Apoptozole

Alias: Apoptosis Activator VII; Apoptozole
Cat No.:V2829 Purity: ≥98%
Apoptozole (formerly known asApoptosis Activator VII) is a potent and selective inhibitor of heat shock protein 70(HSP70) and Hsc70.
Apoptozole Chemical Structure CAS No.: 1054543-47-3
Product category: HSP
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
5mg
10mg
25mg
50mg
100mg
250mg
500mg
Other Sizes
Official Supplier of:
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text
Alternate Text

 

  • Business Relationship with 5000+ Clients Globally
  • Major Universities, Research Institutions, Biotech & Pharma
  • Citations by Top Journals: Nature, Cell, Science, etc.
Top Publications Citing lnvivochem Products
Purity & Quality Control Documentation

Purity: ≥98%

Product Description

Apoptozole (formerly known as Apoptosis Activator VII) is a potent and selective inhibitor of heat shock protein 70(HSP70) and Hsc70. It can cause apoptosis and inhibits the ATPase domain of Hsp70 and Hsc70, with Kd values of 0.14 μM and 0.21 μM, respectively.causes caspase-dependent apoptosis by preventing HSP70 from interacting with APAF-1; this interaction is unaffected by HSP70's interactions with ASK1, JNK, BAX, and AIF. Nevertheless, in aqueous environments, apoptozole may clump together and interact non-specifically with HSP70 proteins, possibly producing inconsistent results and false positives.

Biological Activity I Assay Protocols (From Reference)
Targets
HSP70 ( Kd = 0.14 μM ); HSC70 ( Kd = 0.21 μM )
ln Vitro

In vitro activity: Apoptozole, which binds to Hsc70 and Hsp70 with Kds of 0.21 and 0.14 μM, respectively, is an inhibitor of Hsc70 and Hsp70. In P19 cells, apoptosis is induced by apoptozole (Apoptosis Activator VII; 1 μM). Apoptozole exhibits inhibitory activities against a number of cancer cell lines, with IC50s of 0.22, 0.25, and 0.13 μM for ovarian cancer cells, colon cancer cells, and lung cancer cells, respectively, HCT-15, and A549[1]. While it does not bind to other heat shock protein types like Hsp60, Hsp90, or Hsp40, apoplastozole does bind to the ATPase domain of Hsc70 and Hsp70[2]. With IC50s ranging from 5 to 7 μM, apoptozole (0-15 μM) inhibits the growth of HeLa, MDA-MB-231, and A549 cells. Apoptozole (5 or 10 μM) does not cause AIF-mediated caspase-independent apoptosis in HeLa cells, nor does it affect HSP70's associations with ASK1, JNK, or BAX[3].

ln Vivo
Apoptozole (Apoptosis Activator VII; 4 mg/kg, i.p.) has antitumor properties in A549, RKO (colorectal carcinoma), and HeLa cell xenografts in nude mice[3].
Enzyme Assay
Ammonium heptamolybdate tetrahydrate (5.7% w/v in 6 M HCl), polyvinyl alcohol (2.3% w/v), and malachite green (0.081% w/v) stock solutions are made and kept at 4°C. To make the malachite green reagent, three solutions are combined with water in a 2:1:1:1:2 ratio. A master mixture of the ATPase domain of Hsc70 is prepared in assay buffer (100 mM Tris-HCl, 20 mM KCl, and 6 mM MgCl2, pH 7.4) at a final concentration of 1 mM in order to measure the ATPase activity of Hsc70. In a 96-well plate, aliquots (10 mL) of this mixture are added to each well. Each compound, including apoptozole, is added to this solution in assay buffer, and the plate is then allowed to sit at room temperature for 30 minutes. One milliliter of 4 mM ATP is added to the mixture to initiate the reaction. 200 mM ATP and 1 mM protein in 20 mL of assay buffer are the final concentrations. Each well receives 80 mL of the malachite green reagent after three hours of 37°C incubation. After thoroughly combining the samples, they are incubated for 15 minutes at 37°C. Afterwards, 10 milliliters of 34% sodium citrate are added to halt the nonenzymatic hydrolysis of ATP. The SpectraMax 340 PC 384 is used to measure the absorbance at 620 nm[1].
Cell Assay
In triplicate, 5 × 105 cells are plated in 96-well plates with 0.1 mL of culture media containing 10% FBS. Cells are treated with different concentrations of Apoptozole (0-15 μM) in culture media containing 3% FBS (final volume: 0.2 mL per well) for 24, 48, and 72 hours prior to being treated with MTT. This is done after the 24-hour mark. A UV microplate reader is used to measure absorbance at 570 nm[3].
Animal Protocol
Male naked mice are kept in a pathogen-free environment with regulated humidity and temperature. For the xenograft experiments, tumor cells are injected into 4-week-old mice. Mice are subcutaneously injected with viable A549, RKO, and HeLa cells (5 × 106) in the flank. The A549 and RKO cell xenograft mice are split into two groups at random right away. As a control group, the first group (n = 10) is given a vehicle exclusively. For a duration of two weeks, the second group (n = 10) is administered intraperitoneal injections of Apoptozole (4 mg/kg/day) every other day. Four groups are instantly and randomly assigned to the HeLa cell xenograft mice. The initial group (n = 10) is a control group that only receives vehicles. For two weeks, apoptozole (4 mg/kg/day) is injected intraperitoneally into the second group (n = 10). For two weeks, doxorubicin (15 mg/kg/day) is injected intraperitoneally into the third group (n = 10). For two weeks, intraperitoneal injections of doxorubicin (15 mg/kg/day) and apapozole (4 mg/kg/day) are given to the fourth group (n = 10). Every three days, the tumor volumes of all the mice are determined by measuring their tumors in two dimensions using calipers. The formula for calculating tumor volumes is volume = w × l2/2, where w represents the width of the tumor at its widest point and l is the length perpendicular to w. The average tumor volumes versus time are plotted for each mouse's results[3].
References

[1]. An apoptosis-inducing small molecule that binds to heat shock protein 70. Angew Chem Int Ed Engl. 2008;47(39):7466-9.

[2]. Probing the effect of an inhibitor of an ATPase domain of Hsc70 on clathrin-mediated endocytosis. Mol Biosyst. 2015 Oct;11(10):2763-9.

[3]. A small molecule inhibitor of ATPase activity of HSP70 induces apoptosis and has antitumor activities. Chem Biol. 2015 Mar 19;22(3):391-403.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C33H25F6N3O3
Molecular Weight
625.56
Exact Mass
625.18
Elemental Analysis
C, 63.36; H, 4.03; F, 18.22; N, 6.72; O, 7.67
CAS #
1054543-47-3
Related CAS #
1054543-47-3
Appearance
Solid powder
SMILES
COC1=CC=C(C=C1)C2=C(N(C(=N2)C3=CC(=CC(=C3)C(F)(F)F)C(F)(F)F)CC4=CC=C(C=C4)C(=O)N)C5=CC=C(C=C5)OC
InChi Key
ZIMMTPFXOMAJTQ-UHFFFAOYSA-N
InChi Code
InChI=1S/C33H25F6N3O3/c1-44-26-11-7-20(8-12-26)28-29(21-9-13-27(45-2)14-10-21)42(18-19-3-5-22(6-4-19)30(40)43)31(41-28)23-15-24(32(34,35)36)17-25(16-23)33(37,38)39/h3-17H,18H2,1-2H3,(H2,40,43)
Chemical Name
4-[[2-[3,5-bis(trifluoromethyl)phenyl]-4,5-bis(4-methoxyphenyl)imidazol-1-yl]methyl]benzamide
Synonyms
Apoptosis Activator VII; Apoptozole
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: ~100 mg/mL (159.85 mM)
Water:<1 mg/mL
Ethanol: ~25 mg/mL (~39.96 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.00 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 (4.00 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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.

View More

Solubility in Formulation 3: ≥ 2.5 mg/mL (4.00 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 1.5986 mL 7.9928 mL 15.9857 mL
5 mM 0.3197 mL 1.5986 mL 3.1971 mL
10 mM 0.1599 mL 0.7993 mL 1.5986 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.
/

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.)
+
+
+

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

  • An external file that holds a picture, illustration, etc.
Object name is pone.0140006.g002.jpg

    Competitive displacement curves for ATP-FAM and HSP72 with ATP, ADP, VER-155008 and apoptozole.2015 Oct 12;10(10):e0140006.

  • An external file that holds a picture, illustration, etc.
Object name is pone.0140006.g004.jpg

    Binding isotherms for ATP-FAM, ATP-Cy5, apoptozole-FAM and apoptozole-Cy5 with HSP72.2015 Oct 12;10(10):e0140006.

  • Apoptozole

    SPR sensorgrams of HSP72 interacting with biotinylated apoptozole 4 immobilised on a neutravidin derivatised CM5 sensor chip.2015 Oct 12;10(10):e0140006.
Contact Us Back to top