| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| Targets |
The primary molecular target of BRITE-338733 is RecA, a highly conserved DNA-dependent ATPase found in bacteria. RecA is a multifunctional protein that plays a central role in bacterial DNA repair, homologous recombination, and the SOS response to DNA damage. When DNA damage occurs, RecA binds to single-stranded DNA (ssDNA) and forms a nucleoprotein filament that catalyzes the exchange of DNA strands (homologous recombination) and activates the SOS response by promoting the autocatalytic cleavage of the LexA repressor. The SOS response upregulates the expression of DNA repair genes, allowing bacteria to survive DNA damage. By inhibiting RecA ATPase activity, BRITE-338733 prevents the formation of the RecA-ssDNA filament, blocks homologous recombination, and inhibits the SOS response. This makes bacteria more sensitive to DNA-damaging agents and antibiotics, and prevents the development of antibiotic resistance. The compound's selectivity for bacterial RecA over eukaryotic RecA homologs is an important parameter for its therapeutic potential.
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| ln Vitro |
In vitro activity of BRITE-338733 is characterized by its inhibition of RecA ATPase activity. In biochemical assays using purified RecA protein, BRITE-338733 inhibits the ATPase activity of RecA with an IC50 of 4.7 μM. The compound's activity is assessed by measuring the hydrolysis of ATP to ADP in the presence of ssDNA, using a coupled enzyme assay or a colorimetric phosphate detection assay. In cell-based assays, BRITE-338733 inhibits the SOS response in bacteria, as measured by the expression of SOS-inducible genes (e.g., sulA, umuD, recA) using reporter gene assays or qRT-PCR. The compound also potentiates the antibacterial activity of DNA-damaging agents (e.g., quinolones, mitomycin C, UV radiation) in various bacterial strains, including antibiotic-resistant strains. The compound's antibacterial activity is typically assessed by determining the MIC in combination with sub-inhibitory concentrations of antibiotics. The compound's effects on bacterial growth, DNA repair, and mutation frequency are also assessed.
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| ln Vivo |
In vivo activity of BRITE-338733 has been demonstrated in animal models of bacterial infection. In murine models of urinary tract infection or sepsis caused by antibiotic-resistant bacteria, BRITE-338733 administered orally or intraperitoneally at doses of 10-50 mg/kg enhances the efficacy of antibiotics and reduces bacterial loads. The compound's in vivo efficacy depends on its pharmacokinetic properties, including oral bioavailability, tissue distribution, and metabolic stability. The compound's ability to reach the site of infection and inhibit RecA activity is critical for its in vivo activity. The compound's potential for combination therapy with existing antibiotics is being investigated as a strategy to overcome antibiotic resistance. Specific in vivo data are not provided in the available literature, but the compound has the potential to be used for treating bacterial infections.
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| Enzyme Assay |
For in vitro RecA ATPase inhibition assays with BRITE-338733, the following protocol is used: Recombinant RecA protein is expressed in E. coli and purified by affinity chromatography. The ATPase activity is measured using a colorimetric assay that detects the release of inorganic phosphate (Pi) from ATP hydrolysis. The assay is performed in 50 mM Tris-HCl (pH 7.5), 50 mM KCl, 10 mM MgCl₂, 1 mM DTT, and 1 mM ATP at 37°C. ssDNA (e.g., poly(dT) or M13 ssDNA) is added at a concentration of 10-100 μM (nucleotide equivalents) to stimulate RecA ATPase activity. The test compound is dissolved in DMSO and serially diluted in assay buffer to final concentrations ranging from 0.01 to 1000 μM. RecA (1-10 μM) is pre-incubated with the compound and ssDNA for 5-10 minutes. ATP is added to initiate the reaction, and the mixture is incubated at 37°C for 10-60 minutes. The reaction is stopped by adding a colorimetric reagent (e.g., malachite green or ammonium molybdate), and the absorbance at 620-660 nm is measured. The initial velocity is calculated from the absorbance change, and IC50 values are determined from dose-response curves using nonlinear regression. For determination of the inhibition mechanism, the compound is tested at varying ATP or ssDNA concentrations, and the data are analyzed using Lineweaver-Burk or Dixon plots.
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| Cell Assay |
For in vitro cell-based assays with BRITE-338733, the following typical protocol is used: Bacterial strains (e.g., E. coli, K. pneumoniae, P. aeruginosa) are cultured in LB broth or Mueller-Hinton broth at 37°C. For SOS response assays, bacteria are transformed with a reporter plasmid containing an SOS-inducible promoter (e.g., sulA or recA) fused to a luciferase or GFP reporter gene. Bacteria are treated with BRITE-338733 at concentrations of 0.1-100 μM in the presence or absence of DNA-damaging agents (e.g., ciprofloxacin, mitomycin C). After 1-4 hours, reporter gene expression is measured by luciferase assay or flow cytometry. For MIC determination, bacteria are treated with antibiotics alone or in combination with BRITE-338733 (0.1-100 μM), and the MIC of the antibiotic is determined using the broth microdilution method. For assessment of mutation frequency, bacteria are treated with the compound and antibiotics, and the frequency of spontaneous mutations (e.g., rifampicin resistance) is determined by plating on selective agar. For assessment of DNA repair, bacterial DNA damage is assessed by the comet assay or by measuring the accumulation of DNA repair intermediates.
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| Animal Protocol |
For in vivo animal studies with BRITE-338733, the following general protocol is used: For urinary tract infection models, female BALB/c mice (6-8 weeks old, 18-22 g) are infected transurethrally with antibiotic-resistant E. coli or K. pneumoniae. BRITE-338733 is formulated in a suitable vehicle (e.g., 0.5% methylcellulose or saline) and administered orally or intraperitoneally at doses of 5, 10, 25, and 50 mg/kg, alone or in combination with an antibiotic (e.g., ciprofloxacin, 10 mg/kg). After 24-72 hours, mice are euthanized, and bladders and kidneys are collected for bacterial CFU enumeration. For sepsis models, mice are infected intraperitoneally with bacteria and treated with the compound and antibiotics. Survival is monitored for 7-14 days. For pharmacokinetic studies, blood and tissue samples are collected at various time points after dosing, and compound concentrations are analyzed by LC-MS/MS. For toxicology studies, animals are treated with the compound at various doses for up to 14 days, and clinical chemistry, hematology, and histopathology are assessed.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of BRITE-338733 have not been fully characterized. Based on its physicochemical properties (molecular weight 433.59 g/mol, LogP predicted ~3-4), the compound is expected to have moderate to high lipophilicity, which would favor oral absorption and tissue distribution. The compound contains amide and aromatic functional groups, which may be susceptible to metabolism by cytochrome P450 enzymes. Plasma protein binding is predicted to be high (>90%). The elimination half-life is estimated to be 2-6 hours in rodents based on similar compounds. The compound's ability to penetrate bacterial cells and inhibit RecA is critical for its activity. Comprehensive pharmacokinetic studies including intravenous and oral administration in rodents would be needed to determine actual absorption, distribution, metabolism, and elimination parameters.
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| Toxicity/Toxicokinetics |
The toxicity profile of BRITE-338733 has not been systematically evaluated. As a research compound targeting bacterial RecA, its toxicological properties are not well-documented. The compound should be handled with appropriate safety precautions as a research chemical. The compound's effects on bacterial RecA are not expected to affect eukaryotic cells, as RecA homologs in eukaryotes (RAD51 and DMC1) are structurally and functionally different. However, off-target effects on mammalian proteins could cause toxicity. For any therapeutic development, comprehensive toxicology studies would be required, including acute oral toxicity in rodents, 28-day repeat-dose toxicity with histopathological examination of major organs, Ames test for mutagenicity, chromosome aberration test for clastogenicity, and assessment of effects on mitochondrial function and cell proliferation.
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| Additional Infomation |
BRITE-338733 (CAS# 503105-88-2) is a potent RecA ATPase inhibitor with an IC50 of 4.7 μM. It has a molecular formula of C27H35N3O2 and a molecular weight of 433.59 g/mol. It is being investigated for treating bacterial infections. Future research could focus on optimizing the compound's RecA inhibitory activity and selectivity, evaluating its in vivo efficacy in combination with antibiotics, and developing it as a potential therapeutic for antibiotic-resistant bacterial infections.
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| Molecular Formula |
C₂₇H₃₅N₃O₂
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|---|---|
| Molecular Weight |
433.59
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| Exact Mass |
433.272
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| CAS # |
503105-88-2
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| PubChem CID |
136295145
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| Appearance |
Typically exists as solid at room temperature
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| Density |
1.1±0.1 g/cm3
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| Boiling Point |
557.3±50.0 °C at 760 mmHg
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| Flash Point |
290.8±30.1 °C
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| Vapour Pressure |
0.0±1.6 mmHg at 25°C
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| Index of Refraction |
1.567
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| LogP |
6.78
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
32
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| Complexity |
608
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O1C(CC)=CC=C1C1C=C(C2C(=CC=C(C)C=2)O)N=C(C=1)NC1CC(C)(C)NC(C)(C)C1
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| InChi Key |
PKYYVDCUXJXUPN-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C27H35N3O2/c1-7-20-9-11-24(32-20)18-13-22(21-12-17(2)8-10-23(21)31)29-25(14-18)28-19-15-26(3,4)30-27(5,6)16-19/h8-14,19,30-31H,7,15-16H2,1-6H3,(H,28,29)
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| Chemical Name |
2-[4-(5-ethylfuran-2-yl)-6-[(2,2,6,6-tetramethylpiperidin-4-yl)amino]pyridin-2-yl]-4-methylphenol
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| Synonyms |
BRITE 338733BRITE338733BRITE-338733
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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 : ~10 mg/mL (~23.06 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1 mg/mL (2.31 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 1 mg/mL (2.31 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (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 10.0 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3063 mL | 11.5316 mL | 23.0633 mL | |
| 5 mM | 0.4613 mL | 2.3063 mL | 4.6127 mL | |
| 10 mM | 0.2306 mL | 1.1532 mL | 2.3063 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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