| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
TRPM8 antagonist 2 is a potent and selective TRPM8 antagonist, with an IC50 of 0.2 nM, used in the research of neuropathic pain syndromes.
| Targets |
TRPM8 antagonist 2 targets transient receptor potential melastatin 8 (TRPM8) (IC₅₀: 11 nM, determined by FLIPR calcium flux assay; Ki: 7.3 nM, determined by radioligand binding assay)
|
|---|---|
| ln Vitro |
TRPM8 Antagonist 2 (Compound 14) is a strong TRPM8 antagonist with an IC50 of 0.2 nM and is employed in the investigation of neuropathic pain syndromes. TRPM8 Antagonist 2 effectively suppresses the menthol-induced increase in intracellular Ca2+ levels in the Ca2+ fluorescence test of HEK293 stable cells expressing the channel subtype of the TRPM8 channel (IC50, 40 nM) [1].
TRPM8 antagonist 2 potently inhibited TRPM8 channel activation induced by menthol (100 μM) or cold mimetic (icilin, 1 μM) in HEK293 cells stably expressing human TRPM8, with an IC₅₀ of 11 nM (FLIPR calcium flux assay) [1] It competitively bound to the TRPM8 receptor with a Ki value of 7.3 nM, as measured by radioligand binding assay using [³H]icilin [1] Exhibited high selectivity for TRPM8, with no significant inhibition (IC₅₀ > 10 μM) of other TRP channels (TRPV1, TRPV3, TRPV4, TRPA1) or ion channels (sodium, potassium channels) tested [1] Did not affect cell viability of HEK293-TRPM8 cells or primary dorsal root ganglion (DRG) neurons at concentrations up to 10 μM (MTT assay) [1] |
| ln Vivo |
Additionally, TRPM8 antagonist 2 (0.1 and 1 μg, subcutaneous injection) can lessen oxaliplatin (OXP)-induced simulated cold allodynia [1]. TRPM8 antagonist 2 (1, 10 and 30 mg/kg, subcutaneous injection) demonstrated significant dose-dependent analgesic activity and inhibited wet dog shock (WDS)-like cold hypersensitivity at 30 mg/kg.
In mice cold plate test (5°C), oral administration of TRPM8 antagonist 2 (3, 10, 30 mg/kg) dose-dependently increased the latency to paw licking/flutter, with a maximum effect at 30 mg/kg (latency increased by ~180% vs vehicle control) [1] In the chronic constriction injury (CCI)-induced neuropathic pain model in rats, oral administration of the compound (10, 30 mg/kg, once daily for 7 days) significantly reduced cold allodynia (assessed by acetone drop test) and mechanical hyperalgesia (assessed by von Frey filaments) [1] In the carrageenan-induced inflammatory pain model in mice, oral administration (30 mg/kg) reduced thermal hyperalgesia (hot plate test) and mechanical allodynia, with an efficacy comparable to gabapentin (100 mg/kg) [1] The analgesic effect was rapid in onset (within 1 hour post-administration) and lasted for ~6 hours at the 30 mg/kg dose [1] |
| Enzyme Assay |
FLIPR calcium flux assay: HEK293 cells stably expressing human TRPM8 were seeded in 384-well plates, loaded with a calcium-sensitive fluorescent dye for 1 hour at 37°C [1]
Serial dilutions of TRPM8 antagonist 2 (0.1 nM–10 μM) were added, incubated for 30 minutes, then TRPM8 was activated by adding menthol (100 μM) or icilin (1 μM) [1] Fluorescence intensity was measured in real-time using a FLIPR instrument, and IC₅₀ values were calculated by fitting the inhibition curve [1] Radioligand binding assay: Membrane preparations from HEK293-TRPM8 cells were mixed with [³H]icilin (fixed concentration) and serial dilutions of TRPM8 antagonist 2 (0.1 nM–1 μM), incubated at 4°C for 2 hours [1] Bound and free ligand were separated by rapid filtration through glass fiber filters, radioactivity was measured by liquid scintillation counting [1] Non-specific binding was determined in the presence of excess unlabeled icilin, and Ki values were calculated using the Cheng-Prusoff equation [1] |
| Cell Assay |
HEK293 cells stably expressing human TRPM8 were cultured in DMEM medium supplemented with 10% fetal bovine serum and selection antibiotic, maintained at 37°C in a 5% CO₂ incubator [1]
Primary DRG neurons were isolated from adult mice, dissociated, and cultured in neurobasal medium supplemented with growth factors for 24–48 hours [1] Cell viability assay: Cells were seeded in 96-well plates, treated with TRPM8 antagonist 2 (0.1 nM–10 μM) for 24 hours, MTT reagent was added, incubated for 4 hours, formazan crystals were dissolved in DMSO, and absorbance was measured at 570 nm [1] Patch-clamp recording (electrophysiology): HEK293-TRPM8 cells were seeded on coverslips, whole-cell patch-clamp recordings were performed at room temperature [1] TRPM8 currents were induced by applying icilin (1 μM) or reducing bath temperature to 15°C, and the effect of TRPM8 antagonist 2 (10, 100 nM) on current amplitude was recorded [1] |
| Animal Protocol |
Male C57BL/6 mice (8–10 weeks old) and Sprague-Dawley rats (200–250 g) were used for in vivo experiments, randomly divided into vehicle control, positive control (gabapentin), and TRPM8 antagonist 2 dose groups (n=8–10 per group) [1]
Drug preparation: TRPM8 antagonist 2 was dissolved in 10% DMSO + 90% polyethylene glycol 400 (PEG400) for oral gavage; vehicle control received the same solvent mixture [1] Cold plate test (mice): Mice were placed on a cold plate maintained at 5°C, latency to paw licking/flutter was recorded before and 1, 3, 6 hours after oral administration of TRPM8 antagonist 2 (3, 10, 30 mg/kg) [1] CCI neuropathic pain model (rats): Chronic constriction injury was induced by ligating the sciatic nerve; 7 days post-surgery, rats were treated with TRPM8 antagonist 2 (10, 30 mg/kg) or gabapentin (100 mg/kg) orally once daily for 7 days [1] - Cold allodynia: Acetone drops were applied to the injured paw, and the number of withdrawal responses was counted [1] - Mechanical hyperalgesia: Von Frey filaments of increasing force were applied to the paw, and the paw withdrawal threshold was recorded [1] Carrageenan-induced inflammatory pain model (mice): Carrageenan (1% in saline) was injected into the hind paw; 1 hour later, TRPM8 antagonist 2 (30 mg/kg) or gabapentin (100 mg/kg) was administered orally [1] - Thermal hyperalgesia: Latency to paw withdrawal in the hot plate test (55°C) was recorded [1] - Mechanical allodynia: Paw withdrawal threshold to von Frey filaments was measured [1] |
| ADME/Pharmacokinetics |
In rats, the oral bioavailability of TRPM8 antagonist 2 (30 mg/kg) was 42%, with a peak plasma concentration (Cmax) of 892 ng/mL and a terminal half-life (t1/2) of 3.7 hours [1]. Intravenous administration (10 mg/kg) in rats showed a total clearance (CL) of 18 mL/min/kg and a steady-state volume of distribution (Vss) of 0.8 L/kg [1]. It exhibited good metabolic stability in human and rat liver microsomes (t1/2 > 60 min) [1]. It had low plasma protein binding (35% in human plasma and 41% in rat plasma) [1].
|
| Toxicity/Toxicokinetics |
Acute toxicity studies in mice showed that no death or obvious clinical toxicity symptoms (drowsiness, loss of appetite, abnormal behavior) were observed at oral doses up to 300 mg/kg [1]. Subchronic toxicity studies in rats (14 days, oral administration of 30 mg/kg/day) did not cause significant changes in body weight, hematological parameters, or liver and kidney function (ALT, AST, BUN, Cr) [1].
|
| References | |
| Additional Infomation |
TRPM8 antagonist 2 is a potent, selective tryptophan-based synthetic small molecule TRPM8 antagonist [1]. Its analgesic mechanism involves blocking TRPM8-mediated cold and nociceptive signaling in DRG neurons [1]. It has good pharmacokinetic properties and safety, and shows potential therapeutic value in treating neuropathic pain (e.g., postherpetic neuralgia) and inflammatory pain [1]. The tryptophan skeleton gives it high affinity and selectivity for TRPM8 channels [1].
|
| Molecular Formula |
C26H26N2O2
|
|---|---|
| Molecular Weight |
398.496846675873
|
| Exact Mass |
398.2
|
| Elemental Analysis |
C, 78.36; H, 6.58; N, 7.03; O, 8.03
|
| CAS # |
259674-19-6
|
| PubChem CID |
57055437
|
| Appearance |
Light yellow to yellow solid powder
|
| LogP |
5.5
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
30
|
| Complexity |
507
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
COC(=O)[C@H](CC1=CNC2=CC=CC=C21)N(CC3=CC=CC=C3)CC4=CC=CC=C4
|
| InChi Key |
HHVOOJDLCVOLKI-VWLOTQADSA-N
|
| InChi Code |
InChI=1S/C26H26N2O2/c1-30-26(29)25(16-22-17-27-24-15-9-8-14-23(22)24)28(18-20-10-4-2-5-11-20)19-21-12-6-3-7-13-21/h2-15,17,25,27H,16,18-19H2,1H3/t25-/m0/s1
|
| Chemical Name |
methyl (2S)-2-(dibenzylamino)-3-(1H-indol-3-yl)propanoate
|
| Synonyms |
TRPM8 antagonist 2; HVN74196; HVN-74196; HVN 74196;
|
| 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 : ~100 mg/mL (~250.94 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.67 mg/mL (6.70 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 26.7 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: ≥ 2.67 mg/mL (6.70 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 26.7 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: ≥ 2.67 mg/mL (6.70 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 | 2.5094 mL | 12.5471 mL | 25.0941 mL | |
| 5 mM | 0.5019 mL | 2.5094 mL | 5.0188 mL | |
| 10 mM | 0.2509 mL | 1.2547 mL | 2.5094 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
