| Size | Price | Stock | Qty |
|---|---|---|---|
| 50mg |
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| Other Sizes |
| Targets |
IC50: 0.56 μM (AKR1C3) and 15.1 μM (AKR1C2)[1]
AKR1C3 (also known as 17beta-HSD5) (IC50: 0.56 uM) |
|---|---|
| ln Vitro |
AKR1C3-IN-4's IC50 against AKR1C2 is 15.1 μM [1].
In vitro enzyme assays demonstrate that AKR1C3-IN-4 is a potent and selective inhibitor of aldo-keto reductase 1C3 (AKR1C3) with an IC50 of 0.56 uM. AKR1C3 (also known as type 5 17beta-hydroxysteroid dehydrogenase, 17beta-HSD5) is a key enzyme in the synthesis of androgens (testosterone, androstenedione) and estrogens. Overexpression of AKR1C3 in prostate cancer cells is associated with castration-resistant prostate cancer (CRPC), where intratumoral androgen synthesis drives tumor growth despite castrate levels of circulating androgens. AKR1C3-IN-4 selectively inhibits AKR1C3 over other AKR isoforms (selectivity not specified in public data). No other specific biological activities (e.g., cytotoxicity) are reported. |
| ln Vivo |
No specific in vivo data for AKR1C3-IN-4; however, as a potent and selective AKR1C3 inhibitor, it has potential for in vivo efficacy in xenograft models of castration-resistant prostate cancer (CRPC). In vivo studies would involve administration to tumor-bearing mice (e.g., 22Rv1 or LNCaP xenografts in castrated mice) to assess tumor growth inhibition, reduction of intratumoral androgen levels, and survival. The compound has potential for castrate-resistant prostate cancer research. Pharmacokinetic and toxicity studies would be required for development.
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| Enzyme Assay |
Recombinant human AKR1C3 is expressed in E. coli and purified. The AKR1C3 activity assay measures the reduction of substrates such as 4-androstene-3,17-dione (androstenedione) to testosterone, or the reduction of 9,10-phenanthrenequinone, using NADPH as cofactor. A typical assay uses the fluorogenic substrate 9,10-phenanthrenequinone (15-30 uM) in the presence of NADPH (100-200 uM) in potassium phosphate buffer (pH 6.0-7.0, 100 mM). The reaction is initiated by adding AKR1C3 enzyme (50-200 ng). The decrease in fluorescence (excitation 340 nm, emission 460 nm) or absorbance (340 nm for NADPH oxidation) is monitored for 5-10 minutes at 25degC or 37degC. Alternatively, a LC-MS/MS assay measuring conversion of androstenedione to testosterone can be used for higher specificity. AKR1C3-IN-4 is pre-incubated with the enzyme and NADPH for 5-10 minutes before substrate addition. IC50 values are determined from dose-response curves (IC50 = 0.56 uM). Selectivity against other AKR isoforms (AKR1C1, AKR1C2, AKR1C4) and other 17beta-HSD isoforms (HSD17B1-4) is assessed similarly to confirm selectivity.
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| Cell Assay |
Human prostate cancer cell lines that express AKR1C3 (e.g., 22Rv1, LNCaP, VCaP) are cultured in RPMI-1640 with 10% FBS and antibiotics. Cells are seeded in 96-well plates or 6-well plates and treated with AKR1C3-IN-4 (0.1-100 uM) for 24-72 hours. AKR1C3 activity is measured in cell lysates using the fluorogenic substrate 9,10-phenanthrenequinone or by measuring conversion of androstenedione to testosterone by LC-MS/MS. Intracellular testosterone and androstenedione levels are measured by LC-MS/MS after lipid extraction. Cell proliferation is assessed by MTT, CCK-8, or CellTiter-Glo assays. For androgen receptor (AR) activity studies, cells can be transfected with an AR-responsive reporter (e.g., MMTV-luciferase) and treated with AKR1C3-IN-4 in the presence of androstenedione (10-100 nM) to assess inhibition of AR activation by intratumoral androgen synthesis. Cell viability and apoptosis (Annexin V/PI, caspase-3/7) are assessed. In castration-resistant models, cells are cultured in androgen-depleted medium (charcoal-stripped serum) to mimic the castrate environment. AKR1C3-IN-4 should reduce androgen-dependent proliferation under these conditions. IC50 values are reported as 0.56 uM in biochemical assays; cellular potency may be higher due to compound accumulation.
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| Animal Protocol |
No published in vivo animal study for AKR1C3-IN-4. Based on its AKR1C3 inhibitory activity and potential for castration-resistant prostate cancer (CRPC), a typical protocol would involve establishing subcutaneous tumor xenografts of AKR1C3-expressing human prostate cancer cells (e.g., 22Rv1 or LNCaP) in castrated male immunodeficient mice (e.g., nude mice or NSG mice). Castration is performed surgically (orchidectomy) before tumor implantation or when tumors reach a certain size. When tumors reach 100-200 mm3, mice are randomized and treated with AKR1C3-IN-4 formulated in a suitable vehicle (e.g., 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline). Dosing is once daily by oral gavage or intraperitoneal injection at doses ranging from 1-50 mg/kg for 2-8 weeks. Endpoints: tumor volume, tumor weight, body weight, survival. Plasma and tumor tissue are collected for pharmacokinetic analysis and pharmacodynamic evaluation: AKR1C3 activity in tumor lysates, intratumoral androgen levels (testosterone, androstenedione, dihydrotestosterone) measured by LC-MS/MS, AR target gene expression (PSA, TMPRSS2) by qPCR, tumor proliferation (Ki-67 by IHC), and apoptosis (TUNEL). Plasma testosterone and androstenedione levels are also measured. This protocol is generic and not validated for AKR1C3-IN-4 specifically.
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| ADME/Pharmacokinetics |
No specific pharmacokinetic data for AKR1C3-IN-4. Based on its molecular properties (MW 281.23, LogP ~3-4 from structure), the compound is moderately lipophilic and would be expected to have moderate oral bioavailability. The compound contains a trifluoromethyl group, which may improve metabolic stability. Metabolism would occur in the liver via phase I (oxidation, reduction) and phase II (glucuronidation, sulfation) enzymes. Plasma half-life, Cmax, AUC, clearance, volume of distribution, and protein binding have not been publicly reported. Solubility: DMSO 100 mg/mL (355.58 mM). In vivo formulation: 10% DMSO + 40% PEG300 + 5% Tween80 + 45% saline (≥2.5 mg/mL, 8.89 mM, clear solution). Storage: 4degC, protect from light; in solvent at -80degC for 6 months.
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| Toxicity/Toxicokinetics |
No specific toxicity data for AKR1C3-IN-4. As an AKR1C3 inhibitor, the primary expected on-target toxicity is modulation of steroid hormone synthesis. AKR1C3 is involved in the synthesis of androgens and estrogens in peripheral tissues. Inhibition of AKR1C3 may lead to decreased intratumoral androgen levels in prostate cancer (desired effect), but could also affect androgen and estrogen levels in normal tissues, potentially causing endocrine-related adverse effects (e.g., gynecomastia, hot flashes, fatigue). Off-target inhibition of other AKR isoforms (AKR1C1, AKR1C2) could affect metabolism of other steroids and xenobiotics. In cell-based studies, no significant cytotoxicity has been reported at relevant concentrations. In vivo toxicity studies would assess maximum tolerated dose (MTD), body weight loss, organ toxicity (liver, kidney, reproductive organs), and changes in serum steroid hormone levels. No data are publicly available. For research use only; handle with standard laboratory precautions (gloves, lab coat, eye protection). Not for human use.
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| References | |
| Additional Infomation |
AKR1C3-IN-4 (CAS: 1284180-11-5) is a potent and selective inhibitor of aldo-keto reductase 1C3 (AKR1C3), also known as type 5 17beta-hydroxysteroid dehydrogenase (17beta-HSD5). AKR1C3 is a key enzyme in the synthesis of androgens (testosterone, androstenedione) and estrogens, and its overexpression in prostate cancer cells is associated with castration-resistant prostate cancer (CRPC), where intratumoral androgen synthesis drives tumor growth despite castrate levels of circulating androgens. AKR1C3-IN-4 inhibits AKR1C3 with an IC50 of 0.56 uM. The compound has potential for castration-resistant prostate cancer research. Molecular formula: C14H10F3NO2, molecular weight: 281.23. Also known as 3-((4-(trifluoromethyl)phenyl)amino)benzoic acid. Solubility: DMSO 100 mg/mL (355.58 mM). Storage: 4degC, protect from light. Not approved for clinical use. For research use only.
|
| Molecular Formula |
C14H10F3NO2
|
|---|---|
| Molecular Weight |
281.23
|
| Exact Mass |
281.066
|
| CAS # |
1284180-11-5
|
| PubChem CID |
51346826
|
| Appearance |
White to yellow solid powder
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| Density |
1.395±0.06 g/cm3(Predicted)
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| Boiling Point |
402.8±45.0 °C(Predicted)
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| LogP |
4.22
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
20
|
| Complexity |
338
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
C1=CC(=CC(=C1)NC2=CC=C(C=C2)C(F)(F)F)C(=O)O
|
| InChi Key |
MDZIRNPRVJEHHX-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C14H10F3NO2/c15-14(16,17)10-4-6-11(7-5-10)18-12-3-1-2-9(8-12)13(19)20/h1-8,18H,(H,19,20)
|
| Chemical Name |
3-[4-(trifluoromethyl)anilino]benzoic acid
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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: 100 mg/mL (355.58 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (8.89 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 (8.89 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 25.0 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: ≥ 2.5 mg/mL (8.89 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 | 3.5558 mL | 17.7790 mL | 35.5581 mL | |
| 5 mM | 0.7112 mL | 3.5558 mL | 7.1116 mL | |
| 10 mM | 0.3556 mL | 1.7779 mL | 3.5558 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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