c-Kit (IC50 ~100 nM); Bcr-Abl (IC50 ~100 nM); PDGFR (IC50 ~100 nM)

体外活性：抑制一组酪氨酸和丝氨酸/苏氨酸蛋白激酶的体外试验表明，伊马替尼可有效抑制 v-Abl 酪氨酸激酶和 PDGFR，IC50 分别为 0.6 和 0.1 μM。伊马替尼抑制野生型 c-kit 激酶活性的 SLF 依赖性激活，这些作用的 IC50 约为 0.1 μM，与抑制 PDGFR 所需的浓度相似。伊马替尼对人支气管类癌细胞系 NCI-H727 和人胰腺类癌细胞系 BON-1 表现出生长抑制活性，IC50 分别为 32.4 和 32.8 μM。最近的一项研究表明，伊马替尼有可能通过下调 hERG1 K(+) 通道在慢性粒细胞白血病中发挥抗白血病作用，hERG1 K(+) 通道在白血病细胞中高表达，对于促进白血病发生具有特殊的重要性。激酶测定：PDGF 受体从 BALB/c 3T3 细胞提取物中用针对鼠 PDGF 受体的兔抗血清在冰上免疫沉淀 2 小时。 Protein A-Sepharose 珠用于收集抗原-抗体复合物。免疫沉淀物用 TNET（50 mM Tris，pH 7.5，140 mM NaCl，5 mM EDTA，1% Triton X-100）洗涤两次，用 TNE（50 mM Tris，pH 7.5，140 mM EDTA）洗涤一次，用激酶缓冲液（20 mM Tris，pH 7.5，10 mM MgCl2）。在 4°C 下用 PDGF (50 ng/mL) 刺激 10 分钟后，将不同浓度的药物添加到反应混合物中。 PDGF受体激酶活性通过与10μCi[7-33P]-ATP和1μM ATP在4℃下孵育10分钟来测定。通过 7.5% 凝胶上的 SDS-PAGE 分离免疫复合物。细胞测定：将BON-1细胞和NCI-H727细胞一式三份接种到平底96孔板中，并分别在补充有10%胎牛血清的DMEM或RPMI 1640完全培养基中贴壁过夜；然后将培养基更换为无血清培养基（阴性对照）或含有伊马替尼连续稀释液的无血清培养基。 48小时后（对照培养物未达到汇合），通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物测定法测定代谢活跃细胞的数量，并在吸光度测定中测量吸光度。 Packard Spectra 酶标仪，540 nm。使用以下公式计算生长抑制：抑制率=（1−a/b）×100%，其中a和b分别是处理组和对照组的吸光度值。

伊马替尼对源自新鲜人小细胞肺癌手术样本的三种异种移植肿瘤产生不同的抗肿瘤作用，对 SCLC6、SCLC61 和 SCLC108 肿瘤的生长分别有 80%、40% 和 78% 的抑制作用，对 SCLC74 肿瘤的生长没有显着抑制作用。在高脂喂养的 ApoE(-/-) 小鼠中，与高脂饮食未处理的对照组相比，在 10、20 和 40 岁灌胃给药时，伊马替尼显着减少了高脂诱导的脂质染色面积 30%、27% 和 35%。 mg/kg，分别抑制颈动脉脂质积累

使用兔抗血清对 BALB/c 3T3 细胞提取物中的 PDGF 受体进行免疫沉淀，然后将其置于冰上两小时。使用 Protein A-Sepharose 珠收集抗原-抗体复合物。 TNET（50 mM Tris，pH 7.5、140 mM NaCl、5 mM EDTA、1% Triton X-100）、TNE（50 mM Tris，pH 7.5、140 mM EDTA）和激酶缓冲液（20 mM Tris，pH 7.5， 10 mM MgCl2）是用于洗涤免疫沉淀两次的三种溶液。 PDGF (50 ng/mL) 在 4°C 刺激 10 分钟后，将各种药物浓度添加到反应混合物中。

在添加测试化合物前24小时，将测试的A549细胞以每孔5×10³细胞的密度排列在96孔平底板中。除了不同剂量的甲磺酸伊马替尼（10、100、1000 和 10,000 ng/mL）和其他细胞抑制药物（多西他赛 (DTX) 或伊达比星 (ID)：0.1、1、10、100 ng/mL；顺铂 (CIS) ）：1、10、100、1000 ng/mL），将细胞与两种不同浓度（10 和 100 nM）的 PRI-2191 一起孵育 96 小时。磺胺罗丹明 B (SRB) 测定法用于评估细胞毒性作用。因此，Dmitry Nevozhay 软件 Cheburator 0.4 计算每个单独实验的 IC50 [4]。

Mice: We use female NOD/SCID mice that are 12–16 weeks old and weigh 20–25 g. On Day 0, mice receive a subcutaneous (s.c.) inoculation of A549 tumor cells suspension (5×10⁶ cells in 0.2 mL of Hank's medium per mouse). Following this, they are randomly assigned to groups that receive different combinations of vitamin D analogs and chemotherapeutics. In the corresponding experiments, one of the two experimental protocols is used: 1. After the tumor cells are injected, treatment begins on Day 7 (when the tumors become palpable). For 19 days (from Days 7 to 25), imatinib mesylate is given intraperitoneally (i.p.) at a dose of 75 mg/kg/day. PRI-2191 is given orally or s.c. three times a week (on Days 7, 12, 14, 16, 19, 21, and 23) at a dose of 2 μg/kg/day. 2. After tumor cells are injected, treatment begins on Day 7 (when tumors become palpable). For 13 days (from Days 7-19), imatinib mesylate is given intraperitoneally (i.p.) at a dose of 50 mg/kg/day. PRI-2191 and PRI-2205 are given subcutaneously (s.c.) three times a week (on Days 7, 10, 12, 14, 17, 19, 21, 24, and 26) at doses of 1 or 10 μg/kg/day, respectively. Blood is drawn while the mice are sedated at the conclusion of the trials, and they are then killed.
Rats: In the experiments, male Lewis rats weighing between 270 and 320 g are employed. The Imatinib group (n = 7) receives an intraperitoneal injection of Imatinib mesylate (50 mg/kg), while the vehicle group (n = 7) receives 0.5 mL of 20% DMSO without Imatinib. Preliminary testing reveals that the 25 mg/kg dose slightly improves lung function without reaching statistical significance. Based on previous reports and this result, the intraperitoneal administration of 50 mg/kg was chosen. The animals have a left thoracotomy, and a tiny metallic clamp is used to occlude the left hilum. The occlusion is carried out 20 minutes following the administration of imatinib or the vehicle. Tidal volume (TV) and respiratory rate (RR) are set to 8 mL/kg and 80 breaths/min, respectively, during clamping. The clamp is taken off after 90 minutes of ischemia, and reperfusion is sustained for an additional 120 minutes. The bilateral lung's blood flow and ventilation are restored during reperfusion. The animals in the sham group (n=6) undergo 210 minutes of ventilation, thoracotomy, and heparinization.

[1]. Inhibition of c-kit receptor tyrosine kinase activity by STI 571, a selective tyrosine kinase inhibitor. Blood. 2000 Aug 1;96(3):925-32.

[2]. Sorafenib inhibits imatinib-resistant KIT and platelet-derived growth factor receptor beta gatekeeper mutants. Clin Cancer Res. 2007 Jun 1;13(11):3363-9.

[3]. ARG tyrosine kinase activity is inhibited by STI571.Blood. 2001 Apr 15;97(8):2440-8.

[4]. Vitamin D Analogs Potentiate the Antitumor Effect of Imatinib Mesylate in a Human A549 Lung Tumor Model. Int J Mol Sci. 2015 Nov 13;16(11):27191-207.

[5]. Protective Effects of Imatinib on Ischemia/Reperfusion Injury in Rat Lung. Ann Thorac Surg. 2016 Jul 23. pii: S0003-4975(16)30523-9.

[6]. Targeting invadopodia-mediated breast cancer metastasis by using ABL kinase inhibitors. Oncotarget. 2018 Apr 24;9(31):22158-22183.

C30H35N7O4S

589.71

589.2471238

C, 61.10; H, 5.98; N, 16.63; O, 10.85; S, 5.44

220127-57-1

Imatinib;152459-95-5;N-Desmethyl imatinib;404844-02-6

white to off-white to brownish or yellowish tinged crystalline powder

CC1=C(C=C(C=C1)NC(=O)C2=CC=C(C=C2)CN3CCN(CC3)C)NC4=NC=CC(=N4)C5=CN=CC=C5.CS(=O)(=O)O

YLMAHDNUQAMNNX-UHFFFAOYSA-N

InChI=1S/C29H31N7O.CH4O3S/c1-21-5-10-25(18-27(21)34-29-31-13-11-26(33-29)24-4-3-12-30-19-24)32-28(37)23-8-6-22(7-9-23)20-36-16-14-35(2)15-17-36;1-5(2,3)4/h3-13,18-19H,14-17,20H2,1-2H3,(H,32,37)(H,31,33,34);1H3,(H,2,3,4)

methanesulfonic acid;4-[(4-methylpiperazin-1-yl)methyl]-N-[4-methyl-3-[(4-pyridin-3-ylpyrimidin-2-yl)amino]phenyl]benzamide

STI571; CGP-57148B; ST-1571 Mesylate; CGP 57148; CGP57148; CGP-57148; CGP-57148B; CGP57148B; ; STI-571; STI 571; Imatinib mesylate; Brand name: Gleevec (USA); Glivec (other countries)

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.53 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (3.53 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (3.53 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: Saline: 30 mg/mL

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Solubility in Formulation 5: 100 mg/mL (169.57 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。