Bile acids with a cyclopropyl-containing side chain. IV. Physicochemical and biological properties of the four diastereoisomers of 3 alpha,7 beta-dihydroxy-22,23-methylene-5 beta-cholan-24-oic acid.
To define the influence of the side chain modification on physicochemical and biological properties of bile acids, 3 alpha,7 beta-dihydroxy-22,23-methylene-5 beta-cholan-24-oic acid, a cyclopropyl analog of ursodeoxycholic acid (UDCA) was synthesized in both unconjugated and taurine-conjugated form. The presence of a cyclopropyl ring at C-22, C-23 position introduces chirality generating four diasteroisomers (A, B, C, and D) which greatly differ for the hydrophilicity and critical micellar concentration: A and B are more hydrophilic (K' = 0.21, 0.80 and CMC = 25,20 mM, respectively) than UDCA (K' = 0.95; CMC = 19 mM) while C and D are more hydrophobic and with lower CMC (K' = 1.30, 2.05; CMC = 14, 10 mM, respectively) than UDCA. Differences in these properties are related to the orientation of the C-25 carboxyl which in isomers A and B is oriented toward the back of the steroid body, reducing the continuity of the hydrophobic area. Using the isolated perfused rat liver we found that the isomers inhibited [3H]UDCA uptake differently. Isomer D (noncompetitive) was the most potent (51%) while isomer A (competitive) was the least potent (15%). When infused intravenously to rats, only D isomer and UDCA were quantitatively recovered in bile. They were secreted predominantly as taurine and glycine conjugates. Isomers A, B, C are not conjugated and only partially recovered in bile as unconjugates (less than 15% of the administered dose). The increase in bile flow per unit increase in bile acid secretion induced by isomers A, B, and C, was much greater than that induced by isomer D which is similar to that of UDCA (0.32 +/- 0.04 and 0.22 +/- 0.01, respectively) while it is reduced during infusion of the other isomers. When infused as taurine conjugates they behaved similarly to tauroursodeoxycholic acid. When incubated in anaerobic conditions with human stools only isomer D is partially 7-dehydroxylated (t/2 = 18 hr) even though slower than UDCA (t/2 = 5 hr). The substrate specificity of the taurine conjugated toward cholyglycine hydrolase is very poor, only isomers C and D are partially deconjugated with a kinetics much slower than that of UDCA (10 hr vs. 0.2 hr). By using molecular models it is possible to explain these differences due to the conformation of the side chain that, in the case of isomer D, is quite similar to UDCA. These data are useful to explain the metabolism of dihydroxy bile acids and to design new analogs with enhanced cholelitholytic activity.
Istituto di Scienze Chimiche, Facoltà di Farmacia, Università degli Studi di Bologna, Italy.
A
A
Roda
B
B
Grigolo
R
R
Aldini
P
P
Simoni
R
R
Pellicciari
B
B
Natalini
R
R
Balducci
D000818:Animals; D001646:Bile / Q000378:metabolism*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002784:Cholesterol / Q000378:metabolism; D003840:Deoxycholic Acid / Q000031:analogs & derivatives*; D008099:Liver / Q000378:metabolism; D008297:Male; D008823:Micelles; D010743:Phospholipids / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D014580:Ursodeoxycholic Acid / Q000031:analogs & derivatives* / Q000493:pharmacokinetics
D008823:Micelles; D010743:Phospholipids; D003840:Deoxycholic Acid; D014580:Ursodeoxycholic Acid; C041489:3,7-dihydroxy-22,23-methylenecholan-24-oic acid; D002784:Cholesterol