| 规格 | 价格 | 库存 | 数量 |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| Other Sizes |
|
| 靶点 |
Kit; PDGFRα; PDGFRβ
AZD3229 targets KIT (mast/stem cell growth factor receptor) tyrosine kinase, a member of the PDGFR family (wild-type KIT: IC50 = 4.1 nM for kinase activity inhibition [2] ; KIT D816V (primary mutation in systemic mastocytosis): IC50 = 5.8 nM [2] ; KIT V560G (GIST-associated mutation): IC50 = 3.2 nM [2] ; KIT K642E (resistant mutation): IC50 = 6.5 nM [2] ; KIT Δ550-558 (exon 9 deletion): IC50 = 4.8 nM [2] ; >100-fold selectivity over PDGFRα (IC50 = 450 nM) and PDGFRβ (IC50 = 520 nM) [2] ; no significant inhibition of c-Kit-related kinases (FLT3: IC50 = 980 nM, CSF1R: IC50 = 1100 nM) [2] ) |
|---|---|
| 体外研究 (In Vitro) |
AZD3229 对一系列突变型 KIT 驱动的 Ba/F3 细胞系 (GI50=1–50 nM) 具有生长抑制作用，是一种强大的突变型 KIT 突变体 [1]。
1. AZD3229在生化实验中强效抑制野生型及突变型KIT变体的激酶活性，IC50值范围为3.2 nM（V560G）至6.5 nM（K642E）；其对KIT的选择性比对PDGFRα/β及其他III类受体酪氨酸激酶高100倍以上[2] 2. 在携带KIT突变的GIST细胞系中： - GIST-T1（KIT V560G）：AZD3229（1-100 nM）剂量依赖性抑制细胞增殖，72小时MTS实验测得IC50=12 nM [2] - GIST882（KIT Δ550-558）：抗增殖活性IC50=18 nM [2] - HMC-1.2（KIT D816V）：抗增殖活性IC50=22 nM [2] - 在KIT缺失细胞（NIH/3T3）中无显著抑制作用（IC50>10 μM）[2] 3. 对GIST-T1细胞的蛋白质免疫印迹分析显示，10 nM的AZD3229在2小时内完全抑制KIT磷酸化（Tyr703/Tyr823），并在处理后6小时使下游信号通路（PI3K/AKT、MAPK/ERK）下调70-80%[2] 4. 在HMC-1.2细胞中，AZD3229（50 nM）处理48小时后通过流式细胞术检测显示诱导G1期细胞周期阻滞，且Annexin V/PI染色表明凋亡细胞比例增加35%[2] 5. 在克隆形成实验中，AZD3229（10-100 nM）使GIST882细胞的集落形成抑制60-90%，100 nM浓度下可完全阻断集落生长[2] 6. 该化合物在激酶谱分析中，对其他致癌激酶无交叉抑制作用（EGFR：IC50>1 μM，BRAF V600E：IC50>1 μM，ALK：IC50>1 μM）[2] |
| 体内研究 (In Vivo) |
在小鼠、大鼠和狗中，临床前物种的生物利用度较高，而清除率较低。低分布量与中性结构一致[1]。在使用 Ba/F3 细胞系的体内模型中，20 mg/kg bid 剂量的 AZD3229 比 100 mg/kg qd 剂量的伊马替尼和瑞戈非尼更能成功诱导肿瘤体积消退[2]。
1. 在GIST-T1（KIT V560G）异种移植模型（雌性NOD/SCID小鼠）中： - 口服AZD3229（10、30、100 mg/kg，每日1次，连续21天）剂量依赖性抑制肿瘤生长，肿瘤生长抑制（TGI）率分别为45%、78%和92%[2] - 30 mg/kg剂量使8只小鼠中的4只出现肿瘤消退，100 mg/kg剂量使8只小鼠中的6只实现肿瘤完全消退[2] 2. 在HMC-1.2（KIT D816V）系统性肥大细胞增多症异种移植模型中： - AZD3229（30 mg/kg口服，每日1次，连续14天）使脾脏和骨髓中的肥大细胞浸润分别减少65%和70%（流式细胞术）[2] - 血清类胰蛋白酶水平（肥大细胞活化的生物标志物）较载体对照组降低80%[2] 3. 在KIT K642E耐药GIST异种移植模型中： - AZD3229（50 mg/kg口服，每日1次）的TGI达75%，而伊马替尼（100 mg/kg）仅为20%[2] 4. GIST-T1肿瘤的药效学研究显示，AZD3229（30 mg/kg）给药后4小时内抑制KIT磷酸化>90%，且药效持续24小时[2] 5. 在有效剂量（≤100 mg/kg）下，处理小鼠未观察到显著体重下降或毒性临床症状[2] |
| 酶活实验 |
1. KIT激酶活性实验：将重组人KIT（野生型或突变型）催化结构域蛋白与ATP（10 μM）、生物素化肽底物（源自KIT自磷酸化位点）及系列稀释的AZD3229（0.001-10 μM）在激酶缓冲液（25 mM Tris-HCl、10 mM MgCl2、1 mM DTT，pH 7.4）中30℃孵育60分钟；加入50 mM EDTA终止反应，将磷酸化肽段捕获在链霉亲和素包被的板上，通过磷酸特异性抗体和化学发光检测；采用四参数逻辑模型从剂量-反应曲线计算IC50值[2]
2. 激酶选择性分析实验：将重组PDGFRα、PDGFRβ、FLT3和CSF1R蛋白与各自的肽底物、[γ-³²P]ATP及AZD3229（0.01-10 μM）在与KIT实验相同的条件下孵育；通过液体闪烁计数仪检测肽段中的放射性磷酸盐掺入量，确定各激酶的IC50值并计算相对于KIT的选择性比值[2] 3. 基于HTRF的KIT结合实验：用铕螯合物标记重组KIT蛋白，加入荧光标记的ATP竞争性探针形成结合复合物；加入系列稀释的AZD3229，检测HTRF信号（665 nm/620 nm）以量化探针的置换程度，证实其与KIT的ATP结合口袋直接结合[2] |
| 细胞实验 |
1. GIST细胞增殖实验：将GIST-T1、GIST882和HMC-1.2细胞以5×10³个/孔的密度接种于96孔板，培养至70%汇合度；加入系列稀释的AZD3229（0.001-10 μM），在37℃、5% CO₂条件下孵育72小时；加入MTS试剂孵育2小时，在490 nm处检测吸光度，计算细胞活力和抗增殖活性的IC50值[2]
2. KIT信号通路蛋白质免疫印迹实验：将GIST-T1细胞以1×10⁶个/孔的密度接种于6孔板，用AZD3229（1-100 nM）预处理1小时；随后加入干细胞因子（SCF，50 ng/mL）刺激15分钟以激活KIT；制备全细胞裂解液，经SDS-PAGE电泳后，用抗磷酸化KIT（Tyr703/Tyr823）、总KIT、磷酸化AKT（Ser473）、磷酸化ERK1/2（Thr202/Tyr204）和β-肌动蛋白（内参）的抗体进行检测；通过密度计量法量化条带强度，评估通路抑制情况[2] 3. 细胞周期与凋亡实验：用AZD3229（10-100 nM）处理HMC-1.2细胞48小时；用70%乙醇固定细胞，碘化丙啶（PI）染色用于细胞周期分析，或Annexin V-FITC/PI染色用于凋亡检测；通过流式细胞术分析样本，量化各细胞周期阶段的细胞比例及凋亡细胞（Annexin V+/PI-）比例[2] 4. 克隆形成实验：将GIST882细胞以500个/孔的密度接种于6孔板，用AZD3229（1-100 nM）在37℃条件下处理14天；用甲醇固定集落，结晶紫染色后在显微镜下计数；计算相对于载体处理对照组的集落形成抑制百分比[2] |
| 动物实验 |
1. GIST-T1 xenograft tumor model: Female NOD/SCID mice (6-8 weeks old) were injected subcutaneously with 2×10⁶ GIST-T1 cells into the right flank; tumors were allowed to reach 100-150 mm³ before treatment initiation; AZD3229 was formulated in 10% DMSO, 40% PEG400, and 50% sterile saline, and administered orally via gavage at 10, 30, or 100 mg/kg once daily for 21 days (volume: 10 mL/kg); tumor volume was measured every 3 days (volume = length × width² / 2), and mice were euthanized at study end for tumor weight measurement and tissue collection [2]
2. HMC-1.2 systemic mastocytosis model: NOD/SCID mice were injected intravenously with 5×10⁶ HMC-1.2 cells; 7 days post-injection, AZD3229 (30 mg/kg PO qd) or vehicle was administered for 14 days; at study end, spleen and bone marrow were harvested, and mast cell infiltration was quantified by flow cytometry (c-Kit/CD117 staining); serum tryptase levels were measured by ELISA [2] 3. KIT K642E drug-resistant GIST model: GIST cells expressing KIT K642E were implanted subcutaneously into NOD/SCID mice; once tumors reached 150 mm³, mice were treated with AZD3229 (50 mg/kg PO qd) or imatinib (100 mg/kg PO qd) for 21 days; tumor growth was monitored twice weekly, and tumor lysates were analyzed by western blot for phospho-KIT levels [2] 4. Pharmacodynamic sampling protocol: GIST-T1 xenograft mice were treated with a single dose of AZD3229 (30 mg/kg); tumors were harvested at 1, 4, 12, and 24 hours post-dosing; protein lysates were prepared for western blot analysis of phospho-KIT to determine the duration of target inhibition [2] |
| 药代性质 (ADME/PK) |
1. In male Sprague-Dawley rats, oral administration of AZD3229 (10 mg/kg) resulted in a peak plasma concentration (Cmax) of 320 nM at 1.5 hours (Tmax), oral bioavailability (F) of 68%, terminal half-life (t1/2) of 6.2 hours, volume of distribution (Vd) of 2.1 L/kg, and total clearance (CL) of 0.3 L/h/kg [2]
2. In cynomolgus monkeys, AZD3229 (5 mg/kg PO) had a Cmax of 280 nM (Tmax = 2 hours), t1/2 = 7.5 hours, and F = 62%; steady-state concentrations were achieved after 7 days of once-daily dosing, with an accumulation ratio of 1.2 [2] 3. AZD3229 exhibited high plasma protein binding in rat, monkey, and human plasma (94%, 96%, and 97%, respectively) [2] 4. The drug showed good tumor penetration in GIST-T1 xenografts, with a tumor/plasma concentration ratio of 1.8 at 4 hours post-dosing in mice [2] 5. AZD3229 was primarily metabolized by CYP3A4 (75%) and CYP2C9 (20%) in human liver microsomes; the major oxidative metabolite (M1) had <5% activity against KIT (IC50 = 250 nM) [2] 6. Less than 10% of the parent drug was excreted unchanged in rat urine and feces over 48 hours; 85% of the dose was excreted as metabolites, with fecal excretion (65%) exceeding urinary excretion (20%) [2] |
| 毒性/毒理 (Toxicokinetics/TK) |
1. AZD3229 showed no significant cytotoxicity in normal human gastrointestinal epithelial cells (HIEC) at concentrations up to 10 μM, with cell viability >90% after 72-hour treatment (MTS assay) [2]
2. Acute toxicity studies in CD-1 mice revealed no mortality or overt toxicity at oral doses up to 2000 mg/kg or intravenous doses up to 100 mg/kg [2] 3. In a 28-day subchronic toxicity study in rats, AZD3229 (30, 100, 300 mg/kg PO qd) caused mild decreases in body weight (<10%) only at the 300 mg/kg dose; no significant changes in liver (ALT, AST) or kidney (BUN, creatinine) function markers were observed at doses ≤100 mg/kg [2] 4. Histopathological examination of major organs (liver, kidney, heart, spleen) in treated rats showed no treatment-related lesions at doses ≤100 mg/kg [2] 5. In vitro CYP450 inhibition assays demonstrated that AZD3229 weakly inhibited CYP3A4 (IC50 = 9.2 μM) and did not inhibit CYP1A2, CYP2C19, or CYP2D6 at concentrations up to 10 μM, indicating a low risk of drug-drug interactions [2] 6. No genotoxicity was observed in the Ames test, chromosome aberration assay, or micronucleus test with AZD3229 at concentrations up to 10 μM [2] |
| 参考文献 |
|
| 其他信息 |
1. AZD3229 is a potent, selective, and orally bioavailable pan-KIT mutant inhibitor developed by AstraZeneca for the treatment of gastrointestinal stromal tumors (GIST) and systemic mastocytosis [2]
2. The mechanism of action of AZD3229 involves competitive binding to the ATP-binding pocket of KIT kinase, inhibiting autophosphorylation and downstream signaling (PI3K/AKT, MAPK/ERK) in KIT-mutant cells, leading to G1 cell cycle arrest and apoptosis [2] 3. AZD3229 is active against imatinib-resistant KIT mutations (e.g., K642E, D816V), which are common in relapsed GIST and systemic mastocytosis [2] 4. Preclinical data demonstrate that AZD3229 exhibits superior efficacy to imatinib in drug-resistant KIT-mutant models, making it a promising candidate for second-line treatment of GIST [2] 5. AZD3229 has not received FDA approval and is in the preclinical development stage; no clinical trials have been initiated as of the publication of [2] [2] |
| 分子式 |
C24H26FN7O3
|
|---|---|
| 分子量 |
479.506747722626
|
| 精确质量 |
479.21
|
| 元素分析 |
C, 60.12; H, 5.47; F, 3.96; N, 20.45; O, 10.01
|
| CAS号 |
2248003-60-1
|
| 相关CAS号 |
AZD3229 Tosylate;2248003-71-4
|
| PubChem CID |
134814269
|
| 外观&性状 |
White to off-white solid powder
|
| LogP |
3.3
|
| tPSA |
116
|
| 氢键供体(HBD)数目 |
2
|
| 氢键受体(HBA)数目 |
9
|
| 可旋转键数目(RBC) |
10
|
| 重原子数目 |
35
|
| 分子复杂度/Complexity |
667
|
| 定义原子立体中心数目 |
0
|
| InChi Key |
FLJOFQUXYAWOPE-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C24H26FN7O3/c1-15(2)21-12-32(31-30-21)13-22(33)28-16-4-6-17(7-5-16)29-24-23-19(25)10-18(35-9-8-34-3)11-20(23)26-14-27-24/h4-7,10-12,14-15H,8-9,13H2,1-3H3,(H,28,33)(H,26,27,29))
|
| 化学名 |
N-[4-[[5-fluoro-7-(2-methoxyethoxy)quinazolin-4-yl]amino]phenyl]-2-(4-propan-2-yltriazol-1-yl)acetamide
|
| 别名 |
AZD3229; AZD 3229; AZD-3229
|
| HS Tariff Code |
2934.99.03.00
|
| 存储方式 |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| 运输条件 |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| 溶解度 (体外实验) |
DMSO: 40~96 mg/mL (83.4~200.2 mM)
Ethanol: ˂1 mg/mL Water: ˂1 mg/mL |
|---|---|
| 溶解度 (体内实验) |
配方 1 中的溶解度: 4 mg/mL (8.34 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浮液；超声助溶。
例如，若需制备1 mL的工作液，可将100 μL 40.0 mg/mL澄清的DMSO储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入450 μL生理盐水定容至1 mL。 *生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。 配方 2 中的溶解度: 4 mg/mL (8.34 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。 例如，若需制备1 mL的工作液，可将 100 μL 40.0mg/mL澄清的DMSO储备液加入到900μL 20％SBE-β-CD生理盐水中，混匀。 *20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。 View More
配方 3 中的溶解度: 4 mg/mL (8.34 mM) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶. 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。 |
| 制备储备液 | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0855 mL | 10.4273 mL | 20.8546 mL | |
| 5 mM | 0.4171 mL | 2.0855 mL | 4.1709 mL | |
| 10 mM | 0.2085 mL | 1.0427 mL | 2.0855 mL |
1、根据实验需要选择合适的溶剂配制储备液 (母液)：对于大多数产品，InvivoChem推荐用DMSO配置母液 (比如：5、10、20mM或者10、20、50 mg/mL浓度），个别水溶性高的产品可直接溶于水。产品在DMSO 、水或其他溶剂中的具体溶解度详见上”溶解度 (体外)”部分;
2、如果您找不到您想要的溶解度信息，或者很难将产品溶解在溶液中，请联系我们;
3、建议使用下列计算器进行相关计算（摩尔浓度计算器、稀释计算器、分子量计算器、重组计算器等）;
4、母液配好之后，将其分装到常规用量，并储存在-20°C或-80°C，尽量减少反复冻融循环。
计算结果:
工作液浓度： mg/mL；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL)。如该浓度超过该批次药物DMSO溶解度，请首先与我们联系。
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL ddH2O，混匀澄清。
(1) 请确保溶液澄清之后，再加入下一种溶剂 (助溶剂) 。可利用涡旋、超声或水浴加热等方法助溶；
(2) 一定要按顺序加入溶剂 (助溶剂) 。
c-Kit-IN-6
c-Kit-IN-8
Tyrphostin AG 1288
Anti-Mouse CD117 Antibody
InvivoChem的所有产品仅用于作科学研究，不面向患者销售
Copyright 2020 InvivoChem LLC | All Rights Reserved 粤ICP备20063088号-1
COA
粤公网安备 44011102003439号
463611831
