Discovery of BMS-986251: A Clinically Viable, Potent, and Selective RORγt Inverse Agonist.

pubs.acs.org/acsmedchemlett Letter Discovery of BMS-986251: A Clinically Viable, Potent, and Selective RORγt Inverse Agonist Robert J. Cherney,* Lyndon A. M. Cornelius, Anurag Srivastava, Carolyn A. Weigelt, David Marcoux, James J.-W. Duan, Qing Shi, Douglas G. Batt, Qingjie Liu, Shiuhang Yip, Dauh-Rurng Wu, Max Ruzanov, John Sack, Javed Khan, Jinhong Wang, Melissa Yarde, Mary Ellen Cvijic, Arvind Mathur, Sha Li, David Shuster, Purnima Khandelwal, Virna Borowski, Jenny Xie, Mary Obermeier, Aberra Fura, Kevin Stefanski, Georgia Cornelius, Joseph A. Tino, John E. Macor, Luisa Salter-Cid, Rex Denton, Qihong Zhao, Percy H. Carter, and T. G. Murali Dhar Cite This: ACS Med. Chem. Lett. 2020, 11, 1221−1227 ACCESS Metrics & More Read Online sı Supporting Information * Article Recommendations ABSTRACT: Novel tricyclic analogues were designed, synthesized, and evaluated as RORγt inverse agonists. Several of these compounds were potent in an IL-17 human whole blood assay and exhibited excellent oral bioavailability in mouse pharmacokinetic studies. This led to the identification of compound 5, which displayed dose-dependent inhibition of IL-17F production in a mouse IL-2/IL-23 stimulated pharmacodynamic model. In addition, compound 5 was studied in mouse acanthosis and imiquimod-induced models of skin inflammation, where it demonstrated robust efficacy comparable to a positive control. As a result of this excellent overall profile, compound 5 (BMS-986251) was selected as a clinically viable developmental candidate. KEYWORDS: RORγt, RORc, inverse agonist, IL-17, IL-23R, psoriasis etinoic acid-related orphan receptor γt (RORγt) is a nuclear hormone receptor (NHR) and a member of the RORγ subfamily.1,2 RORγt is expressed in the thymus and is responsible for the differentiation of CD4+T cells into Th17 cells.3 Hence, RORγt plays a significant role in the production of the pro-inflammatory cytokine IL-17 as well as other cytokines (GM-CSF, IL-21, and IL-22).4 Recently, anti-IL-17 biologics have been shown to be clinically effective against autoimmune diseases such as psoriasis.5−8 As these clinical agents are monoclonal antibodies, there is still a need for small molecule oral therapies modulating IL-17. As a result, there has been much interest in small molecule inhibitors of RORγt, including inverse agonists, as a strategy to suppress IL-17.9−18 Herein, we report our continued optimization of tricyclic19 inverse agonists of RORγt, which culminated in the identification of a viable clinical candidate. As shown in Figure 1, we recently19 reported the synthesis and evaluation of the potent tricyclic RORγt inverse agonists 1 and 2. In an effort to optimize the potency and overall profile of these RORγt inverse agonists, we examined the X-ray crystal structure of 219 in RORγt and observed that the cyclohexane ring of 2 came in close proximity to helix 5 of the receptor (Figure 2). This region of helix 5 contains some lipophilic amino acids, including Ala368. We reasoned that a moiety substituted off the C2 or C3 position of the cyclohexane ring (or cyclic sulfone ring of 1) might bring about a favorable interaction with residues of R © 2020 American Chemical Society Figure 1. Our previously reported RORγt inverse agonists. helix 5 and thereby provide an opportunity to improve affinity for RORγt. From this model, the new C2 or C3 vectors did not appear to disrupt the key carboxylate (or sulfone of compound 1) interactions with Arg367 and Arg364. Likewise, the amide carbonyl of 1 or 2 was still able to engage the backbone NH of Phe377. Based on this rationale, we synthesized and evaluated analogues of compound 1 and 2 as outlined in Table 1. These Received: February 5, 2020 Accepted: March 31, 2020 Published: March 31, 2020 1221 https://dx.doi.org/10.1021/acsmedchemlett.0c00063 ACS Med. Chem. Lett. 2020, 11, 1221−1227 ACS Medicinal Chemistry Letters pubs.acs.org/acsmedchemlett Letter Table 2. Mouse PK Data for Select Compoundsa # Cmax (μM) AUC24h (μM*h) C24h (μM) 5 8 10 11 ± 8 12 ± 2 9±1 68 ± 5 72 ± 18 134 ± 10 0.52 ± 0.13 0.78 ± 0.65 2.2 ± 0.47 a Balb/c mice dosed at 10 mg/kg PO. Values are means from three mice. Vehicle: 5% NMP; 76% PEG 400; 19% TPGS. Table 3. Compound 5 Profilea Assay Result RORγt GAL4 EC50 RORα GAL4 EC50 RORβ GAL4 EC50 IL-17 hWB EC50 mouse Th17 EC50 PXR/LXRα/LXRβ EC50b rCYP 1A2/2C8/2C9/2C19 IC50c rCYP 2D6/3A4 BFC IC50 Caco-2 A-B (nm/s) Caco-2 efflux ratio Protein binding % free h/m/r 12 ± 6 nM >10000 nM >10000 nM 24 ± 6 nM 11 ± 2 nM >5000/>7500/>7500 nM >20/16/>20/>20 μM >20/>20 μM 240 nm/s 0.5 1.2/1.6/2.1 a Protein binding: human (h), mouse (m), rat (r). bPXR, pregnane X receptor; LXR, liver X receptor. crCyP, recombinant cytochrome P450. Figure 2. Crystal structure of compound 2 in RORγt (pdb id: 6U25). compounds were assessed in our RORγt inverse agonist assay (RORγt GAL4 EC50, GAL-4 reporter assay in Jurkat cell line),19 in an IL-17 human whole blood assay (IL-17 hWB EC50),19 and in liver microsomes (LM t1/2) to assess stability. As shown previously,19 compound 1 was active in both the GAL-4 assay and the whole blood assay with excellent liver microsome stability. Starting with compounds substituted at the C2 of the cyclic sulfone, the first diastereomer 3 was almost 3-fold more active in the GAL-4 assay than compound 1, indicating that a new interaction with the receptor was possible. The second diastereomer 4 also had excellent GAL-4 activity similar to that of compound 3, but neither compound offered a real advantage over compound 1 as far as whole blood activity or liver microsome stability. For this reason, we shifted focus to our Table 1. Evaluation of Methyl Substituted Analoguesa # R RORγt GAL4 EC50 (nM) IL-17 hWBb EC50 (nM) LM t1/2: h, m, rc (min) 1 2 3 4 5 6 7 8 9 10 11 Figure 1 Figure 1 4-F-Ph 4-F-Ph 4-F-Ph 3-F-Ph 4-F-Ph 3-F-Ph 3-F-Ph 4-F-Ph 4-F-Ph 24 ± 12 7±6 9 ± 6 (2) 9 ± 3 (2) 12 ± 6 (3) 11 ± 3 (2) 39 (1) 7 (1) 85 (1) 7 ± 1 (2) 106 (1) 43 ± 17 19 ± 8 37 ± 19 (6) 44 ± 17 (4) 24 ± 6 (8) 38 ± 9 (6) 44 ± 25 (2) 27 ± 9 (4) NDd 40 ± 15 (3) ND >120, >120, >120 >120, >120, >120 >120, >120, >120 >120, 98, 33 >120, >120, >120 >120, >120, 107 >120, >120, >120 >120, >120, >120 ND >120, >120, 73 ND EC50 values (n) are displayed as ± standard deviation. bHuman whole blood assay (hWB). cLiver microsomes (LM) incubation: human (h), mouse (m), and rat (r). dND = not determined. a 1222 https://dx.doi.org/10.1021/acsmedchemlett.0c00063 ACS Med. Chem. Lett. 2020, 11, 1221−1227 ACS Medicinal Chemistry Letters pubs.acs.org/acsmedchemlett Letter Table 4. Pharmacokinetic Data for Compound 5 in Preclinical Speciesa iv po species dose (mg/kg) iv/po CL (mL min−1 kg−1) Vss (L/kg) t1/2 (h) Cmax (μM) AUC24h (μM h) F (%) mouse rat dog cyno 2/4 2/4 1/1 1/1 2.7 1.3 ± 0.3 0.18 ± 0.04 1.1 ± 0.2 1.9 1.2 ± 0.3 0.5 ± 0.1 2.0 ± 0.4 7.7 11 ± 0.8 36 ± 3 33 ± 4 4.8 ± 0.3 4.7 ± 0.5 6.4 ± 1.0 3.1 ± 0.3 37 64 ± 3.4 120 ± 21 35 ± 3.1 ∼100 94 ∼10 (...truncated)