Discovery of Disubstituted Cyclohexanes as a New Class of CC Chemokine Receptor 2 Antagonists

Abstract

We describe the design, synthesis, and evaluation of novel disubstituted cyclohexanes as potent CCR2 antagonists. Exploratory SAR studies led to the cis-disubstituted derivative 22, which displayed excellent binding affinity for CCR2 (binding IC50 = 5.1 nM) and potent functional antagonism (calcium flux IC50 = 18 nM and chemotaxis IC 50 = 1 nM). Site-directed mutagenesis studies with 22 suggest the compound is binding near the key receptor residue Glu291, however, 22 is not reliant on Glu291 for its binding affinity.

Multiple Binding Sites for Small-Molecule Antagonists at the CC Chemokine Receptor 2

“…All antagonists displaced 125 I-CCL2 from the receptor ( Fig. 3A; Table 2) with affinities similar to previously reported data (Mirzadegan et al, 2000;Brodmerkel et al, 2005;Zou et al, 2007;Buntinx et al, 2008;Cherney et al, 2008;Moree et al, 2008). BMS22, RS504393, and Teijin were also able to displace (Table 2).…”

“…RS504393 and Teijin interact with the highly conserved glutamic acid residue E291, most likely via their basic nitrogen atom (Mirzadegan et al, 2000;Hall et al, 2009). For BMS22, the adjacent T292 was found to be important for binding (Cherney et al, 2008), indicating that it shares the same binding pocket as RS504393 and Teijin. Notably, for INCB3344, no such data have been reported yet, and here we established that it binds to the same site as RS504393, Teijin, and BMS22.…”

“…CCL2 was purchased from PeproTech (Rocky Hill, NJ), and the CCR2 antagonists BMS22 (Cherney et al, 2008), RS504393 (Mirzadegan et al, 2000), and Teijin compound 1 (Moree et al, 2008) were obtained from Tocris Bioscience (Bristol, UK). INCB3344, JNJ-27141491, and CCR2-RA- [R] were synthesized in-house as described previously (Xue et al, 2004;Brodmerkel et al, 2005;Zou et al, 2007;Doyon et al, 2008 …”

“…These antagonists are RS504393 (6-methyl-19-[2-(5-methyl-2-phenyl-4-oxazolyl)ethyl]-spiro[4H-3,1-benzoxazine-4,49-piperidin]-2(1H)-one) (Mirzadegan et al, 2000), amino]-N-[2- [[cis-2-[[4-(methylthio)benzoyl]amino]cyclohexyl]amino]-2-oxoethyl]-5-(trifluoromethyl)benzamide) (Cherney et al, 2008), methyl]-3-pyrrolidinyl]amino]-2-oxoethyl]-3-(trifluoromethyl)benzamide) (Moree et al, 2008) (Brodmerkel et al, 2005), the (R) isomer CCR2-RA-[R] (Bhangoo et al, 2007), and JNJ-27141491 [(S)-3-[3,4-difluorophenyl)-propyl]-5-isoxazol-5-yl-2-thioxo-2,3-dihydro-1H-imidazole-4-carboxyl acid methyl ester] (Fig. 1).…”

MCP-1 binds to oxidized LDL and is carried by lipoprotein(a) in human plasma

“…In brief, MCP-1 and/or nLDL or OxLDL in chemotaxis buffer (RPMI without phenol red, 10 mM HEPES) were placed in the bottom well, and 1 × 10 6 THP-1 cells were added to the insert. In some experiments THP-1 cells were preincubated with 100 nM BMS CCR2 22 (Tocris), a highly specifi c C-C chemokine receptor type 2 (CCR2) antagonist ( 22,23 ), for 30 min before the start of the migration assay, and the antagonist was present in the media for the duration of the assay. The cells were allowed to migrate for 2 h at 37°C, and Fig.…”