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