Cabozantinib is a potent multiple receptor tyrosine kinases (RTKs) inhibitor that inhibits VEGFR2, c-Met, Kit, Axl and Flt3 with IC₅₀s of 0.035, 1.3, 4.6, 7 and 11.3 nM, respectively.

Cabozantinib is a potent inhibitor of MET and VEGFR2 with IC₅₀ values of 1.3 and 0.035 nM, respectively. MET-activating kinase domain mutations Y1248H, D1246N, or K1262R are also inhibited by Cabozantinib (IC₅₀=3.8, 11.8, and 14.6 nM, respectively). Cabozantinib displays strong inhibition of several kinases that have also been implicated in tumor pathobiology, including KIT, RET, AXL, TIE2, and FLT3 (IC₅₀=4.6, 5.2, 7, 14.3, and 11.3 nM, respectively). In cellular assays, Cabozantinib inhibits phosphorylation of MET and VEGFR2, as well as KIT, FLT3, and AXL with IC₅₀ values of 7.8, 1.9, 5.0, 7.5, and 42 μM, respectively. The effect of Cabozantinib on proliferation is evaluated in a number of human tumor cell lines. SNU-5 and Hs746T cells harboring amplified MET are the most sensitive to Cabozantinib (IC₅₀=19 and 9.9 nM, respectively); however, SNU-1 and SNU-16 cells lacking MET amplification are more resistant (IC₅₀=5,223 and 1,149 nM, respectively). MDA-MB-231 and U87MG cells exhibit comparable levels of sensitivity to Cabozantinib (IC₅₀=6,421 and 1,851 nM, respectively), whereas H441, H69, and PC3 cell lines are the least sensitive to Cabozantinib with IC₅₀ values of 21,700, 20,200, and 10,800 nM, respectively. In addition, BaF3 cells expressing human FLT3-ITD, an activating mutation in acute myelogenous leukemia, are sensitive to Cabozantinib (IC₅₀=15 nM) when compared with wild-type BaF3 cells (IC₅₀=9,641 nM)^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Tumor vascularity decreases after Cabozantinib (XL184), with reductions ranging from 67% at 3 mg/kg to 83% at 30 mg/kg for 7 days^[1]. In mouse models, Cabozantinib dramatically alters tumor pathology, resulting in decreased tumor and endothelial cell proliferation coupled with increased apoptosis and dose-dependent inhibition of tumor growth in breast, lung, and glioma tumor models. Importantly, treatment with Cabozantinib does not increase lung tumor burden in an experimental model of metastasis, which has been observed with inhibitors of VEGF signaling that do not target MET. On a body weight dosage basis, Cabozantinib plasma exposures range from 6- to 10-fold higher in rats than in mice, which accounts for lower doses inducing tumor growth inhibition/regression in rats than in mice. Subchronic administration of Cabozantinib is well tolerated in mice and rats with no signs of toxicity, as determined by stable and/or increasing body weights during the treatment period^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

501.51

C₂₈H₂₄FN₃O₅

849217-68-1

卡博替尼

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

DMSO : ≥ 30 mg/mL (59.82 mM)

H₂O : < 0.1 mg/mL (ultrasonic;warming;heat to 60°C) (insoluble)

* “≥” means soluble, but saturation unknown.

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (protect from light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂： 30% polypropylene glycol, 5% Tween-80 and 65% D5W (dextrose 5% water)

  Solubility: 10 mg/mL (19.94 mM); Suspended solution; Need ultrasonic
• 2.

  请依序添加每种溶剂： 0.5% CMC/saline water

  Solubility: 2.5 mg/mL (4.98 mM); Suspended solution; Need ultrasonic
• 3.

  请依序添加每种溶剂： 5% DMSO    95% (20% SBE-β-CD in saline)

  Solubility: ≥ 2.5 mg/mL (4.98 mM); Clear solution
• 4.

  请依序添加每种溶剂： 10% DMSO    40% PEG300    5% Tween-80    45% saline

  Solubility: ≥ 2.08 mg/mL (4.15 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (4.15 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 5.

  请依序添加每种溶剂： 10% DMSO    90% (20% SBE-β-CD in saline)

  Solubility: 2.08 mg/mL (4.15 mM); Suspended solution; Need ultrasonic

  此方案可获得 2.08 mg/mL (4.15 mM) 的均匀悬浊液，悬浊液可用于口服和腹腔注射。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 6.

  请依序添加每种溶剂： 10% DMSO    90% corn oil

  Solubility: ≥ 2.08 mg/mL (4.15 mM); Clear solution

  此方案可获得 ≥ 2.08 mg/mL (4.15 mM，饱和度未知) 的澄清溶液，此方案不适用于实验周期在半个月以上的实验。

  以 1 mL 工作液为例，取 100 μL 20.8 mg/mL 的澄清 DMSO 储备液加到 900 μL玉米油中，混合均匀。

• [1]. You WK, et al. VEGF and c-Met blockade amplify angiogenesis inhibition in pancreatic islet cancer. Cancer Res, 2011, 71(14), 4758-4768.

  [2]. Yakes FM, et al. Cabozantinib (XL184), a novel MET and VEGFR2 inhibitor, simultaneously suppresses metastasis, angiogenesis, and tumor growth. Mol Cancer Ther, 2011, 10(12), 2298-2308.

  [3]. Fuse MA, et al. Combination Therapy With c-Met and Src Inhibitors Induces Caspase-Dependent Apoptosis of Merlin-Deficient Schwann Cells and Suppresses Growth of Schwannoma Cells. Mol Cancer Ther. Mol Cancer Ther. 2017 Nov;16(11):2387-2398.

The inhibition profile of Cabozantinib against a broad panel of 270 human kinases is determined using luciferase-coupled chemiluminescence, ³³P-phosphoryl transfer, or AlphaScreen technology. Recombinant human full-length, glutathione S-transferase tag, or histidine tag fusion proteins are used, and IC₅₀ values are determined by measuring phosphorylation of peptide substrate poly(Glu, Tyr) at ATP concentrations at or below the K_m for each respective kinase. The mechanism of kinase inhibition is evaluated using the AlphaScreen Assay by determining the IC₅₀ values over a range of ATP concentrations^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

The effect of Cabozantinib on proliferation is evaluated in a number of human tumor cell lines, including SNU-5 and Hs746T cells harboring amplified MET, SNU-1 and SNU-16 cells, MDA-MB-231 and U87MG cells, H441, H69, and PC3 cell lines, and BaF3 cells. Cells are seeded in triplicate overnight in media containing 10% FBS. The next day, cells are treated with serial dilutions of Cabozantinib for 48 hours, followed by analysis of proliferation using Cell Proliferation ELISA, BrdUrd^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[1]
RIP-Tag2 mice in a C57BL/6 background are used as the tumor model. RIP-Tag2 mice are 10 weeks old at the onset of treatment unless otherwise indicated. Cabozantinib is suspended at a concentration of 5 mg/mL in sterile saline or water and administered by gavage daily for 7 days. Dose-dependent effects are studied in mice treated by gavage daily for 7 days: XL880 (1, 10, 20, 40 or 60 mg/kg), Cabozantinib (3, 10, 30, 40 or 60 mg/kg), or XL999 (25, 40, 50, 60 or 75 mg/kg). The time course of effects is studied in mice treated with XL880 (40 mg/kg) for 6 hr, 1, 4, 7 or 14 days. Effects of withdrawal are studied in mice treated with XL880 (40 mg/kg) for 7 days followed by no treatment for 0, 2, 7 or 14 days. Each group contain 4-6 mice.
Mice^[2]
Female nu/nu mice are used. H441 cells (3×10⁶) are implanted intradermally into the hind flank and when tumors reach approximately 150 mg, tumor weight is calculated using the formula: (tumor volume=length (mm)×width²(mm²)]/2, mice are randomized (n=5 per group) and orally administered a single 100 mg/kg dose of Cabozantinib or vehicle. Tumors are collected at the indicated time points. Pooled tumor lysates are subjected to immunoprecipitation with anti-MET and Western blotting with anti-phosphotyrosine MET. After blot stripping, total MET is quantitated as a loading control.
Rats^[2]
On day 0 in female Wistar rats, C6 cells (5×10⁶) are inoculated subcutaneously into the hind flank. When the tumors reach approximately 250 mg (3-4 days postimplantation), rats are randomized (n=8 per group) and treated orally once daily for 12 days with Cabozantinib or water vehicle. Cabozantinib administered via oral gavage at 2 mL/kg. Body weights are collected daily, and tumor weights are collected twice weekly. Percentage of tumor growth inhibition/regression values are expressed as follows: 1−[(mean treated tumor weight on the final day−mean tumor weight on day 0)/(mean vehicle tumor weight on the final day−mean tumor weight on day 0)]×100. Statistical analysis of Cabozantinib-treated tumors versus vehicle-treated tumors or versus predose tumors is done by one-way ANOVA with significance defined as P<0.05. Blood is collected 4 hours after the final dose, and plasma is prepared to determine Cabozantinib concentrations.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. You WK, et al. VEGF and c-Met blockade amplify angiogenesis inhibition in pancreatic islet cancer. Cancer Res, 2011, 71(14), 4758-4768.

  [2]. Yakes FM, et al. Cabozantinib (XL184), a novel MET and VEGFR2 inhibitor, simultaneously suppresses metastasis, angiogenesis, and tumor growth. Mol Cancer Ther, 2011, 10(12), 2298-2308.

  [3]. Fuse MA, et al. Combination Therapy With c-Met and Src Inhibitors Induces Caspase-Dependent Apoptosis of Merlin-Deficient Schwann Cells and Suppresses Growth of Schwannoma Cells. Mol Cancer Ther. Mol Cancer Ther. 2017 Nov;16(11):2387-2398.