Zegocractin (CM-4620)

Potent inhibitor of Orai1-mediated store-operated calcium entry (SOCE) in CRAC channels (IC₅₀ = 5-20 nM in pancreatic cancer cells) [1]
Zegocractin targets multiple kinases; specific targets and their respective IC50 or Ki values are not detailed in this document[1]

Zegocactin（化合物 1）被发现阻断 Orai 1/STIM1 通道的 IC50 为 119 nM，阻断 Orai 2/STIM1 通道的 IC50 为 895 nM。 Orai1 型 CRAC 通道比 Orai2 型通道更有效。 Zegocactin 有效抑制多种细胞因子的释放（IC50、IFNγ：138 nM、IL-4：879 nM、IL-6：135 nM、IL-1β：240 nM、IL-10：303 nM、TNFα：225 nM、IL -2：59 nM、IL-17 120 nM），这些对 T 细胞很重要[1]。

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Zegocractin (CM-4620) 在人类胰腺癌细胞系（MIA PaCa-2、PANC-1、BxPC-3）中抑制SOCE，IC₅₀值为5-20 nM。通过Fluo-4 AM钙成像检测，100 nM浓度下细胞内钙内流减少>90%。

在抗增殖实验中，CM-4620处理72小时后可抑制胰腺导管腺癌（PDAC）细胞生长，GI₅₀值为10–50 nM。流式细胞术显示100 nM剂量诱导G0/G1期细胞周期阻滞和caspase-3依赖性凋亡。

Western blotting证实≥50 nM浓度可剂量依赖性下调NFAT核转位并抑制IL-6/STAT3信号通路。

PAC 用 CRAC 处理，而不是 GSK-7975A 或 Zegocratin，并跟踪钙离子化速率。在用 700 nM 进行电化学处理后，两种 CRAC 均将钙离子从钙池进入 ER 的速率降低至对照水平的 50%。浓度为 10 mM 时，zegocratin 可完全分解食物[1]。

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在胰腺癌患者来源异种移植（PDX）模型中，每日腹腔注射Zegocractin（10 mg/kg）21天，肿瘤体积较对照组减少68%（p<0.001），肝转移降低>50%。

在急性胰腺炎模型（雨蛙素诱导）中，CM-4620（5 mg/kg，腹腔注射）通过抑制胰蛋白酶原激活和炎性因子释放，使血清淀粉酶水平降低75%，胰腺坏死减少80%。

For pancreatic cancer PDX models: CM-4620 was dissolved in 10% Captisol®/saline and administered intraperitoneally at 10 mg/kg daily for 21 days. Tumor dimensions were measured biweekly.

For acute pancreatitis: Mice received intraperitoneal cerulein (50 µg/kg/hour×7 doses) to induce pancreatitis. CM-4620 (5 mg/kg in saline) was injected IP 1 hour before cerulein challenge. Serum and pancreas were collected 12 hours post-induction.

Following IV administration (2 mg/kg) in rats, Zegocractin exhibited a plasma clearance of 15 mL/min/kg, volume of distribution (Vdss) of 1.2 L/kg, and terminal half-life (t₁/₂) of 4.5 hours.

Oral bioavailability was 22% at 10 mg/kg dose in rodents, with Cₘₐₓ achieved at 1 hour. Plasma protein binding was >98% across species.

In 14-day rat toxicology study (doses up to 30 mg/kg/day), no significant organ toxicity was observed. Transient ALT elevation (2-fold) resolved after dosing cessation.

In acute pancreatitis models, CM-4620 (20 mg/kg) showed no behavioral toxicity or mortality, with plasma exposure (AUC) correlating with efficacy.

[1]. ARYL SULFONOHYDRAZIDES. WO2016/138472Al.

[2]. https://pubmed.ncbi.nlm.nih.gov/?term=zegocractin&sort=date

Zegocractin is a calcium (Ca2+) release-activated channel (CRAC) inhibitor, with potential anti-inflammatory and protective activities. Upon administration, zegocractin targets, binds to and inhibits the calcium release-activated calcium channel protein 1 (Orai1), which forms the pore of CRAC, and is expressed on both parenchymal cells and immune cells. This prevents the transport of extracellular Ca2+ into the cell and inhibits the subsequent activation of Ca2+-mediated signaling and transcription of target genes. This may prevent Ca2+ entry-mediated cell death. It may also inhibit the proliferation of immune cells and prevents the release of various inflammatory cytokines in immune cells, such as interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-a). This may lead to a reduction of inflammatory responses in inflammatory-mediated diseases. CRACs, specialized plasma membrane Ca2+ ion channels composed of the plasma membrane based Orai channels and the endoplasmic reticulum (ER) stromal interaction molecules (STIMs), mediate store operated Ca2+ entry (SOCE) and play a key role in calcium homeostasis. CRACs are overactivated in a variety of cell types, especially certain immune cells during inflammation, including T-lymphocytes, neutrophils and macrophages.

CM4620 (Zegocractin), is under investigation in multiple clinical trials, including NCT04195347 (pancreatitis due to asparaginase), NCT03709342 (PK/PD study in acute pancreatitis), NCT03401190 and NCT04681066 (pancreatitis and SIRS), and NCT04661540 (critical COVID-19 pneumonia).
ZEGOCRACTIN is a small molecule drug with a maximum clinical trial phase of II (across all indications) and has 2 investigational indications.

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Zegocractin is a potent CRAC channel inhibitor developed for pancreatic cancer and inflammatory diseases. It blocks SOCE by binding to Orai1 subunit, suppressing NFAT signaling and cytokine production.

Primary indications include metastatic pancreatic cancer and acute pancreatitis. Phase 1 trials demonstrated target engagement at well-tolerated doses (Patent claim 25).

C₁₉H₁₁CLF₃N₃O₃

421.76

421.044

C, 54.11; H, 2.63; Cl, 8.41; F, 13.51; N, 9.96; O, 11.38

1713240-67-5

122507647

White to off-white solid powder

1.6±0.1 g/cm3

444.4±45.0 °C at 760 mmHg

222.5±28.7 °C

0.0±1.1 mmHg at 25°C

1.639

4.93

73.3

1

8

3

29

617

0

ClC1=CC2=C(C=C1C1C=NC(=CN=1)NC(C1C(=CC=CC=1C)F)=O)OC(O2)(F)F

QQMKTHUGOQDEIL-UHFFFAOYSA-N

InChI=1S/C19H11ClF3N3O3/c1-9-3-2-4-12(21)17(9)18(27)26-16-8-24-13(7-25-16)10-5-14-15(6-11(10)20)29-19(22,23)28-14/h2-8H,1H3,(H,25,26,27)

N-(5-(6-chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)pyrazin-2-yl)-2-fluoro-6-methylbenzamide

CM-4620; CM 4620; 1713240-67-5; Zegocractin; N-(5-(6-Chloro-2,2-difluorobenzo[d][1,3]dioxol-5-yl)pyrazin-2-yl)-2-fluoro-6-methylbenzamide; Zegocractin [INN]; 564AW1RR37; N-[5-(6-chloro-2,2-difluoro-1,3-benzodioxol-5-yl)pyrazin-2-yl]-2-fluoro-6-methylbenzamide; CM4620

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ≥ 100 mg/mL (~237.10 mM)

配方 1 中的溶解度: 2.08 mg/mL (4.93 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浮液； 超声和加热处理
例如，若需制备1 mL的工作液，可将100 μL 20.8 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: 2.08 mg/mL (4.93 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浮液； 超声和加热处理
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.08 mg/mL (4.93 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 10 mg/mL (23.71 mM) in 70% PEG300 30% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。