Synthesis and biological activity of raltitrexed-carrier conjugates
Languages of publication
Drugs used in chemotherapy give undesirable side effects, e.g., cardiotoxicity, leucopenia, hair loss and others. Covalent binding of a drug with a carrier may change its biodistribution, elimination and/or rate of transformation in the organism. The aim of this work was to synthesize conjugates of anticancer drug - raltitrexed (RTX) with lysozyme, bovine serum albumin (BSA), and dextran T40 and to investigate their cytotoxicity and influence on the cell cycle in comparison with the free drug. Before conjugation RTX was transformed into anhydride by treatment with dicyclohexylcarbodiimide in dimethylformamide. Activated RTX was added into aqueous solution of carriers at different pH (from 8.5 to 10.5) for 3 to 15 min. The reaction was stopped by reducing the pH to 7.0. Maximum yield of the reaction was obtained at pH 10 for BSA as well as for dextran. The highest level of substitution was obtained after 5 min of the reaction. In in vitro experiments on three cell lines: SW707, LoVo and A549, all conjugates tested had up to a few hundred times higher IC50 than the free drug. Interestingly, it was noticed that the conjugates based on dextran and albumin were more cytotoxic than the free drug in the highest concentrations tested (1000 and 10000 ng/ml). The influence of RTX and the conjugates on SW707 cell cycle was studied. RTX blocked the cell cycle mostly in the G0-G1 and S phase and increased the percentage of apoptotic cells. Cells in the G2-M phase were not observed. The conjugates blocked the cell cycle in the S phase and decreased the percentage of cells in the G0-G1 phase.
- Bolling C, Graefe T, Lubbing C, Jankevicius F, Uktveris S, Cesas A, Meyer-Moldenhauer WH, Starkmann H, Weigel M, Burk K, Hanauske AR (2006) Phase II study of MTX-HSA in combination with cisplatin as first line treatment in patients with advanced or metastatic transitional cell carcinoma. Invest New Drugs 24: 521-527.
- Boratynski J, Opolski A, Wietrzyk J, Gorski A, Radzikowski C (2000) Cytotoxic and antitumor effect of fibrinogen-methotrexate conjugate. Cancer Lett 148: 189-195.
- Boratynski J (1984) Colorimetric method for the determination of ketoses using phenol-acetone-boric acid reagent (PABR). Anal Biochem 137: 528-532.
- Budzynska R, Nevozhay D, Kanska U, Jagiello M, Opolski A, Wietrzyk J, Boratynski J (2007) Antitumor activity of mannan-methotrexate conjugate in vitro and in vivo. Oncol Res 16: 415-421.
- Burger AM, Hartung G, Stehle G, Sinn H, Fiebig HH (2001) Pre-clinical evaluation of a methotrexate-albumin conjugate (MTX-HSA) in human tumor xenografts in vivo. Int J Cancer 92: 718-724.
- Ghose T, Norvell ST, Guclu A, Cameron D, Bodurtha A, MacDonald AS (1972) Immunochemotherapy of cancer with chlorambucil carrying antibody. Br Med J 3: 495-499.
- Hartung G, Stehle G, Sinn H, Wunder A, Schrenk HH, Heeger S, Kranzle M, Edler L, Frei E, Fiebig HH, Heene DL, Maier-Borst W, Queisser W (1999) Phase I trial of methotrexate-albumin in a weekly intravenous bolus regimen in cancer patients. Phase I study group of the association for medical oncology of the German cancer society. Clin Cancer Res 5: 753-759.
- Kanska U, Omar MS, Budzynska R, Nevozhay D, Jagiello M, Opolski A, Boratynski J (2005) Antileukemic activity of glycated fibrinogen-methotrexate conjugates. Anticancer Res 25: 2229-2234.
- Kratz F (2008) Albumin as a drug carrier: design of prodrugs, drug conjugates and nanoparticles. J Control Release 132: 171-183.
- Li KL, Clarke SJ, Rivory LP (2003) Quantitation of plasma thymidine by high-performance liquid chromatography - atmospheric pressure chemical ionization mass spectrometry and its application to pharmacodynamic studies in cancer patients. Anal Chim Acta 486: 51-61.
- Mathe G, Loc T, Bernard J (1958) Effect sur la leucemie 1210 de la souris d'une combinaison par diazotation d'A-methopterine et da γ-globulines de hamsters porteurs de cette leucmie par heterograffe. C R Acad Sci (Paris) 246: 1626-1630.
- McGuire JJ (2003) Anticancer antifolates: current status and future directions. Curr Pharm Des 9: 2593-2613.
- Myc A, Kukowska-Latallo J, Cao P, Swanson B, Battista J, Dunham T, Baker JR Jr (2010) Targeting the efficacy of a dendrimer-based nanotherapeutic in heterogeneous xenograft tumors in vivo. Anticancer Drugs 21: 186-192.
- Nevozhay D, Budzynska R, Jagiello M, Kanska U, Omar MS, Opolski A, Wietrzyk J, Boratynski J, Opolski, A (2006a) The effect of the substitution level of some dextran-methotrexate conjugates on their antitumor activity in experimental cancer models. Anticancer Res 26: 2179-2186.
- Nevozhay D, Budzynska R, Kanska U, Jagiello M, Omar MS, Boratynski J, Opolski A (2006b) Antitumor properties and toxicity of dextran-methotrexate conjugates are dependent on the molecular weight of the carrier. Anticancer Res 26: 1135-1143.
- Nevozhay D, Kanska U, Budzynska R, Boratynski J (2007) Current status of research on conjugates and related drug delivery systems in the treatment of cancer and other diseases. Postepy Hig Med Dosw 61: 350-360.
- Riebeseel K, Biedermann E, Loser R, Breiter N, Hanselmann R, Mulhaupt R, Unger C, Kratz F (2002) Polyethylene glycol conjugates of methotrexate varying in their molecular weight from MW 750 to MW 40000: synthesis, characterization, and structure-activity relationships in vitro and in vivo. Bioconjug Chem 13: 773-785.
- Silverstein AM (1999) Paul Ehrlich's passion: the origins of his receptor. Cell Immunol 194: 213-221.
- Skehan P, Storeng R, Scudiero D, Monks A, McMahn J, Vistica D, Warren JT, Bokesch H, Kenney S, Boyd MR (1990) New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst 82: 1107-1112.
- Stehle G, Wunder A, Sinn H, Schrenk HH, Schutt S, Frei E, Hartung G, Maier-Borst W, Heene DL (1997) Pharmacokinetics of methotrexate-albumin conjugates in tumor-bearing rats. Anticancer Drugs 8: 835-844.
- Stehle G, Wunder A, Schrenk HH, Hartung G, Heene DL, Sinn H (1999) Methotrexate-albumin conjugate causes tumor growth delay in Dunning R3327 HI prostate cancer-bearing rats. Anticancer Drugs 10: 405-411.
- Wilson KS, Malfair Taylor SC (2009) Raltitrexed: optimism and reality. Expert Opin Drug Metab Toxicol 5: 1447-1454.
- Wunder A, Stehle G, Schrenk HH, Hartung G, Heene DL, Maier-Borst W, Sinn H (1998) Antitumor activity of methotrexate-albumin conjugates in rats bearing a Walker-256 carcinoma. Int J Cancer 76: 884-890.
- Yousefi G, Foroutan SM, Zarghi A, Shafaati A (2010) Synthesis and characterization of methotrexate polyethylene glycol esters as a drug delivery system. Chem Pharm Bull (Tokyo) 58: 147-153.
Publication order reference