The competition of drugs to serum albumin in combination chemotherapy:: NMR study

Combination chemotherapy with cyclophosphamide (CM), metotrexate and 5-fluorouracil (FU) is used in treatment of patients with breast carcinoma. Although clinical toxicity of CM combinated with FU is greater than that of CM, the levels were clinically acceptable. The mechanism of competition of CM and FU to bovine serum albumin (BSA) was examined with the use of H-1 and C-13 NMR spectroscopy. The chemical shifts and the linewidth of individual proton and carbon resonances of each drug were measured as a function of the drug/BSA molar ratio in order to analyse the drug-protein interaction and the molecular motion of the drug. The effect of the second drug used in the combination chemotherapy on the analysed NMR parameters is discussed. It was found that FU and CM bind to BSA at molar ratio drug/BSA 160 and 330, respectively. The formation of lev-BSA complex was not confirmed. Whereas it was proved that in the presence of both lev and CM the number of FU molecules bound with BSA increases. It was also observed that FU induces the rising of the affinity between lev and BSA. (c) 2004 Elsevier B.V. All rights reserved.