HIF-1 inhibitor-5 (Compound 16e) is a potent HIF-1 inhibitor (antagonist) with IC50 of 2.38 μM. HIF-1 inhibitor-5 has anti-angiogenic potential.

Physicochemical Properties

Molecular Formula	C28H35NO5
Molecular Weight	465.58
Appearance	Typically exists as solid at room temperature
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)

Biological Activity

Targets	IC50: 2.38 μM (HIF-1)[1]
ln Vitro	Compound 16e, HIF-1 inhibitor-5, inhibits A549 cell migration and invasion [1] at 0–4 μM over 24 hours. VEGF-induced blood vessel development is blocked by HIF-1 inhibitor-5 (0-8 μM; 12 h) [1].
ln Vivo	HIF-1 inhibitor-5 (Compound 16e; 1.25–20 μM; single-dose) prevents mice from developing angiogenesis brought on by VEGF [1].
Cell Assay	Cell Cytotoxicity Assay[1] Cell Types: A549 and HUVEC cells Tested Concentrations: 0-128 μM Incubation Duration: 48 h Experimental Results: demonstrated cytotoxic effects with IC50s of 8.883 μM and 19.599 μM for A549 and HUVEC cells, respectively. Cell Migration Assay [1] Cell Types: A549 cell Tested Concentrations: 0, 2 and 4 μM Incubation Duration: 24 h Experimental Results: The number of migrated A549 cells diminished Dramatically. Cell Invasion Assay[1] Cell Types: A549 cell Tested Concentrations: 0, 2 and 4 μM Incubation Duration: 24 h Experimental Results: Inhibited cell invasion in a concentration-dependent manner.
Animal Protocol	Animal/Disease Models: C57/bl6 female mice, VEGF-induced angiogenesis model[1] Doses: 1.25, 5 and 20 μM Route of Administration: 0.5 mL of matrix glue subcutaneously (sc) injected near the midline of abdomen Experimental Results: diminished the number of blood vessels. Animal/Disease Models: SD rats Doses: 20 mg/kg Route of Administration: intravenous (iv) administration (pharmacokinetic/PK Study) Experimental Results: pharmacokinetic/PK parameters of HIF-1 inhibitor-5 (Compound 16e) in rats after intravenous (iv) administrationa[1] Parameter iv (20 mg/kg) (n = 2) Tmax (h) 0.033 Cmax (μmol/L) 7.06 T1/2 (h) 1.64 CL (mL/h) 2368.50 VZ (L) 5.59 AUC0-t (μmol·h/L) 5.27 AUC0-∞ (μmol •h/L) 8.88 aAfter administration, blood samples were collected at different time (0, 0.083, 0.17, 0.5, 1, 2, 4, 6 h). The combination of compounds in a ratio of 8% ethanol absolute, 4% Tween-80 and 88% normal saline was chosen for the intravenous (iv) injection formulation.
References	[1]. Design, synthesis and evaluation of the novel chalcone derivatives with 2,2-dimethylbenzopyran as HIF-1 inhibitors that possess anti-angiogenic potential. Eur J Med Chem. 2023 Mar 15;250:115171.

Solubility Data

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

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1479 mL	10.7393 mL	21.4786 mL
	5 mM	0.4296 mL	2.1479 mL	4.2957 mL
	10 mM	0.2148 mL	1.0739 mL	2.1479 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.