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The previously reported method for the preparation of decylenediamine-dextran T70 (T70- C10) was improved, and several chemical properties of the polymer support were examined. Methotrexate (MTX) was conjugated to T70-C10 by using 1-ethyl-3- (3-dimethylaminopropyl) - carbodiimide hydrochloride. The T70-C10MTX conjugate (T70-C10MTX) and T70-C10 carrying adsorbed MTX (T70-C10·adsorbed MTX) were dialyzed in order to examine the nature of the adsorption of MTX on T70-C10MTX. Before and after dialysis, the activity of these derivatives was assayed in terms of the binding affinity to dihydrofolate reductase. A more stable T70-C10 was obtained by using more severe reaction conditions in the Schiff's base formation between diaminodecane (C10) and oxidized dextran T70 and in the subsequent reduction. Conjugation was successful and the separation by gel-filtration (Sephadex G50) was good. The chemical properties of T70-C10 and T70-C10MTX were consistent and similar to those of the previous preparations. From the dialysis experiment, T70-C10MTX was estimated to have about 5 to 6% of the inhibitory activity of free MTX.

chemical and pharmaceutical bulletin/chemical and pharmaceutical bulletin

Novel Preparation of Decylenediamine-dextran T70 and Inhibitory Activity toward Dihydrofolate Reductase of Decylenediaminedextran T70-Methotrexate Conjugate