Design, Synthesis, and Activity of Novel N-2,3-Difluorophenyl Cyclobutylamine M4 AChR Positive Allosteric Modulators

doi:10.6023/cjoc202510015

Abstract

Using VU6000918 as the lead compound, employing the Glide docking module of the Schrödinger platform to perform molecular docking and scoring evaluations of the designed target compounds with the allosteric binding sites of the M4 receptor protein, 27 N-2,3-difluorophenyl cyclobutylamine derivatives (V1~V27) were designed and synthesized. The activities of these compounds on M4 receptor cells were evaluated using FLIPR (Fluorescence imaging plate reader) fluorescence detection technology. The results showed that some target compounds exhibited significant activities on M4 receptor cells. Among them, compounds V1 (EC₅₀=0.636 μmol/L), V8 (EC₅₀=0.482 μmol/L), V10 (EC₅₀=0.508 μmol/L), and V13 (EC₅₀=0.621 μmol/L) had higher or comparable activities on M4 receptor cells than the lead compound. Structure-activity relationship (SAR) studies revealed that compounds containing a quinoline ring exhibited higher activity. In particular, compounds with a methyl group at the position 4 of the quinolone showed higher activities than those with other substituents, demonstrating that the introduction of the methyl group can significantly enhance the activity and is worthy of further investigation. Molecular-docking results indicate that these compounds can bind effectively to the allosteric site of the M4 receptor, thereby exerting their potential pharmacological effects. This work provides a guidance for the design of M4 AChR positive allosteric modulators.

Key words: molecular docking, M4 positive allosteric modulator, N-2,3-difluorophenyl cyclobutylamine, synthesis, activity

Cite this article

Min Liu, Yikun Chen, Luanyuan Tian, Haifeng Zhou, Hao Zhou, Qixing Liu. Design, Synthesis, and Activity of Novel N-2,3-Difluorophenyl Cyclobutylamine M4 AChR Positive Allosteric Modulators[J]. Chinese Journal of Organic Chemistry, 2026, 46(5): 2138-2148.

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Compd.	Docking score/(kJ∙mol^－1)	Compd.	Docking score/(kJ∙mol^－1)	Compd.	Docking score/(kJ∙mol^－1)
V1	－31.9	V11	－29.0	V21	－27.4
V2	－32.0	V12	－24.6	V22	－27.9
V3	－32.0	V13	－33.4	V23	－33.0
V4	－31.9	V14	－30.9	V24	－35.2
V5	－29.9	V15	－37.0	V25	－34.5
V6	－31.4	V16	－34.5	V26	－26.4
V7	－29.7	V17	－28.3	V27	－30.0
V8	－31.7	V18	－34.7	VU6000918	－22.7
V9	－24.3	V19	－28.5	VU0467154	－25.6
V10	－29.8	V20	－32.1

Compd.	Docking score/(kJ∙mol^－1)	Compd.	Docking score/(kJ∙mol^－1)	Compd.	Docking score/(kJ∙mol^－1)
V1	－31.9	V11	－29.0	V21	－27.4
V2	－32.0	V12	－24.6	V22	－27.9
V3	－32.0	V13	－33.4	V23	－33.0
V4	－31.9	V14	－30.9	V24	－35.2
V5	－29.9	V15	－37.0	V25	－34.5
V6	－31.4	V16	－34.5	V26	－26.4
V7	－29.7	V17	－28.3	V27	－30.0
V8	－31.7	V18	－34.7	VU6000918	－22.7
V9	－24.3	V19	－28.5	VU0467154	－25.6
V10	－29.8	V20	－32.1

Compd.	EC₅₀/(μmol∙L^－1)	Compd.	EC₅₀/(μmol∙L^－1)	Compd.	EC₅₀/(μmol∙L^－1)
V1	0.636±0.02	V11	2.840±0.02	V21	>30
V2	2.652±0.02	V12	1.450±0.02	V22	>30
V3	3.033±0.02	V13	0.621±0.02	V23	>30
V4	>30	V14	2.376±0.02	V24	>30
V5	1.888±0.02	V15	1.999±0.02	V25	>30
V6	1.094±0.02	V16	>30	V26	>30
V7	0.979±0.02	V17	>30	V27	13.930±0.02
V8	0.482±0.02	V18	>30	VU0467154	0.636±0.02
V9	4.000±0.02	V19	>30	VU6000918	0.120±0.02
V10	0.508±0.02	V20	>30

Compd.	EC₅₀/(μmol∙L^－1)	Compd.	EC₅₀/(μmol∙L^－1)	Compd.	EC₅₀/(μmol∙L^－1)
V1	0.636±0.02	V11	2.840±0.02	V21	>30
V2	2.652±0.02	V12	1.450±0.02	V22	>30
V3	3.033±0.02	V13	0.621±0.02	V23	>30
V4	>30	V14	2.376±0.02	V24	>30
V5	1.888±0.02	V15	1.999±0.02	V25	>30
V6	1.094±0.02	V16	>30	V26	>30
V7	0.979±0.02	V17	>30	V27	13.930±0.02
V8	0.482±0.02	V18	>30	VU0467154	0.636±0.02
V9	4.000±0.02	V19	>30	VU6000918	0.120±0.02
V10	0.508±0.02	V20	>30

新型N-2,3-二氟苯基环丁胺类M4 AChR正向变构调节剂的设计、合成与活性