Microbial Metabolite; isomer of Lithocholic acid

At a concentration of 0.01 percent, isolitocholic acid does not significantly impede CF5 and M120 spore germination and outgrowth, but at a higher concentration of 0.1%, it does. Isolithocholic acid (0.00003%) significantly lowers the toxin activity of strains CF5, BI9, M120, and 630 and inhibits the growth of CD196, M68, CF5, and BI9. With the exception of R20291 and M120, isolithocholic acid (0.0003%) causes all strains to exhibit a considerable drop in toxin activity[3].

In comparison to rats on a regular diet, the high fat diet (HFD) group's fecal Isolithocholic acid levels clearly decreased starting on day 28[4].

We have examined the mechanism for the bacterial transformation of chenodeoxycholic acid and lithocholic acid into the corresponding 3 beta-hydroxy epimers with the use of 3 alpha- and 3 beta-tritiated bile acids. The 3-oxo bile acids were transformed into the 3 alpha- (85%) and 3 beta- (15%) hydroxy bile acids after 20-hr incubation with Clostridium perfringens. Approximately 75% radioactivity was recovered in the aqueous medium when [3 beta-3H]chenodeoxycholic acid or [3 beta-3H]lithocholic acid was incubated with the bacteria, and approximately 15% of radioactivity in the bile acid fraction was associated with the 3 alpha-position of the iso-bile acids. When [3 beta-3H]chenodeoxycholic acid was incubated with unlabeled 3-oxo-5 beta-cholanoic acid, tritiated litho- and iso-lithocholic acids were recovered. These results can be explained only when a 3-oxo intermediate is postulated, and the 3 beta-hydrogen in the bile acids is transferred by the bacterial coenzyme (NAD+ or NADP+) to the 3 alpha-position in the iso-bile acids during the reduction of the 3-oxo compounds [1].

[1]. Transformation of bile acids into iso-bile acids by Clostridium perfringens: possible transport of 3 beta-hydrogen via the coenzyme. Hepatology. 1985;5(6):1126-1131.

[2]. The metabolism of lithocholic acid and lithocholic acid-3-alpha-sulfate by human fecal bacteria. Lipids. 1982;17(7):477-482.

[3]. Inhibition of spore germination, growth, and toxin activity of clinically relevant C. difficile strains by gut microbiota derived secondary bile acids. Anaerobe. 2017;45:86-100.

[4]. Alterations of Bile Acids and Gut Microbiota in Obesity Induced by High Fat Diet in Rat Model. J Agric Food Chem. 2019;67(13):3624-3632.

Isolithocholic acid is a monohydroxy-5beta-cholanic acid with a beta-hydroxy substituent at position 3. The 3beta-hydroxy epimer of lithocholic acid. It has a role as a human metabolite, a rat metabolite and a xenobiotic metabolite. It is a bile acid, a 3beta-hydroxy steroid, a monohydroxy-5beta-cholanic acid and a C24-steroid. It is a conjugate acid of an isolithocholate.
A bile acid formed from chenodeoxycholate by bacterial action, usually conjugated with glycine or taurine. It acts as a detergent to solubilize fats for absorption and is itself absorbed. It is used as cholagogue and choleretic.

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The changing epidemiology of Clostridium difficile infection over the past decades presents a significant challenge in the management of C. difficile associated diseases. The gastrointestinal tract microbiota provides colonization resistance against C. difficile, and growing evidence suggests that gut microbial derived secondary bile acids (SBAs) play a role. We hypothesized that the C. difficile life cycle; spore germination and outgrowth, growth, and toxin production, of strains that vary by age and ribotype will differ in their sensitivity to SBAs. C. difficile strains R20291 and CD196 (ribotype 027), M68 and CF5 (017), 630 (012), BI9 (001) and M120 (078) were used to define taurocholate (TCA) mediated spore germination and outgrowth, growth, and toxin activity in the absence and presence of gut microbial derived SBAs (deoxycholate, isodeoxycholate, lithocholate, isolithocholate, ursodeoxycholate, ω-muricholate, and hyodeoxycholate) found in the human and mouse large intestine. C. difficile strains varied in their rates of germination, growth kinetics, and toxin activity without the addition of SBAs. C. difficile M120, a highly divergent strain, had robust germination, growth, but significantly lower toxin activity compared to other strains. Many SBAs were able to inhibit TCA mediated spore germination and outgrowth, growth, and toxin activity in a dose dependent manner, but the level of inhibition and resistance varied across all strains and ribotypes. This study illustrates how clinically relevant C. difficile strains can have different responses when exposed to SBAs present in the gastrointestinal tract. [3]

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Obesity has become a worldwide health issue and has attracted much public attention. In the current study, we aim to elucidate the roles of bile acids and their associations with gut microbiota during obesity development, employing high fat diet (HFD)-induced obesity in a rat model. We collected feces and plasma, liver tissues, and segments of intestinal tissues and a developed bile acids quantification method by employing an ultraperformance liquid chromatography coupled with mass spectrometry detection (UPLC-MS) strategy. We then assessed bile acids fluxes in the biological matrixes collected. We found that, irrespective of dietary regimes, taurine-conjugated bile acids were the dominant species in the liver whereas unconjugated bile acids were in plasma. However, HFD caused slight increases in the total bile acids pool and particularly the increases in the levels of deoxycholic acid (DCA) (138.67 ± 37.225 nmol/L in control group, 242.61 ± 43.16 nmol/L in HFD group, p = 0.014) and taurodeoxycholic acid (TDCA) (2.8 ± 0.247 nmol/g in control group, 4.5 ± 0.386 nmol/g in HFD group, p = 0.0018) in plasma and liver tissues, respectively, which were consistent with the increased levels of DCA in intestinal tissues and feces. These changes are correlated to an increase in abundance of genera Blautia, Coprococcus, Intestinimonas, Lactococcus, Roseburia, and Ruminococcus. Our investigation revealed the fluxes of bile acids and their association with gut microbiota during obesity development and explicated unfavorable impact of HFD on health.[4]

C24H40O3

376.57

376.297

C, 76.55; H, 10.71; O, 12.75

1534-35-6

Lithocholic acid;434-13-9;Isoallolithocholic acid;2276-93-9

164853

White to off-white solid powder

1.1±0.1 g/cm3

511.0±23.0 °C at 760 mmHg

276.9±19.1 °C

0.0±3.0 mmHg at 25°C

1.528

6.7

2

3

4

27

574

9

C[C@H](CCC(=O)O)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC[C@H]4[C@@]3(CC[C@@H](C4)O)C)C

SMEROWZSTRWXGI-WFVDQZAMSA-N

InChI=1S/C24H40O3/c1-15(4-9-22(26)27)19-7-8-20-18-6-5-16-14-17(25)10-12-23(16,2)21(18)11-13-24(19,20)3/h15-21,25H,4-14H2,1-3H3,(H,26,27)/t15-,16-,17+,18+,19-,20+,21+,23+,24-/m1/s1

(4R)-4-[(3S,5R,8R,9S,10S,13R,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoic acid

Isolithocholic acid; 1534-35-6; beta-Lithocholic acid; 3-Epilithocholic acid; beta-Lithocholanic acid; (4R)-4-[(3S,5R,8R,9S,10S,13R,14S,17R)-3-hydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoic acid; 3beta-Hydroxy-5beta-cholan-24-oic Acid; Iso-LCA;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: 28.57 mg/mL (75.87 mM)

Solubility in Formulation 1: ≥ 2.86 mg/mL (7.59 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 28.6 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.86 mg/mL (7.59 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 28.6 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.86 mg/mL (7.59 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 28.6 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)