Bractoppin
目录号 : GC62128
Bractoppin 是一种串联 BRCT（串联 BRCT，BRCA1），由人乳腺癌和卵巢癌抑制蛋白 (BRCA1) tBRCT 结构域识别磷酸肽的药物样抑制剂，在体外选择性抑制底物结合的纳摩尔活性，IC50 为0.074 77777#181;M。
Cas No.:2290527-07-8
Sample solution is provided at 25 µL, 10mM.
;)
,-1-7$$$37316672.jpg');)
;)
;)
$$$36806353.jpg');)
;33(7)%EF%BC%9A546-561$$$37156877.jpg');)
;)
;)
;)
%201124-1138.$$$35908550.jpg');)
%20766-780.$$$37100057.jpg');)
%20418-432.$$$36893736.jpg');)
%EF%BC%9A-240-255.$$$35108512.jpg');)
533-545$$$36543525.jpg');)
%201-13.$$$36600053.jpg');)
;)
Quality Control & SDS
- View current batch:
- Purity: >99.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Kinase experiment [1]: | |
|
Preparation Method |
For the direct binding assay, 10μl of labeled protein at a final concentration of 20nM was mixed with 10μl of test compound( Bractoppin ) and incubated on ice for 10 min. For the competitive assay, 10μl of labeled protein at a final concentration of 20nM was mixed with 2μM cognate peptide substrate at the EC80 concentration determined by prior titration using a 16-point serial dilution by direct-binding MST. |
|
Reaction Conditions |
0-105nM Bractoppin |
|
Applications |
Bractoppin is a 414-Da compound, with nanomolar potency in displacing cognate BACH1 phosphopeptide substrate from the BRCA1 tBRCT as measured by MST（ IC50 0.074 μM）. Its predicted binding mode to BRCA1 tBRCT reveals favorable hydrophobic interactions in the hydrophobic cavity, as well as a T-shaped, pi-pi stacking interaction with Phe1662, that together significantly contribute toward its activity. Indeed, substitution of the phenyl ring at R1 in Bractoppin with a 4-pyridyl group (CCBT2082) decreased activity by 5-fold by affecting the stacking interaction with Phe1662. |
| Cell experiment [2]: | |
|
Cell lines |
HEK293 cells |
|
Preparation Method |
HEK293 cells stably harboring plasmids for Tet-inducible expression of tBRCT domains were seeded. Compound( Bractoppin )addition or inducible expression of BRCA1 tBRCT were performed as indicated, when cells were 30% confluent, before exposure to 1Gy irradiation. |
|
Reaction Conditions |
1-100uM Bractoppin for 7 days |
|
Applications |
In cells, Bractoppin inhibits substrate recognition detected by Förster resonance energy transfer, and diminishes BRCA1 recruitment to DNA breaks, in turn suppressing damage-induced G2 arrest and assembly of the recombinase, RAD51. |
|
References: [1]. Periasamy J, Kurdekar V, et,al.Targeting Phosphopeptide Recognition by the Human BRCA1 Tandem BRCT Domain to Interrupt BRCA1-Dependent Signaling. Cell Chem Biol. 2018 Jun 21;25(6):677-690.e12. doi: 10.1016/j.chembiol.2018.02.012. Epub 2018 Mar 29. PMID: 29606576; PMCID: PMC6015222. |
|
Bractoppin is a tandem BRCT (tandem BRCT, BRCA1) delivered by human breast and ovarian cancer suppressor protein (BRCA1) tBRCT domain recognizes a drug-like inhibitor of phosphopeptide that selectively inhibits nanomolar activity of substrate binding in vitro with an IC50 of 0.074 µM[1].
Bractoppin has nanomolar potency in displacing cognate BACH1 phosphopeptide substrate from the BRCA1 tBRCT. Its predicted binding mode to BRCA1 tBRCT reveals favorable hydrophobic interactions in the hydrophobic cavity, as well as a T-shaped, pi-pi stacking interaction with Phe1662, that together significantly contribute toward its activity. Indeed, substitution of the phenyl ring at R1 in Bractoppin with a 4-pyridyl group (CCBT2082) decreased activity by 5-fold by affecting the stacking interaction with Phe1662[1].
In cells, Bractoppin inhibits substrate recognition detected by FÖrster resonance energy transfer, and diminishes BRCA1 recruitment to DNA breaks, in turn suppressing damage-induced G2 arrest and assembly of the recombinase, RAD51[1].
References:
[1]. Periasamy J, Kurdekar V, et,al. Targeting Phosphopeptide Recognition by the Human BRCA1 Tandem BRCT Domain to Interrupt BRCA1-Dependent Signaling. Cell Chem Biol. 2018 Jun 21;25(6):677-690.e12. doi: 10.1016/j.chembiol.2018.02.012. Epub 2018 Mar 29. PMID: 29606576; PMCID: PMC6015222.
Bractoppin 是一种串联 BRCT（串联 BRCT，BRCA1），由人乳腺癌和卵巢癌抑制蛋白 (BRCA1) tBRCT 结构域识别磷酸肽的药物样抑制剂，在体外选择性抑制底物结合的纳摩尔活性，IC50 为0.074 77777#181;M[1]。
Bractoppin 在从 BRCA1 tBRCT 中置换同源 BACH1 磷酸肽底物方面具有纳摩尔效力。它与 BRCA1 tBRCT 的预测结合模式揭示了疏水腔中有利的疏水相互作用，以及与 Phe1662 的 T 形 pi-pi 堆叠相互作用，这些共同显着促进了其活性。实际上，用 4-吡啶基取代 Bractoppin 中 R1 的苯环 (CCBT2082) 通过影响与 Phe1662[1] 的堆积相互作用，使活性降低了 5 倍。
在细胞中，Bractoppin 抑制由 FÖ 检测到的底物识别；rster 共振能量转移，并减少 BRCA1 募集到 DNA 断裂，进而抑制损伤诱导的 G2 停滞和重组酶 RAD51[1] 的组装.
| Cas No. | 2290527-07-8 | SDF | |
| 分子式 | C25H23FN4O | 分子量 | 414.47 |
| 溶解度 | DMSO : 250 mg/mL (603.18 mM; Need ultrasonic) | 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。 |
||
| 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 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 2.4127 mL | 12.0636 mL | 24.1272 mL |
| 5 mM | 0.4825 mL | 2.4127 mL | 4.8254 mL |
| 10 mM | 0.2413 mL | 1.2064 mL | 2.4127 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。
Bractoppin, a BRCA1 carboxy-terminal domain (BRCT) inhibitor, suppresses tumor progression in ovarian borderline tumor organoids
Biochem Biophys Res Commun 2023 Jan 1;638:76-83.PMID:36442235DOI:10.1016/j.bbrc.2022.11.063.
Borderline ovarian tumors are a special class of ovarian tumors between benign and malignant, which are not sensitive to traditional chemotherapy regimens, and the development of target drugs is limited due to the lack of cell lines. Tumor organoids can well preserve the genetic characteristics of the primary tumor, but there are only a few reports of application in borderline tumors. In this study, we successfully generated 13 ovarian borderline tumor organoids and tested the antitumor activity of Bractoppin, a BRCA1 carboxy-terminal domain (BRCT) inhibitor. Bractoppin promotes organoid apoptosis. Mechanistically, Bractoppin can inhibit organoid cell cycle progression, inhibit the repair of DSB damage and promote tumor cell apoptosis. In addition, Bractoppin can also promote the apoptosis of ovarian cancer cell lines and inhibit the HR and NHEJ repair ability of tumor cells. We demonstrate the value of ovarian borderline tumor organoids in the exploration of molecular therapy drugs, and Bractoppin may be a valuable small molecule drug in the treatment of BOT.
Structure-Guided Synthesis and Evaluation of Small-Molecule Inhibitors Targeting Protein-Protein Interactions of BRCA1 tBRCT Domain
ChemMedChem 2019 Sep 18;14(18):1620-1632.PMID:31334915DOI:10.1002/cmdc.201900300.
The tandem BRCT domains (tBRCT) of BRCA1 engage phosphoserine-containing motifs in target proteins to propagate intracellular signals initiated by DNA damage, thereby controlling cell cycle arrest and DNA repair. Recently, we identified Bractoppin, the first small-molecule inhibitor of the BRCA1 tBRCT domain, which selectively interrupts BRCA1-mediated cellular responses evoked by DNA damage. Here, we combine structure-guided chemical elaboration, protein mutagenesis and cellular assays to define the structural features responsible for Bractoppin's activity. Bractoppin fails to bind mutant forms of BRCA1 tBRCT bearing K1702A, a key residue mediating phosphopeptide recognition, or F1662R or L1701K that adjoin the pSer-recognition site. However, the M1775R mutation, which engages the Phe residue in the consensus phosphopeptide motif pSer-X-X-Phe, does not affect Bractoppin binding, confirming a binding mode distinct from the substrate phosphopeptide binding. We explored these structural features through structure-guided chemical elaboration and characterized structure-activity relationships (SARs) in biochemical assays. Two analogues, CCBT2088 and CCBT2103 were effective in abrogating BRCA1 foci formation and inhibiting G2 arrest induced by irradiation of cells. Collectively, our findings reveal structural features underlying the activity of a novel inhibitor of phosphopeptide recognition by the BRCA1 tBRCT domain, providing fresh insights to guide the development of inhibitors that target protein-protein interactions.
Targeting Phosphopeptide Recognition by the Human BRCA1 Tandem BRCT Domain to Interrupt BRCA1-Dependent Signaling
Cell Chem Biol 2018 Jun 21;25(6):677-690.e12.PMID:29606576DOI:10.1016/j.chembiol.2018.02.012.
Intracellular signals triggered by DNA breakage flow through proteins containing BRCT (BRCA1 C-terminal) domains. This family, comprising 23 conserved phosphopeptide-binding modules in man, is inaccessible to small-molecule chemical inhibitors. Here, we develop Bractoppin, a drug-like inhibitor of phosphopeptide recognition by the human BRCA1 tandem (t)BRCT domain, which selectively inhibits substrate binding with nanomolar potency in vitro. Structure-activity exploration suggests that Bractoppin engages BRCA1 tBRCT residues recognizing pSer in the consensus motif, pSer-Pro-Thr-Phe, plus an abutting hydrophobic pocket that is distinct in structurally related BRCT domains, conferring selectivity. In cells, Bractoppin inhibits substrate recognition detected by Förster resonance energy transfer, and diminishes BRCA1 recruitment to DNA breaks, in turn suppressing damage-induced G2 arrest and assembly of the recombinase, RAD51. But damage-induced MDC1 recruitment, single-stranded DNA (ssDNA) generation, and TOPBP1 recruitment remain unaffected. Thus, an inhibitor of phosphopeptide recognition selectively interrupts BRCA1 tBRCT-dependent signals evoked by DNA damage.