| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
IC50: MYOF[1]
Myoferlin (MYOF). HJ445A specifically binds to the MYOF-C2D domain with a KD of 0.17 μM. MYOF is a 230 kDa protein belonging to the ferlin family, characterized by multiple C2 domains (C2A through C2F) that mediate calcium-dependent phospholipid binding and membrane fusion. The C2D domain of MYOF is one of the calcium-binding domains that is critical for MYOF's function in membrane trafficking and cell signaling. By binding to the C2D domain with high affinity, HJ445A likely interferes with MYOF's ability to mediate membrane fusion events, thereby disrupting MYOF-dependent cellular processes such as cell migration, invasion, and EMT. MYOF is overexpressed in various cancers and has been implicated in tumor progression, metastasis, and poor prognosis. |
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| ln Vitro |
HJ445A exhibits potent antiproliferative activity against gastric cancer cells with IC50 values of 0.16 μM in MGC803 cells and 0.14 μM in MKN45 cells. These sub-micromolar IC50 values indicate that HJ445A is a highly potent inhibitor of gastric cancer cell proliferation. In addition to inhibiting proliferation, HJ445A also inhibits colony formation in MKN45 cells in a concentration-dependent manner, suggesting that it can suppress the long-term clonogenic survival of cancer cells. The mechanism of action is related to MYOF inhibition: by binding to the MYOF-C2D domain, HJ445A disrupts MYOF-mediated signaling pathways that drive cancer cell proliferation and survival. HJ445A also hinders gastric cancer cell migration by reversing the epithelial-mesenchymal transition (EMT) process, a key step in cancer metastasis.
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| ln Vivo |
HJ445A demonstrates superior antitumor effects in vivo. In preclinical models of gastric cancer, HJ445A effectively inhibits tumor growth and progression. The compound's excellent water solubility (approximately 170-fold improvement compared to compound 6y) facilitates in vivo administration and ensures adequate bioavailability. HJ445A hinders gastric cancer cell migration by reversing the EMT process, which is a critical mechanism by which cancer cells acquire invasive and metastatic properties. By inhibiting MYOF and reversing EMT, HJ445A not only suppresses primary tumor growth but also reduces the metastatic potential of cancer cells. These in vivo efficacy data support the potential of HJ445A as a therapeutic agent for gastric cancer and other MYOF-overexpressing tumors.
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| Enzyme Assay |
Surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) assays are used to measure the binding affinity of HJ445A to the recombinant MYOF-C2D domain. In SPR, the recombinant MYOF-C2D protein is immobilized on a sensor chip, and varying concentrations of HJ445A are injected over the chip surface. The binding response is measured in real-time as resonance units, and association (ka) and dissociation (kd) rate constants are determined from the sensorgrams. The equilibrium dissociation constant (KD) is calculated as kd/ka, yielding the KD value of 0.17 μM for HJ445A binding to MYOF-C2D. In ITC, the binding affinity is determined by measuring the heat released or absorbed upon binding of HJ445A to the MYOF-C2D domain, providing both the binding stoichiometry and thermodynamic parameters (ΔH, ΔS).
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| Cell Assay |
Gastric cancer cell lines (MGC803 and MKN45) are treated with HJ445A at graded concentrations (typically ranging from nanomolar to micromolar) for 72 hours. Cell proliferation is measured using standard colorimetric assays such as CCK-8 (Cell Counting Kit-8) or MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). The absorbance is read at 450 nm (CCK-8) or 540-570 nm (MTT), and IC50 values are calculated from dose-response curves. Colony formation assays are performed in soft agar or by plating cells at low density and allowing them to grow for 10-14 days to assess long-term anti-proliferative effects. Colonies are stained (e.g., with crystal violet) and counted to determine the inhibition of clonogenic survival. Additional assays may include assessment of cell migration (wound healing or transwell assays) and EMT markers (E-cadherin, N-cadherin, vimentin) by Western blotting.
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| Animal Protocol |
Mouse xenograft models bearing gastric cancer tumors (MGC803 or MKN45 cells implanted subcutaneously) are administered HJ445A via oral or intraperitoneal routes at various dosages. Tumor volume is measured regularly using calipers (length × width²/2), and body weight is monitored to assess tolerability. Treatment is typically continued for 2-4 weeks or until tumors reach a predetermined size. At study endpoint, tumors are collected for histopathological analysis (e.g., H&E staining) and biomarker evaluation (e.g., immunohistochemistry for proliferation markers like Ki-67, EMT markers, and apoptosis markers like cleaved caspase-3) to confirm in vivo antitumor efficacy and target engagement. The superior water solubility of HJ445A facilitates formulation and ensures consistent drug exposure in vivo.
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| ADME/Pharmacokinetics |
Expected to have favorable oral bioavailability due to significantly improved water solubility (~170-fold enhancement compared to compound 6y). Good aqueous solubility is a key determinant of oral absorption and bioavailability, as it facilitates dissolution in the gastrointestinal tract and subsequent permeation across the intestinal epithelium. Metabolic stability is likely mediated via hepatic CYP450 enzymes with moderate clearance. Standard pharmacokinetic parameters (Cmax, Tmax, AUC, t1/2, clearance, volume of distribution) are typically determined in rodents following both intravenous and oral administration at relevant dose levels. Formulation in suitable vehicles (e.g., DMSO/PEG400/saline mixtures or aqueous buffers) takes advantage of the compound's excellent solubility. Detailed PK data specific to HJ445A are not extensively reported in the available literature.
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| Toxicity/Toxicokinetics |
Predicted to have an acceptable safety profile in preclinical species at therapeutic doses based on selective MYOF targeting. MYOF is primarily expressed in muscle and certain cancer cells, and its normal function is involved in membrane trafficking and repair. Inhibition of MYOF may affect normal tissue homeostasis, particularly in tissues with high MYOF expression. Standard toxicology studies in rodents are required to assess acute and repeat-dose toxicity, identify potential target-related effects, and establish the therapeutic window. As a research compound, detailed toxicology data are not publicly available. The compound is intended for research use only and not for human therapeutic applications.
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| References | |
| Additional Infomation |
HJ445A is a research-grade small-molecule MYOF inhibitor for cancer research. Molecular formula: C24H27N7O2, molecular weight: 445.52. Soluble in DMSO (100 mg/mL). It is typically stored at -20°C for long-term stability. Synonyms: HJ445-A, H-J445-A. The compound demonstrates excellent water solubility (~170-fold improvement compared to compound 6y), which is a significant advantage for in vivo studies. HJ445A is used in studies investigating the role of myoferlin in cancer pathogenesis, particularly in gastric cancer, and represents a promising tool for validating MYOF as a therapeutic target. For research use only, not for human therapeutic use.
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| CAS # |
3032441-59-8
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| Appearance |
White to off-white solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO :~100 mg/mL (~224.46 mM; with sonication)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (5.61 mM) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one),clear solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared,you can add 100 μL of 25.0 mg/mL clear DMSO stock solution and add it to 400 μL PEG300 and mix well. Then add 50 μL Tween-80 to the above system and mix well. Then continue to add 450 μL of physiological saline to make up to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (5.61 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one),clear solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared,you can add 100 μL of 25.0 mg/mL clear DMSO stock solution and add it to 900 μL of 20% SBE-β-CD saline solution and mix well. 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. View More
Solubility in Formulation 3: 2.5 mg/mL (5.61 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one),clear solution; with ultrasonication. |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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