BTZ043, also known as 8-Nitro-benzothiazinones (BTZs), is a potent inhibitor of decaprenyl-phosphoribose-epimerase (DprE1) with MIC values of of 2.3 nM and 9.2 nM for M. tuberculosis H37Rv and Mycobacterium smegmatis, respectively. It has potential to be used as a antimycobacterial agent that kill Mycobacterium tuberculosis by blocking arabinan synthesis. The inhibition of BTZ-resistant DprE1 followed the trend observed in the MIC measurements, with the C387G mutant being more resistant to inhibition by PyrBTZ01, PyrBTZ02, and BTZ043 (7- to 9-fold increases in IC50) than the C387S mutant (2.5- to 4-fold increases in IC50). Structure-activity relationship (SAR) studies revealed the 8-nitro group of the BTZ scaffold to be crucial for the mechanism of action, which involves formation of a semimercaptal bond with Cys387 in the active site of DprE1. BTZ043 presented favorable in vitro absorption-distribution-metabolism-excretion/toxicity (ADME/T) and in vivo pharmacokinetic profiles. BTZ043 did not show efficacy in a mouse model of acute tuberculosis, suggesting that BTZ-mediated killing through DprE1 inhibition requires a combination of both covalent bond formation and compound potency.
Physicochemical Properties
| Molecular Formula | C17H16F3N3O5S | |
| Molecular Weight | 431.39 | |
| Exact Mass | 431.076 | |
| Elemental Analysis | C, 47.33; H, 3.74; F, 13.21; N, 9.74; O, 18.54; S, 7.43 | |
| CAS # | 1161233-85-7 | |
| Related CAS # | BTZ043 Racemate;957217-65-1 | |
| PubChem CID | 42609849 | |
| Appearance | Solid powder | |
| Density | 1.7±0.1 g/cm3 | |
| Boiling Point | 547.6±60.0 °C at 760 mmHg | |
| Flash Point | 285.0±32.9 °C | |
| Vapour Pressure | 0.0±1.5 mmHg at 25°C | |
| Index of Refraction | 1.666 | |
| LogP | 2 | |
| Hydrogen Bond Donor Count | 0 | |
| Hydrogen Bond Acceptor Count | 9 | |
| Rotatable Bond Count | 1 | |
| Heavy Atom Count | 29 | |
| Complexity | 720 | |
| Defined Atom Stereocenter Count | 1 | |
| SMILES | S1C2C(=C([H])C(C(F)(F)F)=C([H])C=2C(N=C1N1C([H])([H])C([H])([H])C2(C([H])([H])C1([H])[H])OC([H])([H])[C@]([H])(C([H])([H])[H])O2)=O)[N+](=O)[O-] |
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| InChi Key | GTUIRORNXIOHQR-VIFPVBQESA-N | |
| InChi Code | InChI=1S/C17H16F3N3O5S/c1-9-8-27-16(28-9)2-4-22(5-3-16)15-21-14(24)11-6-10(17(18,19)20)7-12(23(25)26)13(11)29-15/h6-7,9H,2-5,8H2,1H3/t9-/m0/s1 | |
| Chemical Name | (S)-2-(2-methyl-1,4-dioxa-8-azaspiro[4.5]decan-8-yl)-8-nitro-6-(trifluoromethyl)-4H-benzo[e][1,3]thiazin-4-one | |
| Synonyms |
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| 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 |
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| 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 | DprE1 | ||
| ln Vitro | BTZ043, also known as 8-Nitro-benzothiazinones (BTZs), is a potent inhibitor of decaprenyl-phosphoribose-epimerase (DprE1) with MIC values of of 2.3 nM and 9.2 nM for M. tuberculosis H37Rv and Mycobacterium smegmatis, respectively. It can display nanomolar bactericidal activity against Mycobacterium tuberculosis in vitro. The inhibition of BTZ-resistant DprE1 followed the trend observed in the MIC measurements, with the C387G mutant being more resistant to inhibition by PyrBTZ01, PyrBTZ02, and BTZ043 (7- to 9-fold increases in IC50) than the C387S mutant (2.5- to 4-fold increases in IC50). Structure-activity relationship (SAR) studies revealed the 8-nitro group of the BTZ scaffold to be crucial for the mechanism of action, which involves formation of a semimercaptal bond with Cys387 in the active site of DprE1. BTZ043 presented favorable in vitro absorption-distribution-metabolism-excretion/toxicity (ADME/T) and in vivo pharmacokinetic profiles. BTZ043 did not show efficacy in a mouse model of acute tuberculosis, suggesting that BTZ-mediated killing through DprE1 inhibition requires a combination of both covalent bond formation and compound potency. | ||
| ln Vivo | At the used concentrations, four weeks of BTZ043 treatment reduces the bacterial burden in the spleen and lungs by one and two logs, respectively. Further findings imply that BTZ043's effectiveness is dependent on time as opposed to dose. After a month, acute (5 g/kg) and chronic (25 and 250 mg/kg) toxicology studies in uninfected mice demonstrate that there are no negative anatomical, behavioral, or physiological effects, even at the highest dose tested. | ||
| Enzyme Assay | BTZ043, also known as 8-Nitro-benzothiazinones (BTZs), is a potent inhibitor of decaprenyl-phosphoribose-epimerase (DprE1) with MIC values of of 2.3 nM and 9.2 nM for M. tuberculosis H37Rv and Mycobacterium smegmatis, respectively. | ||
| Cell Assay | The MIC of BTZ043 against M. tuberculosis H37Rv and Mycobacterium smegmatis are 1 ng/mL (2.3 nM) and 4 ng/mL (9.2 nM), respectively. The in vitro activity of BTZ043 against 30 Nocardia brasiliensis isolates is also tested. The MIC50 and MIC90 values for BTZ043 are 0.125 and 0.25 μg/mL. The MIC for N. carnea ATCC 6847 is 0.003μg/mL, for N. transvalensis ATCC 6865 is 0.003μg/mL, for N. brasiliensis NCTC10300 is 0.03 μg/mL, and for N. brasiliensis HUJEG-1 is 0.125μg/mL. The MIC value for M. tuberculosis H37Rv is 0.000976 μg/mL. The MIC value of BTZ-043 is >64 μg/mL for Escherichia coli ATCC 25922 and S. aureus ATCC 2921. | ||
| Animal Protocol |
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| References |
[1]. PLoS Negl Trop Dis.2015 Oct 16;9(10):e0004022 [2]. Antimicrob Agents Chemother.2015 Aug;59(8):4446-52. [3]. Science. 2009 May 8;324(5928):801-4. |
Solubility Data
| Solubility (In Vitro) | DMSO : 13.3~22 mg/mL ( 30.83~50.99 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (5.80 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. 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 to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume 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 (5.80 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension 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 to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (5.80 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. 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 to 900 μL of corn oil and mix evenly. Solubility in Formulation 4: 10% DMSO+40% PEG300+5% Tween-80+45% Saline: 2.5 mg/mL (5.80 mM) (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3181 mL | 11.5904 mL | 23.1809 mL | |
| 5 mM | 0.4636 mL | 2.3181 mL | 4.6362 mL | |
| 10 mM | 0.2318 mL | 1.1590 mL | 2.3181 mL |