Physicochemical Properties
| Molecular Formula | C22H15CL3N2O2 |
| Molecular Weight | 445.725702524185 |
| Exact Mass | 444.019 |
| CAS # | 212135-62-1 |
| PubChem CID | 10789506 |
| Appearance | White to off-white solid powder |
| LogP | 6.6 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 3 |
| Rotatable Bond Count | 4 |
| Heavy Atom Count | 29 |
| Complexity | 616 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | ClC1C=C(C(C)=CC=1C(C#N)C1C=CC(=CC=1)Cl)NC(C1C=C(C=CC=1O)Cl)=O |
| InChi Key | RPTKRVXJNWPJLU-UHFFFAOYSA-N |
| InChi Code | InChI=1S/C22H15Cl3N2O2/c1-12-8-16(18(11-26)13-2-4-14(23)5-3-13)19(25)10-20(12)27-22(29)17-9-15(24)6-7-21(17)28/h2-10,18,28H,1H3,(H,27,29) |
| Chemical Name | 5-chloro-N-[5-chloro-4-[(4-chlorophenyl)-cyanomethyl]-2-methylphenyl]-2-hydroxybenzamide |
| 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) |
Biological Activity
| Targets | SPAK[1] |
| ln Vitro | ZT-1a decreases the phosphorylation of the Na-K-2Cl cotransporter (NKCC1) and increases the K-Cl cotransporter (KCC) through SPS1-associated proline/alanine-rich kinase (SPAK) [1]. ZT-1a inhibits the phosphorylation of NKCC1 p-Thr203/207/212 by 72±5.2% at 1 μM ZT-1a, and the phosphorylation of KCC site 1/2 in HEK-293 by 65-77% at 3 μM ZT-1a [1]. At 3–10 μM ZT-1a, there is a 70±3.8% inhibition on the phosphorylation of SPAK at Ser373 [1]. Ten micrograms of ZT-1a inhibits NKCC1, but increases KCC3 activity [1]. |
| ln Vivo | In vivo, ZT-1a (10–100 mg/kg) prevents SPAK-dependent phosphorylation of the cation-Cl-cotransporter (CCC)[1]. |
| Animal Protocol |
Animal/Disease Models: Naive mice[1] Doses: 10, 30, 50, and 100 mg/kg Route of Administration: intraperitoneal (ip) administration Experimental Results: Inhibited SPAK-dependent cation-Cl- cotransporters (CCC) phosphorylation in vivo. |
| References |
[1]. Modulation of Brain cation-Cl- Cotransport via the SPAK Kinase Inhibitor ZT-1a. Nat Commun. 2020 Jan 7;11(1):78. |
Solubility Data
| Solubility (In Vitro) | DMSO: 100 mg/mL (224.35 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.67 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 (4.67 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (4.67 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2435 mL | 11.2176 mL | 22.4351 mL | |
| 5 mM | 0.4487 mL | 2.2435 mL | 4.4870 mL | |
| 10 mM | 0.2244 mL | 1.1218 mL | 2.2435 mL |