BSH-IN-1 is a recombinant enteric bacterial bile salt hydrolase (BSHs) potent and covalent inhibitor of B. longum BSH (Gram-positive (Gram+)) and B. theta BSH (Gram-negative (Gram+) ) with IC50s of 108 and 427 nM respectively.

Physicochemical Properties

Molecular Formula	C25H41FO3
Molecular Weight	408.589651823044
Exact Mass	408.303
CAS #	2553217-91-5
Related CAS #	Gut restricted-7;2553218-46-3
PubChem CID	145713829
Appearance	White to yellow solid powder
LogP	5.5
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	5
Heavy Atom Count	29
Complexity	623
Defined Atom Stereocenter Count	10
SMILES	FCC(CC[C@@H](C)[C@H]1CC[C@H]2[C@@H]3[C@@H](C[C@@H]4C[C@@H](CC[C@]4(C)[C@H]3CC[C@@]21C)O)O)=O
InChi Key	XDVGNIBMZHDZLL-GDQMVUAZSA-N
InChi Code	InChI=1S/C25H41FO3/c1-15(4-5-18(28)14-26)19-6-7-20-23-21(9-11-25(19,20)3)24(2)10-8-17(27)12-16(24)13-22(23)29/h15-17,19-23,27,29H,4-14H2,1-3H3/t15-,16+,17-,19-,20+,21+,22-,23+,24+,25-/m1/s1
Chemical Name	(5R)-5-[(3R,5S,7R,8R,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]-1-fluorohexan-2-one
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

ln Vitro	Additionally, BSH-IN-1 (Compound 7) inhibits Gram-positive bacteria (Lactobacillus plantarum WCFS1, C. perfringens ATCC 13124, and Bifidobacterium adolescentis L2-32) as well as Gram-negative bacteria (B. theta VPI-5482, Bacteroides fragilis ATCC 25285 and Bacteroides vuLgatus ATCC 8482). In expanding bacterial cultures, BSH-IN-1 (Compound 7) is also a strong BSH inhibitor, with IC50s of 237 nM and, for Gram-positive and Gram-negative bacteria, B. adolescentis and B. theta, respectively, 1070 nM[1].
ln Vivo	BSH-IN-1 (Compound 7) can be intestinally confined and suppresses BSH activity in C57BL/6 mice after a single dose of 10 mg/kg by gavage[1].
Animal Protocol	Animal/Disease Models: C57BL/6 mice[1] Doses: 10 mg/kg Route of Administration: Single gavage Experimental Results: Inhibited gut bacterial BSH activity and modulated the bile acid pool in vivo while not Dramatically affected the gut bacterial community.
References	[1]. Development of a covalent inhibitor of gut bacterial bile salt hydrolases.Nat Chem Biol. 2020 Mar;16(3):318-326.

Solubility Data

Solubility (In Vitro)

DMSO: 25 mg/mL (61.19 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.12 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 (6.12 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 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 (6.12 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 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	2.4474 mL	12.2372 mL	24.4744 mL
	5 mM	0.4895 mL	2.4474 mL	4.8949 mL
	10 mM	0.2447 mL	1.2237 mL	2.4474 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.