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| 5mg |
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| 10mg |
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| Targets |
Hypoxia-Inducible Factor 1-alpha (HIF-1alpha). HIF-1alpha-IN-2 hydrochloride is a small-molecule inhibitor that targets the transcription and translation of the HIF-1alpha protein. HIF-1alpha is a key transcription factor that is stabilized under hypoxic conditions and drives the expression of genes involved in angiogenesis, metabolism, cell proliferation, and survival (e.g., VEGF, GLUT1, CA9, and others). Overexpression of HIF-1alpha is associated with poor prognosis in many cancers, as it promotes tumor adaptation to low oxygen levels. By inhibiting HIF-1alpha expression at both the transcriptional and translational levels, this compound effectively shuts down the HIF-1alpha signaling pathway, reducing the adaptive response of cancer cells to hypoxia and potentially enhancing the efficacy of other therapies.
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| ln Vitro |
In vitro, HIF-1alpha-IN-2 hydrochloride is a potent inhibitor of HIF-1alpha, demonstrating IC50 values of 28 nM in MDA-MB-231 human breast cancer cells and 15 nM in MiaPaCa-2 human pancreatic cancer cells. It suppresses HIF-1alpha expression by blocking its gene transcription and inhibiting its protein translation. In cell-based assays, treatment with this compound leads to a dose-dependent reduction in HIF-1alpha protein levels under hypoxic conditions, as measured by Western blot. This results in decreased expression of HIF-1alpha target genes, including vascular endothelial growth factor (VEGF) and glucose transporter 1 (GLUT1). Consequently, it reduces cancer cell proliferation, migration, and invasion under hypoxic conditions. The compound is effective at low nanomolar concentrations, indicating high potency. In cell viability assays, it reduces the viability of various cancer cell lines with IC50 values in the low micromolar range, correlating with its HIF-1alpha inhibitory activity.
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| ln Vivo |
No specific in vivo data are available for HIF-1alpha-IN-2 hydrochloride. However, based on its potent in vitro HIF-1alpha inhibitory activity and anticancer effects, it has the potential for in vivo efficacy. In mouse xenograft models of human cancers, a compound with a similar mechanism would likely be administered intraperitoneally (i.p.) or intravenously (i.v.) at doses of 10-50 mg/kg daily or every other day for 2-4 weeks. Endpoints would include tumor volume reduction, HIF-1alpha expression in tumor lysates by Western blot, and inhibition of angiogenesis (CD31 staining). The compound could also reduce VEGF levels in plasma and tumor tissues. However, as of 2026, such studies have not been published for this specific compound. It is currently a research tool for in vitro studies and requires further validation for in vivo applications.
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| Enzyme Assay |
For non-cellular binding assays, a direct binding assay for HIF-1alpha-IN-2 is not standard because its mechanism involves inhibition of transcription and translation, not direct binding to the HIF-1alpha protein itself. However, researchers can study its effect on HIF-1alpha gene transcription using a cell-free transcription assay or a DNA-binding activity assay. For a HIF-1alpha DNA-binding ELISA (TransAM assay): Nuclear extracts are prepared from cells treated with the compound. These extracts are added to a 96-well plate coated with an oligonucleotide containing the hypoxia response element (HRE). Bound HIF-1alpha is detected using a primary antibody against HIF-1alpha, followed by an HRP-conjugated secondary antibody and a colorimetric substrate. Absorbance is measured at 450 nm. The compound reduces HIF-1alpha DNA-binding activity. Alternatively, a HIF-1alpha translation inhibition assay can be performed in a cell-free system using rabbit reticulocyte lysates and in vitro transcribed HIF-1alpha mRNA. The compound is added at various concentrations (0.1-1000 nM), and newly synthesized HIF-1alpha protein is quantified by Western blot or by incorporation of 35S-methionine.
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| Cell Assay |
For cellular assays, use human cancer cell lines such as MDA-MB-231 (breast) or MiaPaCa-2 (pancreatic). Seed cells in 6-well plates (2-5 × 10^5 cells/well) or 96-well plates (5-10 × 10^3 cells/well) in DMEM or RPMI-1640 with 10% FBS at 37degC, 5% CO2. For hypoxic conditions, incubate cells in a hypoxic chamber (1% O2, 5% CO2, 94% N2) or treat with a hypoxia-mimicking agent (e.g., CoCl2, 100-200 uM, or desferrioxamine, 100-200 uM) for 24-48 hours. Treat cells with HIF-1alpha-IN-2 hydrochloride (0.1-1000 nM) for 24-48 hours. For Western blot analysis, lyse cells in RIPA buffer with protease/phosphatase inhibitors, separate lysates (20-40 ug protein) by SDS-PAGE, and immunoblot with anti-HIF-1alpha antibody (detects the ~120 kDa band) and anti-beta-actin as loading control. HIF-1alpha levels under normoxia are very low; hypoxic induction is required. For cell viability assays, use MTT, CCK-8, or CellTiter-Glo after 48-72 hours of treatment. For VEGF secretion, collect supernatants after 48 hours and measure by ELISA. The compound should reduce HIF-1alpha levels and VEGF secretion in a dose-dependent manner. The IC50 for HIF-1alpha inhibition is 28 nM (MDA-MB-231) and 15 nM (MiaPaCa-2). All experiments should be performed in triplicate wells with at least three independent experiments. Control: DMSO (≤0.1%). Positive control: known HIF-1alpha inhibitors such as PX-478 or digoxin.
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| Animal Protocol |
No specific in vivo protocol is available for HIF-1alpha-IN-2 hydrochloride. For a planned in vivo study, use female athymic nude mice (6-8 weeks old, 18-22 g). Inject MDA-MB-231 or MiaPaCa-2 cells subcutaneously (5 × 10^6 cells in 100 uL PBS) into the right flank. When tumors reach 100-150 mm3, randomize mice into treatment groups (n=8-10 per group). Dissolve HIF-1alpha-IN-2 hydrochloride in a suitable vehicle (e.g., 10% DMSO, 40% PEG400, 5% Tween 80 in water) and administer intraperitoneally (i.p.) at doses of 10, 25, and 50 mg/kg once daily for 3-4 weeks. A vehicle control group receives the same volume of vehicle. For pharmacodynamic assessment, collect tumors from a subset of mice after 2 weeks of treatment and analyze HIF-1alpha and VEGF levels by Western blot and ELISA. Tumor volume is measured every 2-3 days with digital calipers (volume = width2 × length / 2). Body weight is monitored as a toxicity indicator. At study termination (when vehicle tumors reach 1500-2000 mm3), euthanize mice, excise tumors, weigh them, and fix in formalin for histology (H&E, CD31 for angiogenesis). All animal procedures must be approved by the IACUC.
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| ADME/Pharmacokinetics |
No specific pharmacokinetic (PK) data are available for HIF-1alpha-IN-2 hydrochloride. As a small-molecule inhibitor (MW not specified but typical of drug-like molecules), it is expected to have moderate to good oral bioavailability (20-60%) and be metabolized by hepatic cytochrome P450 enzymes (likely CYP3A4 and CYP2D6). The plasma half-life in rodents is expected to be 2-6 hours, supporting once- or twice-daily dosing. The compound likely has moderate to high plasma protein binding (70-95%) and distributes to tissues, including tumors, as needed for anticancer efficacy. The hydrochloride salt enhances solubility for oral and intraperitoneal administration. For a PK study, administer the compound to male Sprague-Dawley rats (n=3-5 per time point) intravenously (1-5 mg/kg) and orally (10-50 mg/kg). Collect blood at 0, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours, and quantify concentrations by LC-MS/MS. Calculate PK parameters (AUC, Cmax, Tmax, t1/2, CL, Vd, F%) using non-compartmental analysis. This information is not publicly available.
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| Toxicity/Toxicokinetics |
No specific toxicity data are available for HIF-1alpha-IN-2 hydrochloride. In vitro, the compound is not cytotoxic to cancer cells at its effective concentrations (nM range), but at higher concentrations (>10 uM), it may cause nonspecific cell death in certain cell lines. In vivo toxicity studies have not been reported. Given its mechanism of HIF-1alpha inhibition, potential on-target toxicities may include impaired adaptation to hypoxia, which could affect wound healing, exercise tolerance, and cardiovascular function. However, these are speculative. No genotoxicity, carcinogenicity, or reproductive toxicity studies have been performed. The hydrochloride salt is not associated with additional toxicity. Standard laboratory safety precautions (gloves, lab coat, eye protection) should be used. The compound is for research use only and is not approved for human use. This product is not intended for therapeutic applications.
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| References | |
| Additional Infomation |
Hypoxia-Inducible Factor 1-alpha (HIF-1alpha) is a transcription factor that plays a central role in cellular adaptation to low oxygen conditions. Under normoxia, HIF-1alpha is hydroxylated by prolyl hydroxylases (PHDs) and targeted for proteasomal degradation. Under hypoxia, it stabilizes, translocates to the nucleus, dimerizes with HIF-1beta, and activates the transcription of genes involved in angiogenesis, glycolysis, cell survival, and invasion. HIF-1alpha is overexpressed in many solid tumors and is associated with poor prognosis, metastasis, and resistance to chemotherapy and radiation. Therefore, HIF-1alpha is a validated anticancer target. HIF-1alpha-IN-2 hydrochloride is a potent, small-molecule inhibitor that suppresses HIF-1alpha expression at both the transcriptional and translational levels. It represents a novel chemical tool for studying the role of HIF-1alpha in cancer biology and for preclinical drug development. As of 2026, no HIF-1alpha inhibitor has received FDA approval for cancer therapy, though several agents (e.g., PX-478, digoxin, acriflavine) are in preclinical or clinical development. HIF-1alpha-IN-2 hydrochloride is for research use only and is not approved for human therapy.
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| Molecular Formula |
C21H20CLN3OS
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| Molecular Weight |
397.92
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| Appearance |
Off-white to light yellow solid powder
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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 : PEG300 ：Tween 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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5131 mL | 12.5653 mL | 25.1307 mL | |
| 5 mM | 0.5026 mL | 2.5131 mL | 5.0261 mL | |
| 10 mM | 0.2513 mL | 1.2565 mL | 2.5131 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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