Identification and characterization of a selective radioligand for ELOVL6.
ELOVL6, a member of the elongation of very long-chain fatty acids (ELOVL) family, has recently been identified as the rate-limiting enzyme for the elongation of palmitoyl-CoA. ELOVL6 deficient mice are protected from high-fat diet induced insulin resistance, suggesting that ELOVL6 might be a promising target for the treatment of metabolic disorders. Despite the increasing interest in Elovl6 as a therapeutic target, the lack of chemical tools for this enzyme has limited further elucidation of the biochemical and pharmacological properties of ELOVL6. We have identified Compound-A, a potent inhibitor for ELOVL6, by screening our company library and subsequently optimizing hit compounds. Compound-A potently inhibited human and mouse ELOVL6 and displayed >100-fold greater selectivity for ELOVL6 over other ELOVL family members. Consistent with its potent and selective inhibitory activity toward ELOVL6, [(3)H]Compound-A bound to ELOVL6 with high affinity while showing no specific binding to other ELOVL enzymes. The observation that [(3)H]Compound-A bound to ELOVL6 in a palmitoyl-CoA-dependent manner in the absence of malonyl-CoA and NADPH suggests that Compound-A might recognize an enzyme-substrate complex, e.g. an acyl-enzyme intermediate. Collectively, these observations demonstrate that Compound-A and its tritiated form are useful tools for biochemical and pharmacological characterization of ELOVL6.
Shimamura
Ken
K
Department of Metabolic Disorder, Tsukuba Research Institute, Banyu Pharmaceutical Co., Ltd., Okubo 3, Tsukuba, Ibaraki 300-2611, Japan.
Takahashi
Hidekazu
H
Kitazawa
Hidefumi
H
Miyamoto
Yasuhisa
Y
Nagumo
Akira
A
Tang
Cheng
C
Dean
Dennis
D
Nagase
Tsuyoshi
T
Sato
Nagaaki
N
Tokita
Shigeru
S
D000123:Acetyltransferases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D000081031:Fatty Acid Elongases; D005227:Fatty Acids / Q000032:analysis; D015870:Gene Expression; D022781:Hepatocytes / Q000737:chemistry / Q000201:enzymology; D006801:Humans; D007211:Indoles / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D020128:Inhibitory Concentration 50; D007527:Isoenzymes; D007700:Kinetics; D008024:Ligands; D051379:Mice; D008861:Microsomes / Q000201:enzymology; D015394:Molecular Structure; D010069:Oxadiazoles / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D019308:Palmitic Acid / Q000378:metabolism; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011869:Radioligand Assay; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D012044:Regression Analysis; D021241:Spectrometry, Mass, Electrospray Ionization
C544762:3-(3-cyclohexyl-1-(2-(dimethylamino)-2-oxoethyl)-6-(4-methyl-5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)-1H-indol-2-yl)-N,N-dimethylbenzamide; D000214:Acyl Coenzyme A; C000630955:ELOVL6 protein, human; C000630968:Elovl6 protein, mouse; D004791:Enzyme Inhibitors; D005227:Fatty Acids; D007211:Indoles; D007527:Isoenzymes; D008024:Ligands; D010069:Oxadiazoles; D011993:Recombinant Fusion Proteins; D019308:Palmitic Acid; D000123:Acetyltransferases; D000081031:Fatty Acid Elongases