MAC-545496 is novel and potent inhibitor of GraR (glycopeptide-resistance-associated protein R) with nanomolar activity. MAC-545496 could reverse β-lactam resistance in the community-acquired MRSA USA300 strain. MAC-545496 is an antivirulence agent alone; it attenuates MRSA virulence in Galleria mellonella larvae. MAC-545496 inhibits biofilm formation and abrogates intracellular survival in macrophages. Mechanistic characterization revealed MAC-545496 to be a nanomolar inhibitor of GraR, a regulator that responds to cell-envelope stress and is an important virulence factor and determinant of antibiotic resistance. MAC-545496 has value as a research tool to probe the GraXRS regulatory system and as an antibacterial lead series of a mechanism to combat drug-resistant Staphylococcal infections.
Physicochemical Properties
| Molecular Formula | C18H18CLN5O3S |
| Molecular Weight | 419.885221004486 |
| Exact Mass | 419.081 |
| Elemental Analysis | C, 51.49; H, 4.32; Cl, 8.44; N, 16.68; O, 11.43; S, 7.64 |
| CAS # | 838810-96-1 |
| Related CAS # | 838810-96-1;MAC-545496 HCl; |
| PubChem CID | 2983566 |
| Appearance | Solid powder |
| LogP | 4.2 |
| Hydrogen Bond Donor Count | 2 |
| Hydrogen Bond Acceptor Count | 6 |
| Rotatable Bond Count | 3 |
| Heavy Atom Count | 28 |
| Complexity | 584 |
| Defined Atom Stereocenter Count | 0 |
| InChi Key | AFOKREIUUQFDNW-UHFFFAOYSA-N |
| 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) |
| Chemical Name | N-((5-chloropyridin-2-yl)carbamothioyl)-3-nitro-4-(piperidin-1-yl)benzamide |
| Synonyms | MAC-545496; MAC 545496; MAC545496; |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets | Glycopeptide-resistance-associated protein R (GraR) |
| 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]. |
| 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]. |
| References |
[1]. Discovery of an antivirulence compound that reverses β-lactam resistance in MRSA. Nat Chem Biol. 2019 Nov 25. |
Solubility Data
| Solubility (In Vitro) | DMSO : ~250 mg/mL (~595.39 mM) |
| 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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently. (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 |