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CHM-1

Alias: CHM-1 CHM1CHM 1 NSC-656158 NSC656158 NSC 656158
Cat No.:V10037 Purity: ≥98%
CHM-1 is a microtubule destabilizer that can inhibit tubulin polymerization.
CHM-1
CHM-1 Chemical Structure CAS No.: 154554-41-3
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
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Product Description
CHM-1 is a microtubule destabilizer that can inhibit tubulin polymerization. CHM-1 displays potent and specific anti-tumor effects against human liver cancer mitosis. CHM-1 induces G2-M phase arrest in human hepatoma cells by activating Cdc2 kinase activity, thereby inducing cell growth inhibition and apoptosis.
Biological Activity I Assay Protocols (From Reference)
ln Vitro
In HA22T, Hep3B, and HepG2 cells, CHM-1 (0-100 μM; 24 hours) significantly inhibits growth in a concentration-dependent manner, with HA22T cells showing the strongest effect (IC50 = 0.75 μM)[1]. In HA22T cells, CHM-1 (0-10 μM; 24 hours) dramatically boosts cyclin B1's binding to Cdc2 [1].
ln Vivo
The growth of HA22T tumors is dose-dependently inhibited by CHM-1 (10 mg/kg; IP) [1].
Cell Assay
Cell Viability Assay[1]
Cell Types: HA22T, Hep3B and HepG2 Cell
Tested Concentrations: 0-100 μM
Incubation Duration: 24 hrs (hours)
Experimental Results: Induction of cell cycle G2-M arrest, followed by apoptosis.

Western Blot Analysis [1]
Cell Types: HA22T Cell
Tested Concentrations: 0-10 μM
Incubation Duration: 24 hrs (hours)
Experimental Results: Induced changes in the expression and phosphorylation status of G2-M regulatory factors in human hepatocellular carcinoma cells.
Animal Protocol
Animal/Disease Models: Male severe combined immunodeficiency mice (HA22T) [1]
Doses: 10 mg/kg
Route of Administration: intraperitoneal (ip) injection
Experimental Results: Induced dose-dependent HA22T tumor growth inhibition.
References

[1]. CHM-1, a novel synthetic quinolone with potent and selective antimitotic antitumor activity against human hepatocellular carcinoma in vitro and in vivo. Mol Cancer Ther. 2008 Feb;7(2):350-60.

[2]. CHM-1, a novel microtubule-destabilizing agent exhibits antitumor activity via inducing the expression of SIRT2 in human breast cancer cells. Chem Biol Interact. 2018 Jun 1;289:98-108.

[3]. CHM-1, a new vascular targeting agent, induces apoptosis of human umbilical vein endothelial cells via p53-mediated death receptor 5 up-regulation. J Biol Chem. 2010 Feb 19;285(8):5497-506.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C16H10FNO3
Molecular Weight
283.25
Exact Mass
283.064
CAS #
154554-41-3
PubChem CID
375860
Appearance
Off-white to light yellow solid powder
LogP
3.062
Hydrogen Bond Donor Count
1
Hydrogen Bond Acceptor Count
5
Rotatable Bond Count
1
Heavy Atom Count
21
Complexity
466
Defined Atom Stereocenter Count
0
InChi Key
ZMYDAPJHGNEFGQ-UHFFFAOYSA-N
InChi Code
InChI=1S/C16H10FNO3/c17-11-4-2-1-3-9(11)12-6-14(19)10-5-15-16(21-8-20-15)7-13(10)18-12/h1-7H,8H2,(H,18,19)
Chemical Name
6-(2-fluorophenyl)-[1,3]dioxolo[4,5-g]quinolin-8(5H)-one
Synonyms
CHM-1 CHM1CHM 1 NSC-656158 NSC656158 NSC 656158
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)
Solubility Data
Solubility (In Vitro)
DMSO : ~5 mg/mL (~17.65 mM)
H2O : < 0.1 mg/mL
Solubility (In Vivo)
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

Injection Formulations
(e.g. IP/IV/IM/SC)
Injection Formulation 1: DMSO : Tween 80: Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution 50 μL Tween 80 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO 400 μLPEG300 50 μL Tween 80 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).
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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO 100 μLPEG300 200 μL castor oil 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10: 10 : 80 (i.e. 100 μL Ethanol 100 μL Cremophor 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH 400 μLPEG300 50 μL Tween 80 450 μL Saline)


Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).
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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders


Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 3.5305 mL 17.6523 mL 35.3045 mL
5 mM 0.7061 mL 3.5305 mL 7.0609 mL
10 mM 0.3530 mL 1.7652 mL 3.5305 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.

Calculator

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

  • Calculate the Mass of a compound required to prepare a solution of known volume and concentration
  • Calculate the Volume of solution required to dissolve a compound of known mass to a desired concentration
  • Calculate the Concentration of a solution resulting from a known mass of compound in a specific volume
An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
  • Enter 350.26 in the Molecular Weight (MW) box
  • Enter 10 in the Concentration box and choose the correct unit (mM)
  • Enter 5 in the Volume box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
  • Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
  • Enter 25 into the Concentration (End) box and select the correct unit (mM)
  • Enter 25 into the Volume (End) box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
  • To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.
Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
  • Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.
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Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

  • Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
  • Click the “Calculate” button
  • The answer appears in the Volume (to add to vial) box
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • CHM-1 on cell growth and cell cycle progression in human hepatocellular carcinoma cells. A, chemical structure of CHM-1. B, HA22T, Hep3B, and HepG2 cells were treated with the indicated concentrations of CHM-1 for 24 h (left). MRC5 and AML12 cells were cultured for 24 h in the presence of CHM-1 or doxorubicin (0-100 μmol/L; right). Then, the cell viability was determined using MTT assay. C, time effect of CHM-1 on cell cycle distribution. HA22T cells were treated with vehicle (DMSO) or CHM-1 (1 μmol/L) for the indicated times. D, after 24-h treatment with vehicle or CHM-1 (1 μmol/L), terminal deoxynucleotidyl transferase–mediated dUTP nick end labeling assay was used to detect apoptotic effect (left). Then, HA22T cells were treated with indicated concentrations of CHM-1 to determine apoptosis using Cell Death ELISAPLUS kit (right). Mean ± SE of five independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001, compared with the respective control group.
  • CHM-1 on microtubule assembly in vitro and in vivo. A, cells were incubated with vehicle (DMSO) or 3 μmol/L CHM-1 for 4 h and treated with 3 μmol/L CHM-1, 3 μmol/L colchicine, 3 μmol/L paclitaxel, or 3 μmol/L vincristine for 24 h. Left, 4′,6-diamidino-2-phenylindole; middle, microtubule network; right, merged microtubule network and 4′,6-diamidino-2-phenylindole. B, tubulin proteins (>99% purity) were suspended in G-PEM buffer plus glycerol in the absence (DMSO) or presence of 3, 10, or 30 μmol/L CHM-1, 10 μmol/L paclitaxel, or 10 μmol/L vincristine. C, after 24-h treatment with vehicle or 3 μmol/L antimitotic agents, cytosolic (S, soluble) and cytoskeletal (P, polymerized tubulin) tubulin fractions were separated and followed by immunoblotting with antibody against β-tubulin. D, after 24-h treatment with vehicle or CHM-1, HA22T cells were harvested and lysed for the detection of MAP4, stathmin, and GAPDH protein expression.
  • CHM-1 on caspase cascade and inhibitor of apoptosis proteins in human hepatocellular carcinoma cells. A, HA22T cells were treated with DMSO (as control), the indicated concentrations of CHM-1, or staurosporine (Stau; 1 μmol/L) for 24 h. Cell extracts were then subjected to Western blotting using anti-caspase-3, anti-caspase-6, anti-caspase-7, anti-caspase-8, and anti-caspase-9 antibodies. B, HA22T cells were incubated in the absence or presence of CHM-1 (3 μmol/L) for the indicated time. Cells were lysed for the detection of cIAP1, cIAP2, XIAP, survivin, and GAPDH protein expression. C, caspase protease activity in HA22T cell lysates were assayed by spectrophometric detection. D, HA22T cells were incubated in 1 μmol/L CHM-1 with or without z-VAD-fmk (50 μmol/L) for 24 h. Then, the cell viability was determined using MTT assay. Mean ± SE of five independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001, compared with the respective control group.
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