Mtb F1FO-ATP synthase

Bedaquiline 可抑制 TDR 结核分枝杆菌菌株的生长，其 MIC 值范围为 0.125 至 0.5 mg/L[1]。贝达喹啉的 MIC50 和 MIC90 值分别为 0.03 和 16 mg/L，在缓慢生长的分枝杆菌 (SGM) 中对鸟分枝杆菌具有最强的活性。在快速生长的分枝杆菌 (RGM) 中，脓肿分枝杆菌亚种的 MIC50 和 MIC90 值分别为 0.13 和 >16 mg/L，脓肿分枝杆菌 (M.脓肿) 和脓肿分枝杆菌亚种。马西林 (M. Massiliense) 似乎比偶发分枝杆菌更容易受到贝达喹啉的影响。贝达喹啉对 NTM 物种具有中等的体外活性[2]。贝达喹啉对结核分枝杆菌，包括耐多药结核分枝杆菌的体外活性非常好[3]。

BDQ在斑马鱼脓肿分枝杆菌感染模型中非常有效。值得注意的是，很短的处理时间就足以保护受感染的幼虫免受脓肿分枝杆菌诱导的杀死。脓肿和脊髓数量的减少证实了这一点，脓肿和脊髓被认为是受感染斑马鱼的主要病理生理体征。[7]

胞内ATP定量 使用96孔平板测定细胞内ATP水平，如先前对结核分枝杆菌所述。脓肿分枝杆菌暴露于BDQ或阿米卡星（阴性对照），并在32°C下孵育180分钟。将25微升脓肿分枝杆菌培养物与等体积的BacTiter-Glo试剂在96孔平底白色板中混合，并在黑暗中孵育5分钟。使用BioTek Cytation 3多模读取器检测发光，并使用GraphPad Prism 6软件绘制所获得的值。[7]

药物敏感性测试。[7] 根据CLSI指南，使用指数生长期含有5×106CFU/ml的接种物，基于CaMHB中肉汤微量稀释法测定MIC。将细菌（100μl）接种在96孔板中，并将最高浓度的2μl药物添加到含有两倍体积的细菌悬浮液（200μl）的第一孔中。然后进行两次系列稀释，并在30°C下与药物孵育3-5天。通过目视检查和在560nm处的吸光度来记录MIC，以确认目视记录。实验分三次独立进行，一式三份。 时间杀伤分析。[7] 建立微量滴定板用于MIC测定。在暴露于不同药物浓度0、24、48、72和96小时后，对细菌悬浮液进行连续稀释。在30°C下培养4天后计数CFU。

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.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Data from two women taking bedaquiline and one of their breastfed infants indicate that exposure of the infant to the drug via breastmilk is substantial, with one infant having a therapeutic serum level. The clinical consequences of this exposure are unknown. The drug could protect the infant from multidrug-resistant tuberculosis, or could result in adverse effects. If bedaquiline is required by the mother, it is not a reason to discontinue breastfeeding. Monitor breastfed infants for adverse reactions, such as inadequate weight gain, liver toxicity, nausea, arthralgia, headache, hemoptysis, and chest pain.
◉ Effects in Breastfed Infants
A woman who was co-infected with HIV and rifampin-resistant tuberculosis took bedaquiline (dosage not stated) as part of her antituberculosis regimen, which consisted of pyrazinamide and other unnamed drugs. At the 1-month follow-up, the infant was small and not gaining weight well, but the mother was nauseated from her medication regimen and had also lost weight. Six months later after completion of the mother’s therapy, her infant’s weight was increasing, following the normal trajectory of the growth chart, and reaching her developmental milestones.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

[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 fumarate is a fumarate salt prepared from equimolar amounts of bedaquiline and fumaric acid. It is used in combination therapy 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 contains a bedaquiline(2+).
Bedaquiline Fumarate is the fumarate salt form of bedaquiline, 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.
See also: Bedaquiline (has active moiety).

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Drug Indication
Sirturo is indicated for use as part of an appropriate combination regimen for pulmonary multidrug resistant tuberculosis (MDR TB) in adults and adolescent patients (12 years to less than 18 years of age and weighing at least 30 kg) when an effective treatment regimen cannot otherwise be composed for reasons of resistance or tolerability.  Consideration should be given to official guidance on the appropriate use of antibacterial agents.
Treatment of multi-drug-resistant tuberculosis

C36H35BRN2O6

671.59

670.167

C, 64.38; H, 5.25; Br, 11.90; N, 4.17; O, 14.29

845533-86-0

Bedaquiline;843663-66-1;(Rac)-Bedaquiline;654655-80-8

24812732

White to off-white solid powder

6.842

120.19

3

8

10

45

834

2

OC(/C=C/C(O)=O)=O.BrC1=CC=C(N=C(OC)C([C@H]([C@@](C2=CC=CC3=C2C=CC=C3)(O)CCN(C)C)C4=CC=CC=C4)=C5)C5=C1

ZLVSPMRFRHMMOY-WWCCMVHESA-N

InChI=1S/C32H31BrN2O2.C4H4O4/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;5-3(6)1-2-4(7)8/h4-17,20-21,30,36H,18-19H2,1-3H3;1-2H,(H,5,6)(H,7,8)/b;2-1+/t30-,32-;/m1./s1

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

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

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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

DMSO : ~100 mg/mL ( ~148.9 mM )
Ethanol : ~4 mg/mL

配方 1 中的溶解度: ≥ 2.75 mg/mL (4.09 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 27.5 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: 2.75 mg/mL (4.09 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 27.5mg/mL澄清的DMSO储备液加入到900μL 20％SBE-β-CD生理盐水中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.75 mg/mL (4.09 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 27.5 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: ≥ 2.75 mg/mL (4.09 mM) (饱和度未知) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 5 中的溶解度: ≥ 2.75 mg/mL (4.09 mM) (饱和度未知) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 6 中的溶解度: 配方 1 中的溶解度: ≥ 2.8 mg/mL (4.1 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，您可以取100 μL 28 mg/mL DMSO 储备液加入到400 μL PEG300中，混匀（澄清溶液）；然后再向上述溶液中加入50 μL Tween 80，混匀（澄清溶液）；最后向上述溶液中加入450 μL 生理盐水，混匀（澄清溶液）。
生理盐水的配制：将0.9 g氯化钠溶解于100 mL ddH ₂ O中，得到澄清溶液。

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配方 2 中的溶解度： ≥ 2.8 mg/mL (4.1 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，如果要制备1 mL工作液，则可以取100 μL 28 mg/mL DMSO储备液并添加到900 μL 20% SBE-β-CD 生理盐水溶液，充分混合（澄清溶液）。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: ≥ 2.8 mg/mL (4.1 mM) (饱和度未知) in 10% DMSO + 90% Corn oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，如果要制备1 mL工作液，则可以取100 μL 28 mg/mL DMSO储备液并添加到900 μL 玉米油，混合均匀（澄清溶液）。

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配方 4 中的溶解度: ≥ 2.8 mg/mL (4.1 mM) (饱和度未知) in 5% DMSO + 40% PEG300 + 5% Tween80 + + 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。

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配方 5 中的溶解度: ≥ 2.8 mg/mL (4.1 mM) (饱和度未知) in 5% DMSO + 95% (20% SBE-β-CD in saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。