VEGFR2 (IC50 = 80 nM); PDGFRβ (IC50 = 2 nM)

体外活性：Sunitinib 还有效抑制 Kit 和 FLT-3。舒尼替尼是 VEGFR2 (Flk1) 和 PDGFRβ 的有效 ATP 竞争性抑制剂，Ki 分别为 9 nM 和 8 nM，对 VEGFR2 和 PDGFR 的选择性比 FGFR-1、EGFR、Cdk2、Met、IGFR 高 10 倍以上。 1、Abl 和 src。在表达 VEGFR2 或 PDGFRβ 的血清饥饿 NIH-3T3 细胞中，舒尼替尼抑制 VEGF 依赖性 VEGFR2 磷酸化和 PDGF 依赖性 PDGFRβ 磷酸化，IC50 分别为 10 nM 和 10 nM。 Sunitinib 抑制 VEGF 诱导的血清饥饿 HUVEC 增殖，IC50 为 40 nM，并抑制 PDGF 诱导的过度表达 PDGFRβ 或 PDGFRα 的 NIH-3T3 细胞增殖，IC50 分别为 39 nM 和 69 nM。 Sunitinib 抑制野生型 FLT3、FLT3-ITD 和 FLT3-Asp835 的磷酸化，IC50 分别为 250 nM、50 nM 和 30 nM。 Sunitinib 抑制 MV4;11 和 OC1-AML5 细胞的增殖，IC50 分别为 8 nM 和 14 nM，并以剂量依赖性方式诱导细胞凋亡。激酶测定：舒尼替尼针对 VEGFR2 (Flk-1) 和 PDGFRβ 的 IC50 值是使用含有 RTK 完整胞质结构域的谷胱甘肽 S-转移酶融合蛋白测定的。用于定量 VEGFR2 (Flk-1) 和 PDGFRβ 转磷酸化活性的生化酪氨酸激酶测定在用肽底物聚预涂（20 μg/孔，在 PBS 中；在 4 °C 下孵育过夜）的 96 孔微量滴定板中进行。谷氨酸、酪氨酸 (4:1)。添加 1-5% (w/v) BSA 的 PBS 溶液可封闭多余的蛋白质结合位点。纯化的 GST 融合蛋白在杆状病毒感染的昆虫细胞中产生。然后将 GST-VEGFR2 和 GST-PDGFRβ 添加到含有 2 倍浓度激酶稀释缓冲液的微量滴定孔中，缓冲液由 100 mM HEPES、50 mM NaCl、40 μM NaVO4 和 0.02% (w/v) BSA 组成。 GST-VEGFR2 或 GST-PDGFRβ 的最终酶浓度为 50 ng/mL。随后将 25 μL 稀释的舒尼替尼添加到每个反应孔中，以产生适合每种酶的一系列抑制剂浓度。通过在 MnCl2 溶液中添加不同浓度的 ATP 来启动激酶反应，使得最终 ATP 浓度跨越酶的 Km，并且 MnCl2 的最终浓度为 10 mM。将板在室温下孵育 5-15 分钟，然后添加 EDTA 终止反应。然后用TBST洗涤板3次。将兔多克隆抗磷酸酪氨酸抗血清按 1:10,000 稀释在含有 0.5% (w/v) BSA、0.025% (w/v) 脱脂奶粉和 100 μM NaVO4 的 TBST 中添加到孔中，并在 37° 下孵育 1 小时C。然后用TBST洗涤板3次，然后添加与辣根过氧化物酶缀合的山羊抗兔抗血清（在TBST中1:10,000稀释）。将板在 37°C 下孵育 1 小时，然后用 TBST 洗涤 3 次。添加 2,2'-连氮基-二-[3-乙基苯并噻唑啉磺酸]作为底物后，对每孔中磷酸酪氨酸的量进行定量。细胞测定：在添加舒尼替尼和 FL（50 ng/mL；仅限 FLT3-WT 细胞）之前，将细胞在含有 0.1% FBS 的培养基中饥饿过夜。培养 48 小时后，使用 Alamar Blue 测定或台盼蓝细胞活力测定来测量增殖。添加舒尼替尼 24 小时后，通过蛋白质印迹法检测聚（ADP-核糖）聚合酶 (PARP) 的裂解或 caspase-3 的水平来测量细胞凋亡。

与体内对 VEGFR2 或 PDGFR 磷酸化和信号传导的实质性和选择性抑制一致，舒尼替尼（20-80 mg/kg/天）对包括 HT-29 在内的多种肿瘤异种移植模型表现出广泛且有效的剂量依赖性抗肿瘤活性、A431、Colo205、H-460、SF763T、C6、A375 或 MDA-MB-435。舒尼替尼以 80 毫克/公斤/天的剂量给药 21 天，使八只小鼠中的六只肿瘤完全消退，在治疗结束后 110 天的观察期内肿瘤没有重新生长。舒尼替尼的第二轮治疗对于第一轮治疗期间未完全消退的肿瘤仍然有效。舒尼替尼治疗可显着降低肿瘤 MVD，SF763T 神经胶质瘤肿瘤减少约 40%。 SU11248 治疗可完全抑制表达荧光素酶的 PC-3M 异种移植物的额外肿瘤生长，尽管肿瘤大小没有减小。舒尼替尼治疗（20 mg/kg/天）可显着抑制皮下 MV4;11 (FLT3-ITD) 异种移植物的生长，并延长 FLT3-ITD 骨髓移植模型的存活期。

舒尼替尼针对 PDGFRβ 和 VEGFR2 (Flk-1) 的 IC50 值是通过使用包含整个 RTK 胞质结构域的谷胱甘肽 S-转移酶融合蛋白来确定的。为了测量 VEGFR2 (Flk-1) 和 PDGFRβ 的转磷酸化活性，在已预涂（PBS 中 20 μg/孔）并与肽底物一起孵育的 96 孔微量滴定板中进行生化酪氨酸激酶测定聚-Glu,Tyr (4:1) 在 4 °C 下过夜。在 PBS 中添加 1-5% (w/v) BSA 可阻断多余的蛋白质结合位点。感染杆状病毒的昆虫细胞产生纯化的 GST 融合蛋白。然后在微量滴定孔中填充 2 倍浓度激酶稀释缓冲液中的 GST-VEGFR2 和 GST-PDGFRβ，该缓冲液含有 40 μM NaVO₄、50 mM NaCl、100 mM HEPES 和 0.02%（w/ v) BSA。 50 ng/mL 是 GST-VEGFR2 或 GST-PDGFRβ 的最终酶浓度。为了创建适合每种酶的抑制剂浓度范围，将 25 μL 稀释的舒尼替尼添加到每个反应孔中。将 MnCl₂ 溶液与不同浓度的 ATP 混合以启动激酶反应。 MnCl₂ 的最终浓度为 10 mM，最终 ATP 浓度跨越酶的 Km。让板在室温下静置五到十五分钟后，通过添加 EDTA 停止反应。然后用TBST洗板3次。将兔多克隆抗磷酸酪氨酸抗血清以 1:10,000 稀释度添加到含有 0.025% (w/v) 脱脂奶粉、0.5% (w/v) BSA 和 100 μM NaVO₄ 的 TBST 孔中后，将孔在 37°C 下孵育一小时。 TBST 洗涤 3 次后，用与辣根过氧化物酶缀合的山羊抗兔抗血清（1:10,000 稀释于 TBST）接种平板。 37°C 孵育一小时后，用 TBST 清洗板 3 次。添加 2,2'-连氮基-二-[3-乙基苯并噻唑啉磺酸]作为底物后，即可对每孔中磷酸酪氨酸的量进行定量。

在添加 FL（50 ng/mL；仅 FLT3-WT 细胞）和舒尼替尼之前，将细胞在含有 0.1% FBS 的培养基中饥饿过夜。培养 48 小时后，使用台盼蓝细胞活力测定或 Alamar Blue 测定评估增殖。添加舒尼替尼 24 小时后，使用蛋白质印迹法对细胞凋亡进行定量，以确定 caspase-3 水平或聚（ADP-核糖）聚合酶 (PARP) 裂解。

Mice: The mice used are female nu/nu (8–12 weeks old, 25 g). In short, on day 0, mice receive a subcutaneous injection of 3-5×10⁶ tumor cells into the hind flank region. Once the tumors have grown to the indicated average size, the mice bearing the tumors are treated daily with SU11248 administered orally as a carboxymethyl cellulose suspension or as a citrate buffered (pH 3.5) solution. Tumor growth is assessed using tumor volume measurements taken twice a week. When tumors in animals receiving vehicle treatment reach an average size of 1000 mm³ or are determined to negatively impact the animals' quality of life, studies are usually stopped. Rats: There are forty 200–230 g female Sprague-Dawley rats used. Five to ten animals per group are fed freely. Under 2% isoflurane gas anesthesia, 1×10⁴ Walker 256 cells are injected into the left abdominal mammary fat pad. Rats are weighed every day, and they are gavaged with 30 mg/kg of sunitinib malate or 5 mg/kg of figolimod in olive oil. Calipers are used to measure the tumours. Before the tumors become ulcerated, the animals are put to sleep and killed with an intracardiac injection of ketamine (50 mg/mL). Rats are dissected to look for intestinal, liver, kidney, or lung metastases.

Absorption, Distribution and Excretion
Absorption
Maximum plasma concentrations (Cmax) of sunitinib are generally observed between 6 and 12 hours (Tmax) following oral administration. Food has no effect on the bioavailability of sunitinib. Sunitinib may be taken with or without food. The pharmacokinetics were similar in healthy volunteers and in the solid tumor patient populations tested, including patients with GIST and RCC.

Route of Elimination
Sunitinib is metabolized primarily by the cytochrome P450 enzyme, CYP3A4, to produce its primary active metabolite, which is further metabolized by CYP3A4. Elimination is primarily via feces. In a human mass balance study of [14C]sunitinib, 61% of the dose was eliminated in feces, with renal elimination accounting for 16% of the administered dose.

Volume of Distribution
2230 L (apparent volume of distribution, Vd/F)

Clearance
34 - 62 L/h [Total oral clearance]

Following oral administration, peak plasma concentrations of sunitinib generally occur within 6-12 hours. Food has no effect on bioavailability of sunitinib.

Steady-state concentrations of sunitinib and its primary active metabolite are achieved within 10 to 14 days. By Day 14, combined plasma concentrations of sunitinib and its active metabolite ranged from 62.9 - 101 ng/mL. No significant changes in the pharmacokinetics of sunitinib or the primary active metabolite were observed with repeated daily administration or with repeated cycles in the dosing regimens tested.

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Sunitinib and its primary active metabolite are 95 and 90% bound to human plasma proteins in vitro, respectively.

The apparent volume of distribution (Vd/F) for sunitinib was 2230 L. In the dosing range of 25 - 100 mg, the area under the plasma concentration-time curve (AUC) and Cmax increase proportionately with dose.

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Metabolism / Metabolites
Sunitinib is metabolized primarily by the cytochrome P450 enzyme, CYP3A4, to produce its primary active metabolite, which is further metabolized by CYP3A4.

Sunitinib is metabolized principally by cytochrome P-450 (CYP) isoenzyme 3A4 to several metabolites. The main circulating metabolite, an N-desethyl derivative, has been shown to be equipotent to sunitinib in biochemical and cellular assays; this metabolite accounts for approximately 23-37% of total plasma concentrations of the drug and also is metabolized by CYP3A4.

Sunitinib and its primary active metabolite were the major drug-related compounds identified in plasma, urine, and feces, representing 91.5%, 86.4% and 73.8% of radioactivity in pooled samples, respectively.

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Biological Half-Life
Following administration of a single oral dose in healthy volunteers, the terminal half-lives of sunitinib and its primary active metabolite are approximately 40 to 60 hours and 80 to 110 hours, respectively.
Following oral administration of a single dose in healthy volunteers, the terminal half-life of sunitinib or its primary active metabolite is approximately 40-60 or 80-110 hours, respectively.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of sunitinib during breastfeeding. Because sunitinib and its metabolite are over 90% bound to plasma proteins, the amount in milk is likely to be low. However, one of its metabolites has a half-life of up to 110 hours, and might accumulate in the infant. The manufacturer recommends that breastfeeding be discontinued during sunitinib therapy and for at least 4 weeks after the last dose.
◉ Effects in Breastfed Infants
Relevant published information was not found as of the revision date.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

[1]. J Med Chem . 2003 Mar 27;46(7):1116-9.

[2]. Clin Cancer Res . 2003 Jan;9(1):327-37.

[3]. Blood . 2003 May 1;101(9):3597-605.

[4]. Mol Cancer Ther . 2003 Oct;2(10):1011-21.

[5]. Blood . 2004 Dec 15;104(13):4202-9.

[6]. Mol Cancer Ther . 2006 Oct;5(10):2522-30.

[7]. EMBO J . 2011 Mar 2;30(5):894-905.

Sunitinib Malate is the orally bioavailable malate salt of an indolinone-based tyrosine kinase inhibitor with potential antineoplastic activity. Sunitinib blocks the tyrosine kinase activities of vascular endothelial growth factor receptor 2 (VEGFR2), platelet-derived growth factor receptor b (PDGFRb), and c-kit, thereby inhibiting angiogenesis and cell proliferation. This agent also inhibits the phosphorylation of Fms-related tyrosine kinase 3 (FLT3), another receptor tyrosine kinase expressed by some leukemic cells.
An indole and pyrrole derivative that inhibits VEGFR-2 and PDGFR BETA RECEPTOR TYROSINE KINASES. It is used as an antineoplastic agent for the treatment of GASTROINTESTINAL STROMAL TUMORS, and for treatment of advanced or metastatic RENAL CELL CARCINOMA.
See also: Sunitinib (has active moiety).

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Drug Indication
Gastrointestinal stromal tumour (GIST)Sutent is indicated for the treatment of unresectable and/or metastatic malignant gastrointestinal stromal tumour (GIST) in adults after failure of imatinib mesilate treatment due to resistance or intolerance. Metastatic renal cell carcinoma (MRCC)Sutent is indicated for the treatment of advanced/metastatic renal cell carcinoma (MRCC) in adults. Pancreatic neuroendocrine tumours (pNET)Sutent is indicated for the treatment of unresectable or metastatic, well-differentiated pancreatic neuroendocrine tumours with disease progression in adults. Experience with Sutent as first-line treatment is limited (see section 5. 1).

C26H33FN4O7

532.56

532.233

C, 58.64; H, 6.25; F, 3.57; N, 10.52; O, 21.03

341031-54-7

Sunitinib;557795-19-4; Sunitinib Malate;341031-54-7;Sunitinib-d10;1126721-82-1;Sunitinib-d4;1126721-79-6; 342641-94-5; 1275588-72-1 (mesylate) ; 1126641-10-8; 1327155-72-5 (HCl); 1221149-36-5 (acetate); 1332306-95-2 (oxalate)

6456015

Yellow solid powder

1.3600 g/mL at 25 °C(lit.)

156 °C(lit.)

189-191°C

163 °F

n20/D 1.455(lit.)

2.77

172.06

6

9

10

38

765

1

FC1C([H])=C([H])C2=C(C=1[H])/C(/C(N2[H])=O)=C(\[H])/C1=C(C([H])([H])[H])C(C(N([H])C([H])([H])C([H])([H])N(C([H])([H])C([H])([H])[H])C([H])([H])C([H])([H])[H])=O)=C(C([H])([H])[H])N1[H].O([H])[C@]([H])(C(=O)O[H])C([H])([H])C(=O)O[H]

LBWFXVZLPYTWQI-IPOVEDGCSA-N

InChI=1S/C22H27FN4O2.C4H6O5/c1-5-27(6-2)10-9-24-22(29)20-13(3)19(25-14(20)4)12-17-16-11-15(23)7-8-18(16)26-21(17)28;5-2(4(8)9)1-3(6)7/h7-8,11-12,25H,5-6,9-10H2,1-4H3,(H,24,29)(H,26,28);2,5H,1H2,(H,6,7)(H,8,9)/b17-12-;/t;2-/m.0/s1

N-[2-(diethylamino)ethyl]-5-[(Z)-(5-fluoro-2-oxo-1H-indol-3-ylidene)methyl]-2,4-dimethyl-1H-pyrrole-3-carboxamide;(2S)-2-hydroxybutanedioic acid

sunitinib; SU-11248; SU 11248; Sutent; SU011248 L-malate salt; PHA-290940AD; sunitinib L-malate; Sunitinib malate [USAN]; SU010398; SU11248; SU011248; trade name: Sutent

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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 (4.69 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 (4.69 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 (4.69 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 储备液添加到 900 μL 玉米油中并混合均匀。

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配方 4 中的溶解度: 4% DMSO+30% PEG 300+ddH2O: 2mg/mL

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配方 5 中的溶解度: 10 mg/mL (18.78 mM) in 100 mM citrate buffer (这些助溶剂从左到右依次添加，逐一添加), 悬浮液； 需要超声波并用HCl调节pH至5。

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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。