Synthesis and Structure–Activity Relationship Studies of Benzimidazole-4,7-dione-Based P2X3 Receptor Antagonists as Novel Anti-Nociceptive Agents
Abstract
:1. Introduction
2. Results and Discussions
2.1. Library Screening: Discovery of New Scaffold for hP2X3 Receptor Antagonist
2.2. General Synthesis for Benzimidazole-4,7-dione Derivatives
2.3. Hit Optimization: Structure–Activity and Structure–Property Relationship (SAR and SPR) Studies
2.3.1. The Importance of Halide Substitutions
2.3.2. Halide-Substituted Aniline Derivatives
2.3.3. Synthesis and Biological Evaluation of Piperidine Derivatives at hP2X3R
2.4. Pharmacokinetic and hERG Channel Studies of Compound 14h
2.5. Antiallodynic Effect of Compound 14h in Intravenous Administration of Animal Pain Models
3. Materials and Methods
3.1. Chemistry
3.2. General Procedure
3.3. Synthesis of Compounds
3.4. Cell-Based Assay: Measurement of Calcium Influx by Fluorescence-Based Fluo-4 Dye Uptake in HEK293 Cells Expressing hP2X2/3 and hP2X3 Receptors
3.5. Pharmacokinetic Studies
3.6. Neuropathic Pain Model and Behavioral Assessment
3.7. Statistical Analysis of Antagonistic Activities and Behavioral Assessments of Pain Models
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
References
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No. | R1 | R2 | R3 | R4 | hP2X3R | Metabolic Stability | ||
---|---|---|---|---|---|---|---|---|
IC50 (nM) | Mouse (%) | Rat (%) | Human (%) | |||||
KCB-77033 | CH3 | H | F | F | 1030 ± 73 | 5.2 ± 1.6 | 3.4 ± 0.1 | 9.0 ± 1.5 |
10a | CH3 | H | H | F | 4910 ± 821 | 18.4 ± 2.7 | 26.2 ± 3.4 | 43.1 ± 4.7 |
10b | CH3 | H | H | Cl | 3720 ± 731 | - | - | - |
11a | CF3 | H | H | F | 1020 ± 111 | 95.7 ± 17.2 | >99 | 81.4 ± 1.9 |
11b | CF3 | H | H | Cl | 825 ± 70 | - | - | - |
11c | CF3 | H | F | F | 834 ± 117 | - | - | - |
No. | R1 | R2 | R3 | hP2X3R |
---|---|---|---|---|
IC50 (nM) | ||||
11a | H | H | F | 1020 ± 111 |
11b | H | H | Cl | 825 ± 70 |
11c | H | F | F | 834 ± 117 |
11d | H | Cl | Cl | 808 ± 75 |
11e | Cl | H | F | 660 ± 113 |
11f | H | Cl | F | 703 ± 96 |
11g | F | H | Cl | 526 ± 141 |
11h | H | F | Cl | 541 ± 103 |
11i | H | Br | F | 1040 ± 221 |
11j | H | F | Br | 782 ± 139 |
11k | I | H | Cl | 774 ± 166 |
No. | R | hP2X3R |
---|---|---|
IC50 (nM) | ||
14a | methoxycarbonyl | 979 ± 104 |
14b | propionyl | 921 ± 117 |
14c | benzoyl | 864 ± 112 |
14d | 4-fluorobenzoyl | 450 ± 59 |
14e | 4-chlorobenzoyl | 481 ± 20 |
14f | 4-bromobenzoyl | 804 ± 91 |
14g | 3,4-dichlorobenzoyl | 577 ± 66 |
14h | 3,4-difluorobenzoyl | 375 ± 33 |
14i | 3-chloro-4-fluorobenzoyl | 505 ± 51 |
14j | 4-cyanobenzoyl | 4440 ± 480 |
14k | 4-trifluoromethylbenzoyl | >10,000 |
Property | Condition | Parameter | 14h |
---|---|---|---|
Metabolic Stability (Microsomal Fr.) | Human | % remaining after 30 min | >99% |
Rat | >99% | ||
Mouse | 92.8% | ||
CYP450 inhibition | 1A2 | Compound 10 μM | 6.78% |
2C9 | 79.8% | ||
2C19 | 25.6% | ||
2D6 | 7.74% | ||
3A4 | 55.4% | ||
BBB-PAMPA | UV/vis | logPe | <−6.000 |
spectrophotometry | |||
hERG Channel | ligand-binding assay | Compound 10 μM | <1% |
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Bae, J.; Kim, Y.-O.; Han, X.; Yoon, M.-H.; Kim, W.-M.; Kim, Y.-C. Synthesis and Structure–Activity Relationship Studies of Benzimidazole-4,7-dione-Based P2X3 Receptor Antagonists as Novel Anti-Nociceptive Agents. Molecules 2022, 27, 1337. https://doi.org/10.3390/molecules27041337
Bae J, Kim Y-O, Han X, Yoon M-H, Kim W-M, Kim Y-C. Synthesis and Structure–Activity Relationship Studies of Benzimidazole-4,7-dione-Based P2X3 Receptor Antagonists as Novel Anti-Nociceptive Agents. Molecules. 2022; 27(4):1337. https://doi.org/10.3390/molecules27041337
Chicago/Turabian StyleBae, Jinsu, Yeo-Ok Kim, Xuehao Han, Myung-Ha Yoon, Woong-Mo Kim, and Yong-Chul Kim. 2022. "Synthesis and Structure–Activity Relationship Studies of Benzimidazole-4,7-dione-Based P2X3 Receptor Antagonists as Novel Anti-Nociceptive Agents" Molecules 27, no. 4: 1337. https://doi.org/10.3390/molecules27041337