Cat. No.: HY-107830

Methyl cholate

Name: Methyl cholate
Brand: MedChemExpress
SKU: HY-107830
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Methyl cholate is a bile acid analog and a specific inhibitor of TcdB toxin from Clostridioides difficile. Methyl cholate exerts a stronger selective inhibitory effect on TcdB than on TcdA. Methyl cholate induces conformational stabilization by binding to a unique site of TcdB, thereby blocking the binding of the toxin to host receptors and its self-processing process. Methyl cholate effectively protects human fibroblasts from TcdB-induced cytopathic effects. Methyl cholate exhibits dose-dependent anti-hepatic fibrosis activity in both cellular and zebrafish models, and significantly reduces the expression levels of α-SMA and COL-I. Methyl cholate is suitable for in-depth research in the fields of Clostridioides difficile infection and hepatic fibrosis.

For research use only. We do not sell to patients.

Purity : 99.81%
CAS No.: 1448-36-8
Formula: C₂₅H₄₂O₅
Molecular Weight:422.60

Storage:

Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 2 years , -20°C, 1 year

Methyl cholate Chemical Structure

CAS No.: 1448-36-8

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Top Publications Citing

Use of MCE Products

Fluorescent Dye

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- In Vitro
- In Vivo
Chemical Information
Solvent & Solubility
- In Vitro
- In Vivo
Protocols (5)
Purity & Documentation
References
Help & FAQs
Classifications
Bioz

Biological Activity

Description

All Endogenous Metabolite Isoforms :

In Vitro

Methyl cholate (0.1-100 μM; 10 min pre-incubation, 3.5 hr combined exposure) dose-dependently protects human IMR-90 fibroblasts from TcdB-induced cell rounding with an IC₅₀ of 11 μM, but does not protect against TcdA-induced cell rounding^[1].
Methyl cholate (0.8-200 μM; 30 min pre-incubation, 3 hr reaction) dose-dependently inhibits IP₆-induced autoprocessing of full-length Clostridium difficile TcdB, but does not affect autoprocessing of the isolated GTD-CPD truncated construct^[1].
Methyl cholate (40 μM; 45 min pre-incubation with TcdB, 45 min incubation with cells) potently inhibits Clostridium difficile TcdB binding to Vero cells, reducing bound toxin to ~10% of vehicle control levels^[1].
Methyl cholate (0.1-100 μM; 3 hr combined exposure) dose-dependently protects human IMR-90 fibroblasts from TcdB-induced necrosis with an IC₅₀ of 4.3 μM^[1].
Methyl cholate (80.64 μg/mL; 48 h) inhibits HSC-T6 hepatic stellate cell proliferation with an IC₅₀ of 80.64 μg/mL after 48 h of incubation^[2].
Methyl cholate (low to high dose range; duration sufficient to alter protein expression) dose-dependently reduces α-SMA protein expression, a marker of hepatic stellate cell activation, in HSC-T6 cells^[2].
Methyl cholate (duration sufficient to alter protein expression) increases PIK3-α protein expression in HSC-T6 hepatic stellate cells^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.

Western Blot Analysis^[1]

View Protocol

Cell Line:

Vero cells
Concentration:

40 μM
Incubation Time:

45 min pre-incubation with TcdB; 45 min incubation with cells
Result:

Reduced TcdB binding to Vero cells to levels similar to the neutralizing monoclonal antibody bezlotoxumab, with only ~10% of TcdB remaining bound relative to vehicle controls.

Cell Viability Assay^[2]

View Protocol

Cell Line:

HSC-T6 hepatic stellate cells
Concentration:

80.64 μg/mL
Incubation Time:

48 h
Result:

Inhibited HSC-T6 cell proliferation in a dose-dependent manner, with a calculated half-maximal inhibitory concentration (IC₅₀) of 80.64 μg/mL after 48 h of treatment.

In Vivo

Methyl cholate (6.25-25 μM; water immersion; 3 days after exposure to Thioacetamide (HY-Y0698)) exerts dose-dependent antifibrotic activity in a thioacetamide (TAA)-induced liver fibrosis model of zebrafish larvae, and 25 μM methyl cholate significantly reduces hepatic type Ⅰ collagen levels^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:

Zebrafish larvae (2 days post-fertilization)^[2]
Dosage:

6.25 μM, 12.5 μM, 25 μM
Administration:

aqueous immersion; post 3 days of TAA exposure
Result:

Reduced liver intercellular gaps, cellular enlargement, and vacuolation in a dose-dependent manner relative to the TAA-induced hepatic fibrosis group.
Markedly decreased liver collagen-I levels compared to the fibrosis group (25 μM dose).

Chemical Information

CAS No. 1448-36-8
Appearance Solid
Molecular Weight 422.60
Formula C₂₅H₄₂O₅
Color White to off-white
SMILES

O[C@H]1[C@@]2([H])[C@@]3([H])[C@@]([C@]([C@H](C)CCC(OC)=O)([H])CC3)([C@@H](O)C[C@]2([H])[C@]4(C)[C@](C[C@H](O)CC4)([H])C1)C
Structure Classification
- Steroids
Initial Source
- Microorganisms
Aspergillus niger
Shipping

Room temperature in continental US; may vary elsewhere.

Storage

Powder	-20°C	3 years
	4°C	2 years
In solvent	-80°C	2 years
	-20°C	1 year

Solvent & Solubility

In Vitro:

DMSO : 100 mg/mL (236.63 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

View the Complete Stock Solution Preparation Table

Preparing Stock Solutions

View the Complete Stock Solution Preparation Table

Concentration Solvent Mass

1 mg

5 mg

10 mg

Concentration

Solvent

Mass

1 mM

2.3663 mL

11.8315 mL

23.6630 mL

5 mM

0.4733 mL

2.3663 mL

4.7326 mL

10 mM

0.2366 mL

1.1832 mL

2.3663 mL

Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.

Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Molarity Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass Mass

Concentration Concentration

Volume Volume

Molecular Weight Molecular Weight

Dilution Calculator

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

Concentration _(start) Concentration _(start)

Volume _(start) Volume _(start)

Concentration _(final) Concentration _(final)

Volume _(final) Volume _(final)

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

Stock Solution Working Solution

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline

Solubility: ≥ 2.5 mg/mL (5.92 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)

Solubility: ≥ 2.5 mg/mL (5.92 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

Protocol 3

Add each solvent one by one: 10% DMSO 90% Corn Oil

Solubility: ≥ 2.5 mg/mL (5.92 mM); Clear solution

This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

Dosage Dosage
mg/kg
Animal Weight (Per Animal) Animal Weight (Per Animal)
g
Dosing Volume (Per Animal) Dosing Volume (Per Animal)
μL
Number of Animals Number of Animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

Please enter your animal formula composition:

%
DMSO +

Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
%
+
+
%
Tween-80 + +
%
Saline +

The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).

Working solution concentration: 0.22 mg/mL

Protocols

All Histomorphology Molecular Biology Signal Pathway Related

Fibrosis/Collagen Morphometry
Fibrosis and collagen morphometry is based on the quantitative visualization of fibrillar collagen deposition in tissue sections using histochemical stains such as Sirius Red (Picrosirius Red) or Masson's trichrome, followed by image-based or polarization-enhanced analysis to estimate collagen proportional area as a surrogate of extracellular matrix accumulation during fibrotic remodeling. Sirius Red combined with polarized light microscopy enhances detection of collagen fibers due to birefringence properties, enabling more specific visualization of collagen type I and III fibrils compared to conventional bright-field histology, while whole-section or region-restricted digital morphometry reduces field-selection bias in fibrosis assessment. Alternative quantitative approaches include second harmonic generation (SHG) and two-photon excited fluorescence microscopy, which enable label-free detection of fibrillar collagen and have been validated against histological staining and biochemica
Collagen: Sirius Red Staining
Sirius Red or picrosirius red staining is a histochemical method for visualizing collagen-rich extracellular matrix in tissue sections, and collagen fibers are detected as red-stained structures under bright-field microscopy with enhanced birefringence under polarized light. Picrosirius red is useful for assessing total collagen organization, distribution, and fibrosis burden, but polarized color should not be interpreted as a definitive collagen type I versus type III readout because color is affected by fiber orientation, thickness, and packing.
RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
Research Protocol for Infectious Diseases
Infectious-disease experiments test how pathogens interact with host barriers, innate immune receptors, inflammatory signaling, pathogen replication, and tissue injury; pattern-recognition receptors such as TLRs, RIG-I-like receptors, NOD-like receptors, and inflammasomes detect microbial molecules and activate NF-κB, interferon, and cytokine responses. The central hypothesis is that infection severity reflects the balance between pathogen burden and host response: protective inflammation restricts pathogen growth, whereas excessive or mislocalized inflammation contributes to tissue damage and disease phenotype. Unresolved questions include which host pathways are protective versus pathogenic, why some infection models fail to translate to human disease, and which combined readouts best predict clinically relevant infection outcomes.
Connective Tissue: Masson's Trichrome/Collagen Trichrome Staining
Masson’s Trichrome (collagen/trichrome staining) is a histological technique that differentially stains tissue compartments using sequential acidic dyes to distinguish collagen from muscle and cytoplasmic components based on dye affinity and tissue permeability differences, enabling visualization of fibrosis and connective tissue architecture in histological sections. The classical formulation typically uses Weigert's iron hematoxylin for nuclear staining, Biebrich scarlet-acid fuchsin for cytoplasm and muscle, and aniline blue (or light green variants) for collagen, producing a characteristic blue/green collagen signal contrasted against red cytoplasm and dark nuclei. The staining principle relies on selective displacement of smaller dye molecules by larger anionic dyes in collagen-rich regions under controlled acidified conditions, which enhances collagen-specific dye retention. This property makes the method widely used for fibrosis assessment in organs such as heart, liver, lung, a

Purity & Documentation

Select Batch:

Purity: 99.98%

Assay: 99.98%

ee.: 99.98%

Data Sheet (288 KB)
SDS (393 KB)
- English - EN (393 KB)
- Français - FR (393 KB)
- Deutsch - DE (393 KB)
- Norwegian - NO (393 KB)
- Español - ES (393 KB)
- Swedish - SV (393 KB)
- Italian - IT (393 KB)
- Korean - KR (393 KB)
- Portuguese - PT (393 KB)
COA (2659 KB)
Elemental Analysis Report (2659 KB)
HNMR (2659 KB)
CNMR (2659 KB)
RP-HPLC (2659 KB)
NP-HPLC (2659 KB)
MS (2659 KB)
LCMS (2659 KB)
IR (2659 KB)
GC (2659 KB)
OR (2659 KB)
SDS-PAGE (2659 KB)
SEC-HPLC (2659 KB)
Other Report (2659 KB)
Handling Instructions (2659 KB)

References

[1]. Tam J, et al. Small molecule inhibitors of Clostridium difficile toxin B-induced cellular damage. Chem Biol. 2015;22(2):175-185. [Content Brief]

[2]. Deng K, Li M, Xiang L, et al. Integrated UHPLC-Q-exactive orbitrap HRMS and serum pharmacochemistry for the investigation of anti-hepatic fibrosis effect of Baoganning Decoction[J]. Phytomedicine, 2025, 137: 156363.

Help & FAQs

How should lyophilized recombinant proteins be reconstituted and stored?

1. Before opening the cap, centrifuge the vial at 13000 rpm for 20-30 seconds. This step will ensure that any lyophilized powder that may have adhered to the cap or walls is collected at the bottom of the vial, minimizing the risk of product loss.

Complete Stock Solution Preparation Table

Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg
DMSO	1 mM	2.3663 mL	11.8315 mL	23.6630 mL	59.1576 mL
	5 mM	0.4733 mL	2.3663 mL	4.7326 mL	11.8315 mL
	10 mM	0.2366 mL	1.1832 mL	2.3663 mL	5.9158 mL
	15 mM	0.1578 mL	0.7888 mL	1.5775 mL	3.9438 mL
	20 mM	0.1183 mL	0.5916 mL	1.1832 mL	2.9579 mL
	25 mM	0.0947 mL	0.4733 mL	0.9465 mL	2.3663 mL
	30 mM	0.0789 mL	0.3944 mL	0.7888 mL	1.9719 mL
	40 mM	0.0592 mL	0.2958 mL	0.5916 mL	1.4789 mL
	50 mM	0.0473 mL	0.2366 mL	0.4733 mL	1.1832 mL
	60 mM	0.0394 mL	0.1972 mL	0.3944 mL	0.9860 mL
	80 mM	0.0296 mL	0.1479 mL	0.2958 mL	0.7395 mL
	100 mM	0.0237 mL	0.1183 mL	0.2366 mL	0.5916 mL

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Methyl cholate

All Endogenous Metabolite Isoforms