Cl-amidine

Catalog No.S8141 Batch:S814102

Technical Data

Formula

C₁₄H₁₉ClN₄O₂.C₂HF₃O₂

Molecular Weight

424.8

CAS No.

1043444-18-3

Solubility (25°C)*

In vitro

DMSO

85 mg/mL (200.09 mM)

Water

43 mg/mL (101.22 mM)

Ethanol

Insoluble

In vivo (Add solvents to the product individually and in order)

Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	4.25mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 85 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	saline	43.0mg/ml	Taking the 1 mL working solution as an example, add 43 mg of this product to 1 ml of physiological saline (0.9% NaCL solution), mix evenly to make it clear, The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

Cl-amidine is an irreversible pan-peptidylarginine deiminase (PAD) inhibitor with IC50 values of 5.9 ± 0.3 μM, 0.8 ± 0.3 μM, 6.2 ± 1.0 μM for PAD4, PAD1 and PAD3, respectively. Cl-amidine induces apoptosis.

Targets

PAD1 ^[1] (Cell-free assay)	PAD4 ^[1] (Cell-free assay)	PAD3 ^[1] (Cell-free assay)
0.8 μM	5.9 μM	6.2 μM

In vitro

Cl-amidine antagonizes the PAD4-mediated enhancement of the the p300GBD-GRIP1 interaction in a dose-dependent manner. The inhibitory effect of this compound is not a nonspecific one but is targeted at the active PAD4 enzyme^[1]. Cl-amidine increases p53 expression in CD45 positive immune cells. It triggers the differentiation and apoptosis of multiple cancer cell lines that are p53+/+ and p53−/− (e.g., HL60, HT29, TK6, and U2-OS cells). Cl-amidine induces the expression of p53 and several downstream target genes including the cyclin dependent kinase inhibitor p21, GADD45, and the proapoptotic protein PUMA in U2-OS osteosarcoma cells^[2].

In vivo

Cl-amidine treatment inhibits NZM(New Zealand mixed 2328) NET(neutrophil extracellular trap) formation in vivo and significantly alters circulating autoantibody profiles and complement levels while reducing glomerular IgG deposition. Further, Cl-amidine increases the differentiation capacity of bone marrow endothelial progenitor cells, improves endothelium-dependent vasorelaxation, and markedly delays time to arterial thrombosis induced by photochemical injury. Cl-amidine delays thrombosis development in NZM mice. It inhibits PADs in mice without significant toxicity and improves disease phenotypes in animal models of inflammatory arthritis and inflammatory bowel disease^[3]. And It is shown to reduce disease severity in mouse models of ulcerative colitis and RA^[2].

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines CV-1 cells Concentrations 0-200 μM Incubation Time 40 h Method CV-1 cells are transiently transfected with plasmids encoding a luciferase reporter construct, p300GBD fused to the Gal4 DNA binding domain, the p300 binding domain of GRIP1 (i.e., the AD1 domain) fused to the VP16 activation domain (AD), and either wild-type PAD4 or the catalytically defective C645S mutant. Cl-amidine (0-200 μM) is then added to the cell culture medium and incubated for 40 h. Cell extracts are then prepared, and the luciferase activity present in these extracts is quantified.
Animal Study:^{^[4]}	Animal Models DSS mouse model of colitis (genetic background：C57BL/6 mice) Dosages 75 mg/kg (i.p); 5, 25, and 75 mg/kg(oral) Administration by oral gavage or i.p

Cell Assay:^{^[1]}

Cell lines
CV-1 cells
Concentrations
0-200 μM
Incubation Time
40 h
Method
CV-1 cells are transiently transfected with plasmids encoding a luciferase reporter construct, p300GBD fused to the Gal4 DNA binding domain, the p300 binding domain of GRIP1 (i.e., the AD1 domain) fused to the VP16 activation domain (AD), and either wild-type PAD4 or the catalytically defective C645S mutant. Cl-amidine (0-200 μM) is then added to the cell culture medium and incubated for 40 h. Cell extracts are then prepared, and the luciferase activity present in these extracts is quantified.

Animal Study:^{^[4]}

Animal Models
DSS mouse model of colitis (genetic background：C57BL/6 mice)
Dosages
75 mg/kg (i.p); 5, 25, and 75 mg/kg(oral)
Administration
by oral gavage or i.p

References

[4] Chumanevich AA, et al. Am J Physiol Gastrointest Liver Physiol. 2011, 300(6):G929-38.

+ Expand

Selleck's Cl-amidine has been cited by 25 publications

Moxibustion-mediated alleviation of synovitis in rats with rheumatoid arthritis through the regulation of NLRP3 inflammasome by modulating neutrophil extracellular traps [ Heliyon, 2024, 10(1):e23633]	PubMed: 38187290
Metformin potentiates nephrotoxicity by promoting NETosis in response to renal ferroptosis [ Cell Discov, 2023, 9(1):104]	PubMed: 37848438
Metformin potentiates nephrotoxicity by promoting NETosis in response to renal ferroptosis [ Cell Discov, 2023, 9(1):104]	PubMed: 37848438
Neutrophil extracellular traps as a unique target in the treatment of chemotherapy-induced peripheral neuropathy [ EBioMedicine, 2023, 90:104499]	PubMed: 36870200
Fasting-mimicking diet alleviates inflammatory pain by inhibiting neutrophil extracellular traps formation and neuroinflammation in the spinal cord [ Cell Commun Signal, 2023, 21(1):250]	PubMed: 37735678
Fasting-mimicking diet alleviates inflammatory pain by inhibiting neutrophil extracellular traps formation and neuroinflammation in the spinal cord [ Cell Commun Signal, 2023, 21(1):250]	PubMed: 37735678
Constraint-induced movement therapy alleviates motor impairment by inhibiting the accumulation of neutrophil extracellular traps in ischemic cortex [ Neurobiol Dis, 2023, 179:106064]	PubMed: 36878327
Histones released by NETosis enhance the infectivity of SARS-CoV-2 by bridging the spike protein subunit 2 and sialic acid on host cells [ Cell Mol Immunol, 2022, 10.1038/s41423-022-00845-6]	PubMed: 35273357
Extracellular traps from activated vascular smooth muscle cells drive the progression of atherosclerosis [ Nat Commun, 2022, 13(1):7500]	PubMed: 36473863
NET-Triggered NLRP3 Activation and IL18 Release Drive Oxaliplatin-Induced Peripheral Neuropathy [ Cancer Immunol Res, 2022, 10(12):1542-1558]	PubMed: 36255412

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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