Biochemical Properties of TAK-828F, a Potent and Selective Retinoid-Related Orphan Receptor Gamma t Inverse Agonist

Abstract

Background/Aims: Retinoid-related orphan receptor gamma t (RORγt) is a master regulator of T helper 17 cells that plays a pivotal role in the production of inflammatory cytokines including interleukin (IL)-17. Therefore, RORγt has attracted much attention as a target receptor for the treatment of inflammatory diseases including rheumatoid arthritis, multiple sclerosis, inflammatory bowel diseases, and psoriasis. This study aims to characterize TAK-828F, a potent and selective RORγt inverse agonist. Methods: The biochemical properties of TAK-828F were evaluated using Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) binding assay, surface plasmon resonance (SPR) biosensor assay, cofactor recruitment assay, reporter assay, and IL-17 expression assay. Results: TR-FRET binding assay and SPR biosensor assay revealed rapid, reversible, and high affinity binding of TAK-828F to RORγt. The cofactor recruitment assay showed that TAK-828F inhibited the recruitment of steroid receptor coactivator-1 to RORγt. Furthermore, TAK-828F inhibited the transcriptional activity of human and mouse RORγt with selectivity against human RORα and RORβ. TAK-828F also suppressed IL-17 production in Jurkat cells, overexpressing human RORγt. Conclusion: These favorable properties will be of advantage in the evaluation of TAK-828F in clinical studies for inflammatory diseases. Furthermore, these findings demonstrate that TAK-828F could serve as a pharmacological tool for further studies of RORγt and inflammatory diseases.

Discovery of [cis-3-({(5R)-5-[(7-Fluoro-1,1-dimethyl-2,3-dihydro-1H-inden-5-yl)carbamoyl]-2-methoxy-7,8-dihydro-1,6-naphthyridin-6(5H)-yl}carbonyl)cyclobutyl]acetic Acid (TAK-828F) as a Potent, Selective, and Orally Available Novel Retinoic Acid Receptor-Related Orphan Receptor γt Inverse Agonist

“…We also tested the selectivity against other nuclear receptors in a panel of cellular reporter assays (Invitrogen) and found that compound 10 was a selective RORγt inverse agonist with >800fold selectivity over the other 20 nuclear receptors in both agonist and antagonist modes. 25 In order to determine if our RORγt inverse agonists could modulate the cytokine production through the suppression of differentiation and activation of Th17 and Th1/17 cells, we evaluated the in vivo PD efficacy in the mouse IL-23-induced cytokine expression model. Compound 10 showed robust and dose-dependent inhibition of IL-17A expression after oral administration at 0.3, 1, and 3 mg/kg b.i.d.…”

“…2-(6-Methoxypyridin-2-yl)ethanamine (25). Hydrazine monohydrate (24.9 mL, 514 mmol) was added to a solution of 24 (29.0 g, 103 mmol) in EtOH (300 mL) at room temperature.…”

Agonist Lock Touched and Untouched Retinoic Acid Receptor-Related Orphan Receptor-γt (RORγt) Inverse Agonists: Classification Based on the Molecular Mechanisms of Action

“…Compound 15 (TAK-828F) has potent RORγt inverse agonistic activity, excellent selectivity, and an improved PK/PD profile (LLE, 4.68; IC 50 = 0.002 μM in a binding assay; IC 50 = 0.006 μM in a reporter gene assay; ED 80 = 0.5 mg/kg b.i.d. in IL-17A PD model, F = 96% and AUC 0–8h , 10076 μg·h/mL). − Compound 15 reduces the production of Th17-related cytokines (IL-17, IL-17F, and IL-22) at 100 nM without affecting IFN-γ production in whole blood and shows a dose-dependent pharmacological effect in a naive T cell transfer mouse colitis model . In the EAE model, compound 15 displayed a 50% reduction in the clinical score upon oral dosing at 1 mg/kg b.i.d.…”

Small molecule inhibitors of RORγt for Th17 regulation in inflammatory and autoimmune diseases

“…10 ) [ 150 ]. The regulator suppressed the recruitment of SRC1 with IC 50 of 59 nM and blocked the transcription of RORγt with IC 50 of 9.5 nM [ 151 ]. In mouse model of EAE, the administration of TAK-828F (0.3−3 mg/kg) decreased the symptoms of EAE dose-dependently, and further study suggested that it reduced the Th17 cell-mediated inflammatory response via decreasing IL-17A production [ 152 ].…”

“…As summarized in Table 3 [ [122] , [123] , [124] , [125] , [128] , [129] , [130] , [132] , [133] , [134] , [135] , [136] , [137] , [138] , [139] , [140] , [141] , 145 , 146 , [150] , [151] , [152] , [153] , [154] , [159] , [160] , [161] , [162] , [163] , [164] , 166 , 167 , 169 , 170 ], numerous RORγt inhibitors (antagonists and inverse agonists) are actively being researched for the therapy of probable inflammatory disorders due to significance of RORγt in autoimmune and inflammatory diseases. Some of the reported RORγt inhibitors, such as JTE-451 (Retezorogant), BI730357 (Bevurogant), ABBV-157 (Cedirogant), RTA-1701 and AUR101, have also entered clinical trial evaluation ( Table 4 ) [ 121 , 139 , [171] , [172] , [173] , [174] , [175] , [176] , [177] , [178] , [179] , [180] , [181] , [182] , [183] , [184] , [185] , [186] ].…”