1. Cell Cycle/DNA Damage Anti-infection Neuronal Signaling Metabolic Enzyme/Protease Apoptosis
  2. Nucleoside Antimetabolite/Analog SARS-CoV Virus Protease FAAH Ceramidase Glutathione Peroxidase
  3. Carmofur

Carmofur (HCFU) is a rat recombinant acid ceramidase inhibitor with an IC50 of 29 nM. Carmofur is also a protease inhibitor of SARS-CoV-2 main protease (Mpro), fatty acid amide hydrolase (FAAH) and N-acylethanolamine acid amidase (NAAA). Carmofur has anti-cancer, anti-inflammatory and anti-virus activities, and can be used for the study of COVID-19 and acute lung injury (ALI).

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

Carmofur Chemical Structure

Carmofur Chemical Structure

CAS No. : 61422-45-5

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10 mM * 1 mL in DMSO
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Customer Review

Based on 3 publication(s) in Google Scholar

Top Publications Citing Use of Products
  • Biological Activity

  • Purity & Documentation

  • References

  • Customer Review

Description

Carmofur (HCFU) is a rat recombinant acid ceramidase inhibitor with an IC50 of 29 nM. Carmofur is also a protease inhibitor of SARS-CoV-2 main protease (Mpro), fatty acid amide hydrolase (FAAH) and N-acylethanolamine acid amidase (NAAA). Carmofur has anti-cancer, anti-inflammatory and anti-virus activities, and can be used for the study of COVID-19 and acute lung injury (ALI)[1][2][3].

IC50 & Target

IC50: 29 nM (acid ceramidase)[1].
N-Acylethanolamine Acidase (NAAA)[3].

In Vitro

Carmofur (0.3-10 μM; 20 min-1 h) inhibits acid ceramidase (AC) activity in a concentration- and time-dependent manner in human colon cancer SW403 cells[1].
Carmofur (1-100 μM) inhibits he activity of SARS-CoV-2 with an EC50 of 24.3 μM in Vero E6 cells[2].
Carmofur (5 μM; 6 h) inhibits the activities of FAAH and NAAH with IC50 values of 0.11 μM and 0.71 μM, respectively, in HEK293 cells[3].
Carmofur (10 μM; 30 min) has anti-inflammatory activity in Raw264.7 cells[3].

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

Real Time qPCR[3]

Cell Line: Raw264.7
Concentration: 10 μM. After LPS treatment (500 ng/mL; 72 h)
Incubation Time: 30 min
Result: Effectively reduced the mRNA expression of pro-inflammatory cytokines such as IL-1β, IL-6, iNOS, TNF-α.

Western Blot Analysis[3]

Cell Line: Raw264.7
Concentration: 1 μM, 3 μM, 10 μM. After LPS treatment (500 ng/mL; 72 h)
Incubation Time: 30 min
Result: Down-regulated the expression levels of p-p65 and p-κbα proteins in a dose-dependent manner, blocking the nuclear translocation of p65.
In Vivo

Carmofur (10-30 mg/kg; Intraperitoneal injection (I.P.), single dose) inhibits acid ceramidase (AC) activity in various tissues, including lung and cerebral cortex, in male Swiss Webster mice[1].
Carmofur (10 mg/kg; P.O. , single dose) significantly improves the LPS- (5 mg/kg; tracheal perfusion, single dose) (HY-D1056) induced inflammatory response by inhibiting the activities of FAAH and NAAA in in acute lung injury (ALI) mice, promoting the resolution of lung injury[3].

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

Animal Model: Male C57BL/6J mice model of LPS-induced ALI[3]
Dosage: 3 mg/kg, 10 mg/kg
Administration: Oral gavage (p.o.), single dose. After LPS treatment (5 mg/kg; Tracheal perfusion, single dose)
Result: Significantly inhibited MPO activity, which is a marker of neutrophil abundance.
Alleviated alveolar edema and inhibited neutrophil accumulation.
Molecular Weight

257.26

Formula

C11H16FN3O3

CAS No.
Appearance

Solid

Color

White to off-white

SMILES

O=C(N(C(N1)=O)C=C(F)C1=O)NCCCCCC

Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 6 months
-20°C 1 month
Solvent & Solubility
In Vitro: 

DMSO : 83.33 mg/mL (323.91 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : 0.67 mg/mL (2.60 mM; Need ultrasonic)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 3.8871 mL 19.4356 mL 38.8712 mL
5 mM 0.7774 mL 3.8871 mL 7.7742 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, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • Molarity Calculator

  • Dilution Calculator

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

Mass
=
Concentration
×
Volume
×
Molecular Weight *

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

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

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: 2.08 mg/mL (8.09 mM); Suspended solution; Need ultrasonic

    This protocol yields a suspended solution of 2.08 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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: ≥ 2.08 mg/mL (8.09 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 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:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
+
%
Tween-80 +
%
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

Purity: 99.95%

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, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
H2O / DMSO 1 mM 3.8871 mL 19.4356 mL 38.8712 mL 97.1779 mL
DMSO 5 mM 0.7774 mL 3.8871 mL 7.7742 mL 19.4356 mL
10 mM 0.3887 mL 1.9436 mL 3.8871 mL 9.7178 mL
15 mM 0.2591 mL 1.2957 mL 2.5914 mL 6.4785 mL
20 mM 0.1944 mL 0.9718 mL 1.9436 mL 4.8589 mL
25 mM 0.1555 mL 0.7774 mL 1.5548 mL 3.8871 mL
30 mM 0.1296 mL 0.6479 mL 1.2957 mL 3.2393 mL
40 mM 0.0972 mL 0.4859 mL 0.9718 mL 2.4294 mL
50 mM 0.0777 mL 0.3887 mL 0.7774 mL 1.9436 mL
60 mM 0.0648 mL 0.3239 mL 0.6479 mL 1.6196 mL
80 mM 0.0486 mL 0.2429 mL 0.4859 mL 1.2147 mL
100 mM 0.0389 mL 0.1944 mL 0.3887 mL 0.9718 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Help & FAQs
  • 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.

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