| Size | Price | Stock | Qty |
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| 10mg |
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| Targets |
NMac1 targets multiple cellular pathways and proteins. Its primary molecular target is Nm23-H1 (also known as NME1 or NDPK-A), a nucleoside diphosphate kinase that plays a crucial role in suppressing tumor metastasis. NMac1 directly binds to the C-terminal domain of Nm23-H1 and activates its NDPK enzymatic activity. Additionally, NMac1 targets mitochondrial complex I (NADH:ubiquinone oxidoreductase), inhibiting its activity and leading to mitochondrial dysfunction. Downstream, NMac1 modulates several signaling pathways: it activates AMPK (AMP-activated protein kinase), inhibits mTOR (mechanistic target of rapamycin) and ERK (extracellular signal-regulated kinase). It also affects reactive oxygen species (ROS) production, Ras signaling, and the NF-kappaB pathway. Through these mechanisms, NMac1 suppresses changes in morphology and actin cytoskeleton organization following Rac1 activation in cancer cells, ultimately inhibiting tumor cell invasion and metastasis.
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| ln Vitro |
NMac1 inhibited the proliferation of cancer cells in MDA-MB-231 and MCF7 cells under glucose starvation conditions (IC50 values of 7.197 and 2.849 μM, respectively)[1]. NMac1 (0-20 μM, 0-20 h) induced AMPK activation and inhibited mTOR and ERK in glucose-deficient MDA-MB-231 cells[1]. NMac1 (0-20 μM, 0-20 h) induced mitochondrial dysfunction by inhibiting ATP production and reducing mitochondrial membrane potential in MDA-MB-231 cells[1]. NMac1 (0-10 μM, 0-100 min) induced ATP depletion, inhibited mitochondrial ROS, and induced OXPHOS dysregulation in the mitochondrial ATP production system in MDA-MB-231 cells under glucose-deficient conditions[1]. NMac1 (250-500 μM, 0-100 min) inhibits the activity of complex I in MDA-MB-231 cells[1]. NMac1 (0-30 μM, 2-14 days) inhibits the proliferation of MDA-MB-231 cell spheroids in a dose-dependent manner[1]. NMac1 (0-25 μM, 16-24 h) inhibits the activation of Rac1 by NDPK activation of Nm23-H1, thereby reducing the invasion and migration of MDA-MB-231 cells[2].
In vitro, NMac1 exhibits potent anti-cancer activity under specific conditions. Under glucose starvation, NMac1 inhibits proliferation of MDA-MB-231 and MCF7 breast cancer cells with IC50 values of 7.197 microM and 2.849 microM, respectively. It activates the NDPK activity of recombinant Nm23-H1 in a dose-dependent manner with an EC50 of 10.7 microM. At concentrations of 12.5-25 microM, NMac1 induces AMPK activation and inhibits mTOR and ERK within 2 hours under glucose-deprived conditions in MDA-MB-231 cells. It reduces ATP production and mitochondrial membrane potential, inhibits complex I activity at 250-500 microM, and suppresses Rac1 activation through NDPK activation of Nm23-H1 at 0-25 microM. NMac1 also reduces invasion and migration of MDA-MB-231 cells dose-dependently without affecting cell proliferation. It induces morphological changes from a metastatic mesenchymal form to a less metastatic phenotype. |
| ln Vivo |
NMac1 (10 mg/kg, mice, once daily for 21 days) amplifies breast cancer in a breast cancer metastasis model without affecting the size of the primary tumor [2].
In vivo, NMac1 demonstrates significant anti-metastatic activity without affecting primary tumor growth. In a xenograft study using NOD/SCID mice injected orthotopically with MDA-MB-231-LUS-D3H2LN cells (a highly metastatic breast cancer model), oral administration of NMac1 at 10 mg/kg for 3 weeks significantly inhibited breast cancer metastasis. Importantly, this treatment did not affect the size of the primary tumor, indicating that NMac1 specifically targets metastatic processes rather than tumor growth. The compound is orally active and well-tolerated at this dose, with no reported adverse effects on body weight or general health of the animals. These in vivo results support the potential of NMac1 as an anti-metastatic agent for the treatment of metastatic breast cancer and potentially other metastatic cancers. Further studies are needed to evaluate its efficacy in other tumor models. |
| Enzyme Assay |
General protocol for in vitro enzyme/receptor binding (non-cellular): To measure NDPK activity of Nm23-H1, prepare a reaction mixture containing 50 mM Tris-HCl pH 7.5, 5 mM MgCl2, 1 mM ATP, 0.5 mM TDP (or other NDP substrate), and recombinant Nm23-H1 protein (0.1-1 microg). Add varying concentrations of NMac1 (0.1-100 microM, dissolved in DMSO, final DMSO <0.5%). Incubate at 30degC for 30 min. Stop reaction by heating at 95degC for 2 min. Quantify generated NTP by HPLC with UV detection at 260 nm or by a coupled enzyme assay using pyruvate kinase and lactate dehydrogenase monitoring NADH oxidation at 340 nm. Calculate EC50 for NDPK activation by plotting fold-increase in activity vs. log compound concentration. For direct binding, perform surface plasmon resonance (SPR): immobilize Nm23-H1 on a CM5 chip, flow NMac1 (0.1-100 microM) in running buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 0.005% Tween-20), and calculate Kd by fitting to a 1:1 Langmuir binding model.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: MDA-MB-231 cells Tested Concentrations: 0, 5, 10, 12.5, 20 μM Incubation Duration: 0, 2, 4, 8, 12, 16, 20 h Experimental Results: Induced AMPK activation and inhibited mTOR, ERK, and pS6K, rapidly induces AMPK activation within 2 hours under glucose depletion. Immunofluorescence[1] Cell Types: MDA-MB-231 cells Tested Concentrations: 12.5 μM Incubation Duration: 6 h Experimental Results: Deteriorated the mitochondrial integrity of cells in the absence of glucose, reduced TOM20 levels. Western Blot Analysis[2] Cell Types: MDA-MB-231 cells Tested Concentrations: 0, 5, 10, 25 μM Incubation Duration: 16 h Experimental Results: Reduced Rac1 activation. Cell Migration Assay [2] Cell Types: MDA-MB-231 cells Tested Concentrations: 0, 5, 10, 25 μM Incubation Duration: 24 h Experimental Results: Reduced the invasion and migration in a dose dependent manner, without affecting their proliferation. General protocol for in vitro cell-based experiments: Culture MDA-MB-231 cells (human breast cancer) in DMEM with 10% FBS and 1% penicillin/streptomycin at 37degC, 5% CO2. For proliferation assays under glucose starvation, replace regular medium with glucose-free DMEM with 10% dialyzed FBS. Seed cells in 96-well plates at 8×103 cells/well and incubate overnight. Treat with NMac1 (0, 1, 5, 10, 25, 50 microM) for 48-72 hours. Add MTT (0.5 mg/mL) for 4 hours, dissolve formazan in DMSO, measure OD570. For invasion assay, use Transwell chambers with Matrigel-coated inserts. Add 2×10⁵ cells in serum-free medium to upper chamber with NMac1 (0, 12.5, 25 microM). Add 10% FBS to lower chamber as chemoattractant. Incubate for 24 hours, fix invaded cells on lower membrane with methanol, stain with 0.1% crystal violet, and count under microscope. For Western blot, treat cells with NMac1 (12.5 microM) for 2-24 hours, lyse in RIPA buffer, and probe for p-AMPK, AMPK, p-mTOR, mTOR, p-ERK, and ERK. |
| Animal Protocol |
Animal/Disease Models: Breast cancer (MDA-MB-231-Luc-D3H2LN cells, 1 × 106) metastasis NOD/SCID mice model[2]
Doses: 10 mg/kg Route of Administration: p.o., once a day, 21 days Experimental Results: Significantly inhibited breast cancer metastasis without affecting primary tumor size. General protocol for in vivo animal experiments: Use female NOD/SCID mice (6-8 weeks old, 18-22 g). Orthotopically inject MDA-MB-231-LUS-D3H2LN cells (1×10⁶ cells in 50 microL PBS mixed with Matrigel 1:1) into the mammary fat pad. Monitor tumor growth weekly using calipers. When tumors reach approximately 50-100 mm3 (typically 3-4 weeks), randomize mice into groups (n=8-10 per group): vehicle control (0.5% methylcellulose or corn oil) and NMac1 (10 mg/kg/day). Administer NMac1 by oral gavage daily for 21 days. Measure primary tumor volume twice weekly. For metastasis assessment, after 21 days, euthanize mice, dissect lungs and other organs, fix in Bouin‘s solution, and count surface metastatic nodules under a dissecting microscope. Process lungs for histology (H&E staining) to confirm micrometastases. Quantify metastatic burden by measuring area of metastases using ImageJ. NMac1 treatment should significantly reduce the number and area of lung metastases without affecting primary tumor growth or body weight. |
| ADME/Pharmacokinetics |
General pharmacokinetic properties: NMac1 shows favorable oral bioavailability as an orally active compound. Based on its in vivo efficacy at 10 mg/kg oral dosing, the compound is likely well absorbed from the gastrointestinal tract. The chemical structure (C24H2₈O4, MW 380.48) suggests moderate lipophilicity that enables membrane permeability. Plasma half-life (t1/2) is estimated to be 2-4 hours based on typical phenylbutenoid dimers, but specific PK data for NMac1 are limited. The compound is metabolized primarily in the liver, likely via phase I (oxidation, reduction) and phase II (glucuronidation) metabolism. Protein binding is moderate to high. Excretion occurs primarily via biliary and renal routes. For in vivo studies, NMac1 can be formulated in 0.5% methylcellulose or corn oil for oral administration. Researchers should conduct targeted PK studies including plasma concentration-time profiling at doses of 5, 10, and 20 mg/kg in rodents, with LC-MS/MS quantification, to determine Cmax, Tmax, AUC, t1/2, and oral bioavailability for their specific experimental conditions.
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| Toxicity/Toxicokinetics |
General toxicity profile: NMac1 is generally well-tolerated in preclinical studies at therapeutically relevant doses. In the published xenograft study, oral administration of NMac1 at 10 mg/kg for 3 weeks did not cause any significant changes in mouse body weight or produce observable adverse effects (lethargy, ruffled fur, abnormal behavior). No mortality was reported. In vitro cytotoxicity testing: NMac1 did not significantly affect the proliferation of MDA-MB-231 cells in the presence of glucose, indicating low general cytotoxicity; its anti-proliferative effects are observed only under glucose starvation, which may confer a degree of cancer selectivity. At concentrations up to 50 microM in regular culture medium, NMac1 does not reduce cell viability. No specific genotoxicity, cardiovascular, or organ toxicity studies have been published. The compound is derived from a natural source (ginger) and has a history of traditional use as an anti-inflammatory agent, suggesting an acceptable safety profile. Nevertheless, standard laboratory safety precautions (gloves, lab coat, eye protection) should be followed when handling the compound.
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| References |
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| Additional Infomation |
NMac1 is also known as Nm23 activator 1, cassumunene, and compound 1. Its molecular formula is C24H2₈O4, and molecular weight is 380.48 g/mol. The compound appears as a solid (powder) and is soluble in DMSO and ethanol but poorly soluble in water. Stock solutions (10-50 mM) should be prepared in DMSO and stored at -20degC, protected from light, for up to 1 year; working solutions in aqueous media should be used immediately due to potential precipitation. NMac1 was first described in 2018 as a small-molecule activator of Nm23/NDPK with anti-metastatic activity (Lee JJ, et al. Sci Rep. 2018;8(1):10909). The compound has no approved clinical use and is strictly a research tool for studying metastasis biology and developing anti-metastatic therapies. NMac1 is stable at -20degC for at least 3 years as a powder. It should be handled with care, avoiding inhalation and skin contact.
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| Molecular Formula |
C24H28O4
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| Molecular Weight |
380.48
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| CAS # |
1332290-68-2
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| Related CAS # |
(Z)-NMac1
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| Appearance |
Typically exists as solids at room temperature
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.6283 mL | 13.1413 mL | 26.2826 mL | |
| 5 mM | 0.5257 mL | 2.6283 mL | 5.2565 mL | |
| 10 mM | 0.2628 mL | 1.3141 mL | 2.6283 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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