| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Targets |
PIKfyve-IN-3 targets PIKfyve (phosphatidylinositol-3-phosphate 5-kinase), a lipid kinase that catalyzes the phosphorylation of phosphatidylinositol 3-phosphate (PI3P) to generate phosphatidylinositol 3,5-bisphosphate (PI(3,5)P2). PIKfyve is a member of the PI3K-related kinase (PIKK) family and is involved in the regulation of endosomal trafficking, autophagy, lysosomal homeostasis, and vacuole formation. PI(3,5)P2, the product of PIKfyve activity, is a key signaling lipid that regulates membrane trafficking and ion channel function. PIKfyve has been implicated in various diseases, including cancer, neurodegenerative disorders, and infectious diseases. PIKfyve-IN-3 exhibits remarkable interaction with PIKfyve kinase with a Kd of 0.47 nM, indicating extremely high affinity binding. The compound's potent inhibition of PIKfyve makes it a valuable research tool and a potential therapeutic agent.
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| ln Vitro |
In vitro, PIKfyve-IN-3 demonstrates remarkable interaction with PIKfyve kinase with a Kd of 0.47 nM. The compound's high affinity binding indicates potent inhibition of PIKfyve activity. In cell-based studies, PIKfyve-IN-3 would be expected to inhibit the production of PI(3,5)P2, leading to disruption of endosomal trafficking, autophagy, and lysosomal function. These effects can result in antiproliferative and pro-apoptotic effects in cancer cells. The compound inhibits tumor growth in a HeLa xenograft model, confirming its in vitro activity translates to in vivo efficacy. However, specific quantitative data for in vitro cell proliferation inhibition, such as IC50 values, have not been extensively reported in the available literature. The compound's oral activity further supports its potential for therapeutic development.
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| ln Vivo |
In vivo, PIKfyve-IN-3 inhibits tumor growth in a HeLa xenograft model. The compound has oral activity, indicating that it can be administered orally and achieve sufficient systemic exposure to exert its antitumor effects. In the HeLa xenograft model, PIKfyve-IN-3 treatment resulted in inhibition of tumor growth, demonstrating its in vivo efficacy. The compound's mechanism of action involves inhibition of PIKfyve, leading to disruption of endosomal trafficking and autophagy, which are essential for tumor cell survival and proliferation. The oral bioavailability and in vivo efficacy of PIKfyve-IN-3 make it a promising candidate for further preclinical and clinical development as an anticancer agent. However, specific details regarding dosing regimens, tumor growth inhibition percentages, and other quantitative outcomes are limited in the available literature.
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| Enzyme Assay |
In vitro enzyme assay protocols for PIKfyve-IN-3 would typically involve measuring its binding affinity to PIKfyve kinase and its inhibition of PIKfyve enzymatic activity. For binding affinity, surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) can be used to measure the Kd value. A standard protocol would involve immobilizing recombinant PIKfyve kinase on a sensor chip and flowing varying concentrations of PIKfyve-IN-3 over the chip to measure binding kinetics (ka, kd) and calculate Kd. For kinase activity assays, a standard protocol would involve incubating recombinant PIKfyve with varying concentrations of PIKfyve-IN-3 (typically 0.01 nM to 1 μM), phosphatidylinositol 3-phosphate (PI3P) substrate, and ATP in assay buffer at 30°C for a defined period. The production of PI(3,5)P2 is measured using methods such as mass spectrometry, HPLC, or radiolabeled ATP incorporation. IC50 values are determined from concentration-response curves.
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| Cell Assay |
In vitro cell-based assay protocols for PIKfyve-IN-3 would typically involve assessing its effects on PIKfyve signaling, cell proliferation, and autophagy in cultured cancer cells. A standard protocol would involve seeding cancer cells (e.g., HeLa or other cancer cell lines) in multi-well plates and treating them with varying concentrations of PIKfyve-IN-3 (typically 0.1 nM to 10 μM) for 24-72 hours. PIKfyve activity is assessed by measuring PI(3,5)P2 levels by mass spectrometry or by assessing the localization of downstream effectors such as TRPML1 or other lysosomal markers. Autophagy is assessed by Western blot analysis of LC3-II levels or by using autophagy reporter cell lines. Cell proliferation is measured using MTT, CCK-8, or BrdU incorporation assays. Apoptosis can be assessed by Annexin V staining or caspase activity assays. Appropriate controls include vehicle-treated cells.
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| Animal Protocol |
In vivo animal experimental protocols for PIKfyve-IN-3 have been reported in the context of its evaluation in a HeLa xenograft model. A standard protocol would involve implanting HeLa cells subcutaneously in immunodeficient mice (e.g., athymic nude mice), allowing tumors to reach a certain size (e.g., 100-200 mm3), and then administering PIKfyve-IN-3 orally at doses determined from preliminary pharmacokinetic studies. The compound has oral activity, suggesting that oral gavage would be a suitable route of administration. Treatment would typically be administered daily for 2-4 weeks. Endpoints would include tumor volume measurement (caliper measurement 2-3 times per week), tumor weight at necropsy, assessment of PIKfyve inhibition and PI(3,5)P2 levels in tumor tissues, assessment of autophagy markers, and evaluation of apoptosis. Appropriate controls would include vehicle-treated groups.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of PIKfyve-IN-3 indicate that the compound has oral activity, suggesting good oral bioavailability. However, specific PK parameters such as half-life, Cmax, AUC, bioavailability, volume of distribution, and clearance have not been extensively reported in the available literature. The compound's high affinity for PIKfyve (Kd = 0.47 nM) suggests that it can achieve target engagement at low concentrations. The compound's metabolism, protein binding, and routes of elimination remain to be characterized. Further pharmacokinetic studies would be required to fully understand its absorption, distribution, metabolism, and excretion profile. The compound should be stored under recommended conditions to maintain stability.
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| Toxicity/Toxicokinetics |
Toxicological data for PIKfyve-IN-3 have not been extensively reported in the available literature. The compound has been evaluated in preclinical studies, but detailed toxicity data are limited. As a PIKfyve inhibitor, potential on-target toxicities could include effects on lysosomal function and autophagy, which may affect normal tissues. However, the compound's safety profile has not been fully characterized. The compound is intended for research use only and is not approved for human use. Researchers should follow standard safety precautions when handling the compound, including working in a fume hood, wearing appropriate personal protective equipment, and avoiding inhalation, ingestion, or skin contact.
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| References | |
| Additional Infomation |
PIKfyve-IN-3 is a research-grade compound that functions as a high-affinity inhibitor of the lipid kinase PIKfyve, with a Kd of 0.47 nM. The compound exhibits oral activity and inhibits tumor growth in a HeLa xenograft model. PIKfyve-IN-3 is used in research on PIKfyve function, endosomal trafficking, autophagy, and cancer. It has not entered clinical trials and is not approved for any therapeutic indication. Its mechanism of action involves high-affinity binding and inhibition of PIKfyve, leading to disruption of PI(3,5)P2 production, endosomal trafficking, and autophagy, resulting in antiproliferative and antitumor effects. The compound is available exclusively for research purposes and is not intended for diagnostic, therapeutic, or human applications.
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| Molecular Formula |
C21H23N5O
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| Molecular Weight |
361.44
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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 |
| 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 :~10 mg/mL (~27.67 mM; with sonication (<60°C))
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (2.77 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 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 10.0 mg/mL clear DMSO stock solution and add it to 400 μL PEG300 and mix well. Then add 50 μL Tween-80 to the above system and mix well. Then add 450 μL of physiological saline to make 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: ≥ 1 mg/mL (2.77 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 10.0 mg/mL clear DMSO stock solution and add it to 900 μL of 20% SBE-β-CD in saline and mix well. 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: ≥ 1 mg/mL (2.77 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 | 2.7667 mL | 13.8336 mL | 27.6671 mL | |
| 5 mM | 0.5533 mL | 2.7667 mL | 5.5334 mL | |
| 10 mM | 0.2767 mL | 1.3834 mL | 2.7667 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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