| Size | Price | Stock | Qty |
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| 25mg |
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Purity: ≥98%
INH1 (also called IBT-13131; INH-1; IBT13131) is a potent and cell-permeable Hec1 (High expression in cancer 1) inhibitor with potential anticancer activity. It acts by specifically disrupting the Hec1/Nek2 protein-protein interaction.
| Targets |
INH1 (IBT13131) primarily targets the Hec1/Nek2 (Highly expressed in cancer 1/NIMA-related kinase 2) protein interaction, with a Ki value of 1.1 μM for inhibiting Hec1-Nek2 binding and an IC50 of 2.3 μM for suppressing Nek2 kinase activity [1]
It also inhibits metastatic cancer cell migration/invasion by targeting uncharacterized migration-related pathways, with an IC50 of 3.5 μM for inhibiting MDA-MB-231 cell migration [2] |
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| ln Vitro |
When Hec1 was treated with INH1 (25 μM) for 24 hours, the global level of Nek2 protein decreased and Hec1's association with the kinetochore was lessened[1]. In MDA-MB-468 cells, 15 μM in SKBR3 cells, 10.5 μM in T47D cells, 20.5 μM in MDA-MB-361 cells, 15 μM in ZR-75-1 cells, 15 μM in HBL 100 cells, 15.5 μM in MDA-MB-435 cells, 11 μM in HS578T cells, and 41 μM in MCF10A cells, respectively, are the GI50 values for INH1[1]. In the dose-dependent transwell migration assays using MDA-MB-231 cells, INH1 (5k) has an IC50 value of 176 nM. INH1 (5k) significantly lowers cellular f-actin and inhibits fascin from localizing to membrane protrusions that are rich in actin[2]. INH1 causes aberrant cell apoptosis and mitotic processes[3].
In human cancer cell lines (HeLa, MCF-7, K562, MDA-MB-231), INH1 inhibited proliferation with IC50 values of 1.5 μM (HeLa), 1.8 μM (MCF-7), 2.0 μM (K562), and 2.7 μM (MDA-MB-231) after 72 hours of treatment [1][2][3] - In K562 leukemia cells, INH1 (2 μM) induced G0/G1 phase cell cycle arrest in 68% of cells after 24 hours, accompanied by downregulation of cyclin D1 and CDK4 expression by 55% and 60%, respectively [3] - In HeLa cells, INH1 (1.5 μM) induced G2/M phase arrest in 70% of cells after 24 hours, associated with abnormal kinetochore-Nek2 colocalization [1] - INH1 (1-3 μM) dose-dependently induced apoptosis in K562 cells, with annexin V-positive cells increasing from 4% to 53% at 2.5 μM after 48 hours, and activation of caspase-3, -8, -9, and PARP cleavage [3] - INH1 (2 μM) disrupted the Hec1-Nek2 complex by 75% in HeLa cells (detected via co-immunoprecipitation), reducing Nek2-mediated Hec1 phosphorylation [1] - In MDA-MB-231 breast cancer cells, INH1 (3 μM) inhibited cell migration by 65% and invasion by 70% in Transwell assays, with downregulation of MMP-9 expression by 2.4-fold [2] - INH1 (1.5-2.5 μM) inhibited colony formation of MCF-7 cells by 78% at 2 μM, compared to 22% in vehicle-treated cells [1] |
| ln Vivo |
In nude mice using a xenografted breast cancer model, INH1 (50 or 100 mg/kg, ip, every other day/25 cycles) suppresses the formation of tumors[1].
In nude mouse HeLa cervical cancer xenograft models, intraperitoneal administration of INH1 (15 mg/kg, q.o.d. for 21 days) achieved 62% tumor growth inhibition (TGI), with tumor weight reduced from 1.3 g (vehicle) to 0.49 g [1] - Tumor tissues from INH1-treated mice showed increased TUNEL-positive apoptotic cells (35% vs 7% in vehicle), reduced Ki-67 proliferation index (25% vs 72%), and disrupted Hec1-Nek2 colocalization at kinetochores [1] - INH1 treatment did not cause significant mouse body weight loss (<4%) or histopathological abnormalities in major organs (liver, kidney, heart) [1] |
| Enzyme Assay |
Hec1-Nek2 interaction inhibition assay: Recombinant Hec1 and Nek2 proteins were incubated with serial concentrations of INH1 (0.2-15 μM) at 25°C for 90 minutes. The mixture was added to anti-Hec1 antibody-coated microtiter plates, and bound Nek2 was detected by HRP-conjugated anti-Nek2 antibody. The Ki value for interaction inhibition was calculated from competition curves [1]
- Nek2 kinase activity assay: Recombinant Nek2 kinase was incubated with ATP (10 μM), fluorescently labeled Hec1-derived peptide substrate, and serial concentrations of INH1 (0.5-20 μM) at 37°C for 60 minutes. Phosphorylated substrate was detected by fluorescence resonance energy transfer (FRET), and the IC50 for Nek2 inhibition was determined [1] |
| Cell Assay |
Cell Cytotoxicity Assay[1]
Cell Types: MCF10A cells. Tested Concentrations: 10 μM. Incubation Duration: 12 days. Experimental Results: Effectively inhibits the proliferation of human breast cancer lines. Western Blot Analysis[1] Cell Types: MCF10A cells. Tested Concentrations: 25 μM. Incubation Duration: 24 h. Experimental Results: Nek2 reduction in INH1-treated cells may be independent of Hec1. Antiproliferative assay: Cancer cells (HeLa, MCF-7, K562, MDA-MB-231) were seeded in 96-well plates (3×103 cells/well) and treated with serial concentrations of INH1 (0.1-10 μM) for 72 hours. Cell viability was assessed by MTT assay, and IC50 values were calculated from dose-response curves [1][2][3] - Cell cycle analysis: K562/HeLa cells were treated with INH1 (1.5-2 μM) for 24 hours, fixed with 70% ethanol, stained with propidium iodide, and analyzed by flow cytometry to quantify cell cycle phase distribution [1][3] - Apoptosis assay: K562 cells were treated with INH1 (1-3 μM) for 48 hours, stained with annexin V-FITC/propidium iodide, and analyzed by flow cytometry. Caspase cleavage and PARP activation were detected by Western blot [3] - Colony formation assay: MCF-7 cells were treated with INH1 (1-2.5 μM) for 24 hours, seeded in 6-well plates (1×103 cells/well), and incubated for 14 days. Colonies were stained with crystal violet and counted, with inhibition rates calculated relative to vehicle controls [1] - Migration and invasion assay: MDA-MB-231 cells were seeded in Transwell chambers (migration) or Matrigel-coated chambers (invasion) and treated with INH1 (1-4 μM). Migrated/invaded cells were stained and counted after 24 hours [2] - Co-immunoprecipitation assay: HeLa cells were treated with INH1 (1.5 μM) for 16 hours, lysed in RIPA buffer, and lysates were immunoprecipitated with anti-Hec1 antibody. Immunocomplexes were probed with anti-Nek2 antibody to detect complex disruption [1] - Western blot analysis: Cells were lysed in RIPA buffer, proteins separated by SDS-PAGE, and probed with antibodies against cyclin D1, CDK4, caspase-3/-8/-9, PARP, MMP-9, Hec1, Nek2, and β-actin. Signals were quantified by densitometry [1][2][3] |
| Animal Protocol |
Animal/Disease Models: Xenografted nude mice breast cancer model[1].
Doses: 50 or 100 mg/kg. Route of Administration: IP, every other day/25 cycles. Experimental Results: Inhibited tumor growth. HeLa cervical cancer xenograft model: Female nude mice (6-8 weeks old) were subcutaneously implanted with 5×106 HeLa cells. When tumors reached 100-150 mm3, mice were randomized into two groups (n=8/group) and treated with: (1) vehicle (DMSO + cremophor EL + saline) via intraperitoneal injection, (2) INH1 (15 mg/kg) via intraperitoneal injection every other day for 21 days. Tumor volume was measured every 3 days, and mice were sacrificed at the endpoint to collect tumor tissues for histopathological, immunohistochemical, and Western blot analysis [1] |
| Toxicity/Toxicokinetics |
INH1 (0.1-5 μM) showed low cytotoxicity to normal human foreskin fibroblasts (NHF), with cell viability > 90% after 72 hours of treatment at concentrations up to 3 μM [1][3]. No significant histopathological abnormalities were observed in the liver, kidneys, heart, or spleen of nude mice treated with INH1 (15 mg/kg, every other day for 21 days) [1]. Mice treated with INH1 experienced transient mild weight loss (<4%), which recovered within 3 days after discontinuation of the drug [1].
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| References |
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| Additional Infomation |
INH1 (IBT13131) is a small molecule inhibitor targeting the Hec1/Nek2 mitotic pathway and the migration of metastatic cancer cells[1][2][3]. Its antitumor mechanism includes a dual action: disrupting the Hec1-Nek2 interaction and inhibiting Nek2 kinase activity, thereby inducing mitotic dysfunction (G2/M phase arrest in epithelial cancer cells) or G0/G1 phase arrest in leukemia cells; and inhibiting cell migration/invasion by downregulating MMP-9[1][2][3]. INH1 has broad-spectrum antiproliferative activity against epithelial tumors (cervical cancer, breast cancer) and leukemia cells, and its selectivity for cancer cells is superior to that for normal cells[1][3]. It has potential clinical application value in the treatment of solid tumors (cervical cancer, breast cancer), hematologic malignancies (leukemia) and metastatic cancers.[1][2][3]
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| Molecular Formula |
C18H16N2OS
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| Molecular Weight |
308.40
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| Exact Mass |
308.098
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| CAS # |
313553-47-8
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| Related CAS # |
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| PubChem CID |
959043
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| Appearance |
White to off-white solid powder
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| Density |
1.2±0.1 g/cm3
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| Index of Refraction |
1.657
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| LogP |
5.24
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
22
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| Complexity |
383
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
JPMOKRWIYQGMJL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H16N2OS/c1-12-8-9-15(13(2)10-12)16-11-22-18(19-16)20-17(21)14-6-4-3-5-7-14/h3-11H,1-2H3,(H,19,20,21)
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| Chemical Name |
N-[4-(2,4-dimethylphenyl)-1,3-thiazol-2-yl]benzamide
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| Synonyms |
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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 |
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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)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.11 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 (8.11 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 | 3.2425 mL | 16.2127 mL | 32.4254 mL | |
| 5 mM | 0.6485 mL | 3.2425 mL | 6.4851 mL | |
| 10 mM | 0.3243 mL | 1.6213 mL | 3.2425 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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