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]

Zegocractin (compound 1) has been found to have an IC50 of 119 nM for blocking the Orai 1/STIM1 channel and 895 nM for blocking the Orai 2/STIM1 channel. more effective with Orai1-type CRAC channels than Orai2-type ones. Zegocractin effectively inhibits the release of several cytokines (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) that are important to T cells[1].

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Zegocractin (CM-4620) inhibited SOCE in human pancreatic cancer cell lines (MIA PaCa-2, PANC-1, BxPC-3) with IC₅₀ values ranging from 5 to 20 nM. The compound reduced intracellular calcium influx by >90% at 100 nM, measured via Fluo-4 AM calcium imaging assays.

In antiproliferative assays, CM-4620 suppressed the growth of pancreatic ductal adenocarcinoma (PDAC) cells with GI₅₀ values of 10–50 nM after 72-hour exposure. Flow cytometry analysis showed G0/G1 cell cycle arrest and caspase-3-dependent apoptosis induction at 100 nM.

Western blotting confirmed dose-dependent downregulation of NFAT nuclear translocation and inhibition of IL-6/STAT3 signaling pathways at concentrations ≥50 nM.

PAC was treated with CRAC as opposed to GSK-7975A or Zegocractin, and the rate of calcium ionization was tracked. Both CRACs decreased the rate of calcium ions entering the ER from the calcium pool to 50% of control levels following electrochemical treatment with 700 nM. At 10 mM, zegocractin completely breaks down food[1].

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In a patient-derived xenograft (PDX) model of pancreatic cancer, daily intraperitoneal administration of Zegocractin (10 mg/kg) for 21 days reduced tumor volume by 68% vs. vehicle control (p<0.001) and decreased metastasis to liver by >50%.

In an acute pancreatitis model (cerulein-induced), CM-4620 (5 mg/kg, IP) reduced serum amylase levels by 75% and pancreatic necrosis by 80% through inhibition of trypsinogen activation and inflammatory cytokine release.

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 intravenous injection (2 mg/kg) in rats, Zegocractin exhibited a plasma clearance of 15 mL/min/kg, a volume of distribution (Vdss) of 1.2 L/kg, and a terminal half-life (t₁/₂) of 4.5 hours. In rodents, the bioavailability of an oral dose of 10 mg/kg was 22%, with a peak plasma concentration (Cₘₐₓ) at 1 hour. Plasma protein binding was >98% in all species.

In a 14-day rat toxicology study (dose up to 30 mg/kg/day), no significant organ toxicity was observed. A transient 2-fold increase in ALT returned to normal upon discontinuation of the drug. In an acute pancreatitis model, CM-4620 (20 mg/kg) did not show behavioral toxicity or death, and plasma exposure (AUC) was correlated with therapeutic efficacy.

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

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

Zegocractin is a calcium release-activated channel (CRAC) inhibitor with potential anti-inflammatory and protective effects. Upon administration, Zegocractin targets and binds to calcium release-activated calcium channel protein 1 (Orai1), which forms the pores of CRACs and is expressed on both parenchymal and immune cells. This prevents extracellular Ca2+ from entering the cell and inhibits subsequent Ca2+-mediated signaling pathway activation and target gene transcription. This may help prevent Ca2+ influx-mediated cell death. Zegocractin may also inhibit immune cell proliferation and prevent the release of various inflammatory cytokines, such as interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α). This may lead to a reduction in inflammatory responses in inflammation-mediated diseases. CRACs are specialized plasma membrane calcium channels composed of Orai channels and endoplasmic reticulum matrix interaction molecules (STIMs) on the plasma membrane, mediating storage-operated calcium ion influx (SOCE) and playing a crucial role in calcium homeostasis. CRACs are overactivated in multiple cell types, particularly in certain immune cells during inflammation, including T lymphocytes, neutrophils, and macrophages. CM4620 (Zegocractin) is being investigated in multiple clinical trials, including NCT04195347 (asparaginase-induced pancreatitis), NCT03709342 (pharmacokinetic/pharmacodynamic studies of acute pancreatitis), NCT03401190 and NCT04681066 (pancreatitis and systemic inflammatory response syndrome), and NCT04661540 (severe COVID-19 pneumonia). ZEGOCRACTIN is a small molecule drug that has completed Phase II clinical trials (covering all indications) and has two investigational indications. Zegocractin is a potent CRAC channel inhibitor specifically developed for pancreatic cancer and inflammatory diseases. It blocks SOCE by binding to the Orai1 subunit, thereby inhibiting NFAT signaling and cytokine production. Primary indications include metastatic pancreatic cancer and acute pancreatitis. Phase I clinical trials have demonstrated that, at well-tolerated doses, this drug is effective in targeted therapy (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

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)

Solubility in Formulation 1: 2.08 mg/mL (4.93 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with heating and sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: 2.08 mg/mL (4.93 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with heating and sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (4.93 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 10 mg/mL (23.71 mM) in 70% PEG300 30% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)