HSF1A is a cell-permeable and small-molecule activator of heat shock transcription factor 1 (HSF1). HSF1A inhibits TRiC activity without interfering with ATP hydrolysis, binds TRiC subunits both in vivo and in vitro, and shields cells from stress-induced apoptosis. Human HSF1 is activated upon genetic inactivation or depletion of the TRiC complex, and the in vitro direct interaction between purified TRiC and HSF1 is inhibited by HSF1A. Heat shock transcription factor 1 (HSF1) is a transcription factor that has been conserved throughout evolution and shields cells from stress and apoptosis caused by misfolded proteins.
Physicochemical Properties
| Molecular Formula | C21H19N3O2S2 | |
| Molecular Weight | 409.52 | |
| Exact Mass | 409.092 | |
| Elemental Analysis | C, 61.59; H, 4.68; N, 10.26; O, 7.81; S, 15.66 | |
| CAS # | 1196723-93-9 | |
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| PubChem CID | 44472508 | |
| Appearance | White to off-white solid powder | |
| LogP | 6.117 | |
| Hydrogen Bond Donor Count | 1 | |
| Hydrogen Bond Acceptor Count | 5 | |
| Rotatable Bond Count | 6 | |
| Heavy Atom Count | 28 | |
| Complexity | 595 | |
| Defined Atom Stereocenter Count | 0 | |
| SMILES | C1(S(NC2N(C3=CC=CC=C3)N=C(C3SC=CC=3)C=2)(=O)=O)=CC=C(CC)C=C1 |
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| InChi Key | KJTITGSAONQVPY-UHFFFAOYSA-N | |
| InChi Code | InChI=1S/C21H19N3O2S2/c1-2-16-10-12-18(13-11-16)28(25,26)23-21-15-19(20-9-6-14-27-20)22-24(21)17-7-4-3-5-8-17/h3-15,23H,2H2,1H3 | |
| Chemical Name | 4-ethyl-N-(2-phenyl-5-thiophen-2-ylpyrazol-3-yl)benzenesulfonamide | |
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| 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 |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets | HSF1 | |
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| Enzyme Assay | Using biotin-binding buffer (20 mM HEPES, 5 mM MgCl2, 1 mM EDTA, 100 mM KCl, 0.03% NP-40), in addition to protease inhibitors and 1% Trition-X100, protein extracts are produced from mammalian, yeast, and E. Coli cultures. After 4 hours at 4°C incubation with 100 μM HSF1A-Biotin, about 0.5 mg of protein extract is captured with NeutrAvidin Agarose Resin, corresponding with HSF1A-Biotin'sassociatedproteins. Once the proteins have been cleaned in the biotin binding buffer, they are resolved on a 4–20% SDS-PAGE and immunoblotted using 50 μL of biotin elution buffer (100 mM Tris, 150 mM NaCl, 0.1 mM EDTA, and 2 mM D-biotin). In order to analyze purified TRiC and Hsp70, 5 nM protein is incubated for 4 hours at 4°C in biotin-binding buffer+0.5% Triton X-100 with 100 μM biotin or 100 μM HSF1A-Biotin. The protein is then captured using NeutrAvidin Rinse. Different concentrations of Tcp1 (0.5 μM, 1 mM, 2 mM, 3 mM, and 4 mM) in 25 mM Hepes pH 7.5, 150 mM NaCl are incubated with 0.5 μM Biotin or HSF1A-Biotin for 4 hours at 4°C and then captured using NeutrAvidin Resin for NiNTA purified yeast Tcp1[1]. | |
| Cell Assay | PC12 cells seeded into a 96-well plate (5 ×104 cells/well) are treated with increasing concentrations of HSF1A (2, 4, 8, and 12 μM) for 15 hours. Following this, httQ74-GFP expression is stimulated by incubating the cells for 5 days in the presence of 1 µg/mL Doxycycline. The XTT viability assay is used to evaluate cell viability[2]. | |
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| References |
[1]. A direct regulatory interaction between chaperonin TRiC and stress-responsive transcription factor HSF1. Cell Rep. 2014 Nov 6;9(3):955-66. [2]. Modulation of heat shock transcription factor 1 as a therapeutic target for small molecule intervention in neurodegenerative disease. PLoS Biol. 2010 Jan 19;8(1):e1000291. [3]. Doxorubicin attenuates CHIP-guarded HSF1 nuclear translocation and protein stability to trigger IGF-IIR-dependent cardiomyocyte death. Cell Death Dis. 2016 Nov 3;7(11):e2455. [4]. The chaperonin TRiC/CCT is essential for the action of bacterial glycosylating protein toxins like Clostridium difficile toxins A and B. Proc Natl Acad Sci U S A. 2018 Sep 18;115(38):9580-9585. |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.10 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 (6.10 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (6.10 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 | 2.4419 mL | 12.2094 mL | 24.4188 mL | |
| 5 mM | 0.4884 mL | 2.4419 mL | 4.8838 mL | |
| 10 mM | 0.2442 mL | 1.2209 mL | 2.4419 mL |