| 名稱 | Ibrutinib |
| 描述 | Ibrutinib (PCI-32765) is a Bruton's tyrosine kinase (BTK) inhibitor (IC50=0.5 nM) with irreversible and selective properties. Ibrutinib blocks BTK to inhibit the proliferation and survival of B cells, and possesses antitumor activity, which can be used for the treatment of chronic lymphocytic leukemia, among others. |
| 細(xì)胞實(shí)驗(yàn) | CD20+ B and CD3+ T cells were purified by negative selection (RosetteSep, >90% purity) from buffy coat PBMCs and viably frozen in 10% DMSO. Cells were thawed at 37 °C and maintained in growth media (RPMI media containing 10% FCS). B cells were stimulated with goat anti-human IgM F(ab′)2 (10 μg/mL) and T cells were stimulated with anti-CD3/CD28 coated beads at a 1:1 bead/cell ratio. Cells were stained with PE-CD69 and analyzed by flow cytometry, gating on viable lymphocytes. PCI-32765 at concentrations lower than 10 μM did not decrease B- or T-cell viability during the course of the experiment, although PCI-32765 did block the modest survival benefit of anti-IgM stimulation in B cells. For washout experiments, cells were rinsed three times in 10 volumes of growth media, a protocol that was confirmed to completely wash away inhibition of BCR signaling by PCI-29732, a reversible Btk inhibitor [1]. |
| 激酶實(shí)驗(yàn) | In vitro kinase IC50s were measured using 33P filtration binding assay after 1 h incubation of kinase, 33P-ATP, inhibitor, and substrate [0.2 mg/mL poly(EY)(4:1]. Assays were performed at Reaction Biology [1]. |
| 動物實(shí)驗(yàn) | ale DBA/1 mice were immunized with type II collagen plus Freund adjuvant and boosted 21 d later. On a rolling basis, as significant swelling appeared in at least one paw, mice were enrolled and randomized. PCI-32765 or dexamethasone (0.2 mg/kg) was administered orally once per day for 11 d. Arthritis scores (0–5) were assigned to the mice based on the degree and extent of paw swelling. Mouse anti-type II collagen antibody and total IgG levels were measured by ELISA. Female MRL/MpJ-Faslpr mice received PCI-32765 by oral gavage once per day from week 8 through week 20. Proteinuria was monitored weekly. At week 20, serum was collected and analyzed for BUN and mouse anti-dsDNA antibody levels. Kidney histology was scored according to established criteria (26). No drug-induced weight loss was observed at any of the dose levels tested. These studies were carried out at Boulder Biopath according to approved animal care protocols. Results are presented as the mean ± SEM. Statistical significance between groups were evaluated with repeated measures one-way ANOVA or one-way ANOVA using GraphPad Prism with Tukey or Bonferroni multicomparison posttest [1]. |
| 體外活性 | 方法：人 B 細(xì)胞淋巴瘤細(xì)胞 DOHH2 用 Ibrutinib (0.00064-2 μM) 孵育 1 h，再用 anti-IgG F(ab′)2 (30 μg/mL) 刺激 2 min，使用 Western Blot 檢測靶點(diǎn)蛋白表達(dá)水平。
結(jié)果：Ibrutinib 抑制 Btk 的自體磷酸化 (IC50=11 nM)，Btk 的生理底物 PLCγ 的磷酸化 (IC50=29 nM)，和進(jìn)一步的下游激酶 ERK 的磷酸化 (IC50=13 nM)。[1]
方法：原代人類B淋巴細(xì)胞用 Ibrutinib (1-1000 nM) 處理 30 min，然后用 anti-IgM F(ab')2 (10 μg/mL)、anti-CD3/CD28 (5 μg/mL) 或 PMA (0.5 μg/mL) 刺激細(xì)胞 72 h，使用 Cell Titer Glo reagent 檢測細(xì)胞增殖。
結(jié)果：Ibrutinib 劑量依賴性地抑制抗 IgM 刺激的 B 淋巴細(xì)胞增殖 (IC50=8 nM)，但不抑制 PMA 刺激的增殖，PMA 激活PKC途徑。[2] |
| 體內(nèi)活性 | 方法：為檢測體內(nèi)抗炎活性，將 Ibrutinib (3.125-50 ?mg/kg) 灌胃給藥給關(guān)節(jié)炎 DBA/1 小鼠，每天一次，持續(xù)十一天。
結(jié)果：在所有劑量下治療的小鼠中觀察到臨床關(guān)節(jié)炎評分的顯著抑制。在分別以 3.125 和 12.5 mg/kg/天 的劑量治療 9 至 11 天后，疾病的臨床癥狀出現(xiàn)部分和幾乎完全消除。與體內(nèi)抑制 B 細(xì)胞活化一致，抗膠原自身抗體的產(chǎn)生顯著減少，總 IgG 水平適度降低。[1] |
| 存儲條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | 10% DMSO+40% PEG300+5% Tween 80+45% Saline : 8.2 mg/mL (18.62 mM), Suspension. Please add co-solvents sequentially, clarifying the solution as much as possible before adding the next one. Dissolve by heating and/or sonication if necessary. Working solution is recommended to be prepared and used immediately.
Ethanol : < 1 mg/mL (insoluble or slightly soluble)
DMSO : 45 mg/mL (102.16 mM)
H2O : < 1 mg/mL (insoluble or slightly soluble)
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| 關(guān)鍵字 | inhibit | PCI 32765 | Ligands for Target Protein for PROTAC | Target Protein-binding Moiety | PCI32765 | Inhibitor | Btk | Ibrutinib | Bruton tyrosine kinase |
| 相關(guān)產(chǎn)品 | Pelitinib | Deucravacitinib | Gefitinib | Vemurafenib | Ruxolitinib | (+)-JQ-1 | Staurosporine | Nintedanib | Dasatinib | Baricitinib | GSK 3 Inhibitor IX | Delgocitinib |
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