Synthesis and in vitro biological activity of 4alpha-(2-propenyl)-5alpha-cholest-24-en-3alpha-ol: a 24,25-dehydro analog of the hypocholesterolemic agent 4alpha-(2-propenyl)-5alpha-cholestan-3alpha-ol.
4Alpha-(2-propenyl)-5alpha-cholestan-3alpha-ol (LY295427) was previously identified from a Chinese hamster ovary (CHO) cell-based low density lipoprotein receptor/luciferase (LDLR/Luc) assay to be a potent transcriptional activator of the LDL receptor promoter in the presence of 25-hydroxycholesterol. To investigate the effect of the 24,25-unsaturation in the D-ring side chain (desmosterol D-ring side chain) on antagonizing the repressing effect of 25-hydroxycholesterol, 4alpha-(2-propenyl)-5alpha-cholest-24-en-3alpha-ol (17), a 24,25-dehydro analog of LY295427, was thus synthesized from lithocholic acid via the formation of 3alpha-[[(1,1-dimethylethyl)dimethylsilyl]oxy]-4alpha- (2-propenyl)-5alpha-cholan-24-al (15). Test results showed that 17 had an EC30 value of 2.6 microM, comparable to 2.9 microM of LY295427, in the CHO cell-based LDLR/Luc assay in the presence of 25-hydroxycholesterol. Apparently, the built-in 24,25-unsaturation in the D-ring side chain of 17 had added little effect to antagonizing the repressing effect of 25-hydroxycholesterol. In the [1-14C-acetate]cholesterol biosynthesis inhibition assay, 17 at 10 microg/ml (23 microM) has been shown to inhibit the cholesterol biosynthesis in CHO cells by 38% relative to the vehicle control; whereas LY295427 showed no inhibition in the same assay in our previous studies. In contrast to LY295427, the built-in 24,25-unsaturation in the D-ring side chain of 17 has conferred an inhibitory effect on cholesterol biosynthesis in CHO cells. In summary, the observed LDL receptor promoter activity of 17 is related to its ability to prevent 25-hydroxycholesterol from exerting the repressing effect via an undetermined mechanism and, in part, to inhibit the cholesterol biosynthesis.
Lin
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HS
Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285, USA. lin_ho-shen@lilly.com
Rampersaud
A A
AA
Richett
M E
ME
Harper
R W
RW
Beavers
L S
LS
McClure
D B
DB
Gardner
A J
AJ
Eacho
P I
PI
Foxworthy
P S
PS
Gadski
R A
RA
D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology*; D016466:CHO Cells; D002777:Cholestanols / Q000737:chemistry* / Q000494:pharmacology; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology; D006224:Cricetinae; D008156:Luciferases / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D011401:Promoter Regions, Genetic; D011973:Receptors, LDL / Q000235:genetics; D014158:Transcription, Genetic
C121236:4-(2-propenyl)cholest-24-en-3-ol; D000924:Anticholesteremic Agents; D002777:Cholestanols; C107473:LY 295427; D011973:Receptors, LDL; D002784:Cholesterol; D008156:Luciferases