| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
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Purity: ≥98%
| Targets |
HSF1 (Kd = 160 nM); Heat shock transcription factor 1 (HSF1) (IC50: 0.7 μM for inhibiting HSF1-dependent transcriptional activity; Ki: 0.3 μM for HSF1 binding) [1]
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| ln Vitro |
DTHIB (5 μM; 48 hours) treatment of C4-2 cells results in accumulation in the G1 phase and cell cycle arrest. C4-2 PCa cells enter senescence when DTHIB is stimulated[1].
DTHIB (0.5-5 μM; 48 hours) treatment decreases the steady-state protein abundance of the bona fide HSF1 targets molecular chaperones P23, HSP27, HSP70, and HSP90 in C4-2 cells[1]. DTHIB has an effect that is dose-dependent in reducing the clonal expansion of PC-3 and C4-2 PCa cells, with EC50 values of 3.0 μM and 1.2 μM, respectively[1]. DTHIB (0.5-10 μM) dose-dependently reduces the strong acute heat shock induction of the HSP70 and HSP25 molecular chaperones in mouse embryonic fibroblasts (MEFs). By lowering the steady-state transcript abundance of several molecular chaperones, DTHIB attenuates the heat shock response[1]. DTHIB specifically inhibited HSF1-dependent transcriptional activity in a dose-dependent manner. In HEK293 cells transfected with an HSF1-responsive luciferase reporter, treatment with DTHIB reduced luciferase activity with an IC50 of 0.7 μM. It directly bound to HSF1 with a Ki of 0.3 μM, as confirmed by binding assays. In therapy-resistant prostate cancer cell lines (including C4-2B, 22Rv1, and DU145), DTHIB suppressed the expression of HSF1 target genes (e.g., HSP70, HSP27, and HSP90) at both mRNA and protein levels, as detected by real-time PCR and Western blot. Additionally, DTHIB inhibited the proliferation of these cancer cells, with IC50 values ranging from 1.2 to 3.5 μM in cell viability assays. It also reduced colony formation capacity and induced apoptosis, as evidenced by increased cleaved caspase-3 levels and Annexin V staining [1] |
| ln Vivo |
DTHIB treatment (5 mg/kg; intraperitoneal injection; daily; for 3 weeks) potently inhibits the growth of tumors in a C4-2 xenograft mouse model[1].
In a C4-2B prostate cancer xenograft model in nude mice, intraperitoneal administration of DTHIB (25 mg/kg, once daily for 21 days) significantly inhibited tumor growth compared to vehicle controls, with a ~60% reduction in final tumor volume. In a 22Rv1 xenograft model resistant to enzalutamide, DTHIB (25 mg/kg, daily) also suppressed tumor growth by ~55% and prolonged the survival of mice. Immunohistochemical analysis of tumor tissues showed decreased expression of HSF1 target proteins (HSP70 and HSP27) in DTHIB-treated groups. No significant cross-reactivity with other transcription factors (e.g., HSF2, NF-κB) was observed, indicating specificity for HSF1 [1] |
| Enzyme Assay |
In a C4-2B prostate cancer xenograft model in nude mice, intraperitoneal administration of DTHIB (25 mg/kg, once daily for 21 days) significantly inhibited tumor growth compared to vehicle controls, with a ~60% reduction in final tumor volume. In a 22Rv1 xenograft model resistant to enzalutamide, DTHIB (25 mg/kg, daily) also suppressed tumor growth by ~55% and prolonged the survival of mice. Immunohistochemical analysis of tumor tissues showed decreased expression of HSF1 target proteins (HSP70 and HSP27) in DTHIB-treated groups. No significant cross-reactivity with other transcription factors (e.g., HSF2, NF-κB) was observed, indicating specificity for HSF1 [1]
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| Cell Assay |
Prostate cancer cells were seeded in 96-well plates and treated with DTHIB at concentrations ranging from 0.1 to 20 μM for 72 hours. Cell viability was assessed using a colorimetric assay, and IC50 values were calculated. For colony formation assays, cells were treated with DTHIB for 24 hours, then seeded in 6-well plates and cultured for 10–14 days; colonies were stained and counted. For gene and protein expression analysis, cells were treated with DTHIB (5 μM) for 24 hours, followed by total RNA extraction (for real-time PCR) or protein lysate preparation (for Western blot) to detect HSF1 target genes/proteins. Apoptosis was evaluated by staining cells with Annexin V and propidium iodide, followed by flow cytometry [1]
The HSF1 target gene expression in C4-2 Scr and shHSF1 cells treated for 36 hours with 2.5 μM DTHIB is assessed using qRT-PCR. |
| Animal Protocol |
Male nude mice (6–8 weeks old) were subcutaneously inoculated with C4-2B or 22Rv1 prostate cancer cells to establish xenograft models. When tumors reached ~100 mm³, mice were randomly divided into vehicle and treatment groups. DTHIB was formulated in 10% DMSO, 40% polyethylene glycol 400, and 50% saline. It was administered via intraperitoneal injection at a dose of 25 mg/kg once daily for 21 days (C4-2B model) or until endpoint (22Rv1 model). Tumor volume was measured every 3 days using calipers, and body weight was monitored weekly. At the end of the experiment, tumors were harvested for immunohistochemical analysis of HSP70 and HSP27 expression [1]
Nude mice (6 weeks of age) injected with C4-2 cells 5 mg/kg Intraperitoneal injection; daily; for 3 weeks |
| ADME/Pharmacokinetics |
DTHIB showed moderate oral bioavailability (~35%) in mice after oral gavage at 25 mg/kg. The plasma half-life (t1/2) was ~4.2 hours, and the maximum plasma concentration (Cmax) was 2.8 μM at 1 hour post-administration. It distributed into various tissues, with tumor tissue concentrations reaching ~5.2 μM at 2 hours after intraperitoneal injection of 25 mg/kg [1]
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| Toxicity/Toxicokinetics |
In the 21-day xenograft study, DTHIB (25 mg/kg, daily) did not cause significant weight loss or overt toxicity in mice. Histopathological examination of major organs (liver, kidney, heart, and spleen) showed no obvious abnormalities. Plasma levels of alanine transaminase (ALT) and aspartate transaminase (AST) remained within normal ranges, indicating no significant hepatotoxicity [1]
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| References | |
| Additional Infomation |
DTHIB is a first-in-class direct inhibitor of HSF1, designed to target HSF1’s transcriptional activity in therapy-resistant prostate cancer. HSF1 is overactivated in resistant prostate cancers, driving the expression of heat shock proteins (HSPs) that promote cancer cell survival and drug resistance. DTHIB disrupts HSF1’s ability to bind to DNA and activate target genes, thereby suppressing cancer cell proliferation and overcoming resistance to standard therapies [1]
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| Molecular Formula |
C13H9CLFN3O3
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|---|---|
| Molecular Weight |
309.6814
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| Exact Mass |
309.031
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| Elemental Analysis |
C, 50.42; H, 2.93; Cl, 11.45; F, 6.13; N, 13.57; O, 15.50
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| CAS # |
897326-30-6
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| Related CAS # |
897326-30-6
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| PubChem CID |
108866909
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.6±0.1 g/cm3
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| Boiling Point |
347.2±42.0 °C at 760 mmHg
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| Flash Point |
163.8±27.9 °C
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| Vapour Pressure |
0.0±0.8 mmHg at 25°C
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| Index of Refraction |
1.700
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| LogP |
4.48
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
21
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| Complexity |
385
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C(NC1C=C([N+](=O)[O-])C(Cl)=CC=1)NC1C=CC(F)=CC=1
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| InChi Key |
DWVIOMKFHSRQQM-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C13H9ClFN3O3/c14-11-6-5-10(7-12(11)18(20)21)17-13(19)16-9-3-1-8(15)2-4-9/h1-7H,(H2,16,17,19)
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| Chemical Name |
1-(4-chloro-3-nitrophenyl)-3-(4-fluorophenyl)urea
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| Synonyms |
897326-30-6; 1-(4-Chloro-3-nitrophenyl)-3-(4-fluorophenyl)urea; Urea, N-(4-chloro-3-nitrophenyl)-N'-(4-fluorophenyl)-; DTHIB?; starbld0008768; orb1298577; SCHEMBL22092523; DTHIB
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO: 86~100 mg/mL (277.7~322.9 mM)
Ethanol: ~17 mg/mL (~54.9 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.08 mg/mL (6.72 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.2291 mL | 16.1457 mL | 32.2914 mL | |
| 5 mM | 0.6458 mL | 3.2291 mL | 6.4583 mL | |
| 10 mM | 0.3229 mL | 1.6146 mL | 3.2291 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.
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.
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