CK 666
目录号 : GC14060
CK 666是一种可穿透细胞的肌动蛋白相关蛋白Arp2/3复合物抑制剂（IC50=12μM）。
Cas No.:442633-00-3
Sample solution is provided at 25 µL, 10mM.
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CK 666 is a cell-permeable actin-related protein Arp2/3 complex inhibitor (IC50=12μM)[1]. CK 666 binds to Arp2/3 complex, stabilizes the inactive state of the complex, blocking movement of the Arp2 and Arp3 subunits into the activated filament-like (short pitch) conformation[2]. CK 666 is routinely used in studies of cytoskeletal dynamics such as cell migration, phagocytosis and cytokinesis[3][4].
In vitro, CK 666 (100μM; 24h) inhibited proliferation in KYSE-30 esophageal, T24 bladder, MDA-MB-231 breast, FaDu hypopharyngeal and SKOV3 ovarian cancer cells, elevated LC3-II, decreased p62 to promote autophagy, and increased release of CD63⁺exosome-like vesicles[5]. Pretreatment of human lung microvascular endothelial cells (HLMVEC) and pulmonary artery endothelial cells (HPAEC)with CK 666 (50μM) 1h before sphingosine-1-phosphate (S1P)-induction significantly reduced trans-endothelial electrical resistance (TER), attenuated S1P-induced barrier enhancement, and markedly decreased individual lamellipodium depth[6].
In vivo, CK 666 (30mg/kg; i.p.; once 24h and once 2h before ischemia) markedly decreased serum creatinine and blood urea nitrogen levels, reduced renal malondialdehyde (MDA) content, attenuated tubular necrosis, and significantly lowered TUNEL-positive apoptotic cells in a mouse renal ischemia–reperfusion injury model[7].
References:
[1] Hetrick B, Han MS, Helgeson LA, Nolen BJ. Small molecules CK-666 and CK-869 inhibit actin-related protein 2/3 complex by blocking an activating conformational change. Chem Biol. 2013;20(5):701-712.
[2] Nolen BJ, Tomasevic N, Russell A, et al. Characterization of two classes of small molecule inhibitors of Arp2/3 complex. Nature. 2009;460(7258):1031-1034.
[3] Liu CS, Cheung PW, Dinesh A, et al. Actin-related protein 2/3 complex plays a critical role in the aquaporin-2 exocytotic pathway. Am J Physiol Renal Physiol. 2021;321(2):F179-F194.
[4] Efremov YM, Dokrunova AA, Efremenko AV, Kirpichnikov MP, Shaitan KV, Sokolova OS. Distinct impact of targeted actin cytoskeleton reorganization on mechanical properties of normal and malignant cells. Biochim Biophys Acta. 2015;1853(11 Pt B):3117-3125.
[5] Li L, Cai S, Chen J, et al. CK-666 exerts anticancer effects by regulating autophagy, tunneling nanotubes and extracellular vesicles formation. Biomed Pharmacother. 2025;183:117825.
[6] Belvitch P, Brown ME, Brinley BN, et al. The ARP 2/3 complex mediates endothelial barrier function and recovery. Pulm Circ. 2017;7(1):200-210.
[7] Hu Q, Zhao Y, Sun WY, et al. CK-666 protects against ferroptosis and renal ischemia-reperfusion injury through a microfilament-independent mechanism. J Biol Chem. 2024;300(12):107942.
CK 666是一种可穿透细胞的肌动蛋白相关蛋白Arp2/3复合物抑制剂（IC50=12μM）[1]。CK 666通过与Arp2/3复合物结合，稳定其非活化构象，阻断Arp2与Arp3亚基向活化丝状（短螺距）构象的移动[2]。CK 666常被用于细胞迁移、吞噬和胞质分裂等细胞骨架动力学研究[3][4]。
体外实验中，CK 666（100μM；24h）可抑制KYSE-30食管癌细胞、T24膀胱癌细胞、MDA-MB-231乳腺癌细胞、FaDu下咽癌细胞和SKOV3卵巢癌细胞的增殖，上调LC3-II、下调p62以促进自噬，并增加CD63⁺外泌体样囊泡的释放[5]。在鞘氨醇-1-磷酸（S1P）诱导前1h用CK 666（50μM）预处理人肺微血管内皮细胞（HLMVEC）和肺动脉内皮细胞（HPAEC），可显著降低跨内皮电阻（TER），减弱鞘氨醇-1-磷酸（S1P）诱导的屏障增强效应，并显著减小单个板状伪足的深度[6]。
体内实验中，CK 666（30mg/kg；腹腔注射；分别在缺血前24h和2h给药一次）在小鼠肾缺血-再灌注损伤模型中显著降低血清肌酐和尿素氮水平，减少肾组织丙二醛（MDA）含量，减轻肾小管坏死，并显著降低TUNEL阳性凋亡细胞数量[7]。
| Cell experiment [1]: | |
Cell lines | Human pulmonary artery EC (HPAECs) or human lung microvascular EC (HLMVECs) |
Preparation Method | Human pulmonary artery EC (HPAECs) or human lung microvascular EC (HLMVECs) were cultured in Endothelial Basal Medium (EBM)-2 complete medium supplemented with 10% (v/v) fetal bovine serum (FBS) in a humidified incubator with 5% CO2 at 37°C. EC monolayer barrier function was assessed by an electrical cell-substrate impedance sensing system. In this assay, ECs are grown in wells containing gold microelectrodes on the basal surface allowing the measurement of impedance to a small AC electrical current at a frequency of 4000Hz. Once a baseline resistance (TER) was established, the monolayer was treated with vehicle (0.3% DMSO) or CK 666 (50μM) for 1h followed by thrombin (1U/mL) or S1P (1μM) and monitored continuously for up to 12h. Values are expressed as mean resistance, measured in ohms, or normalized resistance compared to baseline values with the means of several wells pooled in each independent experiment. Baseline resistance in HLMVECs was calculated from three independent experiments with 2–4 repeats of each condition. The maximal change in HPAEC normalized resistance as well as the rate of change (normalized resistance/h) following S1P was calculated from seven independent experiments with each experiment composed of 2–4 repeats of each condition. The time to recovery of baseline barrier function following thrombin (measured in minutes) was calculated from nine independent experiments with each experiment composed of 2–4 repeats of each condition. |
Reaction Conditions | 50μM; 1h |
Applications | Pretreatment of human lung microvascular endothelial cells (HLMVEC) and pulmonary artery endothelial cells (HPAEC)with CK 666 (50μM) 1h before sphingosine-1-phosphate (S1P)-induction significantly reduced trans-endothelial electrical resistance (TER). |
| Animal experiment [2]: | |
Animal models | C57BL/6 WT mice |
Preparation Method | C57BL/6 WT mice were intraperitoneally injected with CK 666 (30mg/kg) 24h and 2h before surgery. The mice were anesthetized, and bilateral renal pedicles were clamped for 35min to induce renal ischemia-reperfusion injury, during which the body temperature of mice was maintained on a heat plate at 38°C. The mice in the Sham group underwent the same procedures as those in the renal I/R group except for drug administration and clamping. The mice were sacrificed 24h after surgery. Serum and tissue samples were collected for subsequent testing. |
Dosage form | 30mg/kg; i.p.; once 24h and once 2h before ischemia |
Applications | CK 666 markedly decreased serum creatinine and blood urea nitrogen levels, reduced renal malondialdehyde (MDA) content, attenuated tubular necrosis in a mouse renal ischemia–reperfusion injury model. |
References: | |
| Cas No. | 442633-00-3 | SDF | |
| 化学名 | 2-fluoro-N-(2-(2-methyl-1H-indol-3-yl)ethyl)benzamide | ||
| Canonical SMILES | FC1=CC=CC=C1C(NCCC2=C(C)NC3=CC=CC=C23)=O | ||
| 分子式 | C18H17FN2O | 分子量 | 296.34 |
| 溶解度 | <29.63mg/ml in DMSO; <29.63mg/ml in ethanol | 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。 |
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.3745 mL | 16.8725 mL | 33.745 mL |
| 5 mM | 674.9 μL | 3.3745 mL | 6.749 mL |
| 10 mM | 337.5 μL | 1.6873 mL | 3.3745 mL |
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动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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| % DMSO % % Tween 80 % saline | ||||||||||
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2.
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