ABSTRACT
CCR1 (CC Chemokine receptor 1) is a widely studied G protein-coupled receptor target expressed on multiple types of leukocytes. It is implicated in initiating and exacerbating inflammatory conditions and thus is viewed as a good target for autoimmune and inflammatory therapeutic applications. Numerous CCR1 antagonists have been reported. Although some early CCR1 antagonists lacked the species cross reactivity that made in vivo animal model study difficult, efforts have been made to improve the compound potency in rodents. Recent identification of new and improved CCR1 antagonists has resulted in promising, in vivo efficacy in a variety of animal models of disease. While several early compounds have been withdrawn from clinical trials due to lack of efficacy, work continues to evaluate CCR1 antagonists in preclinical and clinical settings.
Subject(s)
Receptors, CCR1/antagonists & inhibitors , Animals , Drug Design , Humans , Immune System Diseases/drug therapyABSTRACT
We conducted virtual docking studies using GLIDE with modified LXRbeta ligand-binding domain (LBD) on internal compound collection followed by the gene profiling with ArrayPlate mRNA assay. A total of 69 compounds were found to upregulate LXRalpha and certain LXR regulated genes from 1308 compounds selected by virtual screen (hit rate: 5.3%). Compound 4 was shown to significantly induce the expression of LXR target genes such as ABCA1, ABCG1, APOE, SCD-1, and SREBP-1c in THP-1 differentiated macrophages. In vitro binding assay confirmed that 4 binds to both LXRalpha and LXRbeta directly and recruits coactivator peptide SRC-1. It functions as a full LXR agonist in stimulating cholesterol efflux in THP-1 differentiated macrophages and induces lipogenesis in HepG2 cells. This study demonstrates that the combination of virtual screen and high throughput gene profiling is an efficient approach for rapid identification of novel LXR modulators.
Subject(s)
Benzoates/pharmacology , Benzylamines/pharmacology , Computer Simulation , DNA-Binding Proteins/agonists , Gene Expression Profiling , Phenazocine/analogs & derivatives , Receptors, Cytoplasmic and Nuclear/agonists , Sterols/pharmacology , Sulfonamides/pharmacology , Benzoates/chemistry , Benzylamines/chemistry , Binding Sites , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , Dose-Response Relationship, Drug , Hydrocarbons, Fluorinated , Ligands , Liver X Receptors , Models, Molecular , Molecular Structure , Oligonucleotide Array Sequence Analysis/methods , Orphan Nuclear Receptors , Phenazocine/chemistry , Phenazocine/pharmacology , RNA, Messenger/genetics , Receptors, Cytoplasmic and Nuclear/chemistry , Receptors, Cytoplasmic and Nuclear/genetics , Sterols/chemistry , Structure-Activity Relationship , Sulfonamides/chemistryABSTRACT
A hit-to-lead optimization process was carried out on the high throughput screening hit compound 1 resulting in the identification of several potent and selective CCR1 receptor antagonists. Compound 37 shows the best overall profile with IC(50) values of <100 nM in binding and functional assays.
Subject(s)
Piperidines/chemistry , Piperidines/pharmacology , Receptors, CCR1/antagonists & inhibitors , Calcium/metabolism , Cell Line , Cell Movement/drug effects , Chemokine CCL3/metabolism , Chemotaxis/drug effects , Humans , Molecular Structure , Monocytes/cytology , Monocytes/drug effects , Monocytes/metabolism , Piperidines/chemical synthesis , Structure-Activity RelationshipABSTRACT
Structure-activity relationship studies on a series of Boc-indole derivatives as LXR agonists are described. Compound 1 was identified as an LXR agonist through structure-based virtual screening followed by high-throughput gene profiling. Replacement of the indan linker portion in 1 with an open-chain linker resulted in compounds with similar or improved in vitro potency and cellular functional activity. The Boc group at the N-1 position of the indole moiety can be replaced with a benzoyl group. The SAR studies led to the identification of compound 8, a potent LXRbeta agonist with an EC50 of 12 nM in the cofactor recruitment assay.
Subject(s)
DNA-Binding Proteins/agonists , Indoles/chemistry , Indoles/pharmacology , Receptors, Cytoplasmic and Nuclear/agonists , Binding Sites , Cell Line , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Liver X Receptors , Models, Molecular , Molecular Structure , Orphan Nuclear Receptors , Sterol Regulatory Element Binding Protein 1/genetics , Sterol Regulatory Element Binding Protein 1/metabolism , Structure-Activity Relationship , Up-RegulationABSTRACT
A structurally novel liver X receptor (LXR) agonist (1) was identified from internal compound collection utilizing the combination of structure-based virtual screening and high-throughput gene profiling. Compound 1 increased ABCA1 gene expression by eightfold and SREBP1c by threefold in differentiated THP-1 macrophage cell lines. Confirmation of its agonistic activity against LXR was obtained in the co-factor recruitment and reporter transactivation assays. Structure-activity relationship studies on compound 1 are described.
Subject(s)
DNA-Binding Proteins/agonists , Gene Expression Regulation/drug effects , Indoles/pharmacology , Macrophages/drug effects , Monocytes/drug effects , Receptors, Cytoplasmic and Nuclear/agonists , ATP Binding Cassette Transporter 1 , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Cell Line , Gene Expression Regulation/physiology , Humans , Indoles/chemical synthesis , Liver X Receptors , Macrophages/metabolism , Models, Chemical , Monocytes/cytology , Orphan Nuclear Receptors , Sterol Regulatory Element Binding Protein 1/genetics , Sterol Regulatory Element Binding Protein 1/metabolism , Structure-Activity RelationshipABSTRACT
Design and synthesis of a series of 3-amino-4-(2-(2-(4-benzylpiperazin-1-yl)-2-oxoethoxy)phenylamino)cyclobutenedione derivatives as novel CCR1 antagonists are described. Structure-activity relationship studies led to the identification of compound 22, which demonstrated potent binding activity, functional antagonism of CCR1 as well as good species cross-reactivity. In addition, compound 22 also showed desirable pharmacokinetic profiles and was selected for in vivo studies in the mouse collagen-induced arthritis model.
Subject(s)
Arthritis, Experimental/drug therapy , Benzyl Compounds/pharmacology , Cyclobutanes/pharmacology , Receptors, Chemokine/antagonists & inhibitors , Administration, Oral , Animals , Arthritis, Experimental/chemically induced , Benzyl Compounds/chemistry , Binding Sites , Collagen , Cyclobutanes/chemistry , Disease Models, Animal , Drug Design , Male , Mice , Mice, Inbred BALB C , Receptors, CCR1 , Structure-Activity RelationshipABSTRACT
Genomic research on target identification and validation has created a great need for methods that rapidly provide detailed structural information on protein-ligand interactions. We developed a suite of NMR experiments as rapid and efficient tools to provide descriptive structural information on protein-ligand complexes. The methods work with large proteins and in particular cases also without the need for a complete three-dimensional structure. We will show applications with two tetrameric enzymes of 120 and 170 kDa.
