| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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CZC-54252 HCl (CZC54252) is a novel and potent LRRK2 (leucine-rich repeat kinase 2) inhibitor with a potential for Parkinson's disease (PD).
| Targets |
Whereas the 4-chlorodiaminopyrimidine CZC-54252 exhibited good selectivity and potently inhibited binding of only ten human or mouse kinases，none of which have been classified as predictors of genotoxicity or hematopoietic toxicity.[1]
G2019S LRRK2-induced human neuronal injury was attenuated by CZC-25146 with an EC50 of ~4 nM (EC50 CZC-54252 ~1 nM) and fully reversed to wild-type levels by both compounds at concentrations as low as 8 nM (1.6 nM for CZC-54252) (Fig. 4c-d). In human cortical neurons, no overt cytotoxicity was seen in the efficacious concentration range (only at 5 μM for CZC-25146 and ≥1 μM for CZC-54252) .[1] |
|---|---|
| ln Vitro |
Whereas the 4-chlorodiaminopyrimidine CZC-54252 exhibited good selectivity and potently inhibited binding of only ten human or mouse kinases，none of which have been classified as predictors of genotoxicity or hematopoietic toxicity.[1]
G2019S LRRK2-induced human neuronal injury was attenuated by CZC-25146 with an EC50 of ~4 nM (EC50 CZC-54252 ~1 nM) and fully reversed to wild-type levels by both compounds at concentrations as low as 8 nM (1.6 nM for CZC-54252) (Fig. 4c-d). In human cortical neurons, no overt cytotoxicity was seen in the efficacious concentration range (only at 5 μM for CZC-25146 and ≥1 μM for CZC-54252) .[1] CZC-54252 potently inhibits binding of endogenous mouse LRRK2 to an affinity matrix (la-S7) in a chemoproteomics competition assay with an IC50 of approximately 0.19 μM. CZC-54252 is a potent inhibitor of recombinant human wild-type LRRK2 enzyme activity (IC50 = 1.28 nM) and the G2019S mutant LRRK2 (IC50 = 1.85 nM) in a time-resolved fluorescence resonance energy transfer (TR-FRET)-based kinase activity assay.[1] In primary human cortical neurons, CZC-54252 fully rescues G2019S LRRK2-induced neurite defects (reduction in average neurite length and branchpoint counts) at concentrations as low as 1.6 nM. The estimated EC50 for rescuing neurite length defects is approximately 1 nM.[1] CZC-54252 exhibits good kinase selectivity. Quantitative chemoproteomics profiling against 184 different protein kinases and one lipid kinase using Kinobeads matrix showed it potently inhibits the binding of only ten human or mouse kinases within the tested concentration range (10 nM to 2 μM).[1] CZC-54252 does not cause overt cytotoxicity in human cortical neurons at concentrations up to 1 μM over a seven-day treatment period, as assessed by an AlamarBlue assay.[1] |
| Enzyme Assay |
The potency of CZC-54252 against LRRK2 kinase activity was determined using a time-resolved fluorescence resonance energy transfer (TR-FRET) assay. Recombinant human wild-type or G2019S mutant LRRK2 enzyme was incubated with the compound at various concentrations. The kinase reaction was performed in the presence of ATP at a concentration approximating the KM of LRRK2 for ATP (100 μM). The transfer of a phosphate group to a biotinylated peptide substrate was detected using europium-labeled anti-phospho-specific antibody and streptavidin-allophycocyanin. The resulting TR-FRET signal was measured, and dose-response curves were generated to calculate IC50 values.[1]
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| Cell Assay |
Primary rodent cortical neuron toxicity assay: Primary cortical neurons from embryonic rats were transfected at day in vitro (DIV) 14 with plasmids encoding mutant LRRK2 (G2019S or R1441C) and eGFP (as a marker) using a transfection reagent. CZC-54252 or vehicle (DMSO) was added at the time of transfection and maintained in the culture medium. Neuronal injury was assessed on DIV 16 by microscopic examination, defining injured cells as those losing neurites and rounding up. Neuronal death was quantified by a TUNEL assay to detect DNA fragmentation.[1]
Primary human cortical neuron neurite morphology assay: Primary human cortical neurons from fetal tissue were cultured. At DIV 14, neurons were co-transfected with plasmids encoding GFP and various LRRK2 constructs (wild-type, mutants, or empty vector) using a specialized transfection reagent. CZC-54252 or DMSO was added at indicated concentrations at the time of transfection and replenished with medium changes. On DIV 20, cultures were fixed. Automated image acquisition and analysis were performed using a high-content imaging system equipped with a NeuronalProfiling bioapplication to quantify average neurite length and branchpoint counts per neuron.[1] Cytotoxicity assay in human neurons: Cultured primary human cortical neurons were treated with CZC-54252 at various concentrations for seven days. Potential compound-mediated cytotoxicity was assessed using an AlamarBlue assay. Cells were incubated with AlamarBlue reagent for 2 hours, and the resulting fluorescence was measured. Data were normalized to DMSO-treated control wells.[1] |
| ADME/Pharmacokinetics |
CZC-54252 has poor brain permeability. In the pharmacokinetic studies of CZC-25146 and CZC-54252, its brain-plasma ratio was approximately 4%. [1]
Reference [1] did not provide other specific ADME parameters for CZC-54252 (e.g., clearance, volume of distribution, half-life, oral bioavailability). |
| Toxicity/Toxicokinetics |
After seven days of in vitro culture, CZC-54252 at concentrations up to 1 μM did not cause significant cytotoxicity to human cortical neurons. [1]
The selectivity of CZC-54252 indicates that it does not effectively inhibit kinases classified as predictors of genotoxicity or hematopoietic toxicity according to reference [1]. Reference [1] does not provide in vivo toxicity, lethal dose, or detailed toxicokinetic parameters of CZC-54252. |
| References | |
| Additional Infomation |
CZC-54252 is a lead compound based on diaminopyrimidine, discovered and optimized using a chemical proteomics strategy to develop a highly effective and selective LRRK2 inhibitor. [1]
It was discovered by screening a library of kinase-targeting compounds using an affinity matrix (Ia-S7) derived from sunitinib analogs, and subsequently optimized by dot blot power assays and selectivity analysis based on quantitative LC-MS/MS. [1] This study demonstrates that selective inhibition of LRRK2 kinase activity using compounds such as CZC-54252 is sufficient to alleviate neurite lesion damage and cell death induced by mutant LRRK2 (G2019S and R1441C) in rodent and primary human neuronal cultures, thus validating the effectiveness of LRRK2 as a therapeutic target for Parkinson's disease. [1] Due to its poor brain penetration, this compound was primarily used in in vitro models in the initial studies. The authors believe that such intracranial blockade inhibitors may help in studying the function of LRRK2 in peripheral tissues (such as the kidneys and immune cells). [1] |
| Molecular Formula |
C22H26CL2N6O4S
|
|---|---|
| Molecular Weight |
541.448
|
| Exact Mass |
540.111
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| Elemental Analysis |
C, 48.80; H, 4.84; Cl, 13.09; N, 15.52; O, 11.82; S, 5.92
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| CAS # |
1784253-05-9
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| Related CAS # |
CZC-54252;1191911-27-9
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| PubChem CID |
90488950
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| Appearance |
Light yellow to orange solid powder
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| Hydrogen Bond Donor Count |
4
|
| Hydrogen Bond Acceptor Count |
10
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
35
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| Complexity |
737
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ClC1C=NC(=NC=1NC1C=CC=CC=1NS(C)(=O)=O)NC1C=CC(=CC=1OC)N1CCOCC1.Cl
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| InChi Key |
KWCBHUPLQMUKAF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H25ClN6O4S.ClH/c1-32-20-13-15(29-9-11-33-12-10-29)7-8-19(20)26-22-24-14-16(23)21(27-22)25-17-5-3-4-6-18(17)28-34(2,30)31;/h3-8,13-14,28H,9-12H2,1-2H3,(H2,24,25,26,27);1H
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| Chemical Name |
N-[2-[[5-chloro-2-(2-methoxy-4-morpholin-4-ylanilino)pyrimidin-4-yl]amino]phenyl]methanesulfonamide;hydrochloride
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| Synonyms |
CZC-54252 hydrochloride; CZC-54252 HCl; CZC54252 HCl; CZC 54252 HCl; CZC-54252; CZC54252; CZC 54252;
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 : ~33.33 mg/mL (~61.56 mM)
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8469 mL | 9.2345 mL | 18.4689 mL | |
| 5 mM | 0.3694 mL | 1.8469 mL | 3.6938 mL | |
| 10 mM | 0.1847 mL | 0.9234 mL | 1.8469 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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