| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 100mg | |||
| Other Sizes |
| Targets |
Alpha/beta-hydrolase domain containing 6 (ABHD6) [1]
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|---|---|
| ln Vitro |
In mouse neuroblastoma Neuro2A cell membrane proteomes, KT182 potently inhibited ABHD6 activity as measured by gel-based competitive activity-based protein profiling (ABPP), with an IC50 value of 1.7 nM. [1]
In a 2-arachidonoylglycerol (2-AG) hydrolysis assay using recombinant mouse ABHD6 overexpressed in HEK293T cells, KT182 inhibited ABHD6 with an IC50 value of 6.3 nM. [1] In situ treatment of Neuro2A cells with KT182 for 4 hours resulted in potent inhibition of ABHD6, with an IC50 value of 0.3 nM. [1] Quantitative ABPP-SILAC analysis in Neuro2A cells treated with 3 nM KT182 for 4 hours showed that it blocked >90% of ABHD6 activity and displayed negligible cross-reactivity (<50% inhibition) against more than 50 other serine hydrolases detected in the proteome. [1] |
| ln Vivo |
Intraperitoneal (i.p.) administration of KT182 (1 mg kg⁻¹) to mice for 4 hours produced near-complete blockade of ABHD6 activity in both the brain and liver, as measured by gel-based competitive ABPP. [1]
At lower doses (0.5 and 0.1 mg kg⁻¹, i.p., 4 hr), KT182 maintained approximately 80% inhibition of ABHD6 in the liver. [1] KT182 exhibited systemic activity (affecting both brain and liver) and showed good selectivity in vivo, with carboxylesterase 1 (CES1) identified as a minor off-target at 1 mg kg⁻¹. [1] |
| Enzyme Assay |
The activity of ABHD6 was determined using a 2-AG hydrolysis assay. Membrane lysates from HEK293T cells overexpressing recombinant mouse ABHD6 were diluted and pre-treated with DMSO or compound for 30 minutes at 37°C. The reaction was initiated by adding 2-AG substrate and incubated for 30 minutes at 37°C. The reaction was quenched by adding a chloroform:methanol mixture. The organic phase was analyzed by LC-MS to quantify the release of arachidonic acid. [1]
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| Cell Assay |
For in situ potency measurements, Neuro2A cells were treated with varying concentrations of KT182 in serum-free media for 4 hours at 37°C with 5% CO₂. Cells were then washed, lysed by sonication, and centrifuged to generate membrane proteomes. The proteomes were subjected to gel-based competitive ABPP analysis to determine remaining ABHD6 activity. [1]
For ABPP-SILAC selectivity profiling, Neuro2A cells grown in heavy isotopic medium were treated with KT182 (3 nM) for 4 hours, while light medium-grown cells were treated with DMSO. Cells were harvested, lysed, and membrane/soluble fractions were prepared. Proteomes were labeled with a fluorophosphonate-biotin activity-based probe. Heavy and light proteomes were mixed, biotinylated proteins were enriched with avidin beads, digested with trypsin, and analyzed by LC-MS/MS. [1] |
| Animal Protocol |
For in vivo efficacy and selectivity studies, C57Bl/6 mice were injected intraperitoneally (i.p.) with KT182 dissolved in an 18:1:1 (v/v/v) solution of saline/ethanol/PEG40 (ethoxylated castor oil) at a volume of 10 µL per gram of body weight. The compound was administered at doses of 0.1, 0.5, or 1 mg kg⁻¹. After 4 hours, mice were anesthetized, euthanized, and brain and liver tissues were collected. Tissues were homogenized, and membrane fractions were isolated by centrifugation for subsequent competitive ABPP analysis. [1]
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| References | |
| Additional Infomation |
KT182 is an optimized, irreversible (2-phenyl)piperidinyl-1,2,3-triazolamide ABHD6 inhibitor. [1] It was initially developed from a lead compound through structure-activity relationship studies, with a polar hydroxymethyl substituent introduced at the 3-position of the distal benzene ring of the biphenyltriazole group, thereby fine-tuning the selectivity. [1] It can be used as a brain-penetrating chemical probe for functional analysis of ABHD6 in the central nervous system in animal models. [1] In contrast, the related compound KT203 (20) with a carboxylic acid substituent exhibits peripherally restricted activity. [1]
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| Molecular Formula |
C27H26N4O2
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|---|---|
| Molecular Weight |
438.520946025848
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| Exact Mass |
438.205
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| CAS # |
1402612-62-7
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| PubChem CID |
53364491
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
4.4
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
33
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| Complexity |
632
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(N1C=C(C2=CC=C(C3=CC=CC(CO)=C3)C=C2)N=N1)(N1CCCCC1C1=CC=CC=C1)=O
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| InChi Key |
GICNKPZHUCVFNM-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C27H26N4O2/c32-19-20-7-6-10-24(17-20)21-12-14-22(15-13-21)25-18-31(29-28-25)27(33)30-16-5-4-11-26(30)23-8-2-1-3-9-23/h1-3,6-10,12-15,17-18,26,32H,4-5,11,16,19H2
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| Chemical Name |
[4-[4-[3-(hydroxymethyl)phenyl]phenyl]triazol-1-yl]-(2-phenylpiperidin-1-yl)methanone
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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 : ~100 mg/mL (~228.04 mM)
Ethanol : ~11 mg/mL (~25.08 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.70 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 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 (5.70 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 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 (5.70 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 | 2.2804 mL | 11.4020 mL | 22.8040 mL | |
| 5 mM | 0.4561 mL | 2.2804 mL | 4.5608 mL | |
| 10 mM | 0.2280 mL | 1.1402 mL | 2.2804 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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