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

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

目錄號: HY-18012 純度: 99.62%
COA 產(chǎn)品使用指南

Spebrutinib (AVL-292; CC-292) 是共價,高選擇性和口服活性的 Btk 抑制劑,IC50 為0.5 nM。

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Spebrutinib Chemical Structure

Spebrutinib Chemical Structure

CAS No. : 1202757-89-8

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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

    • 實驗參考方法

    • 純度 & 產(chǎn)品資料

    • 參考文獻

    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]

    細胞效力
    (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]
    體外研究
    (In Vitro)

    Spebrutinib (CC-292) 是一種共價、高選擇性、口服活性的 Btk 抑制劑,IC50 值為 0.5 nM。 Spebrutinib 對 Yes、c-Src、Brk、Lyn 和 Fyn 的抑制作用也較弱,IC50 分別為 723 nM、1.729 μM、2.43 μM、4.4 μM 和 7.15 μM。 廣泛的分析表明,Ramos 細胞中 Spebrutinib 劑量反應(yīng)的 Btk 占據(jù)的 EC50 (EC50=6 nM) 與 Spebrutinib 抑制 Btk 激酶的細胞 EC50 (EC50=8 nM) 直接相關(guān)。 此外,Spebrutinib 在 Ramos 細胞中抑制 90% Btk 活性的濃度為 35 nM,而 Btk 占據(jù) 90% 所需的 Spebrutinib 濃度為 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 號
    性狀

    固體

    顏色

    White to khaki

    運輸條件

    Room temperature in continental US; may vary elsewhere.

    儲存方式
    Powder -20°C 3 years
    4°C 2 years
    In solvent -80°C 2 years
    -20°C 1 year
    溶解性數(shù)據(jù)
    細胞實驗: 

    DMSO 中的溶解度 : ≥ 45 mg/mL (106.27 mM; 吸濕的 DMSO 對產(chǎn)品的溶解度有顯著影響,請使用新開封的 DMSO)

    * "≥" means soluble, but saturation unknown.

    配制儲備液
    濃度 溶劑體積 質(zhì)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
    查看完整儲備液配制表

    * 請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液;一旦配成溶液,請分裝保存,避免反復(fù)凍融造成的產(chǎn)品失效。
    儲備液的保存方式和期限:-80°C, 2 years; -20°C, 1 year。-80°C儲存時,請在2年內(nèi)使用, -20°C儲存時,請在1年內(nèi)使用。

    • 摩爾計算器

    • 稀釋計算器

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

    質(zhì)量
    =
    濃度
    ×
    體積
    ×
    分子量 *

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

    This equation is commonly abbreviated as: C1V1 = C2V2

    濃度 (start)

    C1

    ×
    體積 (start)

    V1

    =
    濃度 (final)

    C2

    ×
    體積 (final)

    V2

    動物實驗:

    請根據(jù)您的 實驗動物和給藥方式 選擇適當(dāng)?shù)娜芙夥桨浮?

    以下溶解方案都請先按照 In Vitro 方式配制澄清的儲備液,再依次添加助溶劑:
    ——為保證實驗結(jié)果的可靠性,澄清的儲備液可以根據(jù)儲存條件,適當(dāng)保存;體內(nèi)實驗的工作液,建議您現(xiàn)用現(xiàn)配,當(dāng)天使用;
    以下溶劑前顯示的百分比是指該溶劑在您配制終溶液中的體積占比;如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象,可以通過加熱和/或超聲的方式助溶

    • 方案 一

      請依序添加每種溶劑: 10% DMSO    40% PEG300    5% Tween-80    45% Saline

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

      此方案可獲得 ≥ 2.5 mg/mL(飽和度未知)的澄清溶液。

      1 mL 工作液為例,取 100 μL 25.0 mg/mL 的澄清 DMSO 儲備液加到 400 μL PEG300 中,混合均勻;再向上述體系中加入 50 μL Tween-80,混合均勻;然后再繼續(xù)加入 450 μL 生理鹽水 定容至 1 mL。

      生理鹽水的配制:將 0.9 g 氯化鈉,溶解于 ddH?O 并定容至 100 mL,可以得到澄清透明的生理鹽水溶液。
    • 方案 二

      請依序添加每種溶劑: 10% DMSO    90% Corn Oil

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

      此方案可獲得 ≥ 2.5 mg/mL(飽和度未知)的澄清溶液,此方案實驗周期在半個月以上的動物實驗酌情使用。

      1 mL 工作液為例,取 100 μL 25.0 mg/mL 的澄清 DMSO 儲備液加到 900 μL玉米油中,混合均勻。

    動物溶解方案計算器
    請輸入動物實驗的基本信息:

    給藥劑量

    mg/kg

    動物的平均體重

    g

    每只動物的給藥體積

    μL

    動物數(shù)量

    由于實驗過程有損耗,建議您多配一只動物的量
    請輸入您的動物體內(nèi)配方組成:
    %
    DMSO +
    +
    %
    Tween-80 +
    %
    Saline
    如果您的動物是免疫缺陷鼠或者體弱鼠,建議 DMSO 中的在最后工作液體系中的占比盡量不超過 2%。
    方案所需 助溶劑 包括:DMSO ,均可在 MCE 網(wǎng)站選購。 ,Tween 80,均可在 MCE 網(wǎng)站選購。
    計算結(jié)果
    工作液所需濃度 : mg/mL
    儲備液配制方法 : mg 藥物溶于 μL  DMSO(母液濃度為 mg/mL)。
    您所需的儲備液濃度超過該產(chǎn)品的實測溶解度,以下方案僅供參考,如有需要,請與 MCE 中國技術(shù)支持聯(lián)系。
    動物實驗體內(nèi)工作液的配制方法 : 取 μL DMSO 儲備液,加入 μL  μL ,混合均勻至澄清,再加 μL Tween 80,混合均勻至澄清,再加 μL 生理鹽水。
    連續(xù)給藥周期超過半月以上,請謹慎選擇該方案。
    請確保第一步儲備液溶解至澄清狀態(tài),從左到右依次添加助溶劑。您可采用超聲加熱 (超聲清洗儀,建議頻次 20-40 kHz),渦旋吹打等方式輔助溶解。
    純度 & 產(chǎn)品資料

    純度: 99.62%

    參考文獻
    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.

    參考文獻

    Spebrutinib 相關(guān)分類

    完整儲備液配制表

    * 請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液;一旦配成溶液,請分裝保存,避免反復(fù)凍融造成的產(chǎn)品失效。
    儲備液的保存方式和期限:-80°C, 2 years; -20°C, 1 year。-80°C儲存時,請在2年內(nèi)使用, -20°C儲存時,請在1年內(nèi)使用。

    可選溶劑 濃度 溶劑體積 質(zhì)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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