Discovery of 5-Phenoxy-2-aminopyridine Derivatives as Potent and Selective Irreversible Inhibitors of Bruton’s Tyrosine Kinase
Abstract
:1. Introduction
2. Results and Discussion
2.1. Drug Design
2.2. Synthesis
2.3. Structure–Activity Relationship Analysis
2.4. Molecular Docking Analysis
2.5. Effects on Experimental Models of Hematological Malignancy
3. Materials and Methods
3.1. Chemistry
3.1.1. General Procedures for the Synthesis of Compound 6
3.1.2. Acrylic Acid 3-(6-Amino-5-piperazinylpyridin-3-yloxy)phenyl Ester (6a)
3.1.3. Acrylic Acid 3-[6-Amino-5-(4-benzylpiperazinyl)pyridin-3-yloxy]phenyl Ester (6b)
3.1.4. Acrylic Acid 3-{6-Amino-5-[4-(4-fluorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl Ester (6c)
3.1.5. Acrylic Acid 3-{6-Amino-5-[4-(4-chlorobenzyl)piperazinyl]pyridin-3-yloxy}phenyl Ester (6d)
3.1.6. Acrylic Acid 3-[6-Amino-5-(4-phenethylpiperazinyl)pyridin-3-yloxy]phenyl Ester (6e)
3.1.7. Acrylic Acid 3-(6-Amino-5-{4-[2-(4-chlorophenyl)ethyl]piperazinyl}pyridin-3-yloxy)phenyl Ester (6f)
3.1.8. 4-[5-(3-Acryloyloxyphenoxy)-2-aminopyridin-3-yl]piperazine carboxylic Acid Benzyl Ester (6g)
3.1.9. General Procedures for the Synthesis of Compounds 17 and 18
3.1.10. Acrylic Acid 2-[6-Amino-5-(3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl Ester (17a)
3.1.11. Acrylic Acid 2-[6-Amino-5-(6-benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl Ester (17d)
3.1.12. Acrylic Acid 3-[6-Amino-5-(3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl Ester (18a)
3.1.13. Acrylic Acid 3-[6-Amino-5-(6-propoxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl Ester (18b)
3.1.14. Acrylic Acid 3-[6-Amino-5-(6-cyclopropylmethoxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl Ester (18c)
3.1.15. Acrylic Acid 3-[6-Amino-5-(6-benzyloxy-3,4-dihydro-1H-isoquinolin-2-yl)pyridin-3-yloxy]phenyl Ester (18d)
3.1.16. Acrylic Acid 3-{6-Amino-5-[6-(4-fluorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl Ester (18e)
3.1.17. Acrylic Acid 3-{6-Amino-5-[6-(4-chlorobenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl Ester (18f)
3.1.18. Acrylic Acid 3-{6-Amino-5-[6-(4-methoxybenzyloxy)-3,4-dihydro-1H-isoquinolin-2-yl]pyridin-3-yloxy}phenyl Ester (18g)
3.2. BTK Kinase Assay
3.2.1. ADP-Glo Assay
3.2.2. HotSpot Kinase Assay
3.3. Molecular Docking Analysis
3.4. In Vitro Growth Inhibition
3.5. In Vivo Xenograft Model
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
FDA | Food and Drug Administration |
JAK3 | Janus kinase 3 |
EGFRK | Epidermal growth factor receptor kinase |
ATP | Adenosine triphosphate |
SRC | Proto-oncogene tyrosine-protein kinase Src |
Boc | tert-Butyloxycarbonyl protecting group |
TFA | Trifluoroacetic acid |
DCM | Dichloromethane |
DMF | Dimethylformamide |
TEA | Triethylamine |
PPA | Polyphosphoric acid |
THF | Tetrahydrofuran |
DMSO | Dimethyl sulfoxide |
ADP | Adenosine diphosphate |
IC50 | The half maximal inhibitory concentration |
Bn | Benzyl |
Me | Methyl |
Et | Ethyl |
Pr | Propyl |
Ac | Acetyl |
GI50 | The half maximal growth inhibitory concentration |
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Compound | R1 | % Inhibition 1 | IC50 (μM) 2 |
---|---|---|---|
6a | H | 14 | ND 3 |
6b | Bn | 85 | 3.17 |
6c | p-FBn | 83 | ND |
6d | p-ClBn | 92 | 1.54 |
6e | PhEt | 89 | 5.01 |
6f | p-ClPhEt | 52 | ND |
6g | benzyloxycarbonyl | 77 | ND |
Ibrutinib | 0.0002 |
Compound | Position | R3 | % Inhibition 1 | IC50 (μM) 2 |
---|---|---|---|---|
17a | 2 | H | 98 | 0.63 |
17d | 2 | OBn | 98 | 0.46 |
18a | 3 | H | 98 | 2.20 |
18b | 3 | OnPr | 99 | 0.97 |
18c | 3 | OCH2cPr | 96 | 1.20 |
18d | 3 | OBn | 97 | 0.10 |
18e | 3 | Op-FBn | 53 * | 8.53 |
18f | 3 | Op-ClBn | 98 | 1.00 |
18g | 3 | Op-OMeBn | 92 * | 0.19 |
Kinase | % Inhibition 1 | ||
---|---|---|---|
17d | 18d | 18g | |
BMX/ETK | 98.7 ± 0.6 | 95.5 ± 0.1 | 96.0 ± 0.1 |
EGFRK | 87.0 ± 0.3 | 60.5 ± 4.4 | 33.5 ± 0.5 |
ERBB2 | 43.4 ± 1.0 | 82.3 ± 1.6 | 78.8 ± 0.7 |
ERBB4 | 95.0 ± 0.1 | 96.2 ± 0.7 | 95.0 ± 0.1 |
ITK | 75.1 ± 0.0 | 52.8 ± 1.8 | 46.0 ± 2.3 |
JAK3 | 64.8 ± 0.3 | 46.2 ± 4.2 | 77.1 ± 3.0 |
TEC | 84.1 ± 0.2 | 71.3 ± 1.0 | 66.4 ± 0.9 |
TXK | 88.8 ± 0.1 | 37.5 ± 3.3 | 49.7 ± 2.5 |
Compound | GI50 (μM) 1 | |||
---|---|---|---|---|
Raji 2 | Ramos 2 | MRC-5 3 | MCF10A 3 | |
17d | 16.1 ± 0.5 | 12.5 ± 0.1 | 28.4 ± 4.0 | 33.4 ± 2.2 |
18g | 25.4 ± 1.4 | 16.2 ± 1.2 | >100 | >100 |
Ibrutinib | 9.5 ± 0.2 | 6.3 ± 4.8 | 40.4 ± 0.9 | 4.6 ± 1.1 |
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Lee, E.; Cho, H.; Lee, D.K.; Ha, J.; Choi, B.J.; Jeong, J.H.; Ryu, J.-H.; Kang, J.S.; Jeon, R. Discovery of 5-Phenoxy-2-aminopyridine Derivatives as Potent and Selective Irreversible Inhibitors of Bruton’s Tyrosine Kinase. Int. J. Mol. Sci. 2020, 21, 8006. https://doi.org/10.3390/ijms21218006
Lee E, Cho H, Lee DK, Ha J, Choi BJ, Jeong JH, Ryu J-H, Kang JS, Jeon R. Discovery of 5-Phenoxy-2-aminopyridine Derivatives as Potent and Selective Irreversible Inhibitors of Bruton’s Tyrosine Kinase. International Journal of Molecular Sciences. 2020; 21(21):8006. https://doi.org/10.3390/ijms21218006
Chicago/Turabian StyleLee, Eun, Hyewon Cho, Da Kyung Lee, JuHyun Ha, Byeong Jo Choi, Ji Hye Jeong, Jae-Ha Ryu, Jong Soon Kang, and Raok Jeon. 2020. "Discovery of 5-Phenoxy-2-aminopyridine Derivatives as Potent and Selective Irreversible Inhibitors of Bruton’s Tyrosine Kinase" International Journal of Molecular Sciences 21, no. 21: 8006. https://doi.org/10.3390/ijms21218006