Brefeldin A
(Synonyms: 布雷非德菌素 A; BFA; Cyanein; Decumbin) 目录号 : GC17683
布雷菲定A（BFA）是一种真菌大环内酯，能够有效、可逆地抑制细胞内囊泡形成和内质网（ER）与高尔基体之间的蛋白质运输。
Cas No.:20350-15-6
Sample solution is provided at 25 µL, 10mM.
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| Cell experiment [1]: | |
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Cell lines |
Mouse primary bone marrow-derived macrophages (BMDMs) from the femurs of C57BL/6 or Nlrp-/- mice |
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Preparation Method |
Quantification of IL-6 and IL-1β in the culture supernatants of mouse BMDMs untreated (Unt) or treated with LPS in the presence of Golgi-Plug, Brefeldin A (2 or 5 ug/ ml), or MCC950 for 3 h followed by ATP treatment |
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Reaction Conditions |
2 or 5 ug/ ml Brefeldin A for 3 h |
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Applications |
Inhibition of vesicle trafficking by Brefeldin A between ER and golgi attenuates IL-1β production from BMDMs upon stimulation with NLRP3 agonists |
| Animal experiment [2]: | |
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Animal models |
Female BALB/ C mice (20 ± 2 g, 5-6 weeks) |
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Preparation Method |
HepG2 tumor-bearing nude mice were injected with M- Brefeldin A daily for 14 days( intravenous injection) |
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Dosage form |
5 mg/kg and 10 mg/kg Brefeldin A for 14 days |
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Applications |
M- Brefeldin A 10 mg/kg group showed effective antitumor effect and significantly delayed tumor progression, while M- Brefeldin A 5 mg/kg mice did not show significant inhibitory effect. |
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References: [1]. Hong S, Hwang I, et,al. Brefeldin A-sensitive ER-Golgi vesicle trafficking contributes to NLRP3-dependent caspase-1 activation. FASEB J. 2019 Mar;33(3):4547-4558. doi: 10.1096/fj.201801585R. Epub 2018 Dec 28. PMID: 30592629. |
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Brefeldin A (BFA) is a fungal macrocyclic lactone and a potent, reversible inhibitor of intracellular vesicle formation and protein trafficking between the endoplasmic reticulum (ER) and the Golgi apparatus[1][2].Brefeldin A is an ATPase inhibitor with IC50 value of 0.2 µM[9].Brefeldin A and its analogs are promising inhibitors in drug development due to a number of key features such as apoptosis?inducing properties as well as antitumor, antifungal, and antiviral effects [3,6].Brefeldin A is a CRISPR/Cas9 activator. Brefeldin A inhibits HSV-1 and has anti-cancer activity[4].
Perturbation of ER-Golgi trafficking by brefeldin A (BFA) treatment attenuated nucleotide-binding oligomerization domain-like receptor family, pyrin-domain-containing 3 (NLRP3) inflammasome activation in mouse bone marrow-derived macrophages (BMDMs) [5].ADP-ribosylation of BARS is mediated by formation of a conjugate between Brefeldin A and ADPR. BARS shows BAC binding when incubated with the medium from the brefeldin A-treated CD38+ HeLa cells[3].Brefeldin A induces anchorage-independent cell death in MDA-MB-231 breast cancer cells with an EC50 of 0.016 µg/mL, inhibits the formation of MDA-MB-231 colonies in 3D and 2D cultures and inhibits the migration and MMP 9 activity of MDA-MB-231[2].
In tumor-bearing mice, M-brefeldin A can prolong blood circulation, improve tumor accumulation ability, and show effective inhibition of tumor growth, M-brefeldin A 10 mg/kg group showed effective antitumor effect and significantly delayed tumor progression, while M-brefeldin A 5 mg/kg mice did not show significant inhibitory effect[7]. Mice were treated with the Golgi blocker Brefeldin A. Since most cytokines are processed and secreted via the classical secretion pathway through the Golgi, brefeldin A blocks cytokine secretion, leading to their accumulation within immune cells, which are eventually detected by flow cytometry. Thus, treatment of mice with brefeldin A allows in situ assessment of cytokine production without the use of reporter mice [8].
Brefeldin A（BFA）是一种真菌大环内酯，能够有效、可逆地抑制内质网和高尔基体之间的蛋白质运输和囊泡形成[1][2]。Brefeldin A是一种ATP酶抑制剂，其IC50值为0.2微米[9]。由于具有诱导细胞凋亡、抗肿瘤、抗真菌和抗病毒等多种特性，Brefeldin A及其类似物在药物开发中被广泛应用[3,6]。此外，Brefeldin A还可以作为CRISPR/Cas9激活剂使用，并且对HSV-1具有抑制作用并具有抗癌活性[4]。
在小鼠骨髓源性巨噬细胞（BMDMs）中，使用卟啉菌素A（BFA）处理扰乱了内质网-高尔基体运输，减弱了核苷酸结合寡聚化域类受体家族、含有pyrin域的3号（NLRP3）炎症小体的激活[5]。通过形成卟啉菌素A和ADPR之间的共轭物介导对BARS进行ADP核糖化。当与经过卟啉菌素A处理的CD38+ HeLa细胞培养基一起孵育时，BARS显示出与BAC结合[3]。卟啉菌素A诱导MDA-MB-231乳腺癌细胞无需附着即可死亡，EC50为0.016 µg/mL，并抑制了MDA-MB-231在三维和二维培养中形成集落以及抑制其迁移和MMP 9活性[2]。
在携带肿瘤的小鼠中，M-布雷菲定A可以延长血液循环时间，提高肿瘤积累能力，并显示出有效抑制肿瘤生长的作用。M-布雷菲定A 10毫克/千克组表现出有效的抗肿瘤效果并显著延缓了肿瘤进展，而M-布雷菲定A 5毫克/千克组则没有显示出明显的抑制作用[7]。小鼠接受了高尔基体阻断剂Brefeldin A治疗。由于大多数细胞因子是通过经典分泌途径通过高尔基体加工和分泌的，Brefeldin A会阻止细胞因子分泌，导致它们在免疫细胞内积累，并最终被流式细胞术检测到。因此，在小鼠中使用Brefeldin A治疗可以对细胞因子产生进行原位评估，无需使用报告小鼠[8]。
References:
[1]: Orci L, Tagaya M, et,al. Brefeldin A, a drug that blocks secretion, prevents the assembly of non-clathrin-coated buds on Golgi cisternae. Cell. 1991 Mar 22;64(6):1183-95. doi: 10.1016/0092-8674(91)90273-2. PMID: 2004424.
[2]: Tseng CN, Hong YR, et,al. Brefeldin A reduces anchorage-independent survival, cancer stem cell potential and migration of MDA-MB-231 human breast cancer cells. Molecules. 2014 Oct 29;19(11):17464-77. doi: 10.3390/molecules191117464. PMID: 25356567; PMCID: PMC6271931.
[3]: Wang J, Fang Y, et,al. Erythroleukemia cells acquire an alternative mitophagy capability. Sci Rep. 2016 Apr 19;6:24641. doi: 10.1038/srep24641. PMID: 27091640; PMCID: PMC4835698.
[4]: Yu C, Liu Y, et,al. Small molecules enhance CRISPR genome editing in pluripotent stem cells. Cell Stem Cell. 2015 Feb 5;16(2):142-7. doi: 10.1016/j.stem.2015.01.003. PMID: 25658371; PMCID: PMC4461869.
[5]: Hong S, Hwang I, et,al. Brefeldin A-sensitive ER-Golgi vesicle trafficking contributes to NLRP3-dependent caspase-1 activation. FASEB J. 2019 Mar;33(3):4547-4558. doi: 10.1096/fj.201801585R. Epub 2018 Dec 28. PMID: 30592629.
[6]: Paek SM. Recent Synthesis and Discovery of Brefeldin A Analogs. Mar Drugs. 2018 Apr 18;16(4):133. doi: 10.3390/md16040133. PMID: 29670019; PMCID: PMC5923420.
[7]: Zhang JM, Jiang YY, et,al. Brefeldin A delivery nanomicelles in hepatocellular carcinoma therapy: Characterization, cytotoxic evaluation in vitro, and antitumor efficiency in vivo. Pharmacol Res. 2021 Oct;172:105800. doi: 10.1016/j.phrs.2021.105800. Epub 2021 Aug 4. PMID: 34363949.
[8]: Kovacs SB, Oh C, et,al. Evaluating cytokine production by flow cytometry using brefeldin A in mice. STAR Protoc. 2020 Dec 30;2(1):100244. doi: 10.1016/j.xpro.2020.100244. PMID: 33458706; PMCID: PMC7797915.
[9]: Wierzbicki PM, Kogut-Wierzbicka M, et,al. Protein and siRNA delivery by transportan and transportan 10 into colorectal cancer cell lines. Folia Histochem Cytobiol. 2014;52(4):270-80. doi: 10.5603/FHC.a2014.0035. Epub 2014 Dec 16. PMID: 25511292.
| Cas No. | 20350-15-6 | SDF | |
| 别名 | 布雷非德菌素 A; BFA; Cyanein; Decumbin | ||
| 化学名 | (1S,2E,7S,10E,12R,13R,15S)-12,15-dihydroxy-7-methyl-8-oxabicyclo[11.3.0]hexadeca-2,10-dien-9-one | ||
| Canonical SMILES | CC1CCCC=CC2CC(CC2C(C=CC(=O)O1)O)O | ||
| 分子式 | C16H24O4 | 分子量 | 280.36 |
| 溶解度 | ≥ 4.67mg/mL in DMSO | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.5668 mL | 17.8342 mL | 35.6684 mL |
| 5 mM | 0.7134 mL | 3.5668 mL | 7.1337 mL |
| 10 mM | 0.3567 mL | 1.7834 mL | 3.5668 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Recent Synthesis and Discovery of Brefeldin A Analogs
Mar Drugs2018 Apr 18;16(4):133.PMID: 29670019DOI: 10.3390/md16040133
The recent development of analogs of brefeldin A (BFA), a fungal metabolite, for the improvement of BFA apoptosis-inducing activity is described. BFA has been isolated from various soil or, more recently, marine fungi and has shown versatile beneficial activities. More importantly, the apoptosis-inducing activity of BFA in cancer cells highlights the possibility of further developing this natural product as an anticancer agent. Besides its biological importance, its structural features have also gathered tremendous interest from both medicinal and synthetic chemists. By a medicinal chemistry and total synthesis approach, numerous analogs from BFA have been developed to improve its inferior bioavailability and its antiproliferative ability. In this review, the recent medicinal chemistry efforts in relation to the production of BFA analogs are extensively presented.
Brefeldin A and kifunensine modulate LPS-induced lung endothelial hyperpermeability in human and bovine cells
Am J Physiol Cell Physiol2021 Aug 1;321(2):C214-C220.PMID: 34161151DOI: 10.1152/ajpcell.00142.2021
Endothelial hyperpermeability is the hallmark of acute respiratory distress syndrome (ARDS). Laborious efforts in the investigation of the molecular pathways involved in the regulation of the vascular barrier shall reveal novel therapeutic targets toward that respiratory disorder. Herein, we investigate in vitro the effects of the α-1,2-mannosidase 1 inhibitor kifunensine (KIF) and brefeldin A (BFA) in the lipopolysaccharides (LPS)-induced endothelial breakdown. Our results suggest that BFA opposes the deteriorating effects of KIF [unfolded protein response (UPR) suppressor] toward the lung microvasculature. Since KIF is a UPR suppressor, and brefeldin A is a UPR inducer, we suggest that a carefully devised UPR manipulation may deliver novel therapeutic avenues in diseases related to endothelial barrier dysfunction (e.g., ARDS and sepsis).
Brefeldin A inhibits clathrin-dependent endocytosis and ion transport in Chara internodal cells
Biol Cell2020 Nov;112(11):317-334.PMID: 32648585DOI: 10.1111/boc.202000031
Background: The Characeae are multicellular green algae, which are closely related to higher plants. Their internodal cells are a convenient model to study membrane transport and organelle interactions.
Results: In this study, we report on the effect of brefeldin A (BFA), an inhibitor of vesicle trafficking, on internodal cells of Chara australis. BFA induced the commonly observed agglomeration of Golgi bodies and trans Golgi network into 'brefeldin compartments' at concentrations between 6 and 500 μM and within 30-120 min treatment. In contrast to most other cells, however, BFA inhibited endocytosis and significantly decreased the number of clathrin-coated pits and clathrin-coated vesicles at the plasma membrane. BFA did not inhibit secretion of organelles at wounds induced by puncturing or local light damage but prevented the formation of cellulosic wound walls probably because of insufficient membrane recycling. We also found that BFA inhibited the formation of alkaline and acid regions along the cell surface ('pH banding pattern') which facilitates carbon uptake required for photosynthesis; we hypothesise that this is due to insufficient recycling of ion transporters. During long-term treatments over several days, BFA delayed the formation of complex 3D plasma membranes (charasomes). Interestingly, BFA had no detectable effect on clathrin-dependent charasome degradation. Protein sequence analysis suggests that the peculiar effects of BFA in Chara internodal cells are due to a mutation in the guanine-nucleotide exchange factor GNOM required for recruitment of membrane coats via activation of ADP-ribosylation factor proteins.
Conclusions and significance: This work provides an overview on the effects of BFA on different processes in C. australis. It revealed similarities but also distinct differences in vesicle trafficking between higher plant and algal cells. It shows that characean internodal cells are a promising model to study interactions between seemingly distant metabolic pathways.
Class II Arfs require a brefeldin-A-sensitive factor for Golgi association
Biochem Biophys Res Commun2020 Sep 10;530(1):301-306.PMID: 32828303DOI: 10.1016/j.bbrc.2020.07.001
Arf proteins are small Ras-family GTPases which recruit clathrin and COPI coats to Golgi membranes and regulate components of the membrane trafficking machinery. It is believed membrane association and activity of Arfs is coupled to GTP binding, with GTP hydrolysis required for vesicle uncoating. In humans, four Arf proteins (Arf1, Arf3, Arf4 and Arf5) are Golgi-associated. Conflicting reports have suggested that HA-GFP-tagged Class II ARFs (Arf4 and Arf5) are recruited to membrane independently of the brefeldin A sensitive exchange factor GBF1, suggesting regulation fundamentally different from the Class I Arfs (Arf1, Arf3), or alternately that the GTPase cycle of GFP-tagged Class II Arfs is similar to other Arfs. We show that these results depend on the fluorescent tag, with Arf4-HA-GFP tag resistant to brefeldin, but Arf4-GFP acting similarly to Arf1-GFP in brefeldin-sensitivity and photobleach assays. Arf4-HA-GFP could be partially reverted to the behavior of Arf4-GFP by mutation of two aspartic acids in the HA tag to alanine. Our results, which indicate a high sensitivity of Arf4 to tagging, can explain the discrepancies between previous studies. We discuss the implications of this study for future work with tagged Arfs.
Elucidation of brefeldin A-induced ER and Golgi stress responses in Neuro2a cells
Mol Cell Biochem2021 Oct;476(10):3869-3877.PMID: 34129155DOI: 10.1007/s11010-021-04187-1
Brefeldin A (BFA) disrupts the structure of the Golgi apparatus to trigger ER stress signaling pathways. On the other hand, treatment with BFA induces the activation of CREB3, the protein structure of which is similar to that of ATF6. In this study, we established Neuro2a cells in which three different transcription factors, namely, ATF4, ATF3 and CREB3, were deficient using the CRISPR/Cas9 approach, and we investigated the BFA-induced ER and Golgi stress response in these cells. BFA treatment rapidly induced ATF4, ATF3, Herp and GADD153 protein expression in Neuro2a cells. ATF4-deficient Neuro2a cells exhibited significantly decreased mRNA and protein expression of ATF3 and Herp but not GADD153; however, cells deficient in ATF3 exhibited minimal effects on GADD34, GADD153 and Herp expression. The cleavage of CREB3 in Neuro2a cells was triggered by BFA; however, the expression of several ER and Golgi stress-related factors was hardly influenced by the CREB3 deficiency in these Neuro2a cells. This study shows that CREB3 minimally associates with typical ER stress-inducible responses in Neuro2a cells. Therefore, identification and characterization of the downstream transcriptional targets of CREB3 is required to clarify not only Golgi stress response but also its relationship with ER stress signaling pathways.