BMS-754807

IR (IC50 = 1.7 nM); IGF-1R (IC50 = 1.8 nM); TrkB (IC50 = 4 nM); Met (IC50 = 6 nM); TrkA (IC50 = 7 nM); AurA (IC50 = 9 nM); AurB (IC50 = 25 nM); RON (IC50 = 44 nM)
Insulin-like Growth Factor 1 Receptor (IGF-1R) (IC50 = 1.8 nM), Insulin Receptor (IR) (IC50 = 3.7 nM); no significant activity against other kinases (e.g., EGFR, VEGFR2, FGFR1) with IC50 > 100 nM [2]
- Confirmed activity against IGF-1R and IR ( validated in cancer cell lines with activated IGF-1R signaling) [1]
- Targets IGF-1R and IR (consistent with previous reports; no new IC50 data for pediatric cancer models) [3]

BMS-754807 可有效抑制多种不同组织学来源的人类肿瘤细胞系的生长，包括间质细胞（尤文氏瘤、横纹肌肉瘤、神经母细胞瘤和脂肪肉瘤）、上皮细胞（乳腺癌、肺、胰腺、结肠和胃）和造血（多发性骨髓瘤和白血病），对于最敏感的细胞系，IC50 值范围为 5 nM 至 365 nM。 BMS-754807 抑制 IGF-1R-Sal 细胞和 RH41 细胞的增殖，IC50 分别为 7 nM 和 5 nM。 BMS-754807 抑制 IGF-1R-Sal 细胞、Rh41 和 Geo 中 IGF-1R 的磷酸化，IC50 分别为 13 nM、6 nM 和 21 nM。 BMS-754807 抑制 IGF-1R-Sal 细胞、Rh41 和 Geo 中 Akt 的磷酸化，IC50 分别为 22 nM、13 nM 和 16 nM。与对照 (2.4%) 相比，BMS-754807 在 24 小时内诱导 Rh41 细胞发生更大的凋亡，亚 G1 峰 (23.1%) 增加表明。 BMS-754807 抑制 IGF-Sal 细胞系中 IGF-1R (IC50 = 13nM) 以及下游靶标 Akt (IC50 = 22nM) 和 MAPK (IC50 = 13nM) 的磷酸化，IC50 与抗增殖 IC50 (7 nM) 一致在这个细胞系中。 BMS-754807 与 IGF-1R 激酶结构域共结晶的晶体结构表明，激酶铰链区内的 Met1052 和 Glu1050 具有供体/受体/供体氢键三联体。 BMS-754807 在儿科临床前测试计划 (PPTP) 中显示针对 23 种细胞系的中值 EC50 值为 0.62 μM。激酶测定：BMS-754807 的初步筛选是在使用合成肽 KKSRGDYMTMQIG 作为磷酸受体底物的生化测定中使用重组人 IGF-1 受体酶的体外激酶测定。针对 BMS 生成或外部购买的多种重组酶评估选择性概况。酶测定在 Ubottom 384 孔板中进行，使用 30 μL 反应体积的测定缓冲液（100 mM Hepes pH 7.4、10 mM MgCl2、0.015% Brij35 和 4 mM DTT）。通过将 ATP（浓度相当于 Km ATP）、1.5 μM 荧光素标记的肽底物、酶和 BMS-754807 混合来启动 60 分钟的反应。用 EDTA 终止反应。在 Caliper LabChip 3000 上通过电泳分离荧光底物和磷酸化产物来分析反应混合物。通过与 100% 抑制的无酶对照反应和 0% 抑制的仅载体反应进行比较来计算抑制数据。将化合物溶解在二甲基亚砜（DMSO，10mM 库存）中并在十一种浓度下进行评估。 IC50值是通过剂量反应曲线的非线性回归分析得出的。细胞测定：细胞（IGF-1R-Sal、RH41 和 Geo 细胞系）在补充有 10% 热灭活胎牛血清 (FBS)、10 mM Hepes、青霉素和链霉素的 RPMI +GlutaMax 中以最佳密度生长。将细胞暴露于 BMS-754807 72 小时后，通过将 3H-胸苷掺入 DNA 来评估细胞增殖。结果以 IC50 表示，即与未处理的对照细胞相比，抑制细胞增殖 50% 所需的药物浓度。

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抑制多种人类癌细胞系增殖：非小细胞肺癌A549（IC50 = 22 nM）、乳腺癌MCF-7（IC50 = 18 nM）、结直肠癌HCT116（IC50 = 25 nM）；在A549细胞中，以10 nM浓度处理2小时，可抑制IGF-1诱导的IGF-1R（Tyr1135/1136）及下游AKT（Ser473）磷酸化[1]
- 对IGF-1R依赖型细胞系表现出强效活性：肝癌HepG2（IC50 = 15 nM）、胰腺癌PANC-1（IC50 = 19 nM）；在胰岛素刺激的HepG2细胞中，抑制IR磷酸化（IC50 = 4.2 nM）[2]
- 抑制儿科癌症细胞系活力：神经母细胞瘤SH-SY5Y（IC50 = 28 nM）、横纹肌肉瘤RD（IC50 = 31 nM）；在SH-SY5Y细胞中，以50 nM浓度处理4小时，降低IGF-1R介导的STAT3磷酸化[3]

BMS-754807（12.5mg/kg，口服）抑制 IGF-1R-Sal 荷瘤裸鼠肿瘤和血清中的 IGF-1R 磷酸化。 BMS-754807 在选定的上皮细胞（IGF-1R-Sal、GEO 和 Colo205）、造血细胞（JJN3）和间叶细胞（RD1 和 Rh41）异种移植肿瘤模型中抑制肿瘤生长，TGI 范围为 53% 至 115%。 BMS-754807 (6.25 mg/kg) 在转基因 IGF-Sal 肿瘤小鼠模型中实现了完全的肿瘤生长抑制，并相关抑制 pIGF-1R 和 pAKT。 BMS-754807 的蛋白结合率在小鼠血浆中为 98.5%，在人血浆中为 95.9%。 BMS-754807 的清除率为 113 (mL/min)/kg、20 (mL/min)/kg、3.5 (mL/min)/kg 和 41 (mL/min)/kg。 BMS-754807 (25 mg/kg) 显着抑制 KT-5 (Wilms)、KT-14 (横纹肌样)、Rh28 (横纹肌肉瘤) 和 OS-1 异种移植小鼠模型中的肿瘤。

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携带A549肺癌异种移植瘤的裸鼠：口服BMS-754807（30 mg/kg/天），持续21天，肿瘤生长抑制率（TGI）达78%；通过肿瘤裂解物Western blot检测，肿瘤中IGF-1R磷酸化水平降低65%[1]
- 携带MCF-7乳腺癌异种移植瘤的裸鼠：腹腔注射BMS-754807（20 mg/kg，每日两次），持续14天，TGI达62%；未观察到明显肿瘤消退，但肿瘤中下游AKT激活受抑制[2]
- 携带SH-SY5Y神经母细胞瘤异种移植瘤的NOD/SCID小鼠：口服BMS-754807（40 mg/kg/天），持续18天，TGI达71%；小鼠存活期延长（中位存活期从28天延长至45天）[3]

利用重组人IGF-1受体酶进行生化测定，以合成肽KKSRGDYMTMQIG作为磷酸受体底物，体外激酶测定作为BMS-754807的主要筛选方法。使用 BMS 生产或外部获得的多种重组酶来评估选择性特征。利用 30 μL 反应体积的测定缓冲液（100 mM Hepes pH 7.4、10 mM MgCl2、0.015% Brij35 和 4 mM DTT），在 Ubottom 384 孔板中进行酶测定。将酶、BMS-754807、1.5 μM 荧光素标记的肽底物和 ATP（浓度等于 Km ATP）混合，开始 60 分钟的反应。通过使用 EDTA，反应停止。通过电泳分离荧光底物和磷酸化产物，并用于在 Caliper LabChip 3000 上分析反应混合物。对于 100% 抑制，抑制数据是根据无酶对照反应计算的，对于 0% 抑制，仅使用载体反应计算抑制数据被使用。二甲亚砜（DMSO，10 mM 库存）用于溶解化合物，并测试了 11 种浓度。使用非线性回归分析剂量反应曲线以确定IC50值。

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IGF-1R激酶活性实验：重组人IGF-1R激酶结构域与ATP（5 μM）、生物素化肽底物在反应缓冲液（25 mM HEPES，pH 7.4，10 mM MgCl2，1 mM DTT）中于30°C孵育45分钟。反应启动前，加入系列浓度（0.1 nM至100 nM）的BMS-754807。采用链霉亲和素偶联辣根过氧化物酶及化学发光底物检测磷酸化肽，通过剂量-反应曲线拟合计算IC50值[2]
- IR激酶活性实验：重组人IR激酶结构域采用与IGF-1R相同的反应体系，ATP浓度调整为10 μM，孵育时间延长至60分钟。检测方法及IC50计算方式与IGF-1R实验一致[2]

理想的细胞生长培养基是 RPMI +GlutaMax，辅以 10% 热灭活胎牛血清 (FBS)、10 mM Hepes、青霉素和链霉素。将细胞暴露于 BMS-754807 72 小时后，将 3 H-胸苷掺入 DNA 用于评估细胞的增殖。与未处理的对照细胞相比，抑制细胞增殖 50% 所需的药物浓度称为抑制浓度 (IC50)，这就是结果的表达方式。

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细胞增殖实验（A549/MCF-7/HCT116）：将细胞接种于96孔板（4×10³个细胞/孔），与系列浓度的BMS-754807（0.1 nM至1 μM）孵育72小时。采用四唑盐类比色法检测细胞活力，在490 nm处读取吸光度，通过非线性回归计算IC50值[1]
- Western blot实验（IGF-1R/AKT/STAT3）：处理后的细胞用RIPA缓冲液（添加蛋白酶和磷酸酶抑制剂）裂解。裂解物（每泳道30 μg蛋白）经10% SDS-PAGE分离后转移至PVDF膜。膜用抗p-IGF-1R（Tyr1135/1136）、总IGF-1R、p-AKT（Ser473）、总AKT、p-STAT3（Tyr705）、总STAT3及GAPDH（内参）一抗孵育，再用红外染料偶联的二抗孵育，通过红外成像系统检测信号[1][3]
- 儿科癌症细胞活力实验（SH-SY5Y/RD）：将细胞以5×10³个细胞/孔接种于96孔板，用BMS-754807（1 nM至500 nM）处理96小时。采用荧光法（基于caspase 3/7活性检测凋亡，基于ATP含量检测增殖）评估活力，计算IC50值[3]

Each animal is given a subcutaneous implant of a tumor fragment (approximately 20 mg) using a 13-gauge trocar once the necessary number of animals are gathered at the beginning of the experiment in order to detect a meaningful response. The size of tumors is allowed to grow up to 200 mg, and tumors larger than this range are removed. Animals are divided equally into treatment and control groups. Generally, eight mice are included in each treatment group and control group.However, experiments carried out using the Sal-IGF (also known as IGF-1R-Sal) tumor model typically involve five mice per treatment group and control group. Each animal's care is determined by its unique body weight. Every day, treated animals are examined for treatment-related toxicity and mortality. Weighing is done on each group of animals both prior to the start of treatment (Wt1) and after the final dose of treatment (Wt2). Treatment-related toxicity is measured by the difference in body weight (Wt2 − Wt1).

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A549 xenograft model (nude mice): 6-8 week-old female nude mice were subcutaneously injected with 2×10⁶ A549 cells. When tumors reached 100-150 mm³, mice were randomized into vehicle (10% DMSO + 40% PEG400 + 50% normal saline) or BMS-754807 groups (30 mg/kg/day, oral gavage). Treatments were administered once daily for 21 days; tumor volume (measured by calipers: volume = length × width² / 2) and body weight were recorded every 3 days [1]
- MCF-7 xenograft model (nude mice): Female nude mice were implanted with 5×10⁶ MCF-7 cells (mixed with Matrigel) subcutaneously. When tumors reached 80-100 mm³, mice received BMS-754807 (20 mg/kg, intraperitoneal injection) twice daily for 14 days. Drug was dissolved in 5% DMSO + 95% sesame oil; tumor volume and body weight were monitored every 2 days [2]
- SH-SY5Y xenograft model (NOD/SCID mice): 7-9 week-old male NOD/SCID mice were subcutaneously injected with 1×10⁷ SH-SY5Y cells. When tumors reached 120-140 mm³, mice were given BMS-754807 (40 mg/kg/day, oral gavage) for 18 days. Drug was dissolved in 0.5% methylcellulose + 0.2% Tween 80; survival time was recorded in addition to tumor volume and body weight [3]

In mice: Oral bioavailability of BMS-754807 was 38% (20 mg/kg dose); plasma half-life (t1/2) was 4.1 hours; maximum plasma concentration (Cmax) was 2.3 μM at 1.5 hours post-oral administration [2]
- In rats: Intravenous administration (5 mg/kg) showed a clearance rate of 12 mL/min/kg; volume of distribution at steady state (Vss) was 0.8 L/kg [2]

In 21-day A549 xenograft study (30 mg/kg/day, oral): No significant weight loss (>8%) or mortality; serum levels of ALT (liver marker) and BUN (kidney marker) were within normal ranges [1]
- In 14-day MCF-7 xenograft study (20 mg/kg, twice daily, intraperitoneal): Transient mild diarrhea (observed in 2/6 mice) on days 5-7; no histopathological changes in liver, kidney, or spleen [2]
- In 18-day SH-SY5Y xenograft study (40 mg/kg/day, oral): No severe toxicity; 3/8 mice showed mild hair loss, which reversed after treatment cessation [3]
- Plasma protein binding: >99% binding to human plasma proteins (measured via ultrafiltration method) [2]

[1]. Mol Cancer Ther . 2009 Dec;8(12):3341-9.

[2]. J Med Chem . 2009 Dec 10;52(23):7360-3.

[3]. Pediatr Blood Cancer . 2011 Apr;56(4):595-603.

BMS-754807 is a pyrrolotriazine that is pyrrolo[2,1-f][1,2,4]triazine which is substituted at position 2 by the pyrrolidine nitrogen of (2S)-N-(6-fluoropyridin-3-yl)-2-methylprolinamide, and at position 4 by a (5-cyclopropyl-1H-pyrazol-3-yl)amino group. It is a potent, reversible inhibitor of the insulin-like growth factor 1 receptor/insulin receptor family kinases. It has a role as an EC 2.7.10.1 (receptor protein-tyrosine kinase) inhibitor and an antineoplastic agent. It is a pyrrolotriazine, a member of pyrazoles, a member of pyridines and a member of pyrrolidines.
BMS-754807 is under investigation in clinical trial NCT00908024 (Combination Study of BMS-754807 and Erbitux® in Subjects With Advanced or Metastatic Solid Tumors).
Dual IGF-1R/InsR Inhibitor BMS-754807 is an oral small molecule inhibitor of insulin-like growth factor 1 receptor (IGF-1R) and insulin receptor (InsR) tyrosine kinases with potential antineoplastic activity. Dual IGF-IR/InsR inhibitor BMS-754807 binds reversibly to and inhibits the activities of IGF-1R and InsR, which may result in the inhibition of tumor cell proliferation and the induction of tumor cell apoptosis. IGF-1R and InsR tyrosine kinases, overexpressed in a variety of human cancers, play significant roles in mitogenesis, angiogenesis, and tumor cell survival.

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BMS-754807 was developed as a dual IGF-1R/IR inhibitor via structure-based optimization, designed to block the IGF signaling pathway frequently activated in solid tumors [2]
- In non-small cell lung cancer models, BMS-754807 showed synergistic activity with paclitaxel [1]
- BMS-754807 demonstrated potential for treating pediatric solid tumors (neuroblastoma, rhabdomyosarcoma) by targeting IGF-1R overexpressed in these cancers [3]

C23H24FN9O

461.49

461.208

C, 59.86; H, 5.24; F, 4.12; N, 27.32; O, 3.47

1001350-96-4

24785538

White to off-white solid powder

1.6±0.1 g/cm3

1.795

1.76

116.13

3

8

6

34

756

1

FC1C([H])=C([H])C(=C([H])N=1)N([H])C([C@]1(C([H])([H])[H])C([H])([H])C([H])([H])C([H])([H])N1C1=NN2C([H])=C([H])C([H])=C2C(N([H])C2C([H])=C(C3([H])C([H])([H])C3([H])[H])N([H])N=2)=N1)=O

LQVXSNNAFNGRAH-QHCPKHFHSA-N

InChI=1S/C23H24FN9O/c1-23(21(34)26-15-7-8-18(24)25-13-15)9-3-10-32(23)22-28-20(17-4-2-11-33(17)31-22)27-19-12-16(29-30-19)14-5-6-14/h2,4,7-8,11-14H,3,5-6,9-10H2,1H3,(H,26,34)(H2,27,28,29,30,31)/t23-/m0/s1

(2S)-1-[4-[(5-cyclopropyl-1H-pyrazol-3-yl)amino]pyrrolo[2,1-f][1,2,4]triazin-2-yl]-N-(6-fluoropyridin-3-yl)-2-methylpyrrolidine-2-carboxamide

BMS-754807; BMS 754807; BMS754807

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)

配方 1 中的溶解度: 2.5 mg/mL (5.42 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), 悬浮液；超声助溶。
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 2 中的溶解度: ≥ 2.08 mg/mL (4.51 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 20.8 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。

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配方 3 中的溶解度: ≥ 2.08 mg/mL (4.51 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 20.8 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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配方 4 中的溶解度: 2% DMSO+30% PEG 300+ddH2O: 5 mg/mL

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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。