Despite a massive industry endeavor to develop RORγ-modulators for autoimmune disorders, there has been no indication of efforts to target the close family member RORα for similar indications. This ...may be due to the misconception that RORα is redundant to RORγ, or the inherent difficulty in cultivating tractable starting points for RORα. RORα-selective modulators would be useful tools to interrogate the biology of this understudied orphan nuclear receptor.
The goal of this research effort was to identify and optimize synthetic ligands for RORα starting from the known LXR agonist T0901317.
Fourty-five analogs of the sulfonamide lead (1) were synthesized and evaluated for their ability to suppress the transcriptional activity of RORα, RORγ, and LXRα in cell-based assays. Analogs were characterized by 1H-NMR, 13C-NMR, and LC-MS analysis. The pharmacokinetic profile of the most selective RORα inverse agonist was evaluated in rats with intraperitoneal (i.p.) and per oral (p.o.)dosing.
Structure-activity relationship studies led to potent dual RORα/RORγ inverse agonists as well as RORα-selective inverse agonists (20, 28). LXR activity could be reduced by removing the sulfonamide nitrogen substituent. Attempts to improve the potency of these selective leads by varying substitution patterns throughout the molecule proved challenging.
The synthetic RORα-selective inverse agonists identified (20, 28) can be utilized as chemical tools to probe the function of RORα in vitro and in vivo.
The synthesis and optimization of a series of small molecule Rev-erbα agonists is described. Compounds with good cell-based potency and efficacy and good mouse exposure provides for in vivo probes to ...interrogate the function of Rev-erbα in animal models of disease.
The structure–activity relationship study of a small molecule Rev-erbα agonist is reported. The potency and efficacy of the agonists in a cell-based assay were optimized as compared to the initial lead. Modest mouse pharmacokinetics coupled with an improved in vitro profile make 12e a suitable in vivo probe to interrogate the functions of Rev-erbα in animal models of disease.
Quinazolines
3 and
13a were discovered as novel c-jun N-terminal kinase (JNK) inhibitors with good brain penetration and pharmacokinetic (PK) properties. Compound
13a is considered a potential ...candidate for in vivo evaluation.
Quinazoline
3 was discovered as a novel c-jun N-terminal kinase (JNK) inhibitor with good brain penetration and pharmacokinetic (PK) properties. A number of analogs which were potent both in the biochemical and cellular assays were discovered. Quinazoline
13a was found to be a potent JNK3 inhibitor (IC
50
=
40 nM), with >500-fold selectivity over p38, and had good PK and brain penetration properties. With these properties,
13a is considered a potential candidate for in vivo evaluation.
The design and synthesis of a novel series of c-jun N-terminal kinase (JNK) inhibitors is described. The development of the 4-(pyrazol-3-yl)-pyridine series was discovered from an earlier pyrimidine ...series of JNK inhibitors. Through the optimization of the scaffold
2, several potent compounds with good in vivo profiles were discovered.
The design and synthesis of a novel series of c-jun N-terminal kinase (JNK) inhibitors is described. The development of the 4-(pyrazol-3-yl)-pyridine series was discovered from an earlier pyrimidine series of JNK inhibitors. Through the optimization of the scaffold
2, several potent compounds with good in vivo profiles were discovered.
A novel series of c-jun N-terminal kinase (JNK) inhibitors were designed and developed from high-throughput-screening lead
1.
A novel series of c-jun N-terminal kinase (JNK) inhibitors were designed ...and developed from a high-throughput-screening hit. Through the optimization of the piperazine amide
1, several potent compounds were discovered. The X-ray crystal structure of
4g showed a unique binding mode different from other well known JNK3 inhibitors.
Amide hydrogen/deuterium exchange is a powerful biophysical technique for probing changes in protein dynamics induced by ligand interaction. The inherent low throughput of the technology has limited ...its impact on drug screening and lead optimization. Automation increases the throughput of H/D exchange to make it compatible with drug discovery efforts. Here we describe the first fully automated H/D exchange system that provides highly reproducible H/D exchange kinetics from 130 ms to 24 h. Throughput is maximized by parallel sample processing, and the system can run H/D exchange assays in triplicate without user intervention. We demonstrate the utility of this system to differentiate structural perturbations in the ligand-binding domain (LBD) of the nuclear receptor PPARγ induced upon binding a full agonist and a partial agonist. PPARγ is the target of glitazones, drugs used for treatment of insulin resistance associated with type II diabetes. Recently it has been shown that partial agonists of PPARγ have insulin sensitization properties while lacking several adverse effects associated with full agonist drugs. To further examine the mechanism of partial agonist activation of PPARγ, we extended our studies to the analysis of ligand interactions with the heterodimeric complex of PPARγ/RXRα LBDs. To facilitate analysis of H/D exchange of large protein complexes, we performed the experiment with a 14.5-T Fourier transform ion cyclotron resonance mass spectrometer capable of measuring mass with accuracy in the ppb range.
PCA is a useful tool for pose normalization in 3D model retrieval, in this paper, we analyze its basic principle, point out its shortcoming and give our solution. Experiments show that our methods ...can enhance the robustness of PCA and align 3D models well.
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•Enantioselective synthesis of 3,3-difluoroproline.•Short scalable synthesis.•Resolution provided both enantiomer in high ee and yield.
An efficient route for the synthesis of ...enantiopure 3,3-difluoroproline on multigram-scale is described herein. The deoxofluorination can be achieved with DAST on the corresponding racemic pyrrolidinone in good yield. Resolution of the racemate by crystallization with D- and L-tyrosine hydrazide provides both enantiomers of 3,3-difluoroproline in high yield and ee%.
We report the development of an efficient method for the conversion of a variety of conjugated alkynyl esters to α-substituted conjugated allenyl esters (racemic) through the use of strong amide ...bases. Substantially improved yields over typical enolate formation conditions were observed with the use of 2 equiv of lithium diisopropylamide. Trapping studies indicate that the second equivalent of base likely leads to the dianion intermediate, which upon addition of methyl iodide, trimethylsilyl chloride, or tributyltin chloride gives mixtures of α-substituted conjugated allenyl and β,γ-alkynyl deconjugated esters. Further optimization revealed that additive salts such as LiCl lead primarily to the allenyl product while the use of HMPA as a cosolvent gives the β,γ-alkynyl deconjugated alkylation product. The role of base, base concentration, and electrophile on product yield and selectivity is also discussed.
Abstract
NR2F6, an orphan member of the nuclear receptor superfamily of ligand-regulated transcription factors, plays an important role in CD4+ T cell differentiation and effector function. As a ...transcriptional repressor, NR2F6 can directly repress expression of the pro-inflammatory cytokines IL-2, IL-17A, IL-21, and IFNγ. Our overexpression data is consistent with this, demonstrating NR2F6 represses IL-17A production in TH17 cells, indicating it also may play a role in autoimmune regulation. Furthermore, genetic experiments have demonstrated the ability of NR2F6 to reduce tumor burden and develop host-protective immunological memory, a consequence of it acting as an immune checkpoint in effector T cells. Collectively, these data suggest that modulation of NR2F6 activity may have important clinical applications for autoimmune and cancer immune therapy. To date, no synthetic or endogenous ligands have been identified that modulate its activity and little is known about its transcriptional function at the molecular level. Here through integrating NR2F6 overexpression and cytokine profiling studies with -omics based approaches, such as RNAseq using T cell specific NR2F6 knockouts, we are working to elucidate NR2F6’s transcriptional function in T cells. Additionally, efforts to identify NR2F6 selective small molecules is underway so we can address how ligand modulation of NR2F6 alters its function. These experiments will help define the molecular mechanisms of NR2F6, which are poorly understood, and determine its niche in cancer immunotherapy and autoimmunity.
Supported by R01 CA225890
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