Mtb F1FO-ATP synthase

TDR M. tuberculosis strains are inhibited in growth by bedaquiline, with MIC values ranging from 0.125 to 0.5 mg/L[2].
With MIC50 and MIC90 values of 0.03 and 16 mg/L, respectively, bedaquiline has the strongest activity against Mycobacterium avium among slowly growing mycobacteria (SGM). With MIC50 and MIC90 values of 0.13 and >16 mg/L, respectively, for both species, Mycobacterium abscessus subsp. abscessus (M. abscessus) and Mycobacterium abscessus subsp. massiliense (M. massiliense) appear to be more susceptible to bedaquiline than Mycobacterium fortuitum among rapidly growing mycobacteria (RGM). Moderate in vitro activity of bedaquiline against NTM species is also demonstrated[3].
In vitro activity of bedaquiline against Mycobacterium tuberculosis, including multidrug-resistant M tuberculosis, is very good[4].

BDQ was highly efficacious in a zebrafish model of M. abscessus infection. Remarkably, a very short period of treatment was sufficient to protect the infected larvae from M. abscessus-induced killing. This was corroborated with reduced numbers of abscesses and cords, considered to be major pathophysiological signs in infected zebrafish. [7]

Intracellular ATP quantification.
Intracellular ATP levels were determined using a 96-well flat-bottom plate, as described previously for M. tuberculosis. M. abscessus was exposed to BDQ or amikacin (negative control) and incubated for 180 min at 32°C. Twenty-five microliters of M. abscessus culture was mixed with an equal volume of the BacTiter-Glo reagent in 96-well flat-bottom white plates and incubated for 5 min in the darkness. Luminescence was detected using a BioTek Cytation 3 multimode reader, and the values obtained were plotted using GraphPad Prism 6 software.[7]

Drug susceptibility testing. [7]
The CLSI guidelines were followed to determine the MICs based on the broth microdilution method in CaMHB using an inoculum containing 5 × 106 CFU/ml in the exponential-growth phase. Bacteria (100 μl) were seeded in 96-well plates, and 2 μl of drug at its highest concentration was added to the first wells containing double the volume of bacterial suspension (200 μl). Twofold serial dilutions were then carried out, and incubation with drugs was performed at 30°C for 3 to 5 days. MICs were recorded by visual inspection and by absorbance at 560 nm to confirm visual recording. Experiments were done in triplicate on three independent occasions. Time-kill assay.[7]
Microtiter plates were set up as for MIC determination. Serial dilutions of the bacterial suspension were plated after 0, 24, 48, 72, and 96 h of exposure to different drug concentrations. CFU were enumerated after 4 days of incubation at 30°C.

Assessment of BDQ efficacy in infected zebrafish. [7]
Rough M. abscessus CIP104536T (ATCC 19977T) carrying pTEC27 (plasmid 30182; Addgene) and expressing the red fluorescent protein tdTomato was prepared and microinjected in zebrafish embryos, according to procedures described earlier. Briefly, mid-log-phase cultures of M. abscessus expressing tdTomato were centrifuged, washed, and resuspended in phosphate-buffered saline (PBS) supplemented with 0.05% Tween 80 (PBS-T). Bacterial suspensions were then homogenized through a 26-gauge needle and sonicated, and the remaining clumps were allowed to settle down for 5 to 10 min. Bacteria were concentrated to an optical density at 600 nm (OD600) of 1 in PBS-T and injected intravenously (≈2 to 5 nl containing 50 to 300 CFU) into the caudal vein in 30-h-postfertilization (hpf) embryos previously dechorionated and anesthetized. To follow infection kinetics and embryo survival, infected larvae were transferred into 24-well plates (2 embryos/well) and incubated at 28.5°C. The CFU numbers in the inoculum were determined by injection of 2 nl of the bacterial suspension in sterile PBS-T and plating on 7H10 with 500 μg/ml hygromycin.

[1]. Bedaquiline susceptibility test for totally drug-resistant tuberculosis Mycobacterium tuberculosis. J Microbiol. 2017 Apr 20.

[2]. TBAJ-876 displays Bedaquiline-like mycobactericidal potency without retaining the parental drug's uncoupler activity. Antimicrob Agents Chemother. 2019 Nov 11.

[3]. Bedaquiline: a novel diarylquinoline for multidrug-resistant tuberculosis. Ann Pharmacother. 2014 Jan;48(1):107-15.

[4]. In Vitro Activity of Bedaquiline against Nontuberculous Mycobacteria in China. Antimicrob Agents Chemother. 2017 Apr 24;61(5).

[5]. Ann Pharmacother.2014 Jan;48(1):107-15.

[6]. Antimicrob Agents Chemother.2017 Apr 24;61(5). pii: e02627-16.
[7]. Antimicrob Agents Chemother. 2017 Nov; 61(11): e01225-17.

Bedaquiline is a quinoline-based antimycobacterial drug used (as its fumarate salt) for the treatment of pulmonary multi-drug resistant tuberculosis by inhibition of ATP synthase, an enzyme essential for the replication of the mycobacteria. It has a role as an antitubercular agent and an ATP synthase inhibitor. It is a member of quinolines, a member of naphthalenes, an organobromine compound, an aromatic ether, a tertiary alcohol and a tertiary amino compound. It is a conjugate base of a bedaquiline(2+).

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I Bedaquiline is a bactericidal antimycobacterial drug belonging to the class of diarylquinoline. The quinolinic central heterocyclic nucleus with alcohol and amine side chains is responsible for bedaquiline-mediated antimycobacterial activity. Although it is closely related to fluoroquinolones, bedaquiline does not affect DNA gyrase; instead, bedaquiline inhibits the c subunit of ATP synthase responsible for synthesizing ATP. Consequently, bedaquiline can be used to treat mycobacterial infection, particularly tuberculosis (TB). Although the current standard of TB treatment of anti-TB drugs for 2 months, including 2 key drugs [isoniazid] and [rifampin], is highly effective, the emergence of multidrug-resistant TB (MDR-TB) to [isoniazid] and [rifampin] has substantially worsened patients outcome. Bedaquiline was approved by the FDA on December 28, 2012, to treat pulmonary MDR-TB, following favorable results in multiple pre-clinical and clinical studies. It is the first drug that was approved in the last 40 years by the FDA for TB unresponsive to current treatments on the market. Currently, bedaquiline is the last-line anti-TB drug and must only be used in an appropriate combination regimen.

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Bedaquiline is a diarylquinoline antimycobacterial drug used in combination with other antituberculosis medications in the treatment of multidrug resistant tuberculosis. The addition of bedaquiline to antituberculosis drug regimens has been linked to an increased rate of transient serum liver test abnormalities during treatment and to several instances of clinically apparent liver injury.

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Bedaquiline is an orally bioavailable diarylquinoline antimycobacterial agent, that can be used in the treatment of pulmonary multi-drug resistant tuberculosis (MDR-TB). Upon oral administration, bedaquiline specifically binds to subunit c of Mycobacterium tuberculosis (M. tuberculosis) adenosine 5'-triphosphate (ATP) synthase, thereby preventing ATP synthase activity. This inhibits ATP synthesis in M. tuberculosis, thereby blocking its energy metabolism and killing M. tuberculosis.

C32H31BRN2O2

555.5

554.16

C, 69.19; H, 5.62; Br, 14.38; N, 5.04; O, 5.76

843663-66-1

Bedaquiline fumarate;845533-86-0;(Rac)-Bedaquiline;654655-80-8;(Rac)-Bedaquiline-d6;2517573-53-2;Bedaquiline impurity 2-d6

5388906

White to yellow solid powder

1.3±0.1 g/cm3

702.7±60.0 °C at 760 mmHg

378.8±32.9 °C

0.0±2.3 mmHg at 25°C

1.666

7.2

45.6Ų

O[C@](CCN(C)C)(C1=C2C=CC=CC2=CC=C1)[C@@H](C3=CC4=CC(Br)=CC=C4N=C3OC)C5=CC=CC=C5

QUIJNHUBAXPXFS-XLJNKUFUSA-N

InChI=1S/C32H31BrN2O2/c1-35(2)19-18-32(36,28-15-9-13-22-10-7-8-14-26(22)28)30(23-11-5-4-6-12-23)27-21-24-20-25(33)16-17-29(24)34-31(27)37-3/h4-17,20-21,30,36H,18-19H2,1-3H3/t30-,32-/m1/s1

(1R,2S)-1-(6-Bromo-2-methoxy-3-quinolyl)-4-dimethylamino-2-(1-naphthyl)-1-phenyl-butan-2-ol

R207910; TMC207; R-207910; TMC-207; R 207910; TMC 207; Bedaquiline; Bedaquiline fumarate; trade name: Sirturo; AIDS-222089; bedaquilina;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : 12.5~33 mg/mL ( 22.50~59.4 mM )

Solubility in Formulation 1: ≥ 0.5 mg/mL (0.90 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 5.0 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.

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Solubility in Formulation 2: 0.5 mg/mL (0.90 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 5.0 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.

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Solubility in Formulation 3: ≥ 0.5 mg/mL (0.90 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 5.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly..

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Solubility in Formulation 4: Solubility in Formulation 1: ≥ 0.5 mg/mL (0.9 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 take 100 μL of 5 mg/mL DMSO stock solution and add to 400 μL of PEG300, mix well (clear solution); Then add 50 μL of Tween 80 to the above solution, mix well (clear solution); Finally, add 450 μL of saline to the above solution, mix well (clear solution).
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.

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Solubility in Formulation 2: 0.5 mg/mL (0.9 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 take 100 μL of 5 mg/mL DMSO stock solution and add to 900 μL of 20% SBE-β-CD in saline, mix well (clear solution).
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.

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Solubility in Formulation 3: 0.5 mg/mL (0.9 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 take 100 μL of 5 mg/mL DMSO stock solution and add to 900 μL of corn oil, mix well (clear solution).

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Solubility in Formulation 4: 1.67mg/ml (3.01mM) in 5% DMSO + 95% Corn oil (add these co-solvents sequentially from left to right, and one by one), clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)