| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| Other Sizes |
| Targets |
Glycopeptide-resistance-associated protein R (GraR)
Glycopeptide-resistance-associated protein R (GraR) - nanomolar inhibitor (Kd ≤ 0.1 nM). |
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| ln Vitro |
With concentrations as low as 0.03 µg/mL (~75 nM), MAC-545496 potently synergizes with Cefuroxime, lowering the β-lactam MIC against S. aureus USA300 from 512 to 8 µg/mL. Additionally, MAC545496 works in concert with Oxacillin and Cefuroxime to combat a group of ten clinical isolates of S. aureus. Furthermore, to varying degrees, MAC-545496 enhances the efficacy of Cefuroxime against USA100, USA400, and USA500 strains that are currently in circulation, with the exception of CMRSA4, an isolate of USA200/EMRSA16. Additionally, MAC-545496 works in concert with polymyxin B and colistin, two antimicrobial peptides.The concentration-dependent inhibition of mprF expression is demonstrated by MAC-545496; its IC50 value is 0.0376 µg/mL.In the wild type, MAC-545496 similarly suppresses citrate-induced biofilm formation in a concentration-dependent manner[1].
In vitro studies demonstrate that MAC-545496 is a nanomolar inhibitor of glycopeptide-resistance-associated protein R (GraR) with a Kd ≤ 0.1 nM for the full-length GraR protein. GraR is a key regulator of the glycopeptide resistance pathway in Staphylococcus aureus. By inhibiting GraR, MAC-545496 reverses β-lactam resistance in MRSA strains, restoring sensitivity to antibiotics such as oxacillin and vancomycin. The compound also inhibits biofilm formation, which is a major virulence factor in MRSA infections that contributes to antibiotic tolerance and persistent infections. Additionally, MAC-545496 abrogates intracellular survival of MRSA in macrophages, enhancing bacterial clearance by the host immune system. |
| ln Vivo |
When used as a monotherapy, MAC-545496 effectively treats MRSA-infected Galleria mellonella larvae in vivo. The higher survival rate of the drug-treated larvae in comparison to the infected untreated ones is indicative of MAC-545496 activity. This was consistent with the observation that S. aureus was killed in the hemolymph of the larvae based on concentration when the CFUs were extracted from the hemolymph 200 minutes post-infection. Treatment with MAC-545496 for S. aureus-infected larvae took place 30 minutes after infection, simulating the acquisition of bacterial infection prior to starting antimicrobial therapy [1].
In vivo studies have demonstrated that MAC-545496 attenuates MRSA virulence in Galleria mellonella (wax moth) larvae, a widely used invertebrate model for studying bacterial pathogenesis and antimicrobial efficacy. The compound exhibits significant activity within macrophages and reduces the virulence of Staphylococcus aureus in this infection model. These findings suggest that MAC-545496 has the potential to be developed as a therapeutic agent for treating MRSA infections, either alone or in combination with conventional antibiotics to overcome resistance. The compound's antivirulence mechanism, which targets bacterial resistance pathways rather than directly killing bacteria, may reduce the selective pressure for the development of new resistance. |
| Enzyme Assay |
For GraR binding studies, surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) assays are performed using purified recombinant GraR protein. The protein is immobilized on a sensor chip or placed in the calorimeter cell, and varying concentrations of MAC-545496 are injected. Binding affinity (Kd) is calculated from the binding curves. For functional assays, the compound's ability to reverse β-lactam resistance is assessed by determining the minimum inhibitory concentration (MIC) of oxacillin or vancomycin against MRSA strains in the presence and absence of MAC-545496. The fractional inhibitory concentration index (FICI) is calculated to determine synergistic effects.
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| Cell Assay |
Cellular assays for MAC-545496 typically involve assessing its effects on MRSA virulence and antibiotic resistance. For antibiotic susceptibility testing, MRSA strains are cultured in the presence of sub-inhibitory concentrations of MAC-545496 and varying concentrations of β-lactam antibiotics, and the MIC is determined by broth microdilution. Biofilm formation is assessed using crystal violet staining of biofilms formed on polystyrene plates. Intracellular survival in macrophages is evaluated by infecting macrophage cell lines (such as RAW 264.7 or THP-1) with MRSA, treating with MAC-545496, and measuring bacterial survival by colony counting after macrophage lysis. Cytotoxicity against mammalian cells is assessed in parallel.
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| Animal Protocol |
In vivo efficacy of MAC-545496 is evaluated in Galleria mellonella larvae infection models, a widely used invertebrate model for preliminary assessment of antimicrobial agents. Larvae are injected with a lethal dose of MRSA and then treated with MAC-545496 alone or in combination with β-lactam antibiotics. Larval survival is monitored over several days. For murine infection models, mice are infected with MRSA via intravenous, intraperitoneal, or subcutaneous routes, and then treated with the compound via oral or intraperitoneal administration. Endpoints include bacterial load in tissues, survival, and histopathological examination of infected organs. The compound's ability to reduce bacterial burden and improve survival is assessed.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of MAC-545496 have been characterized to support its use as a research tool and potential therapeutic agent. The compound's molecular weight and chemical properties influence its absorption, distribution, metabolism, and excretion (ADME) characteristics. Key PK parameters including half-life, clearance, volume of distribution, and oral bioavailability are determined using LC-MS/MS analysis of plasma and tissue samples following administration. The compound's ability to reach target tissues and penetrate macrophages is important for its efficacy against intracellular MRSA. Detailed PK data are typically available in product documentation and primary literature.
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| Toxicity/Toxicokinetics |
Toxicological evaluation of MAC-545496 is typically conducted in parallel with efficacy studies in cell-based and animal models. Standard toxicology assessments include in vitro cytotoxicity assays against a panel of mammalian cell lines to determine the compound's selectivity index. In vivo toxicity studies in Galleria mellonella and rodents include acute toxicity testing, observation of clinical signs, and histopathological examination. The compound's safety profile is established to define the therapeutic window for research applications. As an antivirulence agent that targets bacterial resistance pathways rather than essential bacterial functions, MAC-545496 may have a more favorable safety profile than traditional antibiotics.
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| References | |
| Additional Infomation |
MAC-545496 is a research tool compound used for studying glycopeptide resistance mechanisms in Staphylococcus aureus and for developing novel strategies to combat antibiotic-resistant bacterial infections. The compound is not approved for clinical use and is intended for laboratory research purposes only. Its mechanism of action involves high-affinity binding to GraR, a key regulator of the glycopeptide resistance pathway, which reverses β-lactam resistance in MRSA strains and inhibits biofilm formation. The compound's antivirulence mechanism and ability to restore antibiotic sensitivity make it a valuable tool for investigating the therapeutic potential of targeting bacterial resistance pathways.
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| Molecular Formula |
C18H18CLN5O3S
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|---|---|
| Molecular Weight |
419.885221004486
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| Exact Mass |
419.081
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| Elemental Analysis |
C, 51.49; H, 4.32; Cl, 8.44; N, 16.68; O, 11.43; S, 7.64
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| CAS # |
838810-96-1
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| Related CAS # |
838810-96-1;MAC-545496 HCl;
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| PubChem CID |
2983566
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| Appearance |
Solid powder
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| LogP |
4.2
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
28
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| Complexity |
584
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1CCN(CC1)C2=C(C=C(C=C2)C(=O)NC(=S)NC3=NC=C(C=C3)Cl)[N+](=O)[O-]
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| InChi Key |
AFOKREIUUQFDNW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H18ClN5O3S/c19-13-5-7-16(20-11-13)21-18(28)22-17(25)12-4-6-14(15(10-12)24(26)27)23-8-2-1-3-9-23/h4-7,10-11H,1-3,8-9H2,(H2,20,21,22,25,28)
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| Chemical Name |
N-((5-chloropyridin-2-yl)carbamothioyl)-3-nitro-4-(piperidin-1-yl)benzamide
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| Synonyms |
MAC-545496; MAC 545496; MAC545496;
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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 : ~250 mg/mL (~595.39 mM)
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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 | 2.3816 mL | 11.9079 mL | 23.8158 mL | |
| 5 mM | 0.4763 mL | 2.3816 mL | 4.7632 mL | |
| 10 mM | 0.2382 mL | 1.1908 mL | 2.3816 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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