Abstract
Lyp-1, dokuz amino asitli bir peptid olup, tümör lenf damarlarına özgünlük göstermektedir. Yapılan çalışmalarda, peptidin tümör lenf damarlarında toplandığı ve hücreye internalizasyon özelliği gösterdiği belirlenmiştir. Ayrıca bağlandığı hücrede apoptozu indüklemesi de peptidin diğer önemli bir özelliğidir. Ancak literatürde bu peptide ait miktar tayini yöntemi henüz bulunmamaktadır. Dolayısıyla bu çalışmada, Lyp-1 peptidine ait bir RP-HPLC miktar tayini yöntemi geliştirilmiş ve valide edilmiştir. Validasyon parametreleri doğruluk, kesinlik tekrarlanabilirlik ve tekrar elde edilebilirlik , özgünlük, duyarlılık saptanabilme sınırı ve tayin edilebilme sınırı ve doğrusallık olarak belirlenmiştir. Yöntemin doğruluğu ve kesinliği test edilmiş ve varyasyon katsayıları ICH Validasyon Kılavuzu’na göre % 2’den küçük bulunmuştur. Geliştirilen metot 25 – 200 μg/ml konsantrasyon aralığında doğrusal bulunmuştur. metodun özgünlüğü tasarlanan SMEDDS formülasyonları kullanılarak ile gösterilmiştir. Sonuçta, bu çalışma ile Lyp-1 peptidinin miktar tayini için geçerli bir RP-HPLC metodu geliştirilmiş ve validasyonu başarıyla gerçekleştirilmiştir.
References
- Park, J.H., von Maltzahn, G., Xu, M.J., Fogal, V., Kotamraju, V.R., Ruoslahti, E. : Coop- erative nanomaterial system to sensitize, target, and treat tumors, Proc Natl Acad Sci USA, 107, 981 (2010)
- Laakkonen, P., Porkka, K., Hoffman, J.A., Ruoslahti, E. : A tumor-homing peptide with a targeting specificity related to lymphatic vessels, Nat Med, 8, 751 (2002)
- Laakkonen, P., Akerman, M.E., Biliran, H., Yang, M., Ferrer, F., Karpanen, T. et al. : Antitumor activity of a homing peptide that targets tumor lymphatics and tumor cells, Proc Natl Acad Sci U S A, 101, 9381 (2004)
- Enback, J., Laakkonen, P. : Tumour-homing peptides: tools for targeting, imaging and destruction, Biochem Soc Trans, 35, 780 (2007)
- Laakkonen, P., Zhang, L., Ruoslahti, E. : Peptide targeting of tumor lymph vessels, Ann N Y Acad Sci, 1131, 37 (2008)
- Ruoslahti, E. : Specialization of tumour vasculature, Nat Rev Cancer, 2, 83 (2002).
- Zhang, L., Giraudo, E., Hoffman, J.A., Hanahan, D.,Ruoslahti, E. : Lymphatic zip codes in premalignant lesions and tumors. Cancer Res, 66, 6894 (2006).
- Karmali, P.P., Kotamraju, V.R., Kastantin, M., Black, M., Missirlis, D., Tirrell, M. et al. Targeting of albumin-embedded paclitaxel nanoparticles to tumors. Nanomed: Nano- tech Biol Med, 5, 73 (2009).
- Luo, G., Yu, X., Jin, C., Yang, F., Fu, D., Long, J. et al. : LyP-1-conjugated nanoparticles for targeting drug delivery to lymphatic metastatic tumors. Int J Pharm, 385, 150 (2010).
- Cevik, O., Gursoy, N. Design and Characterization of Tumor Specific Lyp-1 Peptide- Containing SMEDDS for the Lymphatic Targeting of Solid Tumors in Cancer Therapy (2010), AAPS Annual Meeting and Exposition, 14 – 18 November 2010 New Orleans, LA, USA
- Yan Z., W., F., Wen Z., Zhan C., Feng L., Liu Y., Wei X., Xie C., Lu W. LyP-1-conjugated PEGylated liposomes: A carrier system for targeted therapy of lymphatic metastatic tumor, J Control Rel, (2011), doi: 10.1016/j.jconrel.2011.07.034.
- ICH Harmonised Tripartite Guideline, Validation of Analytical Procedures: Text and Methodology Q2 (R1), November, (2005)
- United States Pharmacopoeia 26 – National Formulatory 21, The United States Pharma- copeial Convention, (2003)
- Akerman, M.E., Chan, W.C., Laakkonen, P., Bhatia, S.N., Ruoslahti, E. : Nanocrystal targeting in vivo, Proc Natl Acad Sci USA, 99, 12617, (2002)
Abstract
Lyp-1 is a nine amino acid peptide, which is spesific to tumor lymphatics. It has been shown that the Lyp-1 peptide accumulates in tumor lymphatics, and it could be internalized in the tumor cell. It has also been shown to induce apoptosis in the cell that it binds to. On the other hand, there is not a quantitative analysis method of this peptide in the literature at present. Therefore, a RP-HPLC method has been developed and validated for the quantitative analysis of the Lyp-1 peptide in this study. Analytical validation parameters were accuracy, precision repeatability and reproducibility , specificity, sensitivity limit of detection and limit of quantification and linearity. The accuracy and precision of the method have been tested and the coefficients of variation have been found to be less than 2 % according to ICH Validation Guideline. The method has been found to be linear in the 25 – 200 μg/ml concentration range. The specificity of the method has been determined using the newly developed SMEDDS formulations. In conclusion, a new RP-HPLC method has been developed and validated successfully for the quantitative analysis of the Lyp-1 peptide in this study.
References
- Park, J.H., von Maltzahn, G., Xu, M.J., Fogal, V., Kotamraju, V.R., Ruoslahti, E. : Coop- erative nanomaterial system to sensitize, target, and treat tumors, Proc Natl Acad Sci USA, 107, 981 (2010)
- Laakkonen, P., Porkka, K., Hoffman, J.A., Ruoslahti, E. : A tumor-homing peptide with a targeting specificity related to lymphatic vessels, Nat Med, 8, 751 (2002)
- Laakkonen, P., Akerman, M.E., Biliran, H., Yang, M., Ferrer, F., Karpanen, T. et al. : Antitumor activity of a homing peptide that targets tumor lymphatics and tumor cells, Proc Natl Acad Sci U S A, 101, 9381 (2004)
- Enback, J., Laakkonen, P. : Tumour-homing peptides: tools for targeting, imaging and destruction, Biochem Soc Trans, 35, 780 (2007)
- Laakkonen, P., Zhang, L., Ruoslahti, E. : Peptide targeting of tumor lymph vessels, Ann N Y Acad Sci, 1131, 37 (2008)
- Ruoslahti, E. : Specialization of tumour vasculature, Nat Rev Cancer, 2, 83 (2002).
- Zhang, L., Giraudo, E., Hoffman, J.A., Hanahan, D.,Ruoslahti, E. : Lymphatic zip codes in premalignant lesions and tumors. Cancer Res, 66, 6894 (2006).
- Karmali, P.P., Kotamraju, V.R., Kastantin, M., Black, M., Missirlis, D., Tirrell, M. et al. Targeting of albumin-embedded paclitaxel nanoparticles to tumors. Nanomed: Nano- tech Biol Med, 5, 73 (2009).
- Luo, G., Yu, X., Jin, C., Yang, F., Fu, D., Long, J. et al. : LyP-1-conjugated nanoparticles for targeting drug delivery to lymphatic metastatic tumors. Int J Pharm, 385, 150 (2010).
- Cevik, O., Gursoy, N. Design and Characterization of Tumor Specific Lyp-1 Peptide- Containing SMEDDS for the Lymphatic Targeting of Solid Tumors in Cancer Therapy (2010), AAPS Annual Meeting and Exposition, 14 – 18 November 2010 New Orleans, LA, USA
- Yan Z., W., F., Wen Z., Zhan C., Feng L., Liu Y., Wei X., Xie C., Lu W. LyP-1-conjugated PEGylated liposomes: A carrier system for targeted therapy of lymphatic metastatic tumor, J Control Rel, (2011), doi: 10.1016/j.jconrel.2011.07.034.
- ICH Harmonised Tripartite Guideline, Validation of Analytical Procedures: Text and Methodology Q2 (R1), November, (2005)
- United States Pharmacopoeia 26 – National Formulatory 21, The United States Pharma- copeial Convention, (2003)
- Akerman, M.E., Chan, W.C., Laakkonen, P., Bhatia, S.N., Ruoslahti, E. : Nanocrystal targeting in vivo, Proc Natl Acad Sci USA, 99, 12617, (2002)
Details
| Primary Language | English |
|---|---|
| Journal Section | Research Article |
| Authors | |
| Publication Date | June 1, 2011 |
| Published in Issue | Year 2011 Issue: 2 |