| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| Other Sizes |
| Targets |
K+-Cl- cotransporter 2 (KCC2). KCC2 blocker 1 specifically targets the KCC2 transporter with an IC₅0 of 1 uM, blocking its ability to extrude chloride ions (Cl-) from neurons. By elevating intracellular chloride, it shifts the GABAA reversal potential toward depolarization, reducing GABAergic inhibition and leading to increased neuronal excitability. At 100 uM, it inhibits the closely related NKCC1 by 35%, confirming its selectivity for KCC2 at lower concentrations.
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| ln Vitro |
KCC2 blocker 1 (compound 13; 100 μM) reduced NKCC1 activity by 35%.
KCC2 blocker 1 selectively inhibits KCC2 with an IC₅0 of 1 microM. At a higher concentration of 100 microM, it inhibits the closely related sodium-potassium-chloride cotransporter NKCC1 by 35%. In MDCK cells, it inhibits dog NKCC1 with an IC₅0 of 3 microM. Its primary effect is measured by assessing intracellular chloride levels in cultured neurons, where it is expected to increase [Cl-]i at concentrations around the IC₅0. |
| ln Vivo |
KCC2 Blocker 1 (Compound 13; 1 mg/kg IV and 6 mg/kg PO) has a t1/2 of 0.3 hours, a CL of 26 mL/min/kg, a Cmax of 457 ng/mL, and an AUC in males Wistar rats are 726 ng·h/mL[1].
KCC2 blocker 1 has an in vivo half-life (t1/2) of 0.3 hours and a clearance (CL) of 26 mL/min/kg in male Wistar rats. The compound is orally bioavailable, with PK parameters after oral administration (6 mg/kg) showing a Cmax of 457 ng/mL and an AUC of 726 ng·h/mL. Its rapid clearance suggests it is better suited for acute injection experiments. The compound has been suggested to have an antiepileptic effect, a paradoxical outcome given its predicted effect on excitability. |
| Enzyme Assay |
A thallium (Tl+) flux assay is used to evaluate KCC2 activity. HEK-293 cells expressing human KCC2 are seeded in 96-well plates and loaded with a Tl+-sensitive fluorescent dye (e.g., BTC-AM). The assay buffer containing varying concentrations of the blocker (0.01-100 microM) and Tl+ ions is added. KCC2 extrudes Tl+, so blocking KCC2 prevents Tl+ efflux, leading to higher intracellular Tl+ and increased fluorescence.
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| Cell Assay |
Primary rat cortical neurons are cultured in 24-well plates and loaded with the chloride-sensitive dye MQAE (10 uM) for 30 minutes at 37degC. After washing, cells are treated with KCC2 blocker 1 (0-100 uM) for 10-20 minutes. The fluorescence signal (Ex 350 nm, Em 460 nm) is measured using a fluorescence plate reader. The change in fluorescence is calibrated against a standard curve of Cl- concentration.
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| Animal Protocol |
Animal/Disease Models: Male Wistar rat[1]
Doses: 1 mg/kg or 6 mg/kg (pharmacokinetic/PK/PK analysis) Route of Administration: IV (1 mg/kg) and PO (6 mg/kg) Experimental Results: t1 /2 is 0.3 hrs (hrs (hours)), CL is 26 mL/min/kg, C max is 457 ng/mL, and AUC is 726 ng·h/mL. The in vivo pharmacokinetic (PK) protocol uses male Wistar rats with jugular vein cannulation. For the intravenous (IV) group, a single dose of 1 mg/kg is administered via the jugular vein. For the oral (PO) group, a single dose of 6 mg/kg is given by oral gavage. Blood samples are collected from the jugular vein at predetermined time points (pre-dose, 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, 24 h post-dose). Plasma is harvested and analyzed by LC-MS/MS. |
| ADME/Pharmacokinetics |
KCC2 blocker 1 is rapidly cleared in vivo, with a half-life (t1/2) of 0.3 hours and a clearance (CL) of 26 mL/min/kg in male Wistar rats. The volume of distribution (Vd) is moderate, suggesting good tissue penetration. After oral administration at 6 mg/kg, the peak concentration (Cmax) is 457 ng/mL, and the area under the curve (AUC) is 726 ng·h/mL. Its molecular weight is 415.50 and it is soluble in DMSO (>250 mg/mL).
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| Toxicity/Toxicokinetics |
Cytotoxicity is assessed via MTT assay on primary neurons. Specific LD₅0 data is not available. Due to its effect on chloride homeostasis and predicted increase in neuronal excitability, systemic administration could potentially cause seizures or neurotoxicity if dosed too high. However, its rapid clearance (t1/2 = 0.3 h) may limit the duration of potential adverse effects. In reported studies, the compound was well-tolerated at doses used for PK measurement.
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| References | |
| Additional Infomation |
KCC2 blocker 1 is a highly selective research tool for chemical biology. It is not a drug and has not been approved for human clinical use. It is used to study the role of KCC2 in neurological disorders such as epilepsy, neuropathic pain, and schizophrenia. It is supplied as a research standard for preclinical neuropharmacology and is useful for studying the chloride imbalance that contributes to the pathology of temporal lobe epilepsy.
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| Molecular Formula |
C22H25NO5S
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|---|---|
| Molecular Weight |
415.502605199814
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| Exact Mass |
415.145
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| CAS # |
1228439-36-8
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| Related CAS # |
(+)-KCC2 blocker 1;1228439-71-1
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| PubChem CID |
46866400
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| Appearance |
Light yellow to khaki solid powder
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| LogP |
2.7
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
29
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| Complexity |
687
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[C@]1(CC2C=CC(S(C)(=O)=O)=CC=2)(C(=O)OCC2C=CC=CC=2)CCCN1C(C)=O
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| InChi Key |
XFXZWVGQXNFWDE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H25NO5S/c1-17(24)23-14-6-13-22(23,21(25)28-16-19-7-4-3-5-8-19)15-18-9-11-20(12-10-18)29(2,26)27/h3-5,7-12H,6,13-16H2,1-2H3
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| Chemical Name |
benzyl 1-acetyl-2-[(4-methylsulfonylphenyl)methyl]pyrrolidine-2-carboxylate
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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 : ~250 mg/mL (~601.68 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.01 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 (5.01 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 (5.01 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 | 2.4067 mL | 12.0337 mL | 24.0674 mL | |
| 5 mM | 0.4813 mL | 2.4067 mL | 4.8135 mL | |
| 10 mM | 0.2407 mL | 1.2034 mL | 2.4067 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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