Discovery of Soticlestat, a Potent and Selective Inhibitor for Cholesterol 24-Hydroxylase (CH24H).

Tatsuki Koike,Tsuyoshi Ishii,William Farnaby,Takanobu Kuroita,Maki Miyamoto,Shigeru Kondo,Jason Yano,Etsurou Watanabe,Toshiya Nishi,Haruhi Kamisaki Ando,Masato Yoshikawa

doi:10.1021/acs.jmedchem.1c00864

Tatsuki Koike, Tsuyoshi Ishii + Show 9 more

Open Access

https://doi.org/10.1021/acs.jmedchem.1c00864

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Abstract

Cholesterol 24-hydroxylase (CH24H, CYP46A1), a brain-specific cytochrome P450 (CYP) family enzyme, plays a role in the homeostasis of brain cholesterol by converting cholesterol to 24S-hydroxycholesterol (24HC). Despite a wide range of potential of CH24H as a drug target, no potent and selective inhibitors have been identified. Here, we report on the structure-based drug design (SBDD) of novel 4-arylpyridine derivatives based on the X-ray co-crystal structure of hit derivative 1b. Optimization of 4-arylpyridine derivatives led us to identify 3v ((4-benzyl-4-hydroxypiperidin-1-yl)(2,4'-bipyridin-3-yl)methanone, IC50 = 7.4 nM) as a highly potent, selective, and brain-penetrant CH24H inhibitor. Following oral administration to mice, 3v resulted in a dose-dependent reduction of 24HC levels in the brain (1, 3, and 10 mg/kg). Compound 3v (soticlestat, also known as TAK-935) is currently under clinical investigation for the treatment of Dravet syndrome and Lennox-Gastaut syndrome as a novel drug class for epilepsies.

Highlights

CYP46A1, known as cholesterol 24-hydroxylase (CH24H), is a brain-specific cytochrome P450 (CYP) family enzyme that converts cholesterol to 24S-hydroxycholesterol (24HC) and plays a role in the homeostasis of brain cholesterol.[1]
Soticlestat is currently being investigated as a drug for treatment of Dravet syndrome (DS) and Lennox-Gastaut syndrome (LGS) with a novel mechanism of action.[20−22] In this paper, we report the design, synthesis, and discovery of soticlestat starting from a high-throughput screening (HTS) campaign, followed by further optimization utilizing structurebased drug design (SBDD)
Co-crystal structure of compound 1b with CH24H indicates that CH24H has several residues around the active site such as Gly[369], Ala[474], and heme propionate, which can make further hydrogen bonds with the ligand. In addition to these hydrogen-bonding interactions, we focused on a hydrophobic interaction with a large hydrophobic cavity, in which the steroidal scaffold of cholesterol is placed when acting as a substrate

Summary

■ INTRODUCTION

CYP46A1, known as cholesterol 24-hydroxylase (CH24H), is a brain-specific cytochrome P450 (CYP) family enzyme that converts cholesterol to 24S-hydroxycholesterol (24HC) and plays a role in the homeostasis of brain cholesterol.[1]. 4-Benzylpiperidine derivative 3a (IC50 = 110 nM) and 4benzylpiperazine derivative 3b (IC50 = 93 nM) showed improved CH24H inhibitory activities compared with that of the lead fragment 2a (IC50 = 1900 nM) These results suggested that the benzyl group on the 4-position of piperidine or piperazine ring could access the steroidal cavity to make a hydrophobic interaction in addition to the heme coordination and hydrophobic interaction by the biaryl scaffold. Our design concept has been validated by confirming the four key interactions of compound 3f with CH24H; that is, direct ligation to the heme iron by pyridine nitrogen, hydrophobic interaction with the pocket under Helix F by a tolyl moiety, hydrophobic interaction with the steroidal cavity by a benzyl group, and a hydrogen bond between Gly[369] and the hydroxyl group With these features, compound 3f demonstrated in vitro nanomolar CH24H inhibitory activity for the human CH24H enzyme. This result has demonstrated that compound 3v is a promising CH24H inhibitory agent with dose-dependent 24HC reduction in the brain

■ CONCLUSIONS

■ ACKNOWLEDGMENTS

■ REFERENCES