1. Protein Tyrosine Kinase/RTK
  2. Btk
  3. Spebrutinib

Spebrutinib  (Synonyms: AVL-292; CC-292)

目录号: HY-18012 纯度: 99.62%
COA 产品使用指南

Spebrutinib (AVL-292; CC-292) 是共价,高选择性和口服活性的 Btk 抑制剂,IC50 为0.5 nM。

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Spebrutinib Chemical Structure

Spebrutinib Chemical Structure

CAS No. : 1202757-89-8

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10 mM * 1 mL in DMSO ¥660
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1 mg ¥272
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5 mg ¥600
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10 mg ¥900
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Customer Review

Other Forms of Spebrutinib:

    Spebrutinib purchased from MCE. Usage Cited in: Blood. 2016 Jun 23;127(25):3237-52.  [Abstract]

    Ibrutinib binds to the ATP-binding pocketof HCK and blocks ATP binding. Results from kinase active-site inhibition assaysutilizing an ATP-BTN probe that is used to pull downactive kinases in the presence of Ibrutinib, CC-292, or A419259 in lysates from BCWM.1 WM cells.
    • 生物活性

    • 实验参考方法

    • 纯度 & 产品资料

    • 参考文献

    生物活性

    Spebrutinib (AVL-292; CC-292) is a covalent, orally active, and highly selective with an IC50 of 0.5 nM.

    IC50 & Target

    IC50: <0.5 nM (Btk)[1]

    细胞效力
    (Cellular Effect)
    Cell Line Type Value Description References
    786-0 IC50
    14.9 μM
    Compound: 3
    Cytotoxicity against human 786-O cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human 786-O cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    A549 IC50
    1.98 μM
    Compound: 3
    Cytotoxicity against human A549 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human A549 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    BJ IC50
    27.6 μM
    Compound: 3
    Cytotoxicity against human BJ cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human BJ cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    HEF IC50
    22.6 μM
    Compound: 3
    Cytotoxicity against human HEF cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human HEF cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    HEL IC50
    5.3 μM
    Compound: CC292
    Antiproliferative activity against human HEL cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    Antiproliferative activity against human HEL cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    [PMID: 31843459]
    HeLa IC50
    17.4 μM
    Compound: 3
    Cytotoxicity against human HeLa cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human HeLa cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    HepG2 IC50
    33.5 μM
    Compound: 3
    Cytotoxicity against human HepG2 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human HepG2 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    HT-29 IC50
    18.9 μM
    Compound: 3
    Cytotoxicity against human HT-29 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human HT-29 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    JeKo-1 IC50
    1.3 μM
    Compound: CC292
    Antiproliferative activity against human JeKo1 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    Antiproliferative activity against human JeKo1 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    [PMID: 31843459]
    Jurkat IC50
    6.7 μM
    Compound: 3
    Cytotoxicity against human Jurkat cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human Jurkat cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    K562 IC50
    12.6 μM
    Compound: 3
    Cytotoxicity against human K562 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human K562 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    KARPAS-299 IC50
    20.3 μM
    Compound: 3
    Cytotoxicity against human KARPAS299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human KARPAS299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    MCF-10A IC50
    4.36 μM
    Compound: 3
    Cytotoxicity against human MCF10A cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human MCF10A cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    MCF7 IC50
    29 μM
    Compound: 3
    Cytotoxicity against human MCF7 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human MCF7 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    MDA-MB-231 IC50
    > 40 μM
    Compound: 3
    Cytotoxicity against human MDA-MB-231 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human MDA-MB-231 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    NAMALVA IC50
    3.42 μM
    Compound: Spebrutinib
    Antiproliferative activity against human NAMALWA cells after 72 hrs by CCK-8 assay
    Antiproliferative activity against human NAMALWA cells after 72 hrs by CCK-8 assay
    [PMID: 29146136]
    NCI-H1299 IC50
    38.5 μM
    Compound: 3
    Cytotoxicity against human H1299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human H1299 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    NCI-H3122 IC50
    32.5 μM
    Compound: 3
    Cytotoxicity against human NCI-H3122 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human NCI-H3122 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    OCI-Ly10 IC50
    > 10 μM
    Compound: CC292
    Antiproliferative activity against human OCI-LY10 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    Antiproliferative activity against human OCI-LY10 cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    [PMID: 31843459]
    PC-3 IC50
    22.2 μM
    Compound: 3
    Cytotoxicity against human PC3 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    Cytotoxicity against human PC3 cells assessed as reduction in cell viability at 0.1 to 40 uM incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    Raji IC50
    17.6 μM
    Compound: CC292
    Antiproliferative activity against human Raji cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    Antiproliferative activity against human Raji cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    [PMID: 31843459]
    Raji IC50
    21.6 μM
    Compound: 3
    Antiproliferative activity against human Raji cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
    Antiproliferative activity against human Raji cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    Raji IC50
    25.3 μM
    Compound: 4; AVL-292
    Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
    [PMID: 27994736]
    Raji IC50
    25.3 μM
    Compound: Spebrutinib
    Antiproliferative activity against human Raji cells measured after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Raji cells measured after 48 hrs by CCK-8 assay
    [PMID: 27956037]
    Raji IC50
    25.3 μM
    Compound: Spebrutinib
    Antiproliferative activity against human Raji cells after 72 hrs by CCK-8 assay
    Antiproliferative activity against human Raji cells after 72 hrs by CCK-8 assay
    [PMID: 29146136]
    Raji IC50
    29.4 μM
    Compound: 4
    Antiproliferative activity against human Raji cells over expressing BTK after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Raji cells over expressing BTK after 48 hrs by CCK-8 assay
    [PMID: 28432946]
    Raji IC50
    29.4 μM
    Compound: 4
    Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Raji cells after 48 hrs by CCK-8 assay
    [PMID: 27912175]
    Ramos IC50
    14.2 μM
    Compound: Spebrutinib
    Antiproliferative activity against human Ramos cells after 72 hrs by CCK-8 assay
    Antiproliferative activity against human Ramos cells after 72 hrs by CCK-8 assay
    [PMID: 29146136]
    Ramos IC50
    15.1 μM
    Compound: 4; AVL-292
    Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
    [PMID: 27994736]
    Ramos IC50
    15.1 μM
    Compound: Spebrutinib
    Antiproliferative activity against human Ramos cells measured after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Ramos cells measured after 48 hrs by CCK-8 assay
    [PMID: 27956037]
    Ramos IC50
    18.8 μM
    Compound: 4
    Antiproliferative activity against human Ramos cells over expressing BTK after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Ramos cells over expressing BTK after 48 hrs by CCK-8 assay
    [PMID: 28432946]
    Ramos IC50
    20.9 μM
    Compound: 4
    Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
    Antiproliferative activity against human Ramos cells after 48 hrs by CCK-8 assay
    [PMID: 27912175]
    Ramos IC50
    3.9 μM
    Compound: CC292
    Antiproliferative activity against human Ramos cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    Antiproliferative activity against human Ramos cells assessed as reduction in cell viability incubated for 72 hrs by CCK-8 assay
    [PMID: 31843459]
    Ramos IC50
    5.44 μM
    Compound: 3
    Antiproliferative activity against human Ramos cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
    Antiproliferative activity against human Ramos cells assessed as reduction in cell growth incubated for 48 hrs by MTS assay
    [PMID: 31395509]
    Sf9 IC50
    0.6 nM
    Compound: 4; AVL-292
    Inhibition of recombinant human N-terminal His-tagged BTK expressed in baculovirus infected sf9 cells using poly(4:1 Glu,Tyr) as substrate by ADP-Glo kinase assay
    Inhibition of recombinant human N-terminal His-tagged BTK expressed in baculovirus infected sf9 cells using poly(4:1 Glu,Tyr) as substrate by ADP-Glo kinase assay
    [PMID: 27994736]
    Sf9 IC50
    0.72 nM
    Compound: Spebrutinib
    Inhibition of human recombinant full-length N-terminal His-tagged BTK expressed in baculovirus infected Sf9 insect cells measured after 60 mins by ADP-Glo kinase assay
    Inhibition of human recombinant full-length N-terminal His-tagged BTK expressed in baculovirus infected Sf9 insect cells measured after 60 mins by ADP-Glo kinase assay
    [PMID: 27956037]
    Sf9 IC50
    2.12 nM
    Compound: 3
    Inhibition of human recombinant full length BTK expressed in baculovirus in Sf9 insect cells using Poly (4:1 Glu, Tyr) as substrate incubated for 1 hr by ADP-Glo assay
    Inhibition of human recombinant full length BTK expressed in baculovirus in Sf9 insect cells using Poly (4:1 Glu, Tyr) as substrate incubated for 1 hr by ADP-Glo assay
    [PMID: 31395509]
    Sf9 IC50
    66.5 nM
    Compound: Spebrutinib
    Inhibition of recombinant human N-terminal GST-tagged JAK3 (781 to end residues) expressed in baculovirus infected Sf9 cells using poly (Glu,Tyr) 4:1 as substrate incubated for 60 mins in presence of ATP by ADP-Glo kinase assay
    Inhibition of recombinant human N-terminal GST-tagged JAK3 (781 to end residues) expressed in baculovirus infected Sf9 cells using poly (Glu,Tyr) 4:1 as substrate incubated for 60 mins in presence of ATP by ADP-Glo kinase assay
    [PMID: 31866272]
    体外研究
    (In Vitro)

    Spebrutinib (CC-292) 是一种共价、高选择性、口服活性的 Btk 抑制剂,IC50 值为 0.5 nM。 Spebrutinib 对 Yes、c-Src、Brk、Lyn 和 Fyn 的抑制作用也较弱,IC50 分别为 723 nM、1.729 μM、2.43 μM、4.4 μM 和 7.15 μM。 广泛的分析表明,Ramos 细胞中 Spebrutinib 剂量反应的 Btk 占据的 EC50 (EC50=6 nM) 与 Spebrutinib 抑制 Btk 激酶的细胞 EC50 (EC50=8 nM) 直接相关。 此外,Spebrutinib 在 Ramos 细胞中抑制 90% Btk 活性的浓度为 35 nM,而 Btk 占据 90% 所需的 Spebrutinib 浓度为 39 nM[1]

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Clinical Trial
    分子量

    423.44

    Formula

    C22H22FN5O3

    CAS 号
    性状

    固体

    颜色

    White to khaki

    运输条件

    Room temperature in continental US; may vary elsewhere.

    储存方式
    Powder -20°C 3 years
    4°C 2 years
    In solvent -80°C 2 years
    -20°C 1 year
    溶解性数据
    细胞实验: 

    DMSO 中的溶解度 : ≥ 45 mg/mL (106.27 mM; 吸湿的 DMSO 对产品的溶解度有显著影响,请使用新开封的 DMSO)

    * "≥" means soluble, but saturation unknown.

    配制储备液
    浓度 溶剂体积 质量 1 mg 5 mg 10 mg
    1 mM 2.3616 mL 11.8080 mL 23.6161 mL
    5 mM 0.4723 mL 2.3616 mL 4.7232 mL
    查看完整储备液配制表

    * 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, 2 years; -20°C, 1 year。-80°C储存时,请在2年内使用, -20°C储存时,请在1年内使用。

    • 摩尔计算器

    • 稀释计算器

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    质量
    =
    浓度
    ×
    体积
    ×
    分子量 *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    浓度 (start)

    C1

    ×
    体积 (start)

    V1

    =
    浓度 (final)

    C2

    ×
    体积 (final)

    V2

    动物实验:

    请根据您的 实验动物和给药方式 选择适当的溶解方案。

    以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液,再依次添加助溶剂:
    ——为保证实验结果的可靠性,澄清的储备液可以根据储存条件,适当保存;体内实验的工作液,建议您现用现配,当天使用
    以下溶剂前显示的百分比是指该溶剂在您配制终溶液中的体积占比;如在配制过程中出现沉淀、析出现象,可以通过加热和/或超声的方式助溶

    • 方案 一

      请依序添加每种溶剂: 10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: ≥ 2.5 mg/mL (5.90 mM); 澄清溶液

      此方案可获得 ≥ 2.5 mg/mL(饱和度未知)的澄清溶液。

      1 mL 工作液为例,取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中,混合均匀;再向上述体系中加入 50 μL Tween-80,混合均匀;然后再继续加入 450 μL 生理盐水 定容至 1 mL

      生理盐水的配制:将 0.9 g 氯化钠,溶解于 ddH₂O 并定容至 100 mL,可以得到澄清透明的生理盐水溶液。
    • 方案 二

      请依序添加每种溶剂: 10% DMSO    90% Corn Oil

      Solubility: ≥ 2.5 mg/mL (5.90 mM); 澄清溶液

      此方案可获得 ≥ 2.5 mg/mL(饱和度未知)的澄清溶液,此方案实验周期在半个月以上的动物实验酌情使用。

      1 mL 工作液为例,取 100 μL 25.0 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中,混合均匀。

    动物溶解方案计算器
    请输入动物实验的基本信息:

    给药剂量

    mg/kg

    动物的平均体重

    g

    每只动物的给药体积

    μL

    动物数量

    由于实验过程有损耗,建议您多配一只动物的量
    请输入您的动物体内配方组成:
    %
    DMSO +
    +
    %
    Tween-80 +
    %
    Saline
    如果您的动物是免疫缺陷鼠或者体弱鼠,建议 DMSO 中的在最后工作液体系中的占比尽量不超过 2%。
    方案所需 助溶剂 包括:DMSO ,均可在 MCE 网站选购。 Tween 80,均可在 MCE 网站选购。
    计算结果
    工作液所需浓度 : mg/mL
    储备液配制方法 : mg 药物溶于 μL  DMSO(母液浓度为 mg/mL)。
    您所需的储备液浓度超过该产品的实测溶解度,以下方案仅供参考,如有需要,请与 MCE 中国技术支持联系。
    动物实验体内工作液的配制方法 : 取 μL DMSO 储备液,加入 μL  μL ,混合均匀至澄清,再加 μL Tween 80,混合均匀至澄清,再加 μL 生理盐水
    连续给药周期超过半月以上,请谨慎选择该方案。
    请确保第一步储备液溶解至澄清状态,从左到右依次添加助溶剂。您可采用超声加热 (超声清洗仪,建议频次 20-40 kHz),涡旋吹打等方式辅助溶解。
    纯度 & 产品资料

    纯度: 99.62%

    参考文献
    Cell Assay
    [1]

    Cells are incubated in serum-free RPMI media for 1-1.5 hours. Isolated human B cells are incubated with Spebrutinib at a final concentration of 0.001, 0.01, 0.1 and 1 μM. Ramos cells are incubated with 0.1 nM-3 μM Spebrutinib. Cells are then incubated in the presence of compound for 1 hour at 37°C. Following incubation, cells are centrifuged and resuspended in 100 μL of serum-free RPMI and BCR is stimulated with addition of 5 μg/mL α-human IgM. Samples are centrifuged, washed in phosphate-buffered saline (PBS), and lysed in 100 μL of Cell Extraction Buffer plus 1:10 (v/v) PhosSTOP Phosphatase Inhibitor and 1:10 (v/v) Complete Protease Inhibitor. Antibodies used for immunoblot analysis include P-PLCγ2, PLCγ2 (3871; CST), Syk (2712; CST), P-Syk (2710; CST), Btk, P-Btk, and Tubulin. Membranes are scanned on a Li-Cor Odyssey scanner using infrared fluorescence detection[1].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    参考文献

    完整储备液配制表

    * 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, 2 years; -20°C, 1 year。-80°C储存时,请在2年内使用, -20°C储存时,请在1年内使用。

    可选溶剂 浓度 溶剂体积 质量 1 mg 5 mg 10 mg 25 mg
    DMSO 1 mM 2.3616 mL 11.8080 mL 23.6161 mL 59.0402 mL
    5 mM 0.4723 mL 2.3616 mL 4.7232 mL 11.8080 mL
    10 mM 0.2362 mL 1.1808 mL 2.3616 mL 5.9040 mL
    15 mM 0.1574 mL 0.7872 mL 1.5744 mL 3.9360 mL
    20 mM 0.1181 mL 0.5904 mL 1.1808 mL 2.9520 mL
    25 mM 0.0945 mL 0.4723 mL 0.9446 mL 2.3616 mL
    30 mM 0.0787 mL 0.3936 mL 0.7872 mL 1.9680 mL
    40 mM 0.0590 mL 0.2952 mL 0.5904 mL 1.4760 mL
    50 mM 0.0472 mL 0.2362 mL 0.4723 mL 1.1808 mL
    60 mM 0.0394 mL 0.1968 mL 0.3936 mL 0.9840 mL
    80 mM 0.0295 mL 0.1476 mL 0.2952 mL 0.7380 mL
    100 mM 0.0236 mL 0.1181 mL 0.2362 mL 0.5904 mL
    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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    产品名称:
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    目录号:
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