| Targets |
The primary target of (1S,2R)-bedaquiline is the mycobacterial ATP synthase, specifically subunit c (AtpE) of the F0F1-ATP synthase complex. Bedaquiline inhibits ATP synthase by binding to the oligomeric c-ring of the F0 domain, thereby disrupting ATP synthesis in Mycobacterium tuberculosis. This unique mechanism of action is distinct from other anti-tuberculosis drugs. The stereochemistry is critical for activity, with the (1S,2R) isomer being the active enantiomer and the (1R,2S) isomer being significantly less active.
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| ln Vitro |
In vitro, bedaquiline (the racemate from which this isomer is derived) exhibits potent antimycobacterial activity against Mycobacterium tuberculosis, including multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains. The MIC₉0 (minimum inhibitory concentration for 90% of strains) ranges from 0.015 to 0.12 microg/mL. The compound has a unique mode of action with no cross-resistance to existing anti-tuberculosis drugs. Against replicating and non-replicating M. tuberculosis, bedaquiline shows bactericidal activity. The (1S,2R) isomer is responsible for the majority of the antimycobacterial activity.
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| ln Vivo |
In vivo, bedaquiline has demonstrated efficacy in animal models of tuberculosis. In mouse models of chronic TB infection, oral administration of bedaquiline (25-50 mg/kg) for 4-8 weeks significantly reduces lung and spleen bacterial burden compared to untreated controls. In combination with other anti-TB drugs (e.g., isoniazid, rifampicin, pyrazinamide), bedaquiline shows synergistic or additive activity. Bedaquiline is a cornerstone of MDR-TB treatment regimens in humans, significantly improving treatment outcomes and reducing mortality.
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| Enzyme Assay |
Non-cell-based biochemical assays for this class of compounds target mycobacterial ATP synthase. A standard protocol uses inverted membrane vesicles prepared from Mycobacterium smegmatis expressing M. tuberculosis F0F1-ATP synthase. ATP hydrolysis activity is measured by a coupled enzyme assay: vesicles (20-50 microg protein) are incubated with varying concentrations of test compound (1 nM to 10 microM) in assay buffer (50 mM MOPS, 5 mM MgCl2, pH 7.5) at 37degC. The reaction is initiated by adding 2 mM ATP, and NADH oxidation is monitored at 340 nm for 10-20 minutes. Alternatively, the activity of ATP synthesis can be measured by luciferase-based assays. IC₅0 values are calculated from dose-response curves.
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| Cell Assay |
Cell-based experiments for bedaquiline are conducted using Mycobacterium tuberculosis strains (e.g., H37Rv) cultured in Middlebrook 7H9 broth supplemented with OADC enrichment and 0.05% Tween 80. A standard broth microdilution assay according to CLSI guidelines: cultures are grown to mid-log phase and diluted to approximately 5 × 10⁵ CFU/mL. Serial two-fold dilutions of test compound (0.001-1 microg/mL) are prepared in 96-well plates. After 7-14 days of incubation at 37degC in 5% CO2, the MIC is determined as the lowest concentration that inhibits visible bacterial growth. For time-kill kinetics, samples are plated on 7H10 agar at various time points (0, 1, 3, 5, 7, 10, 14 days) to determine CFU reduction.
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| Animal Protocol |
In vivo animal experiments for bedaquiline are conducted in mouse models of tuberculosis. A standard protocol in BALB/c mice (6-8 weeks old) involves aerosol infection with M. tuberculosis H37Rv (~100 CFU/lung). Starting 2-4 weeks post-infection (when lung burden reaches plateau), bedaquiline is administered orally at doses of 12.5-50 mg/kg once daily for 4-8 weeks. Control groups receive vehicle or standard anti-TB drugs (isoniazid 25 mg/kg + rifampicin 10 mg/kg). Endpoints include lung and spleen CFU counts at sacrifice, histopathological scoring of lung tissue (H&E and acid-fast staining), survival rates, and body weight changes. Drug combination studies are conducted to assess synergistic activity.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for the (1S,2R) isomer are derived from the racemic bedaquiline, which is the clinically approved drug. Bedaquiline has a long terminal elimination half-life of approximately 5.5 months in humans due to slow release from tissue compartments. The time to maximum plasma concentration (Tmax) is approximately 4-6 hours after oral administration. Food increases the oral bioavailability of bedaquiline by approximately 2-fold. Bedaquiline is highly protein bound (>99.9%), primarily to albumin and alpha1-acid glycoprotein. It is metabolized by CYP3A4 to the N-monodesmethyl metabolite, which also has anti-TB activity. This isomer is typically stored at -20degC as a powder, protected from light and moisture. Solubility: soluble in DMSO (2-5 mg/mL) and organic solvents, but poorly soluble in water.
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| Toxicity/Toxicokinetics |
The toxicity profile of (1S,2R)-bedaquiline is derived from the racemic drug. Bedaquiline causes dose-dependent hepatotoxicity in both animal studies and humans, with elevated transaminases being the most common adverse event. In clinical trials, the most frequently reported adverse effects include nausea, arthralgia, headache, and increased amylase. Cardiotoxicity, specifically QTc prolongation (dose-dependent, mean increase of approximately 15-20 ms), is a notable adverse effect requiring ECG monitoring. Hepatotoxicity (Grade 3 or 4 ALT elevation) occurs in approximately 2-4% of patients. Bedaquiline carries a boxed warning for increased mortality in clinical trials. As a research chemical, standard safety precautions apply: handle in a fume hood with appropriate PPE.
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| Additional Infomation |
Additional information: The compound has an IUPAC name of (1S,2R)-1-(6-bromo-2-methoxyquinolin-3-yl)-4-(dimethylamino)-2-(naphthalen-1-yl)-1-phenylbutan-2-ol. It is a white to off-white solid with a purity of ≥98% from commercial suppliers. Synonyms include (1S,2R)-TMC207 and (1S,2R)-R207910. The compound is for research and analytical applications only, not for human or veterinary use. It is available from chemical suppliers as a drug intermediate and impurity reference standard for pharmaceutical quality control. For additional research, the user may refer to the certificate of analysis for batch-specific purity data.
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| Molecular Formula |
C32H31BRN2O2
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|---|---|
| Molecular Weight |
555.50
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| CAS # |
857086-93-2
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| Appearance |
Typically exists as solids at room temperature
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| Synonyms |
Bedaquiline Impurity 16 (Standard); (1S,2R)-1-(6-bromo-2-methoxyquinoline-3-yl)-4-(dimethylamino)-2-(naphth-1-yl)-1- (Standard)
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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) |
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
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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 | 1.8002 mL | 9.0009 mL | 18.0018 mL | |
| 5 mM | 0.3600 mL | 1.8002 mL | 3.6004 mL | |
| 10 mM | 0.1800 mL | 0.9001 mL | 1.8002 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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