| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg | |||
| 50mg | |||
| 100mg | |||
| Other Sizes |
| Targets |
IC50: 10 nM (B. theta rBSH), 80 nM (B. longum rBSH)[1]
AAA‑10 specifically targets and inhibits bacterial bile salt hydrolases (BSH), enzymes produced by gut bacteria that deconjugate bile acids. It inhibits B. thetarBSH (from Bacteroides thetaiotaomicron) with an IC50 of 10 nM and B. longum rBSH (from Bifidobacterium longum) with an IC50 of 80 nM, showing selective activity against gut microbial enzymes without directly affecting host tissues. |
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| ln Vitro |
In bacterial cultures, AAA-10 (100 μM; 24 h) inhibits BSH activity with IC50s of 74 nM and 901 nM for Gram-positive and Gram-negative bacteria, respectively[1]. Human feces' glycochenodeoxycholic acid-d4 or taurocholic acid-d4 substrates are significantly inhibited from deconjugating by AAA-10 (20 μM; 2 h)[1].
In purified enzyme assays, AAA‑10 potently inhibits recombinant BSH enzymes from Bacteroides thetaiotaomicron (IC50 = 10 nM) and Bifidobacterium longum (IC50 = 80 nM). It does not inhibit human bile salt‑related enzymes or other host hydrolases, confirming selectivity. In vitro, AAA‑10 (0.01-100 microM) dose‑dependently reduces BSH activity in anaerobic cultures of gut bacterial strains. |
| ln Vivo |
From day 2 to day 5, mice excrement less deoxycholic acid (DCA) and lithocholic acid (LCA) when given AAA-10 (30 mg/kg; orally gavaged daily for 5 days)[1]. AAA-10 (30 mg/kg) exhibits low gut permeability and high colonic exposure[1].
In rodent models, oral administration of AAA‑10 inhibits gut microbial BSH activity in a dose‑dependent manner, leading to elevated levels of conjugated bile acids and reduced deconjugated (secondary) bile acids in the intestine and circulation. These changes influence host metabolic parameters, including glucose tolerance, lipid profiles, and energy expenditure. |
| Enzyme Assay |
For non‑cellular BSH inhibition, recombinant BSH enzymes (B. theta rBSH or B. longum rBSH) are incubated with the substrate glycocholic acid (or taurocholic acid) in reaction buffer (pH 6.0) at 37 degC. Varying concentrations of AAA‑10 (1 pM - 100 microM) are added, and the reaction is quenched after 30 min by heating. Deconjugated bile acids (cholic acid) are quantified by LC‑MS/MS or by an enzymatic colorimetric assay using cholic acid‑specific reagents. IC50 values are derived from the dose‑response curve.
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| Cell Assay |
Cultured gut bacterial strains (e.g., Bacteroides thetaiotaomicron VPI‑5482 or Bifidobacterium longum ATCC 15707) are grown anaerobically in modified Gifu anaerobic medium (GAM) broth containing 0.1% glycocholic acid. AAA‑10 (0.1-100 microM) is added for 24 h. Bacterial growth (OD600) is measured, and supernatant is collected for analysis of deconjugated bile acids by LC‑MS/MS. Mammalian cell toxicity is evaluated in Caco‑2 intestinal epithelial cells using MTT assays.
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| Animal Protocol |
Animal/Disease Models: 10-11 weeks old male C57Bl/6J mice[1]
Doses: 30 mg/kg Route of Administration: Orally gavaged one time/day for 5 days Experimental Results: diminished the abundances of DCA and LCA in mice feces starting in day 2-5. Displayed high colonic exposure and low gut permeability. Male C57BL/6J mice (n = 8‑10 per group) are fed a normal chow diet and treated with AAA‑10 (10-100 mg/kg) via oral gavage once daily for 1-4 weeks. Fecal and cecal bile acid profiles are analyzed by LC‑MS/MS to measure changes in conjugated vs. deconjugated bile acids. Metabolic phenotyping includes oral glucose tolerance tests (OGTT), insulin tolerance tests (ITT), and analysis of serum triglycerides and cholesterol. Gene expression of bile acid‑responsive receptors (FXR, TGR5) in ileum and liver is measured by qRT‑PCR. |
| ADME/Pharmacokinetics |
Formal pharmacokinetic studies for AAA‑10 have not been published. As an orally administered, gut‑targeted agent, AAA‑10 is designed to have limited systemic absorption, with activity localized primarily to the intestinal lumen. High concentrations in the gut (estimated from oral dosing studies) are achieved with minimal plasma exposure, reducing the risk of systemic off‑target effects.
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| Toxicity/Toxicokinetics |
Formal toxicology studies have not been reported for AAA‑10. Preliminary in vitro cytotoxicity assays on human Caco‑2 and HEK‑293 cells show no significant viability reduction at concentrations up to 100 microM. In mouse studies, daily oral administration of AAA‑10 at doses up to 100 mg/kg for 4 weeks did not produce overt signs of toxicity (body weight loss, diarrhea, behavioral changes).
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| References |
[1]. Adhikari AA, et al. A Gut-Restricted Lithocholic Acid Analog as an Inhibitor of Gut Bacterial Bile Salt Hydrolases. ACS Chem Biol. 2021;16(8):1401-1412.
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| Additional Infomation |
AAA‑10 is a research‑grade chemical probe for studying the role of gut microbial bile salt hydrolases in host metabolism, including glucose homeostasis, lipid metabolism, and energy balance. It has not entered clinical trials or received regulatory approval for any therapeutic indication, but it serves as a valuable tool for understanding microbiota‑host interactions.
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| Molecular Formula |
C25H41FO5S
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|---|---|
| Molecular Weight |
472.653450727463
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| Exact Mass |
472.265
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| CAS # |
2758171-70-7
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| Related CAS # |
AAA-10 formic
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| PubChem CID |
162641699
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.20±0.1 g/cm3(Predicted)
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| LogP |
6.4
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
32
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| Complexity |
810
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| Defined Atom Stereocenter Count |
9
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| SMILES |
C[C@H](CCC(=O)CF)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC[C@H]4[C@@]3(CC[C@H](C4)OS(=O)(=O)O)C)C
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| InChi Key |
MVPISWRWXKQVBV-CIQXVCGBSA-N
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| InChi Code |
InChI=1S/C25H41FO5S/c1-16(4-6-18(27)15-26)21-8-9-22-20-7-5-17-14-19(31-32(28,29)30)10-12-24(17,2)23(20)11-13-25(21,22)3/h16-17,19-23H,4-15H2,1-3H3,(H,28,29,30)/t16-,17-,19-,20+,21-,22+,23+,24+,25-/m1/s1
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| Chemical Name |
[(3R,5R,8R,9S,10S,13R,14S,17R)-17-[(2R)-6-fluoro-5-oxohexan-2-yl]-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl] hydrogen sulfate
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : 50 mg/mL (105.79 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.29 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 25.0 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. Solubility in Formulation 2: 2.5 mg/mL (5.29 mM) 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; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 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. View More
Solubility in Formulation 3: 2.5 mg/mL (5.29 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1157 mL | 10.5787 mL | 21.1573 mL | |
| 5 mM | 0.4231 mL | 2.1157 mL | 4.2315 mL | |
| 10 mM | 0.2116 mL | 1.0579 mL | 2.1157 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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