NVP-BSK805 2HCl

製品コードS2686 別名:BSK805

NVP-BSK805 2HCl化学構造

分子量(MW):563.47

NVP-BSK805 2HClは一種の有効的で、選択性的で、ATP競争性のJAK2阻害剤で、IC50値が0.5 nMになって、JAK2に作用する選択性はJAK1、JAK3とTYK2に作用する選択性より20倍以上がそれぞれ高くなります。

サイズ 価格(税別)  
JPY 101260.00
JPY 44820.00
JPY 61420.00
JPY 161020.00

カスタマーフィードバック(2)

  • Determination of the kynurenine production in HFF cells after IDO induction by IFN-γ (100 U/mL). The cells were incubated for 72 h under normoxia (20% O2) or hypoxia (1% O2) and treated with different amounts of the JAK2 inhibitor BSK805 (0-2 uM).

    PLoS One 2013 8(5), e63301. NVP-BSK805 2HCl purchased from Selleck.

    HEL cells were treated for 3 hours with the indicated concentrations of NVP-BSK805. NVP-BSK805 inhibits Jak2-V617F mediated signal transduction at nanomolar concentrations in intact cells.

    Dr. Catherine Rolvering and Dr. Claude Haan of Université du Luxembour. NVP-BSK805 2HCl purchased from Selleck.

製品安全説明書

JAK阻害剤の選択性比較

生物活性

製品説明 NVP-BSK805 2HClは一種の有効的で、選択性的で、ATP競争性のJAK2阻害剤で、IC50値が0.5 nMになって、JAK2に作用する選択性はJAK1、JAK3とTYK2に作用する選択性より20倍以上がそれぞれ高くなります。
ターゲット
JAK2 [1]
(Cell-free assay)
TYK2 [1]
(Cell-free assay)
JAK3 [1]
(Cell-free assay)
JAK1 [1]
(Cell-free assay)
~0.5 nM 10.76 nM 18.68 nM 31.63 nM
体外試験

NVP-BSK805 is found to potently inhibit JAK2, whereas displaying more than 20-fold selectivity towards JAK1, JAK3, and TYK2. NVP-BSK805 causes half-maximal inhibition of full-length JAK2V617F and JAK2 wild-type enzymes at 0.5 nM. NVP-BSK805 blocks the growth of JAK2V617F cells (Ba/F3) and induces apoptosis with a GI50 at concentrations <100 nM. As constitutive STAT5 phosphorylation in dependent on JAK2, NVP-BSK805 is found to potently suppress STAT5 phophorylation at ≥ 100 nM concentrations in the JAK2 V617F -mutant cell lines, like MB-02. Incubation of SET-2 cells with 150 nM and 1µM of NVP-BSK805, which corresponds to concentration yielding 75% and 95% growth inhibition, respectively, for 24, 48, and 72 hours lead to concentration- and time- dependent induction of apoptosis. These results are evidenced by the detection of cleaved PARP, reduced Bcl-xL expression, and a strong increase in the number of cells with less than 2N DNA content. [1] NVP-BSK805 triggered cell death requires activation of caspase cascades and is overcome by caspase inhibition in both SET-2 and MB-02 cells. NVP-BSK805 modulates the post-translational modification of Bim and levels of Mcl-1 in JAK2V617F cells, SET-2 and MB-02 cells. [2]

体内試験 Oral bioavailability of NVP-BSK805 in mice is estimated to be 45%, while it is 50% in rats. Oral administration of NVP-BSK805 at 150 mg/kg suppresses STAT5 phosphorylation, splenomegaly, and leukemic cell spreading in a Ba/F3 JAK2V617F cell–driven mouse model. NVP-BSK805 suppresses rhEpo-induced STAT5 phosphorylation as well as rhEpo-mediated polycythemia and splenomegaly in BALB/c mice at doses of 25, 50, and 100 mg/kg orally. [1]

お薦めの試験操作(参考用のみ)

キナーゼ試験:[1]
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Enzymatic Assays:

The human JAK2 kinase domain (amino acids 840-1132) is contained in plasmid construct pAcG2TtevJAK2. The plasmid constructs for JAK3 (813-1124), TYK2 (888-1187), and JAK1(866-1154) are designed. The generation of the recombinant baculoviruses with BD BaculoGoldTM Bright linearized DNA, plaque assay, and virus amplification from single plaques is performed according to the manual (BD Biosciences Pharmingen). Janus kinase domains are expressed in Sf9 cells in 400 mL shake flasks with 100 mL ExCell420 culture medium (JRH Biosciences Ltd) with penicillin/streptomycin solution for 48 h at 27°C. Suspension culture cells are infected at a density of 1 × 106 and the multiplicity of infection (MOI) for each virus is optimized for yield of soluble protein. The kinase domain of human JAK2 and of JAK1, JAK3, and TYK2 is expressed at an MOI of 1 and 0.5, respectively. Time of expression at 27°C is 48 hours for JAK2 and 48 hours or 72 hours for JAK1, JAK3, and TYK2. Forty-eight or seventy-two hours post-infection, the cells from a 100 mL expression culture are harvested by centrifugation at 3000 x g for 5 minutes and lysed with 12 mL lysis buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 2 mM EDTA, 1 mM DTT, 1 mM sodium orthovanadate, 1 % Triton X-100, 10 % glycerol, 1 x EDTA-free complete protease inhibitor cocktail (Roche Diagnostics), and 12.5 U/mL Benzonase for 30 minutes at 4°C, followed by centrifugation at 14,000 × g for 45 minutes to pellet insoluble material. For GST-tag affinity purification of kinase domain proteins all steps are performed at 4°C. The cleared lysates are incubated with 0.2 mL of a 50 % slurry of washed Glutathione Sepharose 4B for 2 hours at 4°C, followed by 5 washes with 1 mL of 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1 % Triton X-100, 1 mM DTT, and 10 % glycerol. Bound protein is eluted in 5 aliquots each starting with a 10 minutes incubation with 0.25 mL elution buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.1 % Triton X-100, 1mM DTT, 10 % glycerol, 10 mM reduced L-glutathione). Eluates are concentrated about 5-fold with Amicon Ultra-4 spin columns. After addition of Brij35 to 0.1 % final concentration the protein is snap frozen in small aliquots and stored at -80 °C. In these conditions, kinase activities are stable for at least 6 months. The JAK kinasedomain enzymes are incubated for 30 minutes at room temperature in a medium containing 0.1 μM [γ33P]-ATP, 1 mM MnCl2, 5 mM MgCl2, 30 μM of synthetic peptide substrate EQEDEPEGDYFEWLE, 1 mM DTT, 1 % DMSO, 50 μg/mL BSA, 0.01 % Brij35, and 50 mM Tris-HCl pH 7.5. The ATP concentration is below the Km for all proteins. Curves are fitted by non-linearregression using the logistic equation and the global fit function of XLfit® (model 205). Expression and characterization of full-length wild type and V617F mutant JAK2 as well as kinase assay conditions have been described elsewhere. Kinase selectivity of NVP-BSK805 is assessed in an internal kinase panel: In the Caliper assays, kinase reactions are carried out with peptide substrates that migrate with different velocities in an electrical field when phosphorylated. The peptides carry a fluorescent label in order to allow the detection and quantification of the peptides in a capillary system. Peptide fluorescence intensities are quantified using the LC3000 instrument (Caliper Life Sciences, Hopkinton, MA, USA). Kinase activity is measured by quantifying the amount of ATP remaining in solution following a kinase reaction. In the LanthaScreen™ TR-FRET kinase assays, terbium is used as the lanthanide chelate combined with an antibody directed against the phosphorylated substrate.
細胞試験: [1]
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  • 細胞株: Ba/F3 cells
  • 濃度: 100 nM
  • 反応時間: 72 hours
  • 実験の流れ: The anti-proliferative activity of JAK2 inhibitors is determined by incubating cells for 72 hours with an 8-point concentration range of compound and cell proliferation relative to DMSO-treated cells is measured using the colorimetric WST-1 (Roche Diagnostics GmbH) cell viability readout. Of each triplicate treatment, the mean is calculated and these data are plotted in XLfit 4 (ID Business Solutions, Ltd.) to determine the half-maximal growth inhibition (GI50) values.
    (参考用のみ)
動物試験:[1]
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  • 動物モデル: RhEpo-induced polycythemia model in Female BALB/c mice
  • 製剤: VP-BSK805 is freshly formulated in MP/PEG300/Solutol HS15 (5/80/15%).
  • 投薬量: 50, 75, and 100 mg/kg
  • 投与方法: Once daily oral administration
    (参考用のみ)

溶解度 (25°C)

体外 DMSO 113 mg/mL (200.54 mM)
Ethanol 15 mg/mL (26.62 mM)
Water 3 mg/mL (5.32 mM)
体内 順序で溶剤を入れること:
30% PEG400+0.5% Tween80+5% propylene glycol
30 mg/mL

* 溶解度検測はSelleck技術部門によって行いますので、文献より提供された溶解度と差異がある可能性がありますが、生産工芸と不同ロット(lot)で起きる正常な現象ですから、ご安心ください。

化学情報

分子量 563.47
化学式

C27H28F2N6O.2HCl

CAS No. 1092499-93-8 (free base)
保管
別名 BSK805

便利ツール

モル濃度計算器

モル濃度計算器

解決のために必要とされるマス、ボリュームまたは濃度を計算してください。

マス (g) = 濃度 (mol/L) x ボリューム (L) x 分子量 (g/mol)

モル濃度計算器方程式

  • マス
    濃度
    ボリューム
    分子量

*貯蔵液を準備するとき、常に、オンであるとわかる製品のバッチに特有の分子量を使って、を通してラベルとMSDS/COA(製品ページで利用可能な)。

希釈計算器

希釈計算器

貯蔵液を準備することを要求される希釈剤を計算してください. セレック希釈計算器は、以下の方程式に基づきます:

開始濃度 x 開始体積 = 最終濃度 x 最終体積

希釈の計算式

この方程式は、一般に略語を使われます:C1V1 = C2V2 ( 輸入 輸出 )

  • C1
    V1
    C2
    V2

常に貯蔵液を準備するとき、小びんラベルとMSDS/COA(オンラインで利用できる)で見つかる製品のバッチに特有の分子量を使ってください。

連続希釈計算器方程式

  • 連続希釈剤

  • 計算結果

  • C1=C0/X C1: LOG(C1):
    C2=C1/X C2: LOG(C2):
    C3=C2/X C3: LOG(C3):
    C4=C3/X C4: LOG(C4):
    C5=C4/X C5: LOG(C5):
    C6=C5/X C6: LOG(C6):
    C7=C6/X C7: LOG(C7):
    C8=C7/X C8: LOG(C8):
分子量計算器

分子量计算器

そのモル質量と元素組成を計算するために、合成物の化学式を入力してください:

総分子量:g/mol

チップス: 化学式は大文字と小文字の区別ができます。C10H16N2O2 c10h16n2o2

モル濃度計算器

マス 濃度 ボリューム 分子量

技術サポート

ストックの作り方、阻害剤の保管する方法、細胞実験や動物実験に注意すべきな点を全部含めており、製品を取扱う時よくあった質問に対して取扱説明書でお答えいたします。

Handling Instructions

他の質問がある場合は、お気軽くお問合せください。

  • * 必須

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細胞株 試験類型 濃度 培養時間 溶剤類型 活性叙述 PMID