![Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX
www.icjpir.com
~166~
Available online at www.icjpir.com ISSN: 2349-5448
Intercontinental journal of pharmaceutical
Investigations and Research
ICJPIR |Volume 4 | Issue 1 | Jan – Mar- 2017 Research Article
A new analytical method development and validation for the estimation of
lenvatinib by using RP-HPLC method
K.Kranthi kiran, N.Priyanka, M.Ramakrishna, M.V.Chendrashekar, N.Nagadhurga
Associate Professor & Jogaiah Institute of Technology & Sciences College of Pharmacy,
Kalagampudi. A.P India
Corresponding Author: K. Kranthi Kiran
Email: kothapallikranthikiran@gmail.com
ABSTRACT
A simple and selective LC method is described for the determination of Lenvatinib dosage forms. Chromatographic
separation was achieved on a c18 column using mobile phase consisting of a mixture of Phosphate buffer (KH2PO4):
Acetonitrile (80:20) with detection of 240nm. Linearity was observed in the range 60-140 µg /ml for Lenvatinib (r2
=0.996) for the amount of drug estimated by the proposed methods was in good agreement with the label claim.
The proposed methods were validated. The accuracy of the methods was assessed by recovery studies at three
different levels. Recovery experiments indicated the absence of interference from commonly encountered
pharmaceutical additives. The method was found to be precise as indicated by the repeatability analysis, showing
%RSD less than 2. All statistical data proves validity of the methods and can be used for routine analysis of
pharmaceutical dosage form.
Keywords: Lenvatinib, Reverse phase HPLC.
INTRODUCTION
A drug includes all medicines intended for
internal or external use for or in the diagnosis,
treatment, mitigation or prevention of disease or
disorder in human beings or animals, and
manufactured exclusively in accordance with the
formulae mentioned in authoritative books [1].
Pharmaceutical analysis is a branch of
chemistry involving a process of identification,
determination, quantification, purification and
separation of components in a mixture or
determination of chemical structure of compounds.
There are two main types of analysis – Qualitative
and Quantitative analysis [3-5].
Qualitative analysis is performed to establish
composition of a substance [6-8]. It is done to
determine the presence of a compound or
substance in a given sample or not. The various](https://image.slidesharecdn.com/icjpir-17-110166-192-191217111835/85/A-new-analytical-method-development-and-validation-for-the-estimation-of-lenvatinib-by-using-RP-HPLC-method-1-320.jpg)
![Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX
www.icjpir.com
~167~
qualitative tests are detection of evolved gas, limit
tests, color change reactions, determination of
melting point and boiling point, mass
spectroscopy, determination of nuclear half-life etc
[9, 10].
AIM AND PLAN OF WORK
Aim
To develop new RP HPLC method for the
simultaneous estimation of Lenvatinib in
pharmaceutical dosage form.
PLAN OF WORK
Solubility determination of Lenvatinib various
solvents and buffers.
Determine the absorption maxima of the drug in
UV–Visible region in different solvents/buffers
and selecting the solvents for HPLC method
development.
Optimize the mobile phase and flow rates for
proper resolution and retention times.
Validate the developed method as per ICH
guidelines
METHODOLOGY
Mobile phase
A mixture of 80 volumes of Phosphate
bufferpH4.0 and 20volumes of Acetonitrile was
prepared. The mobile phase was sonicated for
10min to remove gases.
Determination of working wavelength (λmax)
In estimation of drug wavelength maxima is
used.. So this wavelength is used in estimation to
estimate drug accurately.
Preparation of standard stock solution of
LENVATINIB
100mg of LENVITINIB was weighed and
transferred in to 100ml volumetric flask and
dissolved in methanol and then make up to the
mark with methanol and prepare 100 µg /ml of
solution by diluting 1ml to 10ml with methanol.
RESULTS AND DISCUSSIONS
Solubility Studies
These studies are carried out at 25 0
C
Lenvatinib
Freely soluble in methanol,water and phosphate
buffer.
Wavelength determination
In simultaneous estimation of two drugs
isobestic wavelength is used. Isobestic point is the
wavelength where the molar absorptivity is the
same for two substances that are interconvertible.
So this wavelength is used in simultaneous
estimation to estimate both drugs accurately.
Preparation of standard stock solution of
LENVATINIB
100 mg of LENVATINIB was weighed in to
100ml volumetric flask and dissolved in Methanol
and then dilute up to the mark with methanol and
prepare 10 µg /ml of solution by diluting 1ml to
10ml with methanol, wavelength is found to be
240nm.](https://image.slidesharecdn.com/icjpir-17-110166-192-191217111835/85/A-new-analytical-method-development-and-validation-for-the-estimation-of-lenvatinib-by-using-RP-HPLC-method-2-320.jpg)
![Kranthi K K et al, ICJPIR 2017, 4(1), 166-192
www.icjpir.com
~192~
BIBLIOGRAPHY
[1]. Matsui, J.; Funahashi, Y.; Uenaka, T.; Watanabe, T.; Tsuruoka, A.; Asada, M. "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". Clinical Cancer Research. . doi:10.1158/1078-0432.CCR-07-5270. PMID 18765537, 14 (17),
2008, 5459–65.
[2]. Haberfeld, H, ed. Austria-Codex (in German). Vienna: Österreichischer Apothekerverlag. 2015.
[3]. FDA Professional Drug Information for Lenvima.
[4]. http://www.fda.gov/Drugs/InformationOnDrugs/ApprovedDrugs/ucm501070.htm
[5]. Inoue, K. "Oxidative Metabolic Pathway of Lenvatinib Mediated by Aldehyde Oxidase". Drug Metab
Dispos. doi:10.1124/dmd.114.058073. PMID 24914245. 42, 2014, 1326–33.
[6]. Glen, H; D. Boss; T. R. Evans; M. Roelvink; et al. "A phase I dose finding study of E7080 in patients
(pts) with advanced malignancies". Journal of Clinical Oncology, ASCO Annual Meeting Proceedings
Part I. 25(18S), 2007, 14073.
[7]. Clinical trial number NCT01321554 for "A Trial of E7080 in 131I-Refractory Differentiated Thyroid
Cancer" at ClinicalTrials.gov
[8]. "Phase III trial shows lenvatinib meets primary endpoint of progression free survival benefit in treatment
of radioiodine-refractory differentiated thyroid cancer" (PDF). Eisai. 2014.
[9]. U.S. Food and Drug Administration. Hematology/Oncology (Cancer) Approvals & Safety
Notifications. [1]
[10]. European Medicines Agency Summary of the European public assessment report (EPAR) for Lenvima [2]](https://image.slidesharecdn.com/icjpir-17-110166-192-191217111835/85/A-new-analytical-method-development-and-validation-for-the-estimation-of-lenvatinib-by-using-RP-HPLC-method-27-320.jpg)
A new analytical method development and validation for the estimation of lenvatinib by using RP-HPLC method
- 1. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~166~ Available online at www.icjpir.com ISSN: 2349-5448 Intercontinental journal of pharmaceutical Investigations and Research ICJPIR |Volume 4 | Issue 1 | Jan – Mar- 2017 Research Article A new analytical method development and validation for the estimation of lenvatinib by using RP-HPLC method K.Kranthi kiran, N.Priyanka, M.Ramakrishna, M.V.Chendrashekar, N.Nagadhurga Associate Professor & Jogaiah Institute of Technology & Sciences College of Pharmacy, Kalagampudi. A.P India Corresponding Author: K. Kranthi Kiran Email: kothapallikranthikiran@gmail.com ABSTRACT A simple and selective LC method is described for the determination of Lenvatinib dosage forms. Chromatographic separation was achieved on a c18 column using mobile phase consisting of a mixture of Phosphate buffer (KH2PO4): Acetonitrile (80:20) with detection of 240nm. Linearity was observed in the range 60-140 µg /ml for Lenvatinib (r2 =0.996) for the amount of drug estimated by the proposed methods was in good agreement with the label claim. The proposed methods were validated. The accuracy of the methods was assessed by recovery studies at three different levels. Recovery experiments indicated the absence of interference from commonly encountered pharmaceutical additives. The method was found to be precise as indicated by the repeatability analysis, showing %RSD less than 2. All statistical data proves validity of the methods and can be used for routine analysis of pharmaceutical dosage form. Keywords: Lenvatinib, Reverse phase HPLC. INTRODUCTION A drug includes all medicines intended for internal or external use for or in the diagnosis, treatment, mitigation or prevention of disease or disorder in human beings or animals, and manufactured exclusively in accordance with the formulae mentioned in authoritative books [1]. Pharmaceutical analysis is a branch of chemistry involving a process of identification, determination, quantification, purification and separation of components in a mixture or determination of chemical structure of compounds. There are two main types of analysis – Qualitative and Quantitative analysis [3-5]. Qualitative analysis is performed to establish composition of a substance [6-8]. It is done to determine the presence of a compound or substance in a given sample or not. The various
- 2. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~167~ qualitative tests are detection of evolved gas, limit tests, color change reactions, determination of melting point and boiling point, mass spectroscopy, determination of nuclear half-life etc [9, 10]. AIM AND PLAN OF WORK Aim To develop new RP HPLC method for the simultaneous estimation of Lenvatinib in pharmaceutical dosage form. PLAN OF WORK Solubility determination of Lenvatinib various solvents and buffers. Determine the absorption maxima of the drug in UV–Visible region in different solvents/buffers and selecting the solvents for HPLC method development. Optimize the mobile phase and flow rates for proper resolution and retention times. Validate the developed method as per ICH guidelines METHODOLOGY Mobile phase A mixture of 80 volumes of Phosphate bufferpH4.0 and 20volumes of Acetonitrile was prepared. The mobile phase was sonicated for 10min to remove gases. Determination of working wavelength (λmax) In estimation of drug wavelength maxima is used.. So this wavelength is used in estimation to estimate drug accurately. Preparation of standard stock solution of LENVATINIB 100mg of LENVITINIB was weighed and transferred in to 100ml volumetric flask and dissolved in methanol and then make up to the mark with methanol and prepare 100 µg /ml of solution by diluting 1ml to 10ml with methanol. RESULTS AND DISCUSSIONS Solubility Studies These studies are carried out at 25 0 C Lenvatinib Freely soluble in methanol,water and phosphate buffer. Wavelength determination In simultaneous estimation of two drugs isobestic wavelength is used. Isobestic point is the wavelength where the molar absorptivity is the same for two substances that are interconvertible. So this wavelength is used in simultaneous estimation to estimate both drugs accurately. Preparation of standard stock solution of LENVATINIB 100 mg of LENVATINIB was weighed in to 100ml volumetric flask and dissolved in Methanol and then dilute up to the mark with methanol and prepare 10 µg /ml of solution by diluting 1ml to 10ml with methanol, wavelength is found to be 240nm.
- 3. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~168~ Fig: UV-VIS spectrum of LENVATINIB RESULTS The wavelength of maximum absorption (λmax) of the drug, 10 μg/ml solution of the drugs in methanol were scanned using UV-Visible spectrophotometer within the wavelength region of 200–400 nm against methanol as blank. The resulting spectra and the absorption curve shows the isobestic point was found to be 240 nm for the combination. METHOD DEVELOPMENT OF LENVATINIB Trial- 1 Preparation of standard solution Weigh accurately 100 mg of LENVITINIB in 100 ml of volumetric flask and dissolve in 10ml of mobile phase and make up the volume with mobile phase From above stock solution 100 µg/ml of LENVITINIB is prepared by diluting 1ml to 10ml with mobile phase. This solution is used for recording chromatogram. Fig: Chromatogram of LENVATINIB Observation Peak Asymmetry factor for LENVATINIB meet the system suitability requirements. The run time is very correct. Theoretical plates were more than 2000. Hence it is taken for optimization.
- 4. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~169~ Table 1: Optimized chromatographic conditions Mobile phase Phosphate buffer(KH2PO4): Acetonitrile(80:20) Column INERTSIL column,C18(150x4.6 ID) 5µm Flow rate 1.0 ml/min Column temperature Room temperature(20-25o C) Sample temperature Room temperature(20-25o C) Wavelength 240nm Injection volume 20 µl Run time 6 min Retention time About Mobile phase Phosphate buffer(KH2PO4): Acetonitrile(80:20) Assay Preparation of samples for Assay Preparation of standard solution Weigh accurately 100 mg of LENVITINIB in 100 ml of volumetric flask and dissolve in 10ml of mobile phase and make up the volume with mobile phase From above stock solution 100 µg/ml of LENVITINIB is prepared by diluting 1ml to 10ml with mobile phase. This solution is used for recording chromatogram. Preparation of sample solution 20 tablets (each tablet contains 62.5mg of LENVITINIB was weighed and taken into a mortar and crushed to fine powder and uniformly mixed. Tablet stock solutions of LENVITINIB (100μg/ml) were prepared by dissolving weight equivalent to 62.5 mg of LENVITINIB and dissolved in sufficient mobile phase. After that filtered the solution using 0.45-micron syringe filter and Sonicated for 5 min and dilute to 100ml with mobile phase. Further dilutions are prepared in 5 replicates of 100μg/ml of LENVITINIB was made by adding 1 ml of stock solution to 10 ml of mobile phase. Calculation The amount of LENVITINIB present in the formulation by using the formula given below, and results shown in above table Where, AS: Average peak area due to standard preparation AT: Peak area due to assay preparation WS: Weight of LENVITINIB in mg WT: Weight of sample in assay preparation DT: Dilution of assay preparation
- 5. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~170~ Fig: Chromatogram of Assay standard preparation-1 Fig: Chromatogram of Assay standard preparation-2 Fig: Chromatogram of Assay standard preparation-3
- 6. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~171~ Fig: Chromatogram of Assay standard preparation-4 Fig: Chromatogram of Assay standard preparation-5
- 7. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~172~ Fig: Chromatogram of Assay sample preparation-1 Fig: Chromatogram of Assay sample preparation-2
- 8. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~173~ Fig: Chromatogram of Assay sample preparation-3 Fig: Chromatogram of Assay sample preparation-4
- 9. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~174~ Fig: Chromatogram of Assay sample preparation-5 Table: Assay Results LENVITINIB Standard Area Sample Area Injection-1 4430.026 4426.890 Injection-2 4447.469 4417.702 Injection-3 4387.764 4410.326 Injection-4 4376.391 4411.818 Injection-5 4347.941 4442.974 Average Area 6372.840 4426.890 Observation The amount of LENVITINIB present in the taken dosage form was found to be 90% and 110% respectively. VALIDATION Specificity by Direct comparison method There is no interference of mobile phase, solvent and placebo with the analyte peak and also the peak purity of analyte peak which indicate that the method is specific for the analysis of analytes in their dosage form. Preparation of mixed standard solution Weigh accurately 10 mg of LENVATINIB in 100 ml of volumetric flask and dissolve in 10ml of mobile phase and make up the volume with mobile phase From above stock solution 100 µg/ml of LENVATINIB is prepared by diluting 1ml to 10ml with mobile phase. This solution is used for recording chromatogram. Tablet sample 20 tablets (each tablet contains 10mg of LENVATINIB was weighed and taken into a mortar and crushed to fine powder and uniformly
- 10. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~175~ mixed. Tablet stock solutions of LENVATINIB (100μg/ml) were prepared by dissolving weight equivalent to 10 mg of LENVATINIB and dissolved in sufficient mobile phase. After that filtered the solution using 0.45-micron syringe filter and Sonicated for 5 min and dilute to 100ml with mobile phase. Further dilutions are prepared in 5 replicates of 100μg/ml of LENVATINIB was made by adding 1 ml of stock solution to 10 ml of mobile phase. Fig: Blank chromatogram for specificity by using mobile phase Fig: Chromatogram for specificity of LENVATINIB sample
- 11. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~176~ Fig: Chromatogram for Specificity of LENVATINIB standard Observation It is observed from the above data, diluent or excipient peaks are not interfering with the LENVATINIB peaks. Linearity and range Preparation of standard stock solution Standard stock solutions of LENVATINIB (microgram/ml) were prepared by dissolving 10 mg and LENVATINIB dissolved in sufficient mobile phase and dilute to 100 ml with mobile phase. Further dilutions were given in the table Table: Linearity Preparations Preparations Volume from standard stock transferred in ml Volume made up in ml (with mobile phase) Concentration of solution(µg /ml) Preparation 1 0.6 10 60 Preparation 2 0.8 10 80 Preparation 3 1.0 10 100 Preparation 4 1.2 10 120 Preparation 5 1.4 10 140
- 12. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~177~ Fig: Chromatogram of LENVATINIB preparation-1 Fig: Chromatogram of LENVATINIB preparation-2
- 13. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~178~ Fig: Chromatogram of LENVATINIB preparation-3 Fig: Chromatogram of LENVATINIB preparation-4
- 14. Kranthi K K et al, ICJPIR 2017, 4(1), XXX-XXX www.icjpir.com ~179~ Fig: Chromatogram of LENVATINIB for preparation-5 Table: Linearity of LENVATINIB PPM AREA 60 2559.483 80 3414.608 100 4426.89 120 5354.686 140 6043.075 Linearity graph of LENVATINIB y = 6.416x + 16.91 R² = 0.9976 0.000 1000.000 2000.000 3000.000 4000.000 5000.000 6000.000 7000.000 0 20 40 60 80 100 120 140 160 Area Conc Linearity of Lenvatinib
- 15. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~180~ Acceptance criteria The relationship between the concentrations of LENVATINIB should be linear in the specified range and the correlation should not be less than 0.99. Observation The correlation coefficient for linear curve obtained between concentration vs. Area for standard preparations of LENVATINIB is 0.991 is linear in the range examined since all points lie in a straight line and the correlation coefficient is well within limits. Accuracy Accuracy of the method was determined by Recovery studies. To the formulation (pre analyzed sample), the reference standards of the drugs were added at the level of 50%, 100%, 150%. The recovery studies were carried out three times and the percentage recovery and percentage mean recovery were calculated for drug is shown in table. To check the accuracy of the method, recovery studies were carried out by addition of standard drug solution to pre-analyzed sample solution at three different levels 50%, 100%, 150% Fig: Chromatogram of 50% recovery (injection 1) Fig: Chromatogram of 100% recovery (injection 2)
- 16. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~181~ Fig: Chromatogram of 150% recovery (injection 3) Fig: Chromatogram of 50% recovery (injection 1)
- 17. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~182~ Fig: Chromatogram of 100% recovery (injection 2) Fig: Chromatogram of 150% recovery (injection 3) Fig: Chromatogram of 50% recovery (injection 1)
- 18. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~183~ Fig: Chromatogram of 100% recovery (injection 2) Fig: Chromatogram of 150% recovery (injection 3) Acceptance criteria The % recovery of LENVATINIB should lie between 90% and 110%. Table : Recovery results for LENVATINIB Recovery level Accuracy LENVATINIB Amount taken(mcg/ml) Area Average area %Recovery Average % Recovery 75% 100 4198.222 4296.968 100.67 100.5 100 4295.484 100 4397.199 100% 120 5450.032 5464.189 102.86 120 5466.67 120 5476.267 125% 140 6195.931 6187.917 99.05 140 6184.41 140 6183.410
- 19. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~184~ Precision Method precision Prepared sample preparations of LENVATINIB as per test method and injected 6 times in to the column. Acceptance criteria The % Relative standard deviation of Assay preparations of LENVATINIB should be not more than 2.0%. Fig: Chromatogram of precision injection 1
- 20. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~185~ Fig: Chromatogram of precision injection 2 Fig: Chromatogram of precision injection 3
- 21. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~186~ Fig: Chromatogram of precision injection 4 Fig: Chromatogram of precision injection 5
- 22. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~187~ Fig: Chromatogram of precision injection 6 Table: Results for precision of LENVATINIB Lenvatinib S.No. Rt Area 1 3.560 4426.890 2 3.530 4417.702 3 3.533 4410.326 4 3.543 4411.818 5 3.533 4442.974 6 3.537 4413.064 avg 3.5393 4420.462 stdev 0.0111 12.553 %RSD 0.31 0.28 Observation Test results for LENVATINIBwas showing that the %RSD of Assay results are within limits. Robustness Chromatographic conditions variation To demonstrate the robustness of the method, prepared solution as per test method and injected at different variable conditions like using different conditions like Temperature and wavelength. System suitability parameters were compared with that of method precision. Acceptance criteria The system suitability should pass as per the test method at variable conditions.
- 23. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~188~ Fig: Chromatogram of LENVATINIB Robustness (0.8 ml/min) Fig: Chromatogram of LENVATINIB for Robustness (1.2 ml/min)
- 24. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~189~ Fig: Chromatogram of LENVATINIB for Robustness (238nm) Fig: Chromatogram of LENVATINIB for Robustness (242nm) Table: Result of Robustness study Parameter Lenvatinib Retention time(min) Area Flow 0.8ml/min 1.2ml/min 4.383 2.943 5477.973 3702.038
- 25. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~190~ Wavelength 238nm 242nm 3.510 3.517 4496.216 4253.798 Observation From the observation it was found that the system suitability parameters were within limit at all variable conditions. Ruggedness The ruggedness of the method was studied by the determining the analyst to analyst variation by performing the Assay by two different analysts. Acceptance criteria The % Relative standard deviation of Assay values between two analysts should be not more than 2.0%. Fig: Chromatogram of Analyst 01 standard preparation Fig: Chromatogram of Analyst 01 sample preparation
- 26. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~191~ Fig: Chromatogram of Analyst 02 standard preparation Fig: Chromatogram of Analyst 02 sample preparation Table: Results for Ruggedness Lenvatinib %Assay Analyst 01 99.92 Analyst 02 99.33 %RSD 0.27 Observation From the observation the between two analysts Assay values not greater than 2.0%, hence the method was rugged.
- 27. Kranthi K K et al, ICJPIR 2017, 4(1), 166-192 www.icjpir.com ~192~ BIBLIOGRAPHY [1]. Matsui, J.; Funahashi, Y.; Uenaka, T.; Watanabe, T.; Tsuruoka, A.; Asada, M. "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". Clinical Cancer Research. . doi:10.1158/1078-0432.CCR-07-5270. PMID 18765537, 14 (17), 2008, 5459–65. [2]. Haberfeld, H, ed. Austria-Codex (in German). Vienna: Österreichischer Apothekerverlag. 2015. [3]. FDA Professional Drug Information for Lenvima. [4]. http://www.fda.gov/Drugs/InformationOnDrugs/ApprovedDrugs/ucm501070.htm [5]. Inoue, K. "Oxidative Metabolic Pathway of Lenvatinib Mediated by Aldehyde Oxidase". Drug Metab Dispos. doi:10.1124/dmd.114.058073. PMID 24914245. 42, 2014, 1326–33. [6]. Glen, H; D. Boss; T. R. Evans; M. Roelvink; et al. "A phase I dose finding study of E7080 in patients (pts) with advanced malignancies". Journal of Clinical Oncology, ASCO Annual Meeting Proceedings Part I. 25(18S), 2007, 14073. [7]. Clinical trial number NCT01321554 for "A Trial of E7080 in 131I-Refractory Differentiated Thyroid Cancer" at ClinicalTrials.gov [8]. "Phase III trial shows lenvatinib meets primary endpoint of progression free survival benefit in treatment of radioiodine-refractory differentiated thyroid cancer" (PDF). Eisai. 2014. [9]. U.S. Food and Drug Administration. Hematology/Oncology (Cancer) Approvals & Safety Notifications. [1] [10]. European Medicines Agency Summary of the European public assessment report (EPAR) for Lenvima [2]