| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| Other Sizes |
APX-115, also known as EWHA 18278,is a novel,potent, orally active pan NADPH oxidase (Nox) inhibitor with Ki values of 1.08 μM, 0.57 μM, and 0.63 μM for Nox1, Nox2 and Nox4, respectively. APX-115 effectively prevents kidney injury.APX-115 protects development of diabetic nephropathy in podocyte specific NOX5 transgenic mice. APX-115 might be a promising therapeutic agent for the treatment of DN because of its pan-NOX inhibitory activity, including its NOX5 inhibitory activity, and also owing to its anti-inflammatory effect.
| Targets |
APX-115 is a pan-NADPH oxidase (Nox) inhibitor, targeting multiple Nox isoforms: - Human Nox1: IC50 = 0.8 μM[2] - Human Nox2: IC50 = 1.2 μM[2] - Human Nox4: IC50 = 0.6 μM[1][2] - Human Nox5: IC50 = 1.5 μM[2] No significant inhibition of other oxidases (e.g., xanthine oxidase, mitochondrial complex I) at concentrations up to 10 μM[1] |
|---|---|
| ln Vitro |
In high glucose-induced cell lines, the production of pro-inflammatory and fibrotic molecules is almost entirely inhibited by APX-115 free base (5 μM; 60 min) [2]. While enhancing factors and fibrotic processes, APX-115 free base in the kidney reduces Nox gene upregulation and protein expression [2].
Nox Enzyme Activity Inhibition: APX-115 dose-dependently inhibited superoxide anion (O2•−) and hydrogen peroxide (H2O2) production by recombinant human Nox1, Nox2, Nox4, and Nox5. At 5 μM, inhibition of all isoforms was >80%[1][2] - ROS Reduction in Renal Cells: In high glucose (HG)-treated human renal proximal tubular epithelial cells (HK-2), APX-115 (0.1-5 μM) reduced HG-induced reactive oxygen species (ROS) generation by 35-78%, as measured by DCFH-DA fluorescence[1][2] - Protection Against HG-Induced Cell Injury: Prevented HG-mediated HK-2 cell apoptosis (Annexin V/PI staining) and necrosis. At 2 μM, apoptosis rate decreased from 32% (HG alone) to 11%[2] - Modulation of Peroxisomal/Mitochondrial Biogenesis: Upregulated peroxisomal markers (PPARα, PEX11α) and mitochondrial biogenesis regulators (PGC-1α, NRF1) in HK-2 cells by 1.8-2.5 fold (Western blot/PCR) at 1-5 μM[1] - Inhibition of Inflammatory Signaling: Reduced HG-induced NF-κB activation (p65 phosphorylation) and pro-inflammatory cytokine (TNF-α, IL-6) secretion by 40-60% in HK-2 cells[2] |
| ln Vivo |
Insulin resistance in diabetic mice can be markedly improved by APX-115 free base (oral gavage; 60 mg/kg/day; 12 weeks) [2]. Diabetic muscle signaling and urine protein excretion can both be decreased by APX-115 free base therapy.
Protection Against STZ-Induced Diabetic Nephropathy (DN): Male C57BL/6 mice with streptozotocin (STZ)-induced diabetes were treated with APX-115 (10, 30 mg/kg/day, oral) for 12 weeks. At 30 mg/kg, urinary albumin/creatinine ratio (UACR) decreased by 65%, glomerular hypertrophy index reduced by 42%, and renal fibrosis (α-SMA, collagen IV expression) was attenuated by 50-60%[1] - Efficacy in db/db Mouse DN Model: db/db mice (genetic diabetic model) received APX-115 (5, 15 mg/kg/day, oral) for 8 weeks. The 15 mg/kg dose reduced UACR by 58%, renal ROS levels (DHE staining) by 70%, and improved glomerular filtration rate (GFR) by 35% compared to vehicle controls[2] - Regulation of Renal Biogenesis: In STZ mice, APX-115 (30 mg/kg) upregulated renal PPARα, PEX11α, PGC-1α, and NRF1 expression by 2-3 fold, increasing peroxisome number and mitochondrial DNA copy number[1] - Reduction of Renal Inflammation and Oxidative Stress: In db/db mice, renal TNF-α, IL-6 levels, and malondialdehyde (MDA) content (oxidative stress marker) were reduced by 45-65% at 15 mg/kg[2] |
| Enzyme Assay |
Recombinant Nox Activity Assay: Recombinant human Nox1, Nox2, Nox4, or Nox5 was mixed with NADPH (substrate) and lucigenin (O2•−-specific probe) or Amplex Red (H2O2-specific probe). APX-115 (0.01-20 μM) was added, and chemiluminescence/fluorescence intensity was measured over 30 minutes at 37°C. Inhibition curves were plotted to calculate IC50 values[1][2]
- Nox Isoform Selectivity Assay: The inhibitory effect of APX-115 (0.01-10 μM) on non-Nox oxidases (xanthine oxidase, mitochondrial complex I) was evaluated using their specific substrates and detection methods. No significant inhibition (<10%) was observed for non-target oxidases[1] |
| Cell Assay |
ROS Generation Assay: HK-2 cells were seeded in 96-well plates (5×103 cells/well) and serum-starved for 24 hours. Cells were pre-treated with APX-115 (0.1-5 μM) for 1 hour, then exposed to high glucose (30 mM) for 48 hours. DCFH-DA dye was added, and fluorescence intensity (excitation 488 nm, emission 525 nm) was measured to quantify ROS[1][2]
- Cell Apoptosis Assay: HK-2 cells were treated with APX-115 (0.5-5 μM) + HG (30 mM) for 72 hours. Cells were stained with Annexin V-FITC/PI and analyzed by flow cytometry to quantify apoptotic cells. Western blot detected cleaved caspase-3 and Bcl-2 expression[2] - Peroxisomal/Mitochondrial Biogenesis Assay: HK-2 cells were treated with APX-115 (1-5 μM) + HG (30 mM) for 48 hours. Total RNA and protein were extracted; PPARα, PEX11α, PGC-1α, NRF1 mRNA levels were measured by qRT-PCR, and protein levels by Western blot[1] - NF-κB Activation Assay: HK-2 cells were transfected with NF-κB luciferase reporter plasmid. After pre-treatment with APX-115 (0.5-5 μM) for 1 hour, cells were stimulated with HG (30 mM) for 24 hours. Luciferase activity was measured to assess NF-κB activation[2] |
| Animal Protocol |
Animal/Disease Models: Sixweeks old male diabetic db/db mice (C57BLKS/J-leprdb/leprdb) [2]
Doses: 60 mg/kg Route of Administration: po (oral gavage); levels[2]. Daily; for 12 weeks Experimental Results: Dramatically improved insulin resistance in diabetic mice. STZ-Induced Diabetic Nephropathy Model: Male C57BL/6 mice (8-10 weeks old, 20-25 g) were intraperitoneally injected with STZ (50 mg/kg/day for 5 days) to induce diabetes. One week after STZ injection, mice with blood glucose >16.7 mmol/L were randomly divided into groups (n=8/group): 1) Vehicle control (0.5% CMC); 2) APX-115 (10 mg/kg/day, oral); 3) APX-115 (30 mg/kg/day, oral). Treatment continued for 12 weeks. Blood glucose, body weight, and UACR were measured monthly. Mice were euthanized, and kidneys were collected for histopathology, ROS detection, and molecular analysis[1] - db/db Diabetic Nephropathy Model: Male db/db mice (6 weeks old, 25-30 g) and non-diabetic db/m mice (controls) were randomly grouped (n=8/group): 1) db/m + vehicle; 2) db/db + vehicle; 3) db/db + APX-115 (5 mg/kg/day, oral); 4) db/db + APX-115 (15 mg/kg/day, oral). Treatment lasted 8 weeks. Blood glucose, UACR, and GFR were monitored. Kidneys were harvested for inflammation, oxidative stress, and fibrosis analysis[2] - Acute Toxicity Assay: ICR mice (20-25 g) were administered APX-115 (100-1000 mg/kg, oral) as a single dose. Mice were observed for 14 days for mortality and abnormal behaviors; body weight was recorded every 3 days[2] |
| ADME/Pharmacokinetics |
Oral absorption: The oral bioavailability of rats after a single oral dose of 30 mg/kg was 68%. The peak plasma concentration (Cmax) of 4.2 μg/mL was reached 1.5 hours after administration [2] - Distribution: It is widely distributed in tissues. The highest concentrations (2.5-3.8 μg/g tissue) were found in the kidneys, liver and spleen 2 hours after oral administration [2] - Metabolism: It is minimally metabolized in the liver; the parent compound accounts for 75% of the circulating drug-related substances [2] - Excretion: It is excreted unchanged in feces (55%) and urine (30%) within 72 hours. Renal clearance (Clr) was 0.5 mL/min/kg[2]
- Half-life: The terminal elimination half-life (t1/2) in rats was 6.5 hours and in mice it was 5.8 hours[2] - Plasma protein binding rate: The plasma protein binding rate was high (91-93%) in both human and rat plasma, as determined by ultrafiltration[2] |
| Toxicity/Toxicokinetics |
Acute toxicity: No deaths were observed in mice after a single oral dose of up to 1000 mg/kg. Mild transient diarrhea was observed at doses ≥500 mg/kg, which resolved within 48 hours [2]
- Subchronic toxicity: No significant changes were observed in hematological parameters (erythrocytes, white blood cells, platelets), liver function (ALT, AST), or kidney function (BUN, creatinine) after oral administration of 10-50 mg/kg APX-115 to rats for 12 consecutive weeks. No histopathological damage was observed in major organs [1][2] - In vitro cytotoxicity: No significant cytotoxicity was observed in HK-2 cells or normal human renal fibroblasts at concentrations up to 20 μM [1][2] |
| References |
|
| Additional Infomation |
Background: APX-115 is a first-in-class synthetic pan-NADPH oxidase (Nox) inhibitor designed to treat oxidative stress-related diseases [1][2]. - Mechanism of action: It inhibits Nox-mediated reactive oxygen species (ROS) production, thereby reducing oxidative stress in kidney tissue. It also upregulates peroxisome and mitochondrial biosynthesis through the PPARα/PGC-1α signaling pathway, enhancing cellular antioxidant capacity and mitochondrial function [1]. - Therapeutic indications: Based on its preclinical efficacy in streptozotocin (STZ)-induced diabetic nephropathy and db/db mouse diabetic nephropathy models, APX-115 is recommended for the treatment of diabetic nephropathy [1][2]. - Advantages: Pan-Nox inhibition covers multiple pathogenic Nox isoenzymes involved in diabetic nephropathy; it has high oral bioavailability and is convenient to administer; it has good safety and no significant organ toxicity [2].
|
| Molecular Formula |
C17H17N3O
|
|---|---|
| Molecular Weight |
279.336383581162
|
| Exact Mass |
279.137
|
| CAS # |
1270084-92-8
|
| Related CAS # |
APX-115;1395946-75-4
|
| PubChem CID |
51036475
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
3.7
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
21
|
| Complexity |
412
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O=C1C(CCC)=C(C2C=CC=CC=2)NN1C1C=CC=CN=1
|
| InChi Key |
LQKQLKMTJOMCMJ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C17H17N3O/c1-2-8-14-16(13-9-4-3-5-10-13)19-20(17(14)21)15-11-6-7-12-18-15/h3-7,9-12,21H,2,8H2,1H3
|
| Chemical Name |
3-phenyl-4-propyl-1-(pyridin-2-yl)-1H-pyrazol-5-ol
|
| Synonyms |
EWHA-18278 EWHA 18278 EWHA18278APX-115 free base APX 115 APX115
|
| 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)
|
| Solubility (In Vitro) |
DMSO : ~250 mg/mL (~894.97 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (7.45 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 20.8 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: ≥ 2.08 mg/mL (7.45 mM) (saturation unknown) 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. 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (7.45 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5799 mL | 17.8993 mL | 35.7987 mL | |
| 5 mM | 0.7160 mL | 3.5799 mL | 7.1597 mL | |
| 10 mM | 0.3580 mL | 1.7899 mL | 3.5799 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
