| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
EC50: 10.5 μM (Ca2+ current response from TPC2)[1]
TPC2-A1-N targets the two-pore channel 2 (TPC2), a Ca2+ permeable channel located in endolysosomal membranes. The compound acts as a TPC2 agonist, mimicking the physiological effects of NAADP (nicotinic acid adenine dinucleotide phosphate). TPC2-A1-N induces significant Ca2+ responses from TPC2 with an EC50 of 7.8 μM. Its effects can be blocked by several TPC blockers. |
|---|---|
| ln Vitro |
Ancient members of the voltage-gated ion channel superfamily are known as two-pore channels (TPC1-3). The endo-lysosomal system expresses TPCs, which control the trafficking of different payloads. Depending on how it is triggered, TPC2 can mediate various physiological and potentially pathological effects. TPC2's ion selectivity is agonist-dependent rather than fixed. One special type of ion channel that responds to various activating ligands by conducting distinct ions is TPC2. Ca2+ signals are consistently evoked by TPC2-A1-N (10 μM), and the response reaches its plateau more quickly in TPC2-A1-N than in TPC2-A1-P. TPC2-A1-N has an EC50 of 7.8 μM in full concentration-effect relationships for the plateau response in a cell line that is stable in expressing TPC2L11A/L12A. Cells expressing TPC2L11A/L12A are activated by TPC2-A1-N (10 μM) to induce Ca2+ influx through the TPC2 pore, but not TPC2L11A/L12A/L265P. Furthermore, by removing extracellular Ca2+, the known TPC2 blockers Tetrandrine (Tet), Raloxifene (Ral), and Fluphenazine (Flu) can specifically prevent the reactions to TPC2-A1-N[1]. TPC2-A1-N (30 μM) elicits currents using Na+ as the main permeation in endo-lysosomal patch-clamp tests conducted on vacuolin-enlarged endo-lysosomes derived from HEK293 cells transiently expressing human TPC2 (hTPC2), but not in cells expressing TPC1[1]. TPC2-A1-N (30 μM) causes more currents in endo-lysosomes isolated from cells expressing a gain-of-function variation of TPC2 (TPC2M484L) than in the wild-type isoform in endo-lysosomal patch-clamp experiments. TPC2-A1-N has an EC50 value of 0.6 μM[1].
In vitro, TPC2-A1-N is a specific activator that triggers rapid Ca2+ signals in cells stably expressing TPC2 at a concentration of 10 μM. The compound induces Ca2+ influx in TPC2-expressing cells with an EC50 of 7.8 μM. It activates endogenous TPC2 to moderately increase lysosomal pH in untransfected cells in a time-dependent manner. Standard in vitro assays include calcium imaging using fluorescent indicators, patch-clamp electrophysiology, and measurement of lysosomal pH using pH-sensitive dyes. |
| ln Vivo |
In vivo, TPC2-A1-N can be used to probe different functions of TPC2 channels in intact cells. As a TPC2 agonist, it can be used to study the role of TPC2 in endolysosomal signaling, calcium homeostasis, and various cellular processes. However, comprehensive in vivo pharmacological studies are limited. The compound is primarily used as a chemical probe for studying TPC2 function in research settings. It is not intended for therapeutic applications.
|
| Enzyme Assay |
For non-cell-based receptor binding assays, TPC2-A1-N can be evaluated using purified TPC2 protein or membrane preparations from cells expressing TPC2. Binding affinity is assessed using radioligand binding or surface plasmon resonance. However, given that TPC2 is a channel rather than a traditional receptor, functional assays are more commonly used. The compound's ability to activate TPC2-mediated calcium flux can be measured in liposome-based assays or using membrane preparations with fluorescent calcium indicators.
|
| Cell Assay |
For in vitro cellular assays, cells expressing TPC2 (e.g., HEK293 cells stably expressing TPC2) are cultured in appropriate media. Cells are loaded with calcium-sensitive fluorescent dyes such as Fluo-4 AM. After dye loading, cells are treated with various concentrations of TPC2-A1-N, and calcium signals are measured using a fluorescence plate reader or imaging system. EC50 values for calcium responses are calculated from dose-response curves. For lysosomal pH measurements, cells are loaded with pH-sensitive dyes and treated with the compound.
|
| Animal Protocol |
For in vivo animal studies, TPC2-A1-N can be administered to rodents via intraperitoneal or intravenous injection. The compound's effects on endolysosomal signaling and calcium homeostasis can be assessed in various tissues. However, comprehensive in vivo studies are limited. The compound is primarily used as a chemical probe for studying TPC2 function in cell-based assays rather than in whole animals. Dosing regimens would vary depending on the specific experimental objectives.
|
| ADME/Pharmacokinetics |
TPC2-A1-N has a molecular weight of 401.17 and a molecular formula of C17H9Cl2F3N2O2. It is soluble in DMSO at 200 mg/mL. The compound should be stored at low temperature (powder: -20°C for 3 years; in solvent: -80°C for 1 year). It can be formulated for in vivo administration using 10% DMSO + 40% PEG300 + 5% Tween 80 + 45% saline. It is supplied in high purity (99.86%). The compound is for research use only.
|
| Toxicity/Toxicokinetics |
The toxicity profile of TPC2-A1-N has not been extensively reported. As a TPC2 agonist, potential adverse effects may include disruption of endolysosomal calcium homeostasis. The compound is for research use only and not for human consumption. Standard toxicological evaluation would include acute and repeated-dose toxicity studies, as well as assessment of effects on calcium signaling and cellular function. The compound should be handled with appropriate safety precautions.
|
| References | |
| Additional Infomation |
See other relationships...
TPC2-A1-N is a novel lipophilic, membrane-permeable subtype-selective Ca2+ permeable two-pore channel 2 (TPC2) small molecule agonist. It activates endogenous TPC2 to moderately increase lysosomal pH in untransfected cells in a time-dependent manner. TPC2-A1-N induces significant Ca2+ responses from TPC2 with an EC50 of 7.8 μM. It can be used to probe different functions of TPC2 channels in intact cells. It is available for research purposes only. |
| Molecular Formula |
C17H9CL2F3N2O2
|
|---|---|
| Molecular Weight |
401.166772603989
|
| Exact Mass |
399.999
|
| CAS # |
136186-07-7
|
| PubChem CID |
54735328
|
| Appearance |
White to off-white solid powder
|
| LogP |
5.8
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
26
|
| Complexity |
597
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
ClC1C=C(C=C(C=1)/C(=C(\C#N)/C(NC1C=CC(C(F)(F)F)=CC=1)=O)/O)Cl
|
| InChi Key |
FVJSEJNZNQVVDZ-PFONDFGASA-N
|
| InChi Code |
InChI=1S/C17H9Cl2F3N2O2/c18-11-5-9(6-12(19)7-11)15(25)14(8-23)16(26)24-13-3-1-10(2-4-13)17(20,21)22/h1-7,25H,(H,24,26)/b15-14-
|
| Chemical Name |
(Z)-2-cyano-3-(3,5-dichlorophenyl)-3-hydroxy-N-[4-(trifluoromethyl)phenyl]prop-2-enamide
|
| 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 (623.18 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.18 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 (5.18 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 (5.18 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 | 2.4927 mL | 12.4635 mL | 24.9271 mL | |
| 5 mM | 0.4985 mL | 2.4927 mL | 4.9854 mL | |
| 10 mM | 0.2493 mL | 1.2464 mL | 2.4927 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
