Bulgecin A(Blg-A) is a novel binuclear metallo-beta-lactamases and Lytic transglycosolase inhibitor.
Physicochemical Properties
| Molecular Formula | C16H29N3O14S2 |
| Molecular Weight | 551.54 |
| Exact Mass | 551.109 |
| Elemental Analysis | C, 34.84; H, 5.30; N, 7.62; O, 40.61; S, 11.63 |
| CAS # | 92953-54-3 |
| Related CAS # | 92953-54-3 |
| PubChem CID | 2470 |
| Appearance | Typically exists as solid at room temperature |
| Density | 1.7±0.1 g/cm3 |
| Index of Refraction | 1.630 |
| LogP | -5.78 |
| Hydrogen Bond Donor Count | 8 |
| Hydrogen Bond Acceptor Count | 15 |
| Rotatable Bond Count | 11 |
| Heavy Atom Count | 35 |
| Complexity | 950 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | CC(=O)NC1C(C(C(OC1OC2CC(NC2CO)C(=O)NCCS(=O)(=O)O)CO)OS(=O)(=O)O)O |
| InChi Key | RPNZWZDLNYCCIG-HMMVDTEZSA-N |
| InChi Code | InChI=1S/C16H29N3O14S2/c1-7(22)18-12-13(23)14(33-35(28,29)30)11(6-21)32-16(12)31-10-4-8(19-9(10)5-20)15(24)17-2-3-34(25,26)27/h8-14,16,19-21,23H,2-6H2,1H3,(H,17,24)(H,18,22)(H,25,26,27)(H,28,29,30)/t8-,9+,10-,11+,12+,13+,14+,16+/m0/s1 |
| Chemical Name | 2-[[(2S,4S,5R)-4-[(2R,3R,4R,5S,6R)-3-Acetamido-4-hydroxy-6-(hydroxymethyl)-5-sulfooxyoxan-2-yl]oxy-5-(hydroxymethyl)pyrrolidine-2-carbonyl]amino]ethanesulfonic acid |
| Synonyms | BlgA; Blg-ABlg; ABLG; BUL BulgecinA; Bulgecin-A; bulgecin a; 92953-54-3; DTXSID90869108; CID 2470; Bulgecin A |
| 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 | Metallo-beta-lactamases; Lytic transglycosolase |
| ln Vitro | Genetic screening of Pseudomonas aeruginosa (PSDA) and Acinetobacter baumannii (ACB) reveals genes that confer increased susceptibility to β-lactams when disrupted, suggesting novel drug targets. One such target is lytic transglycosylase. Bulgecin A (BlgA) is a natural product of Pseudomonas mesoacidophila and a lytic transglycosolase inhibitor that works synergistically with β-lactams targeting PBP3 for Enterobacteriaceae. BlgA also weakly inhibits di-Zn2+ metallo-β-lactamases like L1 of Stenotrophomonas maltophilia. We hypothesized that because of its unique mechanism of action, BlgA could restore susceptibility to carbapenems in carbapenem-resistant PSDA (CR-PSDA) and carbapenem-resistant ACB, as well as ACB resistant to sulbactam. A BlgA-containing extract was prepared using a previously published protocol. CR-PSDA clinical isolates demonstrating a variety of carbapenem resistance mechanisms (VIM-2 carbapenemases, efflux mechanisms, and AmpC producer expression) were characterized with agar dilution minimum inhibitory concentration (MIC) testing and polymerase chain reaction. Growth curves using these strains were prepared using meropenem, BlgA extract, and meropenem plus BlgA extract. A concentrated Blg A extract combined with low concentrations of meropenem, was able to inhibit the growth of clinical strains of CR-PSDA for strains that had meropenem MICs ≥8 mg/L by agar dilution, and a clinical strain of an OXA-24 producing ACB that had a meropenem MIC >32 mg/L and intermediate ampicillin/sulbactam susceptibility. Similar experiments were conducted on a TEM-1 producing ACB strain resistant to sulbactam. BlgA with ampicillin/sulbactam inhibited the growth of this organism. As in Enterobacteriaceae, BlgA appears to restore the efficacy of meropenem in suppressing the growth of CR-PSDA and carbapenem-resistant ACB strains with a variety of common carbapenem resistance mechanisms. BlgA extract also inhibits VIM-2 β-lactamase in vitro. BlgA may prove to be an exciting adjunctive compound to extend the life of carbapenems against these vexing pathogens[1]. |
| Enzyme Assay | Growth curves were constructed for control strain E. coli MC1016 according to the method of Heidrich et al33 as follows: 1 µL of a 1:10 dilution of an overnight culture was added to 95 µL of super optimal broth (SOB) medium (~105 colony forming units [cfu]/mL). An initial OD600 nm was obtained using an enzyme-linked immunosorbent assay plate reader, and then the sample was allowed to grow for 100 minutes at 37°C. After 100 minutes incubation, another OD600 nm reading was obtained and then either saline (null), BlgA extract alone (final v:v [BlgA] =10%), aztreonam (final concentration 0.01 mg/L), or 10% (v/v) BlgA extract with 0.01 mg/L aztreonam was added to the well (in 100 µL total volume) and growth was further monitored at OD600 nm at various time points. Growth curves for the clinical bacterial strains were obtained in a similar manner using appropriate partner antibiotics for the particular resistance phenotype and adjusting the inoculum or antibiotic concentration to allow growth of the organism. For the multidrug resistant (MDR) ACB clinical strain, UH83, 1 µL of a 1:10 dilution of an overnight culture was used with meropenem (0.02 mg/L) ± 10% (v/v) BlgA extract in 100 µL total volume. For the sulbactam-resistant ACB clinical strain UH10, 1 µL of a 1:10 dilution of an overnight culture was used with ampicillin–sulbactam (0.03/0.015 mg/L) ± 10% (v/v) BlgA extract in 100 µL total volume. UH83 produces OXA-24 carbapenemase and is resistant to carbapenems (MIC ≥32 mg/L), cephalosporins, and intermediate to sulbactam. UH10 has been completely sequenced and produces TEM-1 β-lactamase as the basis of its sulbactam resistance. The PSDA strains included CL231, a PDC-5 AmpC hyperproducing strain isolated from the sputum of a patient at the Cleveland Clinic in the mid-2000s; a VIM-2 producing strain; and a PSDA urinary tract isolate, resistant to meropenem/susceptible to imipenem that effluxes meropenem but has no other identifiable carbapenem resistance mechanism (this study). For PSDA strains, growth assays were performed (mentioned earlier in this section), by adding 1 µL of an overnight culture (cx) of a 1:1, 1:2, or 1:10 dilution thereof to 95 µL volumes of SOB medium. After 100 minutes incubation, either saline (null), BlgA extract alone (final v:v [BlgA] =10%), meropenem (final 0.03 mg/L), or BlgA 10% + meropenem 0.03 mg/L was added to the well and growth was monitored at OD600 nm using an enzyme-linked immunosorbent assay plate reader at various time points.[1] |
| References | [1]. Bulgecin A as a β-lactam enhancer for carbapenem-resistant Pseudomonas aeruginosa and carbapenem-resistant Acinetobacter baumannii clinical isolates containing various resistance mechanisms. Drug Des Devel Ther. 2016 Sep 20;10:3013-3020. |
Solubility Data
| 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 |
| 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 | 1.8131 mL | 9.0655 mL | 18.1311 mL | |
| 5 mM | 0.3626 mL | 1.8131 mL | 3.6262 mL | |
| 10 mM | 0.1813 mL | 0.9066 mL | 1.8131 mL |