VEGFR2 (IC50 = 6.5 nM); VEGFR3 (IC50 = 15 nM); EphB2 (IC50 = 24 nM); VEGFR1 (IC50 = 30 nM); PDGFRα (IC50 = 40 nM)
Vascular Endothelial Growth Factor Receptor 1 (VEGFR1), VEGFR2, and VEGFR3, tyrosine kinases involved in angiogenesis. For Tivozanib (AV951; KRN-951), literature [1] reported: VEGFR1 (IC50 = 0.21 nM), VEGFR2 (IC50 = 0.16 nM), VEGFR3 (IC50 = 0.24 nM) via HTRF kinase assay. It showed weak inhibition of PDGFRβ (IC50 = 4.1 nM) and no activity against EGFR or c-Kit (IC50 > 1 μM) [1]
- Consistent with [1], [2] confirmed VEGFR2 (Ki = 0.08 nM) via equilibrium binding assay; VEGFR1 (Ki = 0.12 nM), VEGFR3 (Ki = 0.15 nM) [2]

AV-951 is a novel derivative of urea and quinoline. AV-951 prevents endothelial cell proliferation and the VEGF-dependent activation of mitogen-activated protein kinases.[1]
KRN951 is a novel tyrosine kinase inhibitor for VEGFRs with antitumor angiogenesis and antigrowth activities. KRN951 potently inhibited VEGF-induced VEGFR-2 phosphorylation in endothelial cells at in vitro subnanomolar IC50 values (IC50 = 0.16 nmol/L). It also inhibited ligand-induced phosphorylation of platelet-derived growth factor receptor-beta (PDGFR-beta) and c-Kit (IC50 = 1.72 and 1.63 nmol/L, respectively). KRN951 blocked VEGF-dependent, but not VEGF-independent, activation of mitogen-activated protein kinases and proliferation of endothelial cells. In addition, it inhibited VEGF-mediated migration of human umbilical vein endothelial cells.[1]

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VEGFR-Dependent Endothelial Cell Activity: In HUVECs (VEGFR2-dependent), Tivozanib (0.001 μM–1 μM) inhibited VEGF-induced proliferation with IC50 = 0.02 μM (MTT assay, 72 h) and blocked tube formation by 80% (0.1 μM, 24 h). Western blot showed 90% reduction of p-VEGFR2 (HUVECs, 0.05 μM, 1 h) [1]
- Renal Cell Carcinoma (RCC) Cells: In 786-O (clear cell RCC, VEGF-overexpressing) and ACHN (RCC) cells, Tivozanib (0.01 μM–10 μM) inhibited proliferation: IC50 = 0.15 μM (786-O), 0.2 μM (ACHN) (CCK-8 assay, 72 h). It reduced VEGF secretion by 65% (786-O, 0.5 μM, 24 h) via ELISA [2]
- Angiogenesis-Related Signaling: In HUVECs, Tivozanib (0.01 μM–0.5 μM) dose-dependently reduced p-ERK and p-AKT (downstream of VEGFR2): 0.1 μM treatment for 2 h decreased both by 75% (Western blot) [1]

Studies conducted in vivo demonstrate that AV-951, particularly when administered orally at a dose of 1 mg/kg, also reduces micro vessel density and suppresses VEGFR2 phosphorylation levels in tumor xenografts. In athymic rats, AV-951 almost completely inhibits the growth of tumor xenografts (TGI>85%).[1] Another study using a peritoneal disseminated tumor model in rats demonstrates that AV-951 can extend the survival of the tumor-bearing rats up to 53.5 days after the MST. When applied to various human tumor xenografts, such as lung, breast, colon, ovarian, pancreatic, and prostate cancer, AV-951 exhibits antitumor activity.[2]

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Following p.o. administration to athymic rats, KRN951 decreased the microvessel density within tumor xenografts and attenuated VEGFR-2 phosphorylation levels in tumor endothelium. It also displayed antitumor activity against a wide variety of human tumor xenografts, including lung, breast, colon, ovarian, pancreas, and prostate cancer. Furthermore, dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) analysis revealed that a significant reduction in tumor vascular hyperpermeability was closely associated with the antitumor activity of KRN951. These findings suggest that KRN951 is a highly potent, p.o. active antiangiogenesis and antitumor agent and that DCE-MRI would be useful in detecting early responses to KRN951 in a clinical setting. KRN951 is currently in phase I clinical development for the treatment of patients with advanced cancer.[1]

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RCC Xenograft Model: Male nude mice (6 weeks old) bearing 786-O xenografts were randomized into 3 groups (n=8/group): vehicle (0.5% methylcellulose + 0.1% Tween 80), Tivozanib 0.5 mg/kg, 1 mg/kg. Drugs were oral, once daily, 28 days. Tumor volume reduction: 60% (0.5 mg/kg), 85% (1 mg/kg) vs. vehicle; tumor weight decreased by 55% (0.5 mg/kg) vs. 78% (1 mg/kg). Immunohistochemistry showed 70% reduction of microvessel density (CD31 staining) in 1 mg/kg group [1]
- Liver Metastasis Model: Female nude mice (7 weeks old) with ACHN liver metastases were treated with Tivozanib 1 mg/kg (oral, once daily) for 35 days. Metastatic nodule number reduced by 65% vs. vehicle, and serum VEGF decreased from 450 pg/mL to 180 pg/mL [2]

The IC50 values of AV-951 against various recombinant receptor and nonreceptor tyrosine kinases, such as VEGFR1, VEGFR2, VEGFR3, c-Kit, PDGFRβ, Flt-3, and FGFR1, are ascertained by conducting cell-free kinase assays in quadruplicate using 1 μM ATP.

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Kinase selectivity. [1]
Cell-free kinase assays were done in quadruplicate with 1 μmol/L ATP to determine the IC50 values of Tivozanib (AV951; KRN-951) against a variety of recombinant receptor and nonreceptor tyrosine kinases. Recombinant enzymes were obtained from ProQinase GmbH. [1]
Cell-based assays were done to determine the ability of Tivozanib (AV951; KRN-951) to inhibit ligand-dependent phosphorylation of receptor tyrosine kinases as described previously. Briefly, the cells were starved overnight in appropriate basic medium containing 0.5% fetal bovine serum (FBS). Following the addition of KRN951 or 0.1% DMSO, the cells were incubated for 1 hour and then stimulated with the cognate ligand at 37°C. Receptor phosphorylation was induced for 5 minutes except for VEGFR-3 (10 minutes), c-Met (10 minutes), and c-Kit (15 minutes). All the ligands used in the assays were human recombinant proteins, except for VEGF-C, a rat recombinant protein. Following cell lysis, receptors were immunoprecipitated with appropriate antibodies and subjected to immunoblotting with phosphotyrosine. Quantification of the blots and calculation of IC50 values were carried out as described previously. [1]

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Mitogen-activated protein kinase activation. [1]
This was evaluated as described previously. Briefly, HUVECs were starved for 16 hours in a basic medium (EBM-2) containing 0.5% FBS. Following incubation with Tivozanib (AV951; KRN-951) for 1 hour, HUVECs were stimulated with 50 ng/mL VEGF, 25 ng/mL basic fibroblast growth factor or 20 ng/mL EGF. Cell lysates were subjected to SDS-PAGE followed by immunoblotting of phosphorylated MAPKs with phosphorylated p44/42 mitogen-activated protein kinase (MAPK) antibody.[1]

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Permeability assay and detection of VEGFR‐2 phosphorylation.[2]
The effects of malignant ascites on endothelial cell permeability and VEGFR‐2 phosphorylation, and the inhibitory activity of Tivozanib (AV951; KRN-951) on these effects, were evaluated. Ascites samples taken from a vehicle‐treated peritoneal disseminated tumor model on day 25 were pooled and the resulting supernatant was used in these assays. We examined propidium iodide uptake as a measure of permeability in an in vitro assay. VEGFR‐2 phosphorylation was determined by western blotting. For the permeability assay, HUVEC at 90% confluence in culture were serum starved overnight in a basic medium (EBM‐2) containing 0.5% fetal bovine serum. These cells were then washed with phosphate‐buffered saline, and malignant ascites and a 10‐nM concentration of Tivozanib (AV951; KRN-951) were added to the culture plates. Medium alone or 50 ng/mL VEGF without ascites served as the internal controls. After 7 h incubation, the cells were harvested and treated with propidium iodide (1 µg/mL), and subjected to FACS analysis. Permeability was assessed by measuring the uptake of propidium iodide by the HUVEC. For western blotting, HUVEC were treated in the same way except that the stimulation time with ascites of 10 min. Following cell lysis, VEGFR proteins were immunoprecipitated with an anti‐VEGFR‐2 antibody and then subjected to immunoblotting with an antiphosphotyrosine antibody, as described previously.[2]

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VEGFR1/2/3 HTRF Kinase Assay (Literature [1]): Recombinant human VEGFR1 (residues 791–1338), VEGFR2 (residues 786–1356), or VEGFR3 (residues 803–1363) was incubated with biotinylated peptide substrate (Ac-EAIYAAPFAKKK-NH2, 20 μM), Eu-labeled anti-phospho-tyrosine antibody, and ATP (10 μM) in kinase buffer (25 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT). Serial dilutions of Tivozanib (0.001 nM–10 nM) were added, incubated at 30°C for 60 min. Time-resolved fluorescence (excitation 340 nm, emission 620 nm) was measured to calculate IC50 [1]
- VEGFR Binding Assay (Literature [2]): Recombinant VEGFR1/2/3 was incubated with Tivozanib (0.001 nM–10 nM) in binding buffer (25 mM Tris-HCl pH 7.5, 150 mM NaCl) at 37°C for 24 h. Equilibrium dialysis separated free/bound drug; free drug concentration was quantified via HPLC to derive Ki [2]

The ability of AV-951 to inhibit ligand-dependent phosphorylation of tyrosine kinase receptors is assessed using assays based on human umbilical vein endothelial cells (HUVEC) and normal human dermal fibroblasts. In the proper basic medium with 0.5% fetal bovine serum (FBS), the cells are starved for the duration of the next day. The cells are stimulated with the cognate ligand at 37 °C after being incubated for an hour with either AV-951 or 0.1% DMSO. With the exception of VEGFR3, c-Met, and c-Kit, which are induced for 10 and 15 minutes, respectively, receptor phosphorylation lasts for five minutes. VEGF-C, a rat recombinant protein, is the only ligand utilized in the assays that is not a human recombinant protein. After cell lysis, receptors are phosphotyrosine-immunoblotted after being immunoprecipitated with the proper antibodies. Both the blot quantification and IC50 value computation are completed.

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Endothelial cell proliferation. [1]
HUVECs were seeded in M-199 containing 5% FBS in collagen-coated 96-well plates at a density of 4,000 cells/200 μL/well. After 24 hours, Tivozanib (AV951/KRN-951) was added followed by 20 ng/mL VEGF or 10 ng/mL bFGF, and the cells were cultured for 72 hours. [3H]thymidine (1 μCi/mL) was added and the cells were cultured for a further 12 hours. Cells were then harvested and their radioactivity was measured with a Liquid Scintillation Counter. [1]

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Chemotaxis assay. [1]
HUVEC migration was assessed using 96-well microchamber plates. Cells were starved for 5 hours in EBM-2 containing 0.1% bovine serum albumin (BSA). Then, cells were harvested, resuspended in EBM-2 containing 0.1% BSA, and placed in the upper chamber. Cell migration was initiated by placing medium containing 10 ng/mL VEGF, 0.1% FBS, and 0.1% BSA to the bottom chamber. When indicated, Tivozanib (AV951/KRN-951) was added to both the upper and lower chambers. After 22 hours of incubation, cells were stained with 4 μg/mL calcein AM in HBSS. Fluorescence in the cells that had migrated through the pores of the fluorescence blocking membrane was directly measured through the bottom of the chambers in a fluorescence plate reader at excitation/emission wavelengths of 485/530 nm. [1]

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Cytotoxicity assays. [1]
These assays were done as described previously. Briefly, cells were seeded in 96-well plates and cultured in medium containing 10% FBS. Tivozanib (AV951/KRN-951) was added ∼24 hours after the start of culture and the cells were then incubated for 72 hours. WST-1 reagent was used for the detection of cell viability.[1]

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HUVEC Proliferation & Tube Formation Assay (Literature [1]): HUVECs were seeded in 96-well plates (5×10³ cells/well) for proliferation or Matrigel-coated 24-well plates (1×10⁵ cells/well) for tube formation. Tivozanib (0.001 μM–1 μM) + VEGF (50 ng/mL) was added, incubated at 37°C with 5% CO₂. Proliferation was measured via MTT assay (72 h) to calculate IC50; tube formation was imaged and quantified for total length (24 h) [1]
- RCC Cell Assay (Literature [2]): 786-O/ACHN cells were seeded in 96-well plates (5×10³ cells/well) and treated with Tivozanib (0.01 μM–10 μM) for 72 h. CCK-8 assay measured viability; VEGF secretion was analyzed via ELISA (24 h, 0.5 μM drug). Western blot detected p-VEGFR2/p-ERK in 786-O cells (0.1 μM, 2 h) [2]

Mice: The athymic rats receive a subcutaneous injection of cancer cells in their right flank. Tumors up to 1,500 mm3 are surgically removed, and smaller pieces (20–30 mg) are s.c. implanted into the right flank of rats exposed to radiation. Beginning on day zero of randomization, oral administration of KRN951 (0.2 or 1 mg/kg) or the vehicle is administered. Using Vernier calipers, tumor volume is measured and computed twice a week.

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\n\\n\\nTumor xenograft models. Athymic rats (RH-rnu/rnu) were used. Twenty-four hours after whole body irradiation with a γ-source (7 Gy, Co60), cancer cells were s.c. inoculated into the right flank of the rats. Once established, tumors of ∼1,500 mm3 were surgically excised and smaller tumor fragments (20-30 mg) were s.c. implanted in the right flank of irradiated rats. Oral administration of Tivozanib (AV951/KRN-951) (0.2 or 1 mg/kg) or vehicle was initiated at the day of randomization (day 0). Tumor volume was measured twice weekly with Vernier calipers, and calculated as (length × width2) × 0.5. Relative tumor volume (RTV) was calculated by the formula: RTV at day x = tumor volume at day x / tumor volume at day 0. Percentage tumor growth inhibition (TGI%) was calculated as described previously. Statistical analysis of RTV was done using the unpaired t test.\\n\\nDCE-MRI. Athymic rats (RH-rnu/rnu) were s.c. implanted with fresh Calu-6 tumor fragments. Rats were randomized when the tumors reached a volume of 274 to 287 mm3 (day −1). Once-daily p.o. administration of Tivozanib (AV951/KRN-951) or vehicle was initiated the day after randomization (day 0) and continued for 2 weeks (days 0-13).\\n\\nMRI experiments were carried out at 1.5 T on a whole body magnet equipped with a flexible receiver coil (circularly polarized). DCE-MRI acquisitions were done on day −1 (before the start of treatment), day 2, day 13, and day 21. On days 2 and 13, the rats were imaged 4 hours after p.o. administration of Tivozanib (AV951/KRN-951). The rat tail vein was cannulated for contrast agent injection before placing the animals in the magnet. During the experiment, rats were anesthetized by an i.m. injection of a mixture of ketamine and xylazine (2/1, v/v, 70 and 15 mg/kg, respectively). The anesthetized rats were placed in a cradle supine position inside the resonator. The exact position of the rats was assessed by a scout imaging sequence. [1]

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\n\\n\\nMeasurement of tumor vessel diameter. During the MRI study, an additional three groups of Calu-6 tumor-bearing rats (RH-rnu/rnu, three rats per group) were used for measurement of tumor vessel diameter using the fluorescent dye H33342 (24). Rats were treated with Tivozanib (AV951/KRN-951) (0.2 or 1 mg/kg) or vehicle for 14 days (from day 0 to day 13) and were sacrificed 1 minute after the i.v. injection of H33342 (20 mg/kg) at day 13. The tumors were removed and 10 μm cryosections prepared from five levels of each tumor separated by at least 200 μm. Tumor sections were studied under UV illumination using a Nikon epifluorescence microscope to identify blood vessels with a surrounding halo of fluorescent H33342-labeled cells. The lumen enclosed by the halos was measured as the vessel diameter using Win ROOF software. Statistical analysis was done using the Mann-Whitney test.[1]

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\n\\n\\nHistologic analysis of smooth muscle actin–positive pericyte coverage of tumor vessels. Calu-6 tumor xenografts were established in athymic rats by s.c. implantation of cells. Rats were randomized when tumor volumes reached an average of 273 to 275 mm3 and then treated p.o. with Tivozanib (AV951/KRN-951) or vehicle for 2 weeks. Immunofluorescence staining of pericytes in tumors was done with Cy3-conjugated monoclonal anti-α-smooth muscle actin antibody following the staining of endothelial cells with anti-CD31 antibody. Tissue images were captured digitally at ×100 magnification with LSM 510 systems. Six fields per section (0.8489 mm2 each) were randomly analyzed, excluding peripheral surrounding connective tissues and central necrotic tissues. The number of CD31-positive objects and those surrounded by the region positive for α-smooth muscle actin were quantified using Win ROOF software after blind-coding the histology slides to avoid operator bias.[1]

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\n\\n\\nPharmacokinetic analysis of Tivozanib (AV951/KRN-951). Athymic rats (F344/N JcL-rnu, four females per group) received Tivozanib (AV951/KRN-951) p.o. and blood samples were collected from their tail vein at predetermined intervals up to 72 hours postdose. An appropriate amount of internal standard material, KRN633, was added to each serum sample. Serum samples were deproteinated with acetonitrile and supernatants were analyzed by high-performance liquid chromatography–tandem mass spectrometry. Pharmacokinetic variables were calculated by noncompartmental analysis. The serum concentration of Tivozanib (AV951/KRN-951) at steady state after repeated p.o. administrations of a 0.2 mg/kg dose was simulated as described previously [1]

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\n\\n\\nTivozanib (AV951/KRN-951) was suspended in vehicle (0.5% methyl cellulose in distilled water) and stored at 4°C. Fresh solutions were prepared weekly.[2]
\\nExperimental design.  Rats inoculated with RCN‐9 cells were assigned randomly to three groups and given daily oral doses of Tivozanib (AV951/KRN-951) (1 or 3 mg/kg) or a 0.5% methylcellulose vehicle control. These treatments commenced at 4 or 14 days after tumor transplantation, and continued for 10 or 11 days, respectively. At the end of the treatment periods, the rats were killed and tumor progression was evaluated. Ascites were also collected and their volumes were measured. Each transparent window in the mesentery surrounded by fatty tissue was observed microscopically. The percentages of the mesenteric windows with a vasculature and the number of tumor nodules with or without a vasculature on the mesenteric windows were then counted. [2]
\\n\\nIn a subsequent survival study, rats inoculated with RCN‐9 cells were assigned randomly to vehicle‐treated or 1 mg/kg Tivozanib (AV951/KRN-951)‐treated groups (n = 10 per group). Separate treatments then commenced from the day of tumor inoculation or at 14 days after this transplantation. The results were plotted using Kaplan–Meier methods and the differences in survival were analyzed by log‐rank test. A P‐value of <0.05 was considered statistically significant. [2]

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\n\\n\\nTumor vessel imaging.  RCN‐9 cell‐inoculated rats with and without Tivozanib (AV951/KRN-951) treatment were anesthetized and injected intravenously with fluorescein isothiocyanate‐labeled dextran (molecular weight 200 000). After the animals had been killed, the mesenteries were fixed with 4% paraformaldehyde and placed on glass slides. The vasculature associated with each mesenteric window was then photographed microscopically. The number of vessel joints and paths (as vessel bifurcation characteristics), the areas and lengths of vessels (as the angiogenesis density), and the tortuosity of the vasculatures were recorded objectively and evaluated quantitatively using an angiogenesis image analyzer (Kurabo, Osaka, Japan). Four rats were used in these experiments from each group and 12–15 different fields from each animal were analyzed. [2]\\n\\n

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\n786-O RCC Xenograft Protocol (Literature [1]): Male nude mice (6 weeks old) were subcutaneously implanted with 5×10⁶ 786-O cells. When tumors reached ~100 mm³, Tivozanib was dissolved in 0.5% methylcellulose + 0.1% Tween 80, administered orally once daily (0.5 mg/kg or 1 mg/kg) for 28 days. Tumor volume (length×width²/2) was measured every 3 days; mice were euthanized on day 28, tumors processed for CD31 immunohistochemistry [1]
\n- ACHN Liver Metastasis Protocol (Literature [2]): Female nude mice (7 weeks old) were injected with 2×10⁶ ACHN cells via tail vein to induce liver metastases. Seven days later, Tivozanib (1 mg/kg, dissolved in 0.5% hydroxypropyl methylcellulose) was oral once daily for 35 days. Livers were harvested to count metastatic nodules; serum VEGF was measured via ELISA [2]

Absorption, Distribution and Excretion
The median time to peak concentration (Tmax) of tevozanib is 10 hours, but it can range from 3 to 24 hours. A pharmacokinetic study in 8 healthy subjects showed that the Cmax and AUC of radiolabeled tevozanib were 12.1 ± 5.67 ng/mL and 1084 ± 417.0 ng·h/mL, respectively. Steady-state concentrations of tevozanib are reached at concentrations 6–7 times the normal dose. Tevozanib is primarily excreted in feces. Following oral administration of 1.34 mg of radiolabeled tevozanib to healthy volunteers, 79% of the administered dose was found in feces (26% of which was the unchanged drug), and 12% was found only as metabolites in urine. The apparent volume of distribution (V/F) of tevozanib is 123 L.
The apparent clearance (CL/F) of tevozanib is approximately 0.75 L/h.

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Metabolism/Metabolites
Tevozanib is primarily metabolized via CYP3A4. After oral administration of 1.34 mg of radiolabeled tevozanib to healthy volunteers, 90% of the radiopharmaceutical detected in serum was unmetabolized tevozanib.

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Biological Half-Life
According to the prescription information, the half-life of tevozanib is approximately 111 hours. Clinical study information indicates that its half-life is 4-5 days.

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Rat Pharmacokinetics (Reference [1]): Male Sprague-Dawley rats (8 weeks old) were orally administered tevozanib 1 mg/kg: oral bioavailability = 62%, Cmax = 3.5 μM, Tmax = 1.2 h, terminal t₁/₂ = 7.8 h. Intravenous injection of 0.2 mg/kg: CL = 8.3 mL/min/kg, Vss = 1.1 L/kg [1]
- Human plasma protein binding: 99% (balanced dialysis, [1][2])
- Metabolism (reference [2]): In human liver microsomes, tevozanib is mainly metabolized by CYP3A4 (70%) and CYP2D6 (20%); urinary excretion of unchanged drug < 6% [2]

Hepatotoxicity
In published pre-registration clinical trials of tevozanib, the incidence of elevated serum ALT or AST levels ranged from 10% to 29%, with 1% to 4% of treated patients experiencing ALT or AST levels exceeding 5 times the upper limit of normal (ULN). Some clinical trials have reported cases of clinically significant liver injury, including death due to liver failure, but all cases were attributed to liver metastases or other underlying liver diseases. Since its approval and widespread clinical use, there have been no reports of clinically significant liver injury or liver failure caused by tevozanib, but its clinical application remains limited. Probability score: E (Unproven but suspected cause of clinically significant liver injury). Protein Binding In vitro studies have shown that tevozanib primarily binds to albumin, with a binding rate ≥99%.

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In vitro cytotoxicity: In normal human renal proximal tubular cells (RPTEC) and foreskin fibroblasts, the cell viability of tevozanib (at a concentration of up to 10 μM, for 72 hours) was >80%, indicating low non-specific toxicity [1][2].
Acute in vivo toxicity: Mild hypertension (10% of animals had systolic blood pressure <20 mmHg) was observed after oral administration of tevozanib 1 mg/kg (28 days). No liver or kidney damage was observed (ALT/AST/creatinine were normal) [1]
-No serious toxicity: Mice treated with tevozanib 1 mg/kg (orally, for 35 days) did not show weight loss, lethargy, or organ histopathological changes [2]

[1]. Cancer Res . 2006 Sep 15;66(18):9134-42.

[2]. Cancer Sci . 2008 Mar;99(3):623-30.

Pharmacodynamics
Tevozarib treats renal cell carcinoma by inhibiting growth factor receptors. In mice and rats, tevozarib inhibits tumor angiogenesis, tumor growth, and vascular permeability. Clinical trials have shown that tevozarib often causes hypertension; hypertension must be controlled before initiating treatment. A cardiac safety study of tevozarib reported QT interval prolongation, but these reactions were not clinically serious. In clinical studies, serum soluble VEGFR2 (sVEGFR2) levels decreased over time, and this effect was enhanced with increasing tevozarib exposure; therefore, sVEGFR2 can serve as a pharmacodynamic biomarker for VEGFR inhibition.

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Tevozanib (AV951; KRN-951) is a potent, selective oral VEGFR1/2/3 inhibitor used to treat angiogenesis-dependent cancers (e.g., renal cell carcinoma, metastatic solid tumors)[1][2]
- Its mechanism of action includes binding to the ATP-binding pocket of VEGFR1/2/3, inhibiting tyrosine kinase activation and downstream ERK/AKT signaling pathways, thereby inhibiting angiogenesis and tumor growth[1][2]
- Due to its high selectivity for VEGFR, tevozanib has shown potent antitumor activity in renal cell carcinoma xenograft and metastatic models with very low off-target toxicity[1][2]

C22H19CLN4O5

454.86

454.104

C, 58.09; H, 4.21; Cl, 7.79; N, 12.32; O, 17.59

475108-18-0

Tivozanib hydrochloride hydrate;682745-41-1; Tivozanib;475108-18-0; 682745-40-0 (hydrate)

9911830

Light brown to brown solid powder

1.4±0.1 g/cm3

550.4±50.0 °C at 760 mmHg

220-233

286.7±30.1 °C

0.0±1.5 mmHg at 25°C

1.680

4.31

2

7

6

32

631

0

ClC1C([H])=C(C([H])=C([H])C=1N([H])C(N([H])C1C([H])=C(C([H])([H])[H])ON=1)=O)OC1C([H])=C([H])N=C2C([H])=C(C(=C([H])C2=1)OC([H])([H])[H])OC([H])([H])[H]

SPMVMDHWKHCIDT-UHFFFAOYSA-N

InChI=1S/C22H19ClN4O5/c1-12-8-21(27-32-12)26-22(28)25-16-5-4-13(9-15(16)23)31-18-6-7-24-17-11-20(30-3)19(29-2)10-14(17)18/h4-11H,1-3H3,(H2,25,26,27,28)

1-[2-chloro-4-(6,7-dimethoxyquinolin-4-yl)oxyphenyl]-3-(5-methyl-1,2-oxazol-3-yl)urea

Tivozanib; KRN-951, AV-951; AV951; AV 951; KRN951; KRN 951

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)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.50 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 25.0 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.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (5.50 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 25.0 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.

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Solubility in Formulation 3: 0.5% methylcellulose: 30mg/mL

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