RET; FGFR4; FGFR2; FGFR3; VEGFR1 (IC50 = 22 nM); VEGFR2 (IC50 = 4 nM); VEGFR3 (IC50 = 5.2 nM); FGFR1 (IC50 = 46 nM); PDGFRα (IC50 = 51 nM); PDGFRβ (IC50 = 39 nM); c-Kit (IC50 = 100 nM)

Lenvatinib mesylate (E7080 mesylate) has IC50 values of 4, 5.2, and 22 nM for VEGFR1/Flt-1, VEGFR2(KDR), and VEGFR3(Flt-4), respectively. With IC50 values of 51, 39, 46, and 100 nM, respectively, lenitinib inhibits PDGFRα, PDGFRβ, FGFR1, and KIT[3].

---

Kinase inhibitory profile of E7080. [4]
The kinase inhibitory profile of E7080 was determined using a cell-free kinase assay (Table 1). E7080 potently inhibited VEGF-R3 kinase activity (IC50, 5.2 nmol/L; Table 1; Supplementary Fig. S1) and VEGF-R2 kinase activity (IC50, 4.0 nmol/L) to a similar extent (Table 1). E7080 also inhibited VEGF-R1, FGF-R1, and PDGF-Rβ kinase, but the inhibitory activity was about 4 to 10 times less potent (Table 1). EGFR kinase was not effectively inhibited with E7080. E7080 showed strong inhibition of phosphorylation of VEGF-R2 (IC50, 0.83 nmol/L) and VEGF-R3 (IC50, 0.36 nmol/L) in HUVECs after stimulation with VEGF and VEGF-C, respectively (Table 1; Fig. 1). These data indicated that E7080 was a potent inhibitor of VEGF-R3 kinase as well as VEGF-R2 kinase. Inhibitory activity of E7080 against VEGF-induced proliferation of HUVEC (IC50, 2.7 nmol/L) was stronger than basic FGF induced (IC50, 410 nmol/L) in HUVEC and PDGF-induced proliferation of L cells (IC50, 340 nmol/L; Table 1). We were not able to determine the IC50 value for VEGF-C–induced cell proliferation because VEGF-C did not stimulate cell proliferation in our assays.

---

E7080 inhibits both angiogenesis and lymphangiogenesis induced by human breast cancer cells. [4]
MDA-MB-231 cell is a human breast adenocarcinoma cell derived from pleural effusion (25). Metastases of MDA-MB-231 cells inoculated into the m.f.p. developed in the regional lymph nodes and distant lung with high frequency (Table 2), whereas those of MDA-MB-435 was developed only in the distant lung (data not shown). ELISA assay of conditioned medium indicated that both tumor cells expressed significant amounts of VEGF, but only MDA-MB-231 produced high amounts of VEGF-C (Table 3), and neither of cell lines produced detectable amounts of VEGF-D. These data suggested that the VEGF/VEGF-R2 and VEGF-C/VEGF-R3 signals might be activated, resulting in metastases to the regional lymph nodes and distant lung in the MDA-MB-231 m.f.p. xenograft model, whereas only the VEGF/VEGF-R2 signal might be activated, resulting in metastasis to the distant lung in the MDA-MB-435 m.f.p. xenograft model. To determine roles of VEGF/VEGF-R2 and VEGF-C/VEGF-R3 signals in metastasis, we examined the effects of an anti-VEGF antibody, bevacizumab (a selective inhibitor of the VEGF signal), and E7080 (a dual inhibitor of VEGF-R2 and VEGF-R3 kinases), on angiogenesis and lymphangiogenesis in two m.f.p. xenograft models. The extent of angiogenesis and lymphangiogenesis was evaluated by staining tumor tissues with anti-CD31 antibody and anti-LYVE-1 antibody, respectively.

Lenvatinib mesylate (E7080 mesylate) (100 mg/kg, p.o.) also significantly inhibits metastasis to both distant lung and regional lymph nodes after treatment is completed, and bevacizumab significantly inhibits local tumor growth at the m.f.p.[3].
Lenvatinib mesylate (E7080 mesylate) has a dose-dependent effect on the H146 tumor, causing tumor regression at 100 mg/kg in the H146 xenograft model and inhibiting the growth of the tumor at 30 and 100 mg/kg (BID, QDx21). Anti-CD31 antibody IHC analysis reveals that lenvatinib at 100 mg/kg reduces microvessel density more than imatinib treatment and anti-VEGF antibody[4].

---

Efficacy of E7080, Imatinib and a VEGF neutralization antibody in H146 xenograft model [4]
To investigate a role of SCF/KIT signaling in tumor angiogenesis, researchers evaluated the effect of E7080, which inhibits both KDR and KIT kinases, VEGF neutralization antibody, which selectively inhibits VEGF signaling, and imatinib, which inhibits KIT kinase alone, using H146 xenograft model. Oral administration of E7080 inhibited the growth of H146 tumor at 30 and 100 mg/kg (BID, QDx21) in a dose-dependent manner and caused tumor regression at 100 mg/kg (Fig. 6a). Treatment with either imatinib at 160 mg/kg (BID, QDx21) or anti-VEGF antibody at 300 and 500 μg per mouse (twice a week) clearly slowed tumor growth but did not cause tumor regression (Fig. 6a). IHC analysis with anti-CD31 antibody (Fig. 6b) showed that E7080 at 100 mg/kg decreased microvessel density more than anti-VEGF antibody and imatinib treatment (Fig. 6c). E7080 might achieve tumor regression as a result of potent antiangiogenic activity based on inhibition of both KIT and VEGF receptor signaling.

---

E7080 inhibits metastasis to both regional lymph nodes and distant lung in the MDA-MB-231 m.f.p. xenograft model. [4]
Next, researchers evaluated the effects of E7080 and bevacizumab on metastases of MDA-MB-231 to the regional lymph nodes and distant lung. Time to develop metastases of MDA-MB-231 was ∼7 weeks. We treated tumor-bearing mice with inhibitors 43 days after inoculation and administered for 56 days (Fig. 4). Both E7080 and bevacizumab significantly inhibited local tumor growth at the m.f.p., and at the end of treatment, RTVs were 0.81 ± 1.00 (for E7080), 5.11 ± 6.54 (for bevacizumab), and 17.4 ± 13.1 (for vehicle; P < 0.05; Fig. 4). E7080 also significantly inhibited metastasis to both regional lymph nodes and distant lung (P < 0.05; Table 2). Metastases to lymph nodes occurred in 0 of 10 mice and to the lung in 0 of 10 mice after E7080 treatment, whereas metastases to both the lymph nodes and lung occurred in 9 of 12 vehicle-treated mice. Bevacizumab also seemed to decrease the incidence of metastases to the lymph nodes (6 of 10) and lung (3 of 10), but this decrease was only significant in the lung (Table 2). These results suggest that bevacizumab was not able to inhibit the VEGF-C/VEGF-R3 signal.

---

E7080 decreased both angiogenesis and lymphangiogenesis of established metastatic nodules of MDA-MB-231 tumor in the lymph nodes. [4]
Researchers observed a significant decrease in both lymphangiogenesis and angiogenesis in the primary MDA-MB-231 tumor with E7080 treatment (Fig. 3). Thus, we evaluated the effect of E7080 on the growth of metastatic nodules, angiogenesis, and lymphangiogenesis within established metastatic nodules in the lymph nodes after resecting the primary tumor at the m.f.p. (Fig. 5A). The primary tumors were resected ∼90 days after inoculation (Fig. 5A) and E7080 was administered beginning 2 weeks after tumor resection for 4 weeks (Fig. 5C). E7080 seemed to inhibit the growth of metastatic nodules (vehicle: 11.8 ± 10.8; E7080: 0.6 ± 0.3; Fig. 5B and C), but it was not a statistical difference because of large variation of RTVs in the vehicle group, although immunohistochemical analysis with anti-CD31 and anti-LYVE-1 antibody (Fig. 6) indicated that E7080 treatment significantly decreased both MVD (vehicle: 94.3 ± 12.6; E7080: 20.3 ± 2.9/mm2; Fig. 6A and C) and LVD (vehicle: 24.7 ± 13.3; E7080: 1.0 ± 0.9/mm2; Fig. 6B and C) within metastatic nodules in the lymph nodes. These results showed that E7080 inhibited both angiogenesis and lymphangiogenesis within established metastatic nodules in lymph nodes in this MDA-MB-231 xenograft model.

Tyrosine kinase assays using recombinant receptor kinase domains are carried out by HTRF (KDR, VEGFR1, FGFR1, c-Met, EGFR) and ELISA (PDGFRβ). In each experiment, four microliters of successive dilutions of E7080 are combined with ten microliters of enzyme, sixteen microliters of poly (GT) solution (250 ng), and ten microliters of ATP solution (1 μM ATP) in a 96-well round plate (final DMSO concentration is 0.1%). No enzyme is introduced to blank wells. Test articles are not added to control wells. Addition of ATP solution to each well starts the kinase reaction. Each well's reaction mixture is mixed with 10 μL of 0.5 M EDTA to halt the reaction after a 30-minute incubation period at 30°C. The reaction mixture is supplemented with dilution buffer appropriate for each kinase assay. The HTRF assay involves transferring 50 μL of the reaction mixture to a 96-well 1/2 area black EIA/RIA plate, adding 50 μL of HTRF solution per well, and measuring the fluorescence of the reaction mixture using a time-resolved fluorescence detector at 620 and 665 nm for emission and 337 nm for excitation. This allows for the determination of kinase activity. For the ELISA, 96-well polystyrene plates coated with avidin are incubated at room temperature for 30 minutes with 50 μL of the reaction mixture. PY20-HRP solution (70 μL/well) is added to the reaction mixture after washing with wash buffer, and it is then incubated at room temperature for 30 minutes. TMB reagent (100 μL/well) is added to each well following washing with wash buffer. One milligram of H3PO4 (100 μL/well) is added to each well after a few minutes (10–30 minutes). The measurement of absorbance at 450 nm using a microplate reader yields the kinase activity.

---

In vitro kinase assay [3]
Tyrosine kinase assays were performed by HTRF (KDR, VEGFR1, FGFR1, c-Met, EGFR) and ELISA (PDGFRβ), using the recombinant kinase domains of receptors. In both assays, 4 μL of serial dilutions of Lenvatinib (E7080) were mixed in a 96-well round plate with 10 μL of enzyme, 16 μL of poly (GT) solution (250 ng) and 10 μL of ATP solution (1 μmol/L ATP) (final concentration of DMSO was 0.1%). In wells for blanks, no enzyme was added. In control wells no test article was added. The kinase reaction was initiated by adding ATP solution to each well. After 30-min incubation at 30°C, the reaction was stopped by adding 0.5 mol/L EDTA (10 μL/well) to the reaction mixture in each well. Dilution buffer adequate to each kinase assay was added to the reaction mixture.
In the HTRF assay, 50 μL of the reaction mixture was transferred to a 96-well 1/2 area black EIA/RIA plate, HTRF solution (50 μL/well) was added to the reaction mixture, and then kinase activity was determined by measurement of fluorescence with a time-resolved fluorescence detector at an excitation wavelength of 337 nm and an emission wavelengths of 620 and 665 nm.
In the ELISA, 50 μL of the reaction mixture was incubated in avidin coated 96-well polystyrene plates at room temperature for 30 min. After washing with wash buffer, PY20-HRP solution (70 μL/well) was added and the reaction mixture was incubated at room temperature for 30 min. After washing with wash buffer, TMB reagent (100 μL/well) was added to each well. After several minutes (10–30 min), 1 mol/L H3PO4 (100 μL/well) was added to each well. Kinase activity was determined by measurement of absorbance at 450 nm with a microplate reader. Kinase inhibitory activities of Lenvatinib (E7080) other than KDR, VEGFR1, FGFR1, c-Met, EGFR and PDGFRβ were examined by ProQinase Company.

---

Cell-free kinase assay/cell phosphorylated assay. [4]
Tyrosine kinase activity was measured by a homogeneous time-resolved fluorescence assay (VEGF-R2, VEGF-R1, fibroblast growth factor-receptor 1 (FGF-R1), and epidermal growth factor receptor) and by ELISA [platelet-derived growth factor (PDGF) receptor β] using the recombinant kinase domains of these receptors. The kinase inhibitory activity of Lenvatinib (E7080) against VEGF-R3 was examined using the technology platform from the ProQinase Co. For cell-free kinase assay, samples were duplicated and two to three separate experiments were done. HUVECs were cultured with serum-free medium containing 0.5% fetal bovine serum for 24 h. Cells were treated with Lenvatinib (E7080), stimulated by either VEGF (20 ng/mL) or VEGF-C (100 ng/mL) for 10 min, and then collected in lysis buffer. To detect VEGF-R2 and phosphorylated VEGF-R2, 10 to 20 μg of cell lysates were electrophoresed. To detect VEGF-R3 and phosphorylated VEGF-R3, 400 to 1,000 μg of cell lysates were immunoprecipitated by anti-VEGF-R3. Immune complexes were solubilized in 60 μL of sample buffer and electrophoresed. The resolved proteins were analyzed by Western blot with the indicated antibodies: for VEGF-R2 and phosphorylated VEGF-R2 and for VEGF-R3 and anti-phosphotyrosine IgG. Immunoreactive bands were visualized by chemiluminescence using the Image Master VDS-CL. The intensity of each band was measured using 1D Image Analysis software. For cell phosphorylated assay, three separate experiments were done.

In a 96-well plate, 1,000 HUVECs (1,000 cells in each well in serum-free medium containing 2% fetal bovine serum) and 5,000 L6 rat skeletal muscle myoblasts (5,000 cells in each well in serum-free DMEM) are added, and the plate is left to incubate overnight. To each well, E7080, VEGF (20 ng/mL) or FGF-2 (20 ng/mL) containing 2% fetal bovine serum, and PDGFβ (40 ng/mL) are added. Following three days of incubation, the WST-1 reagent is used to calculate the ratios of surviving cells. Samples are replicated and three independent experiments are conducted for the proliferation assay.

---

H146 (1.2×10³ cells/50 μL/well) are cultured in 96-well multi-plates with SFM containing 0.5% BSA. Following an overnight culture at 37°C, SFM (150 μL/well) containing 0.5% FBS and various SCF concentrations are added, either with or without various compound concentrations. WST-1 is used to measure the ratios of surviving cells following a 72-hour culture.

---

Flow cytometric (FCM) analysis [3]
FCM analysis was performed according to Funahashi et al.15 Briefly, cells were detached with trypsinization and, after centrifugation, the cell pellet was incubated with either PBS or 1 μg of primary antibody (anti-KIT antibody) for 30 min at 4°C and then, incubated with 50 μL of anti-PE conjugated secondary antibody diluted 1:50 in PBS. Stained cells were analyzed by flow cytometry using a FACS Calibur instrument to quantify staining intensity and results are shown as histograms.

---

Proliferation assay [3]
H146 (1.2 × 103 cells/50 μL/well) in SFM containing 0.5% BSA were cultured in 96-well multi-plates. After overnight culture at 37°C, SFM (150 μL/well) containing 0.5% FBS and several concentrations of SCF were added with or without several concentrations of compound. After culture for 72 hr, the ratios of surviving cells were measured by WST-1.

---

Proliferation assay stimulated with growth factors. HUVECs (1,000 cells in each well in serum-free medium containing 2% fetal bovine serum) and L6 rat skeletal muscle myoblasts (5,000 cells in each well in serum-free DMEM) were dispensed in a 96-well plate and incubated overnight. Lenvatinib (E7080) and either VEGF (20 ng/mL) or FGF-2 (20 ng/mL) containing 2% fetal bovine serum and PDGFβ (40 ng/mL) were added to each well. Cells were incubated for 3 d and then the ratios of surviving cells were measured by WST-1 reagent. For proliferation assay, samples were duplicated and three separate experiments were done [4].

Female BALB/c nude mice
30 & 100 mg/kg
p.o.

---

Clean-room conditions are used to maintain 8–12 week old, 20–25 g female BALB/c nude mice. Mice's flanks are subcutaneously (s.c.) implanted with 6.5×10⁶ H146 tumor cells. Day 1 of the experiment occurs twelve days after the injection when mice are randomized into treatment (n = 6 or n = 5) and control (n = 12) groups. From day one to day twenty-one, lenvatinib, STI571, and VEGF neutralization antibody are given orally twice daily for lenvatinib and STI571 and twice weekly for the antibody. These substances are suspended in 0.5% methylcellulose and saline, respectively. On the designated days, tumor volume is measured and computed. Relative tumor volume (RTV) is a measure of antitumor activity that is calculated as the volume of the tumor on day 1 divided by the tumor volume at indicated days.

---

Tumor xenograft model [3]
Female BALB/c nude mice (8–12 weeks old, 20–25 g), obtained from Charles River (Kanagawa, Japan), were used. Animals were maintained under clean-room conditions. H146 tumor cells (6.5 × 106) were implanted subcutaneously (s.c.) into the flank region of mice. Twelve days after inoculation, mice were randomized into control (n = 12) and treatment (n = 6 or n = 5) groups and this point in time was identified as day 1. Lenvatinib (E7080) and Imatinib, and VEGF neutralization antibody were suspended in 0.5% methylcellulose and saline, respectively, and administered orally twice a day for Lenvatinib (E7080) and Imatinib and twice a week for antibody from day 1 to day 21. Tumor volume was measured on the indicated days and calculated according to the following equation: tumor volume (mm3) = length × (width)2/2. Antitumor activity was shown as a relative tumor volume (RTV = calculated tumor volume at indicated days/volume on day 1).

---

Immunohistochemical analysis of angiogenesis and lymphangiogenesis in m.f.p. xenograft models. [4]
MDA-MB-231 and MDA-MB-435 tumors were removed from mice treated with either Lenvatinib (E7080) (n = 5) or bevacizumab (n = 5) for 1 wk (day 8) and without treatment (n = 5), embedded in OCT compound, frozen on dry ice, and double stained for an endothelial cell marker CD31 (with rat monoclonal anti-mouse CD31, clone MEC13.3) and a lymph endothelial cell marker (with rabbit polyclonal anti-LYVE-1). CD31 and LYVE-1 were visualized by staining with fuchsin and 3,3′-diaminobenzidine, respectively. Microvessel density (MVD) and lymphatic vessel density (LVD) were assessed by counting tumor microvessel and lymph vessel elements (four to five fields per tumor) and calculating tumor microvessel or lymph vessel densities (i.e., number of vessel elements per field). Experiments were duplicated and statistical analysis was done using the Dunnett-type multiple comparison method.

---

Effect of Lenvatinib (E7080) on the primary tumor growth in the m.f.p. and metastases. [4]
MDA-MB-231 cells highly expressing rsGFP were implanted s.c. into the flanks of nude mice. Tumor fragments (17 ± 2 mg) were prepared from 100 to 200 mm3 tumors grown s.c. and then inoculated into the m.f.p. About 2 wk after inoculation, mice were randomized into control (n = 12) and treatment groups (n = 10) at day 1. Either Lenvatinib (E7080) (in water) or bevacizumab (in saline) was administered orally once a day or i.v. twice a week, respectively, from day 1 to day 56. Antitumor activity was shown as a relative tumor volume (RTV = calculated tumor volume/day 1 tumor volume). Tumors expressing rsGFP in the lymph node and lung were detected by a fluorescence imaging detection system after 56 d of treatment. Data include the average with SD for RTV and the ratio of the number of mice bearing metastatic nodules. Experiments were duplicated and statistical analysis was conducted using the Dunnett-type multiple comparison method.

---

Effect of Lenvatinib (E7080) on tumor growth of metastatic nodules in the lymph nodes after resection of the primary tumor. [4]
rsGFP MDA-MB-231 tumor pieces were transplanted and allowed to grow until metastases were noted in the lymph nodes (∼90 d), which were detected by a fluorescence imaging detection system, and then the primary tumors were removed. Eight mice were divided into two groups. Administration of Lenvatinib (E7080) was started 2 wk after resection of the primary tumors (day 1). Lenvatinib (E7080) was administered orally once a day from day 1 to day 28. Statistical analysis was conducted using the Dunnett-type multiple comparison method.

Absorption, Distribution and Excretion
Time to peak plasma concentration occurred from 1 to 4 hours post­dose. Administration with food did not affect the extent of absorption, but decreased the rate of absorption and delayed the median Tmax from 2 hours to 4 hours.
Following administration of a radiolabeled dose, approximately 64% and 25% of the radiolabel were eliminated in the feces and urine, respectively.

---

Metabolism / Metabolites
Lenvatinib is metabolized by CYP3A and aldehyde oxidase.

---

Biological Half-Life
The terminal elimination half­life of lenvatinib is approximately 28 hours.

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
No information is available on the clinical use of lenvatinib during breastfeeding. Because lenvatinib is more than 98% bound to plasma proteins, the amount in milk is likely to be low. However, its half-life is about 28 hours and it might accumulate in the infant. The manufacturer recommends that breastfeeding be discontinued during lenvatinib therapy and for at least 1 week 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]. Lenvatinib versus Bay 43-9006 in first-line treatment of patients with unresectable hepatocellularcarcinoma: a randomised phase 3 non-inferiority trial. Lancet. 2018 Mar 24;391(10126):1163-1173.

[2]. Lenvatinib: A Promising Molecular Targeted Agent for Multiple Cancers. Cancer Control. 2018 Jan-Dec;25(1):1073274818789361.

[3]. E7080, a novel inhibitor that targets multiple kinases, has potent antitumor activities against stem cell factor producing human small cell lung cancer H146, based on angiogenesis inhibition. Int J Cancer. 2008, 122(3), 664-671.

[4]. Multi-kinase inhibitor E7080 suppresses lymph node and lung metastases of human mammary breast tumor MDA-MB-231 via inhibition of vascular endothelial growth factor-receptor (VEGF-R) 2 and VEGF-R3 kinase. Clin Cancer Res. 2008, 14(17),545.

Lenvatinib mesylate is a methanesulfonate salt obtained by reaction of lenvatinib with one molar equivalent of methanesulfonic acid. A multi-kinase inhibitor and orphan drug used (as its mesylate salt) for the treatment of various types of thyroid cancer that do not respond to radioiodine. It has a role as an EC 2.7.10.1 (receptor protein-tyrosine kinase) inhibitor, a fibroblast growth factor receptor antagonist, an orphan drug, a vascular endothelial growth factor receptor antagonist and an antineoplastic agent. It contains a lenvatinib(1+).
Lenvatinib Mesylate is a synthetic, orally available inhibitor of vascular endothelial growth factor receptor 2 (VEGFR2, also known as KDR/FLK-1) tyrosine kinase with potential antineoplastic activity. E7080 blocks VEGFR2 activation by VEGF, resulting in inhibition of the VEGF receptor signal transduction pathway, decreased vascular endothelial cell migration and proliferation, and vascular endothelial cell apoptosis.
See also: Lenvatinib (has active moiety).

---

Drug Indication
Kisplyx is indicated for the treatment of adults with advanced renal cell carcinoma (RCC): in combination with pembrolizumab, as first-line treatment (see section 5. 1). in combination with everolimus, following one prior vascular endothelial growth factor (VEGF)-targeted therapy.
Lenvima is indicated as monotherapy for the treatment of adult patients with progressive, locally advanced or metastatic, differentiated (papillary/follicular/Hürthle cell) thyroid carcinoma (DTC), refractory to radioactive iodine (RAI). Lenvima is indicated as monotherapy for the treatment of adult patients with advanced or unresectable hepatocellular carcinoma (HCC) who have received no prior systemic therapy.

---

Lenvatinib is a member of the class of quinolines that is the carboxamide of 4-{3-chloro-4-[(cyclopropylcarbamoyl)amino]phenoxy}-7-methoxyquinoline-6-carboxylic acid. A multi-kinase inhibitor and orphan drug used (as its mesylate salt) for the treatment of various types of thyroid cancer that do not respond to radioiodine. It has a role as a vascular endothelial growth factor receptor antagonist, an orphan drug, an antineoplastic agent, an EC 2.7.10.1 (receptor protein-tyrosine kinase) inhibitor and a fibroblast growth factor receptor antagonist. It is a member of quinolines, an aromatic ether, a monocarboxylic acid amide, an aromatic amide, a member of monochlorobenzenes, a member of cyclopropanes and a member of phenylureas. It is a conjugate base of a lenvatinib(1+).
Lenvatinib is a receptor tyrosine kinase (RTK) inhibitor that inhibits the kinase activities of vascular endothelial growth factor (VEGF) receptors VEGFR1 (FLT1), VEGFR2 (KDR), and VEGFR3 (FLT4). Lenvatinib also inhibits other RTKs that have been implicated in pathogenic angiogenesis, tumor growth, and cancer progression in addition to their normal cellular functions, including fibroblast growth factor (FGF) receptors FGFR1, 2, 3, and 4; the platelet derived growth factor receptor alpha (PDGFRα), KIT, and RET. These receptor tyrosine kinases (RTKs) located in the cell membrane play a central role in the activation of signal transduction pathways involved in the normal regulation of cellular processes, such as cell proliferation, migration, apoptosis and differentiation, and in pathogenic angiogenesis, lymphogenesis, tumour growth and cancer progression. In particular, VEGF has been identified as a crucial regulator of both physiologic and pathologic angiogenesis and increased expression of VEGF is associated with a poor prognosis in many types of cancers. Lenvatinib is indicated for the treatment of patients with locally recurrent or metastatic, progressive, radioactive iodine (RAI)-refractory differentiated thyroid cancer. Most patients with thyroid cancer have a very good prognosis with treatment (98% 5 year survival rate) involving surgery and hormone therapy. However, for patients with RAI-refractory thyroid cancer, treatment options are limited and the prognosis is poor, leading to a push for the development of more targeted therapies such as lenvatinib.
Lenvatinib is a Kinase Inhibitor. The mechanism of action of lenvatinib is as a Receptor Tyrosine Kinase Inhibitor.
Lenvatinib is orally available multi-kinase inhibitor and antineoplastic agent that is used in treatment of advanced, metastatic medullary thyroid cancer and refractory renal cell carcinoma. Lenvatinib is associated with a modest rate of serum enzyme elevations during treatment and has been implicated to rare instances of clinically apparent, acute liver injury some of which have been fatal.
Lenvatinib is a synthetic, orally available inhibitor of vascular endothelial growth factor receptor 2 (VEGFR2, also known as KDR/FLK-1) tyrosine kinase with potential antineoplastic activity. Lenvatinib blocks VEGFR2 activation by VEGF, resulting in inhibition of the VEGF receptor signal transduction pathway, decreased vascular endothelial cell migration and proliferation, and vascular endothelial cell apoptosis.
See also: Lenvatinib Mesylate (has salt form).

---

Drug Indication
Lenvatinib is indicated for the treatment of the following cancerous conditions: **Differentiated Thyroid Cancer (DTC)** - Treatment of locally recurrent or metastatic, progressive, radioactive iodine-refractory differentiated thyroid cancer **Renal Cell Carcinoma (RCC)** - First-line treatment, in combination with [pembrolizumab], in adult patients with advanced renal cell carcinoma (RCC) - Treatment of advanced renal cell carcinoma, in combination with [everolimus], in adult patients who have previously tried ≥1 anti-angiogenic therapy **Hepatocellular Carcinoma (HCC)** - First-line treatment of patients with unresectable hepatocellular carcinoma **Endometrial Carcinoma** - Treatment of advanced endometrial carcinoma that is not microsatellite instability-high (MSI-H) or mismatch repair deficient (dMMR), in combination with [pembrolizumab], in patients who have experienced disease progression following prior systemic therapy and are not candidates for curative surgery or radiation
FDA Label
Kisplyx is indicated for the treatment of adults with advanced renal cell carcinoma (RCC): in combination with pembrolizumab, as first-line treatment (see section 5. 1). in combination with everolimus, following one prior vascular endothelial growth factor (VEGF)-targeted therapy.
Lenvima is indicated as monotherapy for the treatment of adult patients with progressive, locally advanced or metastatic, differentiated (papillary/follicular/Hürthle cell) thyroid carcinoma (DTC), refractory to radioactive iodine (RAI). Lenvima is indicated as monotherapy for the treatment of adult patients with advanced or unresectable hepatocellular carcinoma (HCC) who have received no prior systemic therapy.
Treatment of all conditions included in the category of malignant neoplasms except haematopoietic and lymphoid tissue neoplasms, papillary thyroid cancer , follicular thyroid cancer and osteosarcoma
Treatment of follicular thyroid cancer , Treatment of osteosarcoma, Treatment of papillary thyroid cancer

---

Mechanism of Action
Lenvatinib is a receptor tyrosine kinase (RTK) inhibitor that inhibits the kinase activities of vascular endothelial growth factor (VEGF) receptors VEGFR1 (FLT1), VEGFR2 (KDR), and VEGFR3 (FLT4). Lenvatinib also inhibits other RTKs that have been implicated in pathogenic angiogenesis, tumor growth, and cancer progression in addition to their normal cellular functions, including fibroblast growth factor (FGF) receptors FGFR1, 2, 3, and 4; the platelet derived growth factor receptor alpha (PDGFRα), KIT, and RET.

---

Background: In a phase 2 trial, lenvatinib, an inhibitor of VEGF receptors 1-3, FGF receptors 1-4, PDGF receptor α, RET, and KIT, showed activity in hepatocellular carcinoma. We aimed to compare overall survival in patients treated with lenvatinib versus sorafenib as a first-line treatment for unresectable hepatocellular carcinoma. Methods: This was an open-label, phase 3, multicentre, non-inferiority trial that recruited patients with unresectable hepatocellular carcinoma, who had not received treatment for advanced disease, at 154 sites in 20 countries throughout the Asia-Pacific, European, and North American regions. Patients were randomly assigned (1:1) via an interactive voice-web response system-with region; macroscopic portal vein invasion, extrahepatic spread, or both; Eastern Cooperative Oncology Group performance status; and bodyweight as stratification factors-to receive oral lenvatinib (12 mg/day for bodyweight ≥60 kg or 8 mg/day for bodyweight <60 kg) or sorafenib 400 mg twice-daily in 28-day cycles. The primary endpoint was overall survival, measured from the date of randomisation until the date of death from any cause. The efficacy analysis followed the intention-to-treat principle, and only patients who received treatment were included in the safety analysis. The non-inferiority margin was set at 1·08. The trial is registered with ClinicalTrials.gov, number NCT01761266. Findings: Between March 1, 2013 and July 30, 2015, 1492 patients were recruited. 954 eligible patients were randomly assigned to lenvatinib (n=478) or sorafenib (n=476). Median survival time for lenvatinib of 13·6 months (95% CI 12·1-14·9) was non-inferior to sorafenib (12·3 months, 10·4-13·9; hazard ratio 0·92, 95% CI 0·79-1·06), meeting criteria for non-inferiority. The most common any-grade adverse events were hypertension (201 [42%]), diarrhoea (184 [39%]), decreased appetite (162 [34%]), and decreased weight (147 [31%]) for lenvatinib, and palmar-plantar erythrodysaesthesia (249 [52%]), diarrhoea (220 [46%]), hypertension (144 [30%]), and decreased appetite (127 [27%]) for sorafenib. Interpretation: Lenvatinib was non-inferior to sorafenib in overall survival in untreated advanced hepatocellular carcinoma. The safety and tolerability profiles of lenvatinib were consistent with those previously observed.[1]

---

Lenvatinib is a small-molecule tyrosine kinase inhibitor that inhibits vascular endothelial growth factor receptor (VEGFR1-3), fibroblast growth factor receptor (FGFR1-4), platelet-derived growth factor receptor α (PDGFRα), stem cell factor receptor (KIT), and rearranged during transfection (RET). These receptors are important for tumor angiogenesis, and lenvatinib inhibits tumor angiogenesis by inhibiting function of these receptors. Phase I trials of lenvatinib were conducted at the same time in Japan, Europe, and the United States, and tumor shrinkage effects were observed in thyroid cancer, endometrial cancer, melanoma, renal cell carcinoma, sarcoma, and colon cancer. Lenvatinib is a promising drug that has shown therapeutic effects against various solid tumors. Adverse events, such as hypertension, proteinuria, diarrhea, and delayed wound healing, can occur with lenvatinib treatment. Managing these adverse events is also important for the use of lenvatinib. In this mini-review article, we outline the current state, toxicity, and future prospects of lenvatinib toward thyroid cancer, hepatocellular carcinoma, renal cell carcinoma, and lung cancer.[2]

---

E7080 is an orally active inhibitor of multiple receptor tyrosine kinases including VEGF, FGF and SCF receptors. In this study, we show the inhibitory activity of E7080 against SCF-induced angiogenesis in vitro and tumor growth of SCF-producing human small cell lung carcinoma H146 cells in vivo. E7080 inhibits SCF-driven tube formation of HUVEC, which express SCF receptor, KIT at the IC(50) value of 5.2 nM and it was almost identical for VEGF-driven one (IC(50) = 5.1 nM). To assess the role of SCF/KIT signaling in tumor angiogenesis, we evaluated the effect of imatinib, a selective KIT kinase inhibitor, on tumor growth of H146 cells in nude mice. Imatinib did not show the potent antitumor activity in vitro (IC(50) = 2,200 nM), because H146 cells did not express KIT. However, oral administration of imatinib at 160 mg/kg clearly slowed tumor growth of H146 cells in nude mice, accompanied by decreased microvessel density. Oral administration of E7080 inhibited tumor growth of H146 cells at doses of 30 and 100 mg/kg in a dose-dependent manner and caused tumor regression at 100 mg/kg. While anti-VEGF antibody also slowed tumor growth, it did not cause tumor regression. These results indicate that KIT signaling has a role in tumor angiogenesis of SCF-producing H146 cells, and E7080 causes regression of H146 tumors as a result of antiangiogenic activity mediated by inhibition of both KIT and VEGF receptor signaling. E7080 may provide therapeutic benefits in the treatment of SCF-producing tumors.[3]

---

Purpose: Vascular endothelial growth factor (VEGF)-C/VEGF-receptor 3 (VEGF-R3) signal plays a significant role in lymphangiogenesis and tumor metastasis based on its effects on lymphatic vessels. However, little is known about the effect of inhibiting VEGF-R3 on lymphangiogenesis and lymph node metastases using a small-molecule kinase inhibitor. Experimental design: We evaluated the effect of E7080, a potent inhibitor of both VEGF-R2 and VEGF-R3 kinase, and bevacizumab on lymphangiogenesis and angiogenesis in a mammary fat pad xenograft model of human breast cancer using MDA-MB-231 cells that express excessive amounts of VEGF-C. Lymphangiogenesis was determined by lymphatic vessel density (LVD) and angiogenesis by microvessel density (MVD). Results: In contrast to MDA-MB-435 cells, which expressed a similar amount of VEGF to MDA-MB-231 cells with an undetectable amount of VEGF-C, only MDA-MB-231 exhibited lymphangiogenesis in the primary tumor. E7080 but not bevacizumab significantly decreased LVD within the MDA-MB-231 tumor. E7080 and bevacizumab decreased MVD in both the MDA-MB-231 and MDA-MB-435 models. E7080 significantly suppressed regional lymph nodes and distant lung metastases of MDA-MB-231, whereas bevacizumab significantly inhibited only lung metastases. E7080 also decreased both MVD and LVD within the metastatic nodules at lymph nodes after resection of the primary tumor. Conclusions: Inhibition of VEGF-R3 kinase with E7080 effectively decreased LVD within MDA-MB-231 tumors, which express VEGF-C. Simultaneous inhibition of both VEGF-R2 and VEGF-R3 kinases by E7080 may be a promising new strategy to control regional lymph node and distant lung metastases.[4]

C22H23CLN4O7S

522.96

522.097

C, 50.53; H, 4.43; Cl, 6.78; N, 10.71; O, 21.42; S, 6.13

857890-39-2

Lenvatinib;417716-92-8

11237762

White to off-white solid powder

5.818

4

8

6

35

727

0

ClC1C([H])=C(C([H])=C([H])C=1N([H])C(N([H])C1([H])C([H])([H])C1([H])[H])=O)OC1C([H])=C([H])N=C2C([H])=C(C(C(N([H])[H])=O)=C([H])C=12)OC([H])([H])[H].S(C([H])([H])[H])(=O)(=O)O[H]

HWLFIUUAYLEFCT-UHFFFAOYSA-N

InChI=1S/C21H19ClN4O4.CH4O3S/c1-29-19-10-17-13(9-14(19)20(23)27)18(6-7-24-17)30-12-4-5-16(15(22)8-12)26-21(28)25-11-2-3-11;1-5(2,3)4/h4-11H,2-3H2,1H3,(H2,23,27)(H2,25,26,28);1H3,(H,2,3,4)

4-[3-chloro-4-(cyclopropylcarbamoylamino)phenoxy]-7-methoxyquinoline-6-carboxamide;methanesulfonic acid

E-7080 mesylate; E7080; E 7080; LENVATINIB MESYLATE; 857890-39-2; lenvatinibMesylate; Lenvima; Lenvatinib mesilate; E7080 MESYLATE; Lenvatinib mesylate [USAN]; UNII-3J78384F61; ER-203492-00 mesylate; Lenvatinib mesylate; Brand name Lenvima

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.08 mg/mL (3.98 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

---

Solubility in Formulation 2: ≥ 2.08 mg/mL (3.98 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

---

View More

Solubility in Formulation 3: ≥ 2.08 mg/mL (3.98 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

---

Solubility in Formulation 4: 0.5% methylcellulose: 30 mg/kg

---

 (Please use freshly prepared in vivo formulations for optimal results.)