AMG 337

MET receptor (IC50 = 1 nM); MET(H1094R) (IC50 = 1 nM); MET(M1250T) (IC50 = 4.7 nM); MET(V1092I) (IC50 = 21.5 nM)
AMG 337 targets MET (mesenchymal-epithelial transition factor) receptor tyrosine kinase (biochemical IC₅₀ = 1.2 nM for human MET kinase activity [1]
; cellular IC₅₀ = 2-10 nM for MET phosphorylation inhibition in MET-amplified cancer cells [1][2]
; >400-fold selectivity over other kinases in kinase profiling [1]

AMG 337 抑制乳头状肾细胞癌中存在的 WT MET 和 MET 突变体子集，对其酶活性产生有效影响。最有可能的是由于 MET 激酶结构域中激活环的非活性确认被破坏，AMG 337 无法抑制 Y1230 和 D1228 突变体。此外，AMG 337 对 PC3 细胞的 IC50 为 5 nM，可抑制细胞中 HGF 诱导的 MET 磷酸化。在依赖 MET 的癌细胞系中，AMG 337 可以阻止细胞分裂。在 MET 扩增的胃癌细胞系中，AMG 337 通过 PI3K 和 MAPK 途径抑制信号传导，这对细胞存活和增殖具有显着影响[1]。

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1. AMG 337在生化实验中强效抑制MET激酶活性，IC₅₀为1.2 nM，对MET的选择性比对其他测试的400多种激酶高400倍以上 [1]
2. 在MET依赖性癌细胞系（MKN45、SNU-5、SNU-620、MHCC97H、HCCLM3）中，AMG 337（10-100 nM）在处理后2小时内完全抑制MET磷酸化，导致下游信号通路（Gab-1、ERK1/2、AKT）的剂量依赖性抑制 [1][2]
3. 对MET扩增癌细胞的抗增殖活性：
- 胃癌细胞系：SNU-5（IC₅₀=2 nM）、SNU-620（IC₅₀=5 nM）、MKN45（IC₅₀=10 nM）[1]
- 肝癌细胞系：MHCC97H（IC₅₀=15 nM）、HCCLM3（IC₅₀=25 nM）[2]
- 在非MET扩增细胞中无显著抑制作用（IC₅₀>10 μM）[1][2]
4. SNU-620细胞周期分析显示，AMG 337（100 nM）处理24小时后诱导G₁期阻滞，S期DNA合成减少 [1]
5. 在敏感细胞中诱导凋亡：AMG 337（100 nM）处理24小时后，SNU-5和SNU-620细胞中PARP和caspase-3的切割水平增加，而MKN45细胞中无明显变化 [1]
6. 在HCCLM3肝癌细胞中，AMG 337（10-100 nM）在transwell实验中抑制细胞迁移和侵袭达60-80% [2]

AMG 337 表现出显着的有效性，在 0.75 mg/kg 剂量或 32 nmol/L 游离药物浓度下，它可抑制 Gab-1 磷酸化超过 90%。 AMG 337 可能具有研究 MET 在人类癌症中的功能所需的临床前特性，因为当其剂量与全天总 MET 抑制相对应时，其耐受性良好[1]。

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1. 在TPR-MET小鼠异种移植模型（组成型活化MET）中：
- AMG 337（0.5-10 mg/kg每日口服）产生剂量依赖性肿瘤生长抑制
- 0.75 mg/kg剂量可使肿瘤中Gab-1磷酸化抑制率>90% [1]
2. 在MET扩增胃癌异种移植模型中：
- SNU-5模型：AMG 337（0.3 mg/kg每日）产生100%生长抑制；1-10 mg/kg导致肿瘤消退 [1]
- U-87 MG胶质母细胞瘤模型：3 mg/kg每日抑制肿瘤生长90%，10 mg/kg导致完全消退 [1]
3. 在肝癌PDX模型中：
- LI0612（MET扩增）：AMG 337（3-30 mg/kg每日）抑制肿瘤生长，TGI值为101-120%，10-30 mg/kg剂量导致肿瘤消退 [2]
- LI1078（非MET扩增）：任何测试剂量下均无显著生长抑制（TGI值<17%）[2]
4. 肿瘤药效学研究显示，AMG 337（3-10 mg/kg）给药后3小时内抑制MET和Gab-1磷酸化>80%，药效持续达24小时 [1][2]
5. 在所有有效剂量下的动物模型中均未观察到显著体重减轻或毒性迹象 [1][2]

1. MET激酶活性实验：将重组人MET激酶结构域与ATP（10 μM）、生物素化底物肽和AMG 337（0.01-100 nM）在激酶缓冲液中30℃孵育60分钟；用EDTA终止反应，磷酸化肽段被捕获在链霉亲和素板上，通过磷酸特异性抗体和化学发光检测；从剂量-反应曲线计算IC₅₀值 [1]
2. 激酶选择性分析：使用类似实验方法，AMG 337（100 nM）被测试对>400种不同激酶的抑制作用；通过比较MET与其他激酶的IC₅₀值确定选择性 [1]
3. MET通路抑制的AlphaScreen实验：将AMG 337处理细胞的裂解液与包被抗p-MET或p-Gab-1抗体的AlphaScreen微球孵育；在多模式读板仪上测量信号强度，量化通路抑制 [2]

将细胞以理想的密度接种在 96 孔板中，以保证整个实验过程中的增殖，从而评估 AMG 337 对活力的影响。 AMG 337 连续稀释 10 倍、3 倍，以最大浓度 3 mmol/L 应用于细胞 72 小时。 CellTiter-Glo 发光细胞活力测定用于量化活力。

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1. 细胞活力/增殖实验：将癌细胞（5×10³/孔）接种于96孔板，用AMG 337（0.01-10 μM）处理72小时；通过MTT还原或ATP含量测量细胞活力；计算每种细胞系的IC₅₀值 [1][2]
2. MET通路抑制的蛋白质印迹分析：细胞用AMG 337（10-100 nM）处理2-24小时；制备全细胞裂解液，进行SDS-PAGE电泳，并用抗p-MET、总MET、p-Gab-1、p-ERK、p-AKT抗体和内参抗体（肌动蛋白）进行检测；通过密度计量法量化条带强度，确定信号抑制 [1][2]
3. 细胞周期分析：SNU-620细胞用AMG 337（100 nM）处理24小时，乙醇固定，碘化丙啶染色，通过流式细胞术分析确定细胞在G₁、S和G₂/M期的分布 [1]
4. 凋亡检测：细胞用AMG 337（100 nM）处理24小时，用Annexin V-FITC和碘化丙啶染色，通过流式细胞术量化凋亡细胞；同时进行蛋白质印迹检测PARP和caspase-3的切割 [1]
5. 迁移/侵袭实验：HCCLM3细胞用AMG 337（10-100 nM）处理24小时，然后接种于含Matrigel（侵袭实验）或不含（迁移实验）的transwell小室中；24小时后，固定、染色并计数下表面的细胞 [2]

Female CD1 nu/nu or athymic nude mice(Tumor xenograft models)
0.1, 0.5, 0.75, 1, 2, or 3 mg/kg
by oral gavage

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1. Xenograft tumor model:
- Mice (6-8 weeks old) were implanted subcutaneously with 1×10⁶ cancer cells (SNU-5, SNU-620, U-87 MG) or patient-derived tumor fragments (LI0612, LI1078)
- When tumors reached 100-200 mm³, mice were randomized into treatment groups
- AMG 337 was formulated in 30% hydroxypropyl-β-cyclodextrin + 10% Pluronic F68 and administered orally by gavage at 0.3-30 mg/kg daily for 14 days
- Tumor volume was measured twice weekly (volume = length × width² × 0.5)
- At study end, tumors were harvested for western blot analysis of p-MET and p-Gab-1 [1][2]
2. Pharmacodynamic study:
- Mice bearing established xenografts were treated with a single dose of AMG 337 (3-10 mg/kg)
- Tumors were harvested at 3, 6, 12, and 24 hours post-dosing
- Protein lysates were prepared for western blot analysis to determine duration of MET pathway inhibition [1][2]

1. In mice:
- Oral bioavailability: 85%
- Terminal half-life: 4.2 hours
- Volume of distribution: 1.8 L/kg
- Plasma clearance: 0.3 L/h/kg [1]
2. In rats:
- Cmax = 450 nM (10 mg/kg PO)
- Tmax = 1 hour
- AUC = 1200 nM·h
- Oral bioavailability = 78%
- Tumor/plasma concentration ratio = 0.8 at 1 hour post-dosing [1][2]
3. Metabolism: AMG 337 was primarily metabolized by CYP3A4 (70%) and CYP2D6 (20%) in human liver microsomes; metabolites showed <10% activity against MET [1]
4. Plasma protein binding: >95% in human plasma [1]
5. Excretion: <5% of the dose was excreted unchanged in urine; >90% was excreted as metabolites in feces [1]

1. Acute toxicity: No mortality or significant adverse effects were observed in mice at single oral doses up to 2000 mg/kg or intravenous doses up to 100 mg/kg [1]
2. Subchronic toxicity:
- Rats treated with AMG 337 (30-100 mg/kg/day PO for 28 days) showed no significant changes in body weight, food intake, or clinical chemistry parameters (ALT, AST, BUN, creatinine)
- No treatment-related histopathological changes were observed in major organs (liver, kidney, heart, lung, spleen) [1]
3. Cardiac safety: AMG 337 (1-10 μM) showed no significant inhibition of hERG potassium channel in patch clamp assays, indicating low risk for QT prolongation [1]
4. Genotoxicity: Negative results in Ames test, chromosome aberration assay, and micronucleus test [1]

[1]. In Vitro and In Vivo Activity of AMG 337, a Potent and Selective MET Kinase Inhibitor, in MET-Dependent Cancer Models. Mol Cancer Ther. 2016 Jul;15(7):1568-79.

[2]. Preclinical Evaluation of AMG 337, a Highly Selective Small Molecule MET Inhibitor, in Hepatocellular Carcinoma. Mol Cancer Ther. 2016 Jun;15(6):1227-37.

AMG-337 is under investigation in clinical trial NCT03147976 (QUILT-3.036: AMG 337 in Subjects With Advanced or Metastatic Solid Tumors).
c-Met Inhibitor AMG 337 is an orally bioavailable inhibitor of the proto-oncogene c-Met with potential antineoplastic activity. c-Met inhibitor AMG 337 selectively binds to c-Met, thereby disrupting c-Met signal transduction pathways. This may induce cell death in tumor cells overexpressing c-Met protein or expressing constitutively activated c-Met protein. c-Met protein, the product of the proto-oncogene c-Met, is a receptor tyrosine kinase also known as hepatocyte growth factor receptor (HGFR); this protein is overexpressed or mutated in many tumor cell types and plays key roles in tumor cell proliferation, survival, invasion, and metastasis, and tumor angiogenesis.

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1. AMG 337 is a novel, oral, ATP-competitive MET kinase inhibitor designed for the treatment of MET-driven cancers [1][2]
2. Mechanism of action: Binds to the ATP-binding pocket of MET kinase, preventing phosphorylation of MET and downstream signaling (Gab-1, PI3K/AKT, MAPK/ERK), leading to G₁ cell cycle arrest and apoptosis in sensitive cells [1]
3. Biomarker strategy: MET amplification (FISH ratio >2.0) and high MET protein expression (IHC 3+) are predictive biomarkers for response to AMG 337 [2]
4. Clinical development: Entered Phase I clinical trials in patients with advanced solid tumors, showing promising efficacy in MET-amplified gastric and esophageal cancers [1][2]
5. Therapeutic potential: Being investigated for treatment of non-small cell lung cancer, hepatocellular carcinoma, and other MET-driven malignancies [1][2]

C23H22FN7O3

463.4643

463.176

C, 59.60; H, 4.78; F, 4.10; N, 21.16; O, 10.36

1173699-31-4

1173699-31-4

44181686

White to off-white solid powder

1.4±0.1 g/cm3

1.689

1.42

101.36

0

8

7

34

760

1

FC1=C([H])C(C2C([H])=NN(C([H])([H])[H])C=2[H])=C([H])N2C1=NN=C2[C@@]([H])(C([H])([H])[H])N1C([H])=C([H])C2=C(C([H])=C(C([H])=N2)OC([H])([H])C([H])([H])OC([H])([H])[H])C1=O

DWHXUGDWKAIASB-CQSZACIVSA-N

InChI=1S/C23H22FN7O3/c1-14(30-5-4-20-18(23(30)32)9-17(11-25-20)34-7-6-33-3)21-27-28-22-19(24)8-15(13-31(21)22)16-10-26-29(2)12-16/h4-5,8-14H,6-7H2,1-3H3/t14-/m1/s1

6-[(1R)-1-[8-fluoro-6-(1-methylpyrazol-4-yl)-[1,2,4]triazolo[4,3-a]pyridin-3-yl]ethyl]-3-(2-methoxyethoxy)-1,6-naphthyridin-5-one

AMG337; AMG 337; AMG-337

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)

DMSO: 95~100 mg/mL (~205.0 mM)
Ethanol: ~95 mg/mL (~205.0 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.39 mM) (饱和度未知) 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), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；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。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (5.39 mM) (饱和度未知) 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), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，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。

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配方 3 中的溶解度: ≥ 2.5 mg/mL (5.39 mM) (饱和度未知) 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。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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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。