VEGFR1 (IC50 = 13 nM); VEGFR2 (IC50 = 4.2 nM); VEGFR3 (IC50 = 46 nM); PDGFRβ (IC50 = 22 nM); Braf (IC50 = 28 nM); VEGFR2 (BRafV600E = 19 nM); Raf-1 (IC50 = 2.5 nM)

Regorafenib 有效抑制 NIH-3T3/VEGFR2 细胞中的 VEGFR2 自磷酸化，IC50 为 3 nM。 Regorafenib 的 IC50 为 90 nM，在 PDGF-BB 刺激后阻断 HAoSMC 中的 PDGFR-β 自磷酸化。罗戈非尼抑制 vegf165 刺激的 HUVEC 增殖，IC50 为 3 nM[1]。瑞戈非尼的 IC50 为 5 μM，以浓度依赖性方式抑制 Hep3B 细胞的生长。 JNK 靶向磷酸化 c-Jun，但不是总 c-Jun，随后在 Hep3B 细胞中被调节剂阿非尼上调[3]。

瑞戈非尼在 10 至 100 mg/kg 剂量下可有效减缓 Colo-205 异种移植物的生长，10 mg/kg 剂量下第 14 天的 TGI 为 75%。瑞戈非尼在 MDA-MB-231 模型中，剂量低至 3 mg/kg 时非常有效，产生 81% 的显着 TGI，在剂量为 10 和 30 mg/kg 时，TGI 上升至 93%，达到肿瘤停滞[1]。

体外测试中使用重组 VEGFR2（鼠类 aa785–aa1367）、VEGFR3（鼠类 aa818–aa1363）、PDGFRβ（aa561–aa1106）、Raf-1（aa305–aa648）和 BRafV600E（aa409–aa765）激酶结构域。在恒定的 1 M 瑞戈非尼浓度下，进行初始体外激酶抑制分析。选择响应激酶，例如 VEGFR1 和 RET，用于计算 50% 抑制浓度 (IC50) 值。使用谷胱甘肽-S-转移酶的重组融合蛋白、TIE2 的胞内结构域和肽生物素-Ahx-EPKDDAYPLYSDFG 作为底物，使用均相时间分辨荧光 (HTRF) 测定来测量 TIE2 激酶抑制。

GIST 882 和 TT 细胞在含有 L-谷氨酰胺的 RPMI 培养基中生长以进行增殖测定，而 MDA-MB-231、HepG2 和 A375 细胞在始终补充有 10% hiFBS 的 DMEM 中生长。胰蛋白酶处理的细胞以每孔 5×104 个细胞的密度接种在含有含 10% FBS 的完全培养基的 96 孔板中，并在 37°C 下生长过夜。添加载体或瑞格非尼（在完全生长培养基中连续稀释至最终浓度在 10 μM 至 5 nM 之间）和 0.2% DMSO，继续孵育 96 小时。使用 CellTitre-GloTM 测量细胞增殖。

Absorption, Distribution and Excretion
Cmax = 2.5 μg/mL; Tmax = 4 hours; AUC = 70.4 μg*h/mL; Cmax, steady-state = 3.9 μg/mL; AUC, steady-state = 58.3 μg*h/mL; The mean relative bioavailability of tablets compared to an oral solution is 69% to 83%.
Approximately 71% of a radiolabeled dose was excreted in feces (47% as parent compound, 24% as metabolites) and 19% of the dose was excreted in urine (17% as glucuronides) within 12 days after administration of a radiolabeled oral solution at a dose of 120 mg.
Regorafenib undergoes enterohepatic circulation with multiple plasma concentration peaks observed across the 24-hour dosing interval.

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Metabolism / Metabolites
Regorafenib is metabolized by CYP3A4 and UGT1A9. The main circulating metabolites of regorafenib measured at steady-state in human plasma are M-2 (N-oxide) and M-5 (N-oxide and N-desmethyl), both of them having similar in vitro pharmacological activity and steady-state concentrations as regorafenib. M-2 and M-5 are highly protein bound (99.8% and 99.95%, respectively). Regorafenib is an inhibitor of P-glycoprotein, while its active metabolites M-2 (N-oxide) and M-5 (N-oxide and N-desmethyl) are substrates of P-glycoprotein.

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Biological Half-Life
Regorafenib, 160 mg oral dose = 28 hours (14 - 58 hours); M2 metabolite, 160 mg oral dose = 25 hours (14-32 hours); M5 metabolite, 160 mg oral dose = 51 hours (32-72 hours);

Hepatotoxicity
In large clinical trials of regorafenib, elevations in serum aminotransferase levels were common, occurring in 39% to 45% of patients, and were greater than 5 times the upper limit of normal (ULN) in 3% to 6%. In addition, there have been several reports of clinically apparent liver injury arising during regorafenib therapy which was often severe and occasionally fatal, estimated to occur in 0.3% of treated subjects. For these reasons, routine monitoring of liver enzymes is recommended. Regorafenib induced liver injury can present in several different patterns or phenotypes. Some patients present within a few days of starting regorafenib with acute hepatic necrosis, high levels of serum aminotransferase and lactic dehydrogenase with mild jaundice, but prolongation of INR and signs of hepatic failure. The injury can be severe but is generally self-limited and recovery is rapid and complete. Other patients present with an acute viral hepatitis like pattern, hepatocelllar (or mixed) serum enzyme elevations and jaundice that can be prolonged and has been fatal in several instances. Autoimmune and immunoallergic features are uncommon. In addition, rare instances of regorafenib associated liver injury have presented with a sinusoidal obstruction-like syndrome or pseudocirrhosis, with marked hepatic nodularity and ascites that eventually improves or resolves. Finally, regorafenib, like other multi-kinase inhibitors [sunitinib, imatinib, sorafenib], has also been associated with episodes of hyperammonemic coma generally arising within a few days or weeks of starting and with rapid reversal upon stopping treatment.
Likelihood score: B (highly likely cause of clinically apparent liver injury).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of regorafenib during breastfeeding. Because regorafenib is 99.5% 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 70 hours, and might accumulate in the infant. The manufacturer recommends that breastfeeding be discontinued during regorafenib therapy and for 2 weeks after the final 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.

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Protein Binding
Regorafenib is highly bound (99.5%) to human plasma proteins.

[1]. Regorafenib (BAY 73-4506): a new oral multikinase inhibitor of angiogenic, stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity. Int J Cancer, 2011, 129(1), 245-255.

[2]. Targeted therapy for metastatic renal cell carcinoma: current treatment and future directions. Ther Adv Med Oncol, 2010, 2(1), 39-49.

[3]. Fluoro-Sorafenib (Regorafenib) effects on hepatoma cells: growth inhibition, quiescence, and recovery. J Cell Physiol, 2013, 228(2), 292-297.

Regorafenib is a pyridinecarboxamide obtained by condensation of 4-[4-({[4-chloro-3-(trifluoromethyl)phenyl]carbamoyl}amino)-3-fluorophenoxy]pyridine-2-carboxylic acid with methylamine. Used for for the treatment of metastatic colorectal cancer in patients who have previously received chemotherapy, anti-EGFR or anti-VEGF therapy. It has a role as an antineoplastic agent, a tyrosine kinase inhibitor and a hepatotoxic agent. It is an aromatic ether, a pyridinecarboxamide, a member of monochlorobenzenes, a member of (trifluoromethyl)benzenes, a member of monofluorobenzenes and a member of phenylureas.
Regorafenib is an orally-administered inhibitor of multiple kinases. It is used for the treatment of metastatic colorectal cancer, advanced gastrointestinal stromal tumours, and hepatocellular carcinoma. FDA approved on September 27, 2012. Approved use of Regorafenib was expanded to treat Hepatocellular Carcinoma in April 2017.
Regorafenib anhydrous is a Kinase Inhibitor. The mechanism of action of regorafenib anhydrous is as a Kinase Inhibitor, and Cytochrome P450 2C9 Inhibitor, and Breast Cancer Resistance Protein Inhibitor, and UGT1A9 Inhibitor, and UGT1A1 Inhibitor.
Regorafenib is an oral multi-kinase inhibitor that is used in the therapy of refractory metastatic colorectal cancer, hepatocellular carcinoma and gastrointestinal stromal tumor. Regorafenib has been associated with frequent serum aminotransferase elevations during therapy and with rare, but sometimes severe and even fatal instances of clinically apparent liver injury.
Regorafenib Anhydrous is the anhydrous form of regorafenib, an orally bioavailable small molecule with potential antiangiogenic and antineoplastic activities. Regorafenib binds to and inhibits vascular endothelial growth factor receptors (VEGFRs) 2 and 3, and Ret, Kit, PDGFR and Raf kinases, which may result in the inhibition of tumor angiogenesis and tumor cell proliferation. VEGFRs are receptor tyrosine kinases that play important roles in tumor angiogenesis; the receptor tyrosine kinases RET, KIT, and PDGFR, and the serine/threonine-specific Raf kinase are involved in tumor cell signaling.
Regorafenib is the hydrate form of regorafenib, an orally bioavailable small molecule with potential antiangiogenic and antineoplastic activities. Regorafenib binds to and inhibits vascular endothelial growth factor receptors (VEGFRs) 2 and 3, and Ret, Kit, PDGFR and Raf kinases, which may result in the inhibition of tumor angiogenesis and tumor cell proliferation. VEGFRs are receptor tyrosine kinases that play important roles in tumor angiogenesis; the receptor tyrosine kinases RET, KIT, and PDGFR, and the serine/threonine-specific Raf kinase are involved in tumor cell signaling.
See also: Regorafenib Monohydrate (active moiety of).

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Drug Indication
Regorafenib is indicated for the treatment of patients with metastatic colorectal cancer (CRC) who have been previously treated with fluoropyrimidine-, oxaliplatin- and irinotecan-based chemotherapy, an anti-VEGF therapy, and, if KRAS wild type, an anti-EGFR therapy. Regorafenib is also indicated for the treatment of patients with locally advanced, unresectable or metastatic gastrointestinal stromal tumour (GIST) who have been previously treated with imatinib mesylate and sunitinib malate. Regorafenib is also indicated for the treatment of patients with hepatocellular carcinoma (HCC) previously treated with sorafenib.
FDA Label
Stivarga is indicated as monotherapy for the treatment of adult patients with: metastatic colorectal cancer (CRC) who have been previously treated with, or are not considered candidates for, available therapies - these include fluoropyrimidine-based chemotherapy, an anti-VEGF therapy and an anti-EGFR therapy; unresectable or metastatic gastrointestinal stromal tumors (GIST) who progressed on or are intolerant to prior treatment with imatinib and sunitinib; hepatocellular carcinoma (HCC) who have been previously treated with sorafenib.
Treatment of all conditions contained in the category of malignant neoplasms (except haematopoietic and lymphoid tissue)

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Mechanism of Action
Regorafenib is a small molecule inhibitor of multiple membrane-bound and intracellular kinases involved in normal cellular functions and in pathologic processes such as oncogenesis, tumor angiogenesis, and maintenance of the tumor microenvironment. In in vitro biochemical or cellular assays, regorafenib or its major human active metabolites M-2 and M-5 inhibited the activity of RET, VEGFR1, VEGFR2, VEGFR3, KIT, PDGFR-alpha, PDGFR-beta, FGFR1, FGFR2, TIE2, DDR2, TrkA, Eph2A, RAF-1, BRAF, BRAFV600E , SAPK2, PTK5, and Abl at concentrations of regorafenib that have been achieved clinically. In in vivo models, regorafenib demonstrated anti-angiogenic activity in a rat tumor model, and inhibition of tumor growth as well as anti-metastatic activity in several mouse xenograft models including some for human colorectal carcinoma.

C₂₁H₁₆CL₂F₄N₄O₃

519.28

518.053

835621-07-3

Regorafenib;755037-03-7;Regorafenib monohydrate;1019206-88-2

11167602

White to off-white solid

6.968

95.84

3

8

5

33

686

0

Cl.O=C(NC1C(F)=CC(OC2C=C(C(NC)=O)N=CC=2)=CC=1)NC1C=C(C(F)(F)F)C(Cl)=CC=1

ACSWJKPZXNIVMY-UHFFFAOYSA-N

InChI=1S/C21H15ClF4N4O3.ClH/c1-27-19(31)18-10-13(6-7-28-18)33-12-3-5-17(16(23)9-12)30-20(32)29-11-2-4-15(22)14(8-11)21(24,25)26;/h2-10H,1H3,(H,27,31)(H2,29,30,32);1H

4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]-3-fluorophenoxy]-N-methylpyridine-2-carboxamide;hydrochloride

BAY-734506 HCl; BAY 734506; BAY734506; Regorafenib HCl. Brand name: Stivarga

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 mg/mL (3.85 mM) in 50% PEG300 50% 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。
*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: 30% PEG400+0.5% Tween80+5% Propylene glycol : 30mg/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;
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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。