Cl-amidine
(Synonyms: N-[(1S)-1-(氨基羰基)-4-[(2-氯-1-亚氨基乙基氨基]丁基]-苯甲酰胺) 目录号 : GC35706
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.
Cas No.:913723-61-2
Sample solution is provided at 25 µL, 10mM.
Cl-amidine (N-α-benzoyl-N5-(2-chloro-1-iminoethyl)-l-ornithine amide), is a PAD4 inactivator with enhanced potency. PAD Inhibitor was proved to exhibit enhanced cell killing in PAD-expressing osteosarcoma cells and block the formation of neutrophil extracellular traps. Cl-amidine, as a PAD inhibitor, could reduce the clinical signs and symptoms of colitis, and induced apoptosis of inflammatory cells. The inhibition of PAD by Cl-amidine markedly suppresses the production of CitH3, and neutralization of CitH3 improves survival markedly in septic mice.[1][2]
Cl-amidine is a more potent inhibitor of PAD which has the IC50 of 5.9 ± 0.3 μM. In vitro experiment indicated that Cl-amidine caused a cell cycle arrest at the G1 phase and miRNA-16 induction through a p53-mediated mechanism. In vivo experiment demonstrated that the enhancement of the p300GBD–GRIP1 interaction mediated by PAD4 was antagonized by Cl-amidine in a dose-dependent manner, and that Cl-amidine treatment caused only a minimal reduction in the efficiency of the interaction, indicating that the inhibitory effect of this compound targeted at the active PAD4 enzyme.[1][3]
References:
[1]. Luo Y, et al. Inhibitors and inactivators of protein arginine deiminase 4: functional and structural characterization. Biochemistry. 2006 Oct 3;45(39):11727-36.
[2].Ting Z, et al. Protective effect of Cl-amidine against CLP-induced lethal septic shock in mice. Sci Rep. 2016; 6: 36696
[3].Cui X, et al. The induction of microRNA-16 in colon cancer cells by protein arginine deiminase inhibition causes a p53-dependent cell cycle arrest. PLoS One. 2013;8(1):e53791.
| Cell experiment [1]: | |
|
Cell lines |
HCT 116 wild-type (WT) colon cancer cells, HCT 116 p53−/− colon carcinoma cells, LS-180 colon cancer cells |
|
Preparation Method |
HCT 116 WT and p53−/− cells were cultured in McCoy's medium supplemented with 10% Newborn Calf Serum, 2 mM glutamine, penicillin (10 U/ml) and streptomycin (10 μg/ml). LS-180 colon cancer cells were cultured in DMEM Medium supplemented with 10% Newborn Calf Serum, 2 mM glutamine, penicillin (10 U/mL) and streptomycin (10 μg/mL). |
|
Reaction Conditions |
1×106 were incubated in 6 well plates in McCoy's 5A medium 1 day before treatment and then treated with 50 µg/mL Cl-amidine. The cells were harvested and fixed with ice cold ethanol. After washing with 1x PBS containing 0.5% BSA twice, the cells were incubated with DNA staining solution consisting of Propidium Iodide (PI; 10 µg/mL) and RNase (0.1 mg/mL) for 30 min at room temperature in the dark. |
|
Applications |
Cl-amidine could lead to a cell cycle arrest at the G1 phase through a p53-mediated mechanism. Cl-amidine could cause miRNA-16 induction in a p53-dependent manner. |
| Animal experiment [2]: | |
|
Animal models |
Male C57BL/6J mice (18–26 g) |
|
Preparation Method |
Mice were injected with intra-peritoneal Cl-amidine dissolved in DMSO, or vehicle DMSO 1 h after cecal ligation and puncture (CLP). DMSO was also injected into mice that were not subjected to CLP (n = 12/group). Mortality was monitored for 10 days after surgeries. |
|
Dosage form |
40 mg/kg |
|
Applications |
Cl-amidine protects mice from sepsis-induced lethality. The inhibition of PAD by Cl-amidine significantly suppresses the CitH3 production, while the CitH3 neutralization markedly improves the survival in septic mice. |
|
References: [1]. Cui X, et al. The induction of microRNA-16 in colon cancer cells by protein arginine deiminase inhibition causes a p53-dependent cell cycle arrest. PLoS One. 2013;8(1):e53791. [2]. Ting Z, et al. Protective effect of Cl-amidine against CLP-induced lethal septic shock in mice. Sci Rep. 2016; 6: 36696. |
|
| Cas No. | 913723-61-2 | SDF | |
| 别名 | N-[(1S)-1-(氨基羰基)-4-[(2-氯-1-亚氨基乙基氨基]丁基]-苯甲酰胺 | ||
| Canonical SMILES | O=C(N[C@H](C(N)=O)CCCNC(CCl)=N)C1=CC=CC=C1 | ||
| 分子式 | C14H19ClN4O2 | 分子量 | 310.78 |
| 溶解度 | Soluble in DMSO | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
||
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 3.2177 mL | 16.0886 mL | 32.1771 mL |
| 5 mM | 0.6435 mL | 3.2177 mL | 6.4354 mL |
| 10 mM | 0.3218 mL | 1.6089 mL | 3.2177 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet