PMID- 11300870 OWN - NLM STAT- MEDLINE DCOM- 20010503 LR - 20191210 IS - 0022-2623 (Print) IS - 0022-2623 (Linking) VI - 44 IP - 6 DP - 2001 Mar 15 TI - Pharmacophore analysis of the nuclear oxysterol receptor LXRalpha. PG - 886-97 AB - A cell-free assay was developed for the orphan nuclear receptor LXRalpha that measures the ligand-dependent recruitment of a peptide from the steroid receptor coactivator 1 (SRC1) to the nuclear receptor. Using this ligand-sensing assay (LiSA), the structural requirements for activation of the receptor by oxysterols and related compounds were studied. The minimal pharmacophore for receptor activation was shown to be a sterol with a hydrogen bond acceptor at C24. 24(S),25-Epoxycholesterol (1), which meets this criterion, is among the most efficacious of the oxysterols and is an attractive candidate as the LXRalpha natural hormone. Cholenic acid dimethylamide (14) showed increased efficacy compared to 1, whereas the unnatural oxysterol 22(S)-hydroxycholesterol (4) was shown to be an antagonist of 1 in the LiSA. The structural requirements for SRC1 recruitment in the LiSA correlated with the transcriptional activity of compounds in a cell-based reporter assay employing LXRalpha-GAL4 chimeric receptors. Site-directed mutagenesis identified Trp(443) as an amino acid critical for activation of LXRalpha by oxysterol ligands. This information was combined with the structure-activity relationship developed from the LiSA to develop a 3D homology model of LXRalpha. This model may aid the design of synthetic drugs targeted at this transcriptional regulator of cholesterol homeostasis. FAU - Spencer, T A AU - Spencer TA AD - Dartmouth College, Hanover, New Hampshire 03755, USA. FAU - Li, D AU - Li D FAU - Russel, J S AU - Russel JS FAU - Collins, J L AU - Collins JL FAU - Bledsoe, R K AU - Bledsoe RK FAU - Consler, T G AU - Consler TG FAU - Moore, L B AU - Moore LB FAU - Galardi, C M AU - Galardi CM FAU - McKee, D D AU - McKee DD FAU - Moore, J T AU - Moore JT FAU - Watson, M A AU - Watson MA FAU - Parks, D J AU - Parks DJ FAU - Lambert, M H AU - Lambert MH FAU - Willson, T M AU - Willson TM LA - eng GR - HL52069/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, U.S. Gov't, P.H.S. PL - United States TA - J Med Chem JT - Journal of medicinal chemistry JID - 9716531 RN - 0 (Cholic Acids) RN - 0 (DNA-Binding Proteins) RN - 0 (Hydroxycholesterols) RN - 0 (Ketocholesterols) RN - 0 (Liver X Receptors) RN - 0 (Orphan Nuclear Receptors) RN - 0 (Receptors, Cytoplasmic and Nuclear) RN - 0 (Receptors, Steroid) RN - 0 (Sterols) RN - 0 (Transcription Factors) RN - 0 (cholenic acid dimethylamide) RN - 17711-16-9 (22-hydroxycholesterol) RN - 68138-65-8 (24,25-epoxycholesterol) RN - 8DUH1N11BX (Tryptophan) RN - 97C5T2UQ7J (Cholesterol) RN - EC 2.3.1.48 (Histone Acetyltransferases) RN - EC 2.3.1.48 (Nuclear Receptor Coactivator 1) SB - IM MH - Amino Acid Sequence MH - Animals MH - Binding Sites MH - Blotting, Western MH - Cell Line MH - Cell Nucleus/metabolism MH - Cell-Free System MH - Chlorocebus aethiops MH - Cholesterol/*analogs & derivatives/chemical synthesis/chemistry/pharmacology MH - Cholic Acids/chemical synthesis/chemistry/pharmacology MH - DNA-Binding Proteins MH - Energy Transfer MH - Fluorescence MH - Histone Acetyltransferases MH - Hydroxycholesterols/chemical synthesis/chemistry/pharmacology MH - Ketocholesterols/chemical synthesis/chemistry/pharmacology MH - Liver X Receptors MH - Models, Molecular MH - Molecular Sequence Data MH - Nuclear Receptor Coactivator 1 MH - Orphan Nuclear Receptors MH - Receptors, Cytoplasmic and Nuclear/*agonists/antagonists & inhibitors MH - Receptors, Steroid/*agonists/antagonists & inhibitors MH - Stereoisomerism MH - Sterols/chemical synthesis/chemistry/*pharmacology MH - Structure-Activity Relationship MH - Transcription Factors/metabolism MH - Tryptophan/chemistry EDAT- 2001/04/13 10:00 MHDA- 2001/05/05 10:01 CRDT- 2001/04/13 10:00 PHST- 2001/04/13 10:00 [pubmed] PHST- 2001/05/05 10:01 [medline] PHST- 2001/04/13 10:00 [entrez] AID - jm0004749 [pii] AID - 10.1021/jm0004749 [doi] PST - ppublish SO - J Med Chem. 2001 Mar 15;44(6):886-97. doi: 10.1021/jm0004749.