We identified three RORγt-specific inhibitors that suppress T helper 17 (Th17) cell responses including Th17 cell-mediated autoimmune disease. We systemically characterized RORγt binding in the presence and absence of drug with corresponding whole-genome transcriptome sequencing. RORγt acts both as a direct activator of Th17 cell signature genes and as a direct repressor of signature genes from other T-cell lineages, with the strongest transcriptional effects on cis-regulatory sites containing the RORα binding motif. RORγt is central in a densely interconnected regulatory network that shapes the balance of T-cell differentiation. The three inhibitors identified here modulated the RORγt-dependent transcriptional network to varying extents and through distinct mechanisms. Whereas one inhibitor displaced RORγt from its target-loci, the two more potent inhibitors affected transcription predominantly without removing DNA-binding. Our work illustrates the power of a system-scale analysis of transcriptional regulation to characterize potential therapeutic compounds that inhibit pathogenic Th17 cells and suppress autoimmunity.

This study develops a model grounded in the contingency theory (i.e., context-structureperformance) applicable to Chinese state-owned enterprises (SOEs). Using data from a sample of 205 industrial SOEs, the study shows that SOE growth performance relative to the industry is positively predicted by formal control, inversely predicted by decentralization, and positively predicted by the interaction of the two. Customer product knowledge utilization, unrelated to growth performance relative to the industry, is positively predicted by formal control and the interaction of formal control with decentralization. Foreign induced industry competitiveness, technological turbulence, size, and production technology routineness are treated as context variables and modeled accordingly.

A novel series of tertiary amines as retinoid-related orphan receptor gamma-t (RORγt) inverse agonists was discovered through agonist/inverse agonist conversion. The level of RORγt inhibition can be enhanced by modulating the conformational disruption of H12 in RORγt LBD. Linker exploration and rational design led to the discovery of more potent indole-based RORγt inverse agonists.KEYWORDS: RORγt, agonists, inverse agonists, Th17 cell differentiation, cocrystal structure, structure-based design R etinoid-related orphan receptor gamma-t (RORγt) is a member of the nuclear receptor superfamily. RORγt is a key regulator of the development and functions of T-helper 17 (Th17) cells which are implicated in the pathology of a variety of human inflammatory and autoimmune disorders. 1,2 The RORγt inhibitors have potential utility in controlling the activity of Th17 cells and can be developed as therapeutic agents for treatment of Th17-related autoimmune diseases. A few small molecule inhibitors of RORγt have been reported in the literature. 3−10 In this paper, we report the discovery of tertiary amines and indoles as potent RORγt inverse agonists using structure-and knowledge-based compound design.A high-throughput screen (HTS) of the GSK in-house compound collection using a RORγ fluorescence resonance energy transfer (FRET) assay 11 resulted in identification of thiazole amide 1 as a RORγt inverse agonist with IC 50 of 1.0 μM. The binding of 1 to the RORγt ligand binding domain (LBD) was confirmed with a thermal shift of 7.1°C in a thermal shift assay. 11 SAR exploration on the left-hand side (LHS) of 1 led to the identification of tertiary amine 2 as a potent RORγt agonist with a EC 50 of 0.02 μM in dual FRET assay (Scheme 1). 12 Dual FRET assay, using the same technology as the FRET assay but without adding a surrogate agonist, only relies on the basal level of RORγt activity and is able to measure both agonists and inverse agonists. Peptide profiling study using dual FRET assay showed that coactivator peptide (e.g., steroid receptor coactivator 1 (SRC1)) was recruited upon binding of 2 to RORγt LBD whereas corepressor peptide (e.g., nuclear receptor corepressor 2 (NCOR2)) was not. 12 Given the structure similarity of 1 and 2, we assume that they adopt a similar binding mode within RORγt LBD despite their difference as agonist and inverse agonist. To understand the binding mode of the chemical series, an in-silico docking study for 2 based on a reported RORγt crystal structure 13 was conducted.A RORγt LBD crystal structure (PDB accession code: 3KYT) was selected and processed for the docking study. A total of 40 poses with the best scores were obtained and visually inspected after docking with Surflex-Dock v2.3 14−16 in Sybyl 8.1, 17 among which the top 10 poses were found to be representative and thus further ranked using MM/GBSA 18−20 affinity scores based on the VSGB2.0 solvent model. 21,22 As a

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A novel series of biaryl amides was identified as RORγt inhibitors through core replacement of a starting hit 1. Structure−activity relationship exploration on the biaryl moiety led to discovery of potent RORγt inhibitors with good oral bioavailability and CNS penetration. Compounds 9a and 9g demonstrated excellent in vivo efficacy in EAE mice dose dependently with once daily oral administration. KEYWORDS: RORγt inhibitor, Th17 cell differentiation, biaryl amides, EAE, multiple sclerosis T helper (Th) 17 cells, a lineage of CD4 + effector T cells characterized by the production of IL-17A and IL-17F, are pathogenic in human autoimmune inflammatory diseases including multiple sclerosis (MS). 1−4 The presence of IL-17 was detected in MS lesions, and Th17 cells were observed in the infiltrations of mouse experimental autoimmune encephalomyelitis (EAE) central nervous system (CNS). 5,6 Differentiation and function of Th17 cells are controlled by the transcription factor retinoic acid receptor-related orphan receptor-gamma-t (RORγt). 7−9,11 It has been shown that the genetic deficiency of RORγt in mice severely impaired Th17 cell differentiation and conferred resistance to EAE. 10 RORγt inhibitors has potential utility in reducing the activity of Th17 cells and therefore can be developed as therapeutic agents for the treatment of Th17 cell mediated autoimmune diseases. 12−18 A few small molecule RORγt inhibitors have been reported in literature. 19 Digoxin, 20 SR1001, 21 and ursolic acid 22 were first reported to inhibit RORγt and ameliorate EAE in mice via intraperitoneal administration. Other small molecular RORγt inhibitors 23−31 were later disclosed. Recently, we reported discovery of thiazole ketone amides (e.g., 2) and thiophene ketone amides (e.g., 3) as novel RORγt inhibitors based on a high throughput screening (HTS) hit 1 (Figure 1). 32 These ketones, especially the thiophene ketones, showed good RORγt activities but were poorly orally bioavailable and lack of CNS penetration that is believed to be important for developing an effective oral MS drug. In this Letter, we report the discovery of novel biaryl amides as first potent, orally bioavailable, and CNS penetrant RORγt inhibitors, which demonstrated EAE in vivo efficacy dose dependently via oral administration.The lack of CNS penetration of thiazole/thiophene ketones was attributed to their ketone moiety as the nonketone thiazole amide 1 is CNS penetrant with a brain-to-blood ratio (Br/Bl) of 1.5 in a mouse CNS study (i.p., 2 mg/kg). 33 Encouraged by