VEGFR2 (IC50 = 40 nM); VEGFR3 (IC50 = 110 nM); EGFR/HER1 (IC50 = 500 nM)

Vandetanib inhibits VEGFR3 and EGFR with IC50 of 110 nM and 500 nM, respectively. Vandetanib exhibits little action against MEK, CDK2, c-Kit, erbB2, FAK, PDK1, Akt, and IGF-1R, with an IC50 above 10 μM. In contrast, it is insensitive to PDGFRβ, Flt1, Tie-2, and FGFR1. Vandetanib has little effect on basal endothelial cell growth but inhibits HUVEC proliferation induced by VEGF, EGF, and bFGF with IC50 values of 60 nM, 170 nM, and 800 nM, respectively. With an IC50 ranging from 2.7 μM (A549) to 13.5 μM (Calu-6) [1], vandetanib inhibits the development of tumor cells. When compared to the Cat S inhibitor LHVS (IC50=0.001 μM) and tested in mouse B cell lines (IC50=1.5±0.4 μM), odanacatib is a mild antigen presentation inhibitor. With lowest inhibitory doses of 1-10 μM and 0.01 μM, respectively, Odanacatib also demonstrated lesser inhibitory effects on the processing of MHC II invariant chain protein Iip10 in mouse splenocytes as compared to LHVS [2]. Vandetanib suppresses the phosphorylation of EGFR in liver cancer cells and VEGFR-2 in HUVEC, as well as cell division [4].

In the H1650 xenograft model, vandetanib (15 mg/kg, po) reduces tumor growth with an IC50 of 3.5±1.2 μM, which is better than gefitinib's anti-tumor efficacy [3]. Vandetanib (50 or 75 mg/kg) reduced the number of intrahepatic metastases, upregulated VEGF, TGF-α, and EGF in tumor tissues, increased tumor cell apoptosis, inhibited tumor growth, and enhanced survival rate in tumor-bearing mice [4]. It also inhibited the phosphorylation of VEGFR-2 and EGFR in tumor tissues.

In 96-well plates coated with a poly(Glu, Ala, Tyr) 6:3:1 random copolymer substrate, vandetanib is incubated with the enzyme, 10 mM MnCl₂, and 2 μM ATP. The next step is to identify phosphorylated tyrosine by sequentially incubating 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), a horseradish peroxidase-linked sheep antimouse immunoglobulin antibody, and a mouse IgG anti-phosphotyrosine 4G10 antibody. To investigate selectivity against tyrosine kinases linked to FGFR1, c-kit, erbB2, IGF-1R, FAK, PDGFRβ, Tie-2, and FGFR1, this methodology is modified. Appropriate ATP concentrations at or slightly below the corresponding Km (0.2–14 μM) were used in all enzyme assays (tyrosine or serine–threonine). Selectivity against serine-threonine kinases (CDK2, AKT, and PDK1) is investigated in 96-well plates using a pertinent scintillation proximity-assay (SPA). The conditions for the CDK2 assays were as follows: 10 mM MnCl₂, 4.5 μM ATP, 0.15 μCi of [γ-³³ P]ATP/reaction, 50 mM HEPES (pH 7.5), 1 mM DTT, 0.1 mM sodium orthovanadate, 0.1 mM sodium fluoride, 10 mM sodium glycerophosphate, 1 mg/mL BSA fraction V, and a retinoblastoma substrate (a portion of the retinoblastoma gene, 792–928, expressed in a glutathione S-transferase expression system; 0.22 μM initial concentration). The reactions are conducted at room temperature for 60 minutes and then quenched for two hours using 150 μL of a solution that contains 0.8 mg/reaction of protein A SPA-polyvinyltoluene beads, 3 μg of rabbit immunoglobulin anti-glutathione S-transferase antibody, and EDTA (62 mM final concentration). After that, the plates are sealed, centrifuged for five minutes at 1200 x g, and counted for thirty seconds using a Microplate scintillation counter.

The MTT assay is modified to measure growth inhibition. In a nutshell, the cells are exposed to either vandetanib or gefitinib for 72 hours after being plated at a density of 2000 cells per well in 96-well plates. Triples of each assay are run. For every medication, the 50% inhibitory concentration (IC50) is calculated using the mean±standard deviation (SD).

Absorption, Distribution and Excretion
Slow absorption—median peak plasma concentration is reached in 6 hours. After multiple doses, vandetanib plasma concentrations can accumulate to approximately 8-fold, reaching steady state after about 3 months. Approximately 69% of the drug is recovered 21 days after a single dose of vandetanib. Of this, 44% is found in feces and 25% in urine. Volume of distribution (Vd) is approximately 7450 liters. Vandetanib binds to human serum albumin and α1-acid glycoprotein, with an in vitro protein binding rate of approximately 90%. After reaching steady state with a once-daily dose of 300 mg vandetanib in colorectal cancer patients, the average protein binding rate in ex vivo plasma samples was 94%. Approximately 69% of the drug is recovered during the 21-day collection period after a single dose of ¹⁴C-vandetanib, with 44% found in feces and 25% in urine. Excretion of this dose is slow, and based on the plasma half-life, it is expected to continue to be excreted after 21 days. Vandetanib is not a substrate of hOCT2 expressed in HEK293 cells. Vandetanib inhibits the uptake of the selective OCT2-labeled substrate 14C-creatinine by HEK-OCT2 cells, with a mean IC50 of 2.1 μg/mL. This value is higher than the vandetanib plasma concentration (0.81 μg/mL) observed after multiple 300 mg doses. Vandetanib inhibits renal excretion of creatinine, which may explain the elevated plasma creatinine levels in subjects treated with vandetanib.
After oral administration of capreressa, absorption is slow, and peak plasma concentrations are typically reached 6 hours (median, range 4–10 hours) after administration. After multiple doses, vandetanib plasma concentrations can accumulate to approximately 8-fold, reaching steady state after approximately 3 months. Food does not affect vandetanib exposure.
The protein binding rate of ¹⁴C-vandetanib in the plasma of mice, rats, rabbits, dogs, and humans is moderate, ranging from 83% to 90%. Following a single oral administration, vandetanib and/or its metabolites exhibit slow but widespread tissue distribution in colored and colorless male rats, consistent with the distribution pattern of lipophilic compounds. Peak concentrations of vandetanib and/or its metabolites are reached in most tissues 6–8 hours post-administration. The radioactive material is prominently distributed in brain tissue. Retention of the radioactive material was observed in colored tissues, indicating its affinity for melanin. Significant radioactive distribution was observed in the milk of lactating rats and in the plasma of lactating pups.
For more complete data on the absorption, distribution, and excretion of vandetanib (8 items in total), please visit the HSDB record page.

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Metabolism/Metabolites
Unmetabolized vandetanib and its metabolites vandetanib N-oxide and N-desmethylvandetanib were detected in plasma, urine, and feces. N-desmethylvandetanib was primarily produced by CYP3A4, while vandetanib N-oxide was primarily produced by the flavin-containing monooxygenases FMO1 and FMO3.
The metabolism of vandetanib appeared similar in the toxicological study species (rats and dogs) as well as in mice and humans. The two major metabolites identified in excrement were N-desmethylvandetanib and vandetanib N-oxide. In mice, a minor metabolite, O-desalkylvandetanib glucuronide, was also identified. Glucuronide conjugates were also detected in human urine. Metabolism and bile excretion appear to be the main pathways of vandetanib clearance in preclinical animal models. In vitro CYP identification studies indicated that CYP3A4 is involved in the formation of N-desmethylvandetanib. Vandetanib-N-oxide is generated by FMO1 and FMO3 (FMO = flavin monooxygenase). These two enzymes are also present in the kidneys, suggesting that renal excretion may contribute to vandetanib clearance. Following oral administration of (14)C-vandetanib, unchanged vandetanib and its metabolites vandetanib-N-oxide and N-demethylvandetanib were detected in plasma, urine, and feces. Glucuronide conjugates were only found as minor metabolites in excretions. N-demethylvandetanib was primarily generated by CYP3A4, while vandetanib-N-oxide was generated by the flavin monooxygenases FMO1 and FMO3. The circulating concentrations of N-demethylvandetanib and vandetanib-N-oxide were approximately 7-17% and 1.4-2.2% of vandetanib, respectively. ...At all time points, the total radioactivity concentration in plasma was higher than that of vandetanib, indicating the presence of circulating metabolites. Unmetabolized vandetanib and two expected metabolites (N-demethylvandetanib and vandetanib-N-oxide) were detected in plasma, urine, and feces. Additionally, a trace metabolite (glucuronide conjugate) was found in urine and feces. Unmodified vandetanib and its N-demethyl and N-oxide metabolites were detected in plasma, urine, and feces.

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Biological half-life
The median half-life is 19 days.
...In patients with medullary thyroid carcinoma (MTC), a 300 mg dose of capreressa is characterized by...a median plasma half-life of 19 days.
...Vandertanib is slowly absorbed and eliminated, with a half-life of approximately 10 days after a single oral dose.

Hepatotoxicity
In large clinical trials of vandetanib, abnormalities in routine liver function tests were common, with elevated serum transaminases occurring in up to half of the patients, and 2% to 5% of patients having transaminase levels more than five times the upper limit of normal (ULN). In premarketing trials of vandetanib for thyroid cancer, no clinically significant liver injury (with jaundice or liver failure) was reported. Since its approval and widespread use, no published reports of hepatotoxicity caused by vandetanib have been found, and the product information does not mention hepatotoxicity. However, many kinase inhibitors used in cancer chemotherapy have been associated with clinically significant liver injury cases, typically occurring within the first 2 to 12 weeks of treatment, manifesting as symptoms such as fatigue, nausea, and jaundice, and elevated serum enzymes in a hepatocellular pattern, but without immune hypersensitivity or autoimmune characteristics. Some tyrosine kinase inhibitors (such as imatinib and nilotinib) have also been considered to be associated with hepatitis B virus reactivation.
Probability Score: E (Unconfirmed, but suspected as a rare cause of clinically significant liver damage).

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Effects during Pregnancy and Lactation
◉ Overview of Use During Lactation
There is currently no information regarding the clinical use of vandetanib during lactation. Because vandetanib binds to plasma proteins at a rate of up to 90%, its concentration in breast milk is likely to be low. However, its half-life is 19 days, and it may accumulate in the infant. The manufacturer recommends discontinuing breastfeeding during vandetanib treatment and for 4 months after the last dose.
◉ Effects on Breastfed Infants
No published information found as of the revision date.
◉ Effects on Lactation and Breast Milk
No published information found as of the revision date.

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Protein Binding
The protein binding rate is approximately 90%.

[1]. ZD6474 inhibits vascular endothelial growth factor signaling, angiogenesis, and tumor growth following oral administration. Cancer Res. 2002 Aug 15;62(16):4645-55.

[2]. Interaction of the EGFR inhibitors gefitinib, vandetanib, pelitinib and neratinib with the ABCG2 multidrug transporter: implications for the emergence and reversal of cancer drug resistance. Biochem Pharmacol. 2012 Aug 1;84(3):260-7.

[3]. Vandetanib is effective in EGFR-mutant lung cancer cells with PTEN deficiency. Exp Cell Res. 2013 Feb 15;319(4):417-23.

[4]. Vandetanib, an inhibitor of VEGF receptor-2 and EGF receptor, suppresses tumor development and improves prognosis of liver cancer in mice. Clin Cancer Res. 2012 Jul 15;18(14):3924-33.

Vandetanib is a quinazoline compound with the chemical name 7-[(1-methylpiperidin-4-yl)methoxy]quinazoline, containing additional methoxy and 4-bromo-2-fluorophenylamino substituents at positions 6 and 4, respectively. It is used to treat patients with locally advanced or metastatic, unresectable, symptomatic or progressive medullary thyroid carcinoma. Vandetanib is a tyrosine kinase inhibitor and an antitumor drug. It is an aromatic ether, secondary amine, quinazoline compound, piperidine compound, organobromine compound, and organofluorine compound. Vandetanib is an orally administered, once-daily kinase inhibitor that inhibits tumor angiogenesis and tumor cell proliferation, showing potential for treating various tumor types. On April 6, 2011, vandetanib was approved by the FDA for the treatment of unresectable locally advanced or metastatic medullary thyroid carcinoma in adults. Vandetanib is a kinase inhibitor. Its mechanism of action is as a protein kinase inhibitor. Vandetanib is a kinase inhibitor. Its mechanism of action is as a protein kinase inhibitor, P-glycoprotein inhibitor, and organic cation transporter 2 inhibitor. Vandetanib is a multi-kinase inhibitor used to treat advanced or metastatic medullary thyroid carcinoma. Transient elevations in serum transaminases are common during vandetanib treatment, but have not been found to be associated with clinically significant acute liver injury with jaundice. Vandetanib is an orally bioavailable 4-phenylaminoquinazoline drug. Vandetanib selectively inhibits the tyrosine kinase activity of vascular endothelial growth factor receptor 2 (VEGFR2), thereby blocking VEGF-stimulated endothelial cell proliferation and migration, and reducing tumor vascular permeability. The drug also blocks the tyrosine kinase activity of epidermal growth factor receptor (EGFR), a receptor tyrosine kinase that mediates tumor cell proliferation, migration, and angiogenesis.

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Drug Indications
Vandertanib is currently approved as an alternative to local treatment for unresectable and metastatic disease. Because vandetanib can prolong the QT interval, it is contraindicated in patients with severe cardiac complications such as congenital long QT syndrome and decompensated heart failure.

FDA Label
Caprelsa is indicated for the treatment of patients with locally advanced or metastatic unresectable, aggressive, and symptomatic medullary thyroid carcinoma (MTC). Caprelsa is indicated for adults, children, and adolescents aged 5 years and older. For patients with unknown RET gene mutations or negative test results, their potential benefit should be considered before developing an individualized treatment plan.
Treatment of Medullary Thyroid Carcinoma

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Mechanism of Action
ZD-6474 is a potent and selective VEGFR (vascular endothelial growth factor receptor), EGFR (epidermal growth factor receptor), and RET (transfection rearrangement) tyrosine kinase inhibitor. Both VEGFR and EGFR-dependent signaling pathways are clinically validated cancer pathways, including non-small cell lung cancer (NSCLC). RET activity plays an important role in certain types of thyroid cancer, and early data on vandetanib in medullary thyroid carcinoma have earned it orphan drug designation from regulatory agencies in the United States and the European Union. In vitro studies have shown that vandetanib inhibits phosphorylation of receptor tyrosine kinases stimulated by epidermal growth factor (EGF) in tumor cells and endothelial cells, as well as phosphorylation of tyrosine kinases stimulated by vascular endothelial growth factor (VEGF) in endothelial cells. In vitro studies have also shown that vandetanib inhibits the tyrosine kinase activity of EGFR and VEGFR family members, RET, BRK, TIE2, and EPH receptors and Src kinase family members. These receptor tyrosine kinases are involved in normal cellular function and pathological processes such as tumorigenesis, metastasis, tumor angiogenesis, and maintenance of the tumor microenvironment. Furthermore, the N-demethyl metabolite of this drug accounts for 7% to 17.1% of vandetanib exposure, and its inhibitory activity against VEGF receptors (KDR and Flt-1) and EGFR is similar to that of the parent compound. Oncogenic transformation of RET (transfection rearrangement) tyrosine kinases is a common feature of medullary thyroid carcinoma (MTC). Vandetanib is an ATP-competitive inhibitor that inhibits RET, epidermal growth factor receptor (EGFR), and vascular endothelial growth factor receptor kinases. This study investigated the mechanism of action of vandetanib in TT and MZ-CRC-1 human MTC cell lines carrying RET mutations, specifically cysteine 634-to-tryptophan (C634W) and methionine 918-to-threonine (M918T). Vandetanib inhibited MTC cell proliferation and phosphorylation of RET, Shc, and p44/p42 mitogen-activated protein kinase (MAPK). RNA interference single-receptor knockdown assays showed that MTC cell proliferation was RET-dependent. Adoptive expression of the vandetanib-resistant V804M RET mutant rescued the proliferation of TT cells treated with vandetanib, indicating that RET is a key target of vandetanib in MTC cells. Following RET inhibition, adoptive activation of EGFR partially rescued TT cell proliferation, the MAPK signaling pathway, and the expression of cell cycle-related genes. This suggests that simultaneous inhibition of RET and EGFR by vandetanib may overcome the risk of MTC cells escaping RET blockade through compensatory overactivation of EGFR. Transfection rearrangement (RET) is widely expressed in neuroblastoma (NB) and contributes to its high metastatic potential and survival. This study aimed to investigate whether vandetanib (a RET inhibitor) inhibits the proliferation, migration, and invasion of neuroblastoma (NB) cells in vitro. We evaluated the effects of vandetanib on the proliferation, apoptosis, cell cycle, and RET phosphorylation levels of SK-N-SH and SH-SY5Y cells in vitro. Transwell cell migration and invasion assays were used to analyze the migration and invasion abilities of NB cells treated with vandetanib. The expression levels of mRNA and protein in NB cells treated with vandetanib were detected by qPCR, Western blotting, and immunofluorescence. The results showed that vandetanib inhibited the proliferation of SK-N-SH and SH-SY5Y cells. At low concentrations, it inhibited proliferation by inducing G1 phase cell cycle arrest, while at high concentrations, it inhibited proliferation by inducing apoptosis. Compared with the control group, vandetanib treatment significantly reduced the migration and invasion abilities of both NB cell lines (p<0.01). Furthermore, our data showed that compared with vector-treated cells, vandetanib-treated NB cell lines exhibited significantly reduced mRNA expression levels of CXC chemokine receptor 4 (CXCR4) and matrix metalloproteinase 14 (MMP14) (p<0.01), and similar results were obtained for protein levels by Western blotting and immunofluorescence analysis. Vandetanib may inhibit the proliferation, migration, and invasion of NB cells in vitro. The potential mechanism by which vandetanib inhibits NB cell migration and invasion may be partly attributed to its ability to suppress the expression of CXCR4 and MMP14 in human NB cells. For more complete data on the mechanism of action of vandetanib (6 items in total), please visit the HSDB record page.

C22H24N4O2FBR.C2HO2F3

589.3773

588.099

338992-53-3

Vandetanib;443913-73-3;Vandetanib hydrochloride;524722-52-9;Vandetanib-d6;1174683-49-8

17973223

Typically exists as solid at room temperature

608.2±0.0 °C at 760 mmHg

321.6±0.0 °C

0.0±0.0 mmHg at 25°C

6.74

2

12

6

37

623

0

FZMIEQCEGTVFBZ-UHFFFAOYSA-N

InChI=1S/C22H24BrFN4O2.C2HF3O2/c1-28-7-5-14(6-8-28)12-30-21-11-19-16(10-20(21)29-2)22(26-13-25-19)27-18-4-3-15(23)9-17(18)24;3-2(4,5)1(6)7/h3-4,9-11,13-14H,5-8,12H2,1-2H3,(H,25,26,27);(H,6,7)

N-(4-bromo-2-fluorophenyl)-6-methoxy-7-[(1-methylpiperidin-4-yl)methoxy]quinazolin-4-amine;2,2,2-trifluoroacetic acid

Vandetanib trifluoroacetate; 338992-53-3; Vandetanib (trifluoroacetate); ZD 6474 trifluoroacetate; ZD-6474 trifluoroacetate; SCHEMBL1614619; FZMIEQCEGTVFBZ-UHFFFAOYSA-N;

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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