AAA-10 is an orally bioactive intestinal bacterial bile salt hydrolase (BSH) inhibitor (antagonist) with IC50s of 10 nM and 80 nM for B. theta rBSH and B. longum rBSH, respectively.

Physicochemical Properties

Molecular Formula	C25H41FO5S
Molecular Weight	472.653450727463
Exact Mass	472.265
CAS #	2758171-70-7
Related CAS #	AAA-10 formic
PubChem CID	162641699
Appearance	Typically exists as solid at room temperature
Density	1.20±0.1 g/cm3(Predicted)
LogP	6.4
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	7
Heavy Atom Count	32
Complexity	810
Defined Atom Stereocenter Count	9
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
InChi Key	MVPISWRWXKQVBV-CIQXVCGBSA-N
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
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
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	IC50: 10 nM (B. theta rBSH), 80 nM (B. longum rBSH)[1]
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].
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].
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.
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.

Solubility Data

Solubility (In Vitro)

DMSO : 50 mg/mL (105.79 mM)

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. 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. 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 corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

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.