2. Metabolic Enzyme/Protease Cytoskeleton
  3. Endogenous Metabolite Collagen
  4. Methyl cholate

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.

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Methyl cholate

Methyl cholate Chemical Structure

CAS No. : 1448-36-8

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Based on 1 publication(s) in Google Scholar

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Methyl cholate Related Antibodies

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Description

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[1][2].

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.

Methyl cholate Related Antibodies

Western Blot Analysis[1]

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]

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).
Molecular Weight

422.60

Formula

C25H42O5
CAS No.
Appearance

Solid

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
Initial Source

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)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.3663 mL 11.8315 mL 23.6630 mL
5 mM 0.4733 mL 2.3663 mL 4.7326 mL
View the Complete Stock Solution Preparation Table

* 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.

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

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

This equation is commonly abbreviated as: C1V1 = C2V2

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In Vivo:

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

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.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • 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.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
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% 
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Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation
References

Complete Stock Solution Preparation Table

* 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.

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 Related Classifications

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  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Methyl cholate1448-36-8Endogenous MetaboliteCollagenJS-1 hepatic stellate cellsClostridium difficile infectionCOL-Ihuman IMR-90 fibroblastshepatic fibrosisTcdAHSC-T6 hepatic stellate cellsα-SMAzebrafish fibrosis modelsClostridium difficile Toxin BInhibitorinhibitorinhibit

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