Synthesis of HPMA Copolymer Containing Adriamycin Bound via an Acid-Labile Spacer and its Activity toward Human Ovarian Carcinoma Cells

Abstract

N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer-adriamycin (ADR) conjugate (P-aconityl-ADR) was synthesized by the attachment of cis-aconityl-ADR to an HPMA copolymer precursor using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) as the condensing agent. The ADR release from the HPMA copolymer conjugate was pH sensitive. After 48 h incubation at pH 5, 6, and 7, the amount of ADR released was 63.4, 9.2, and 2.8% respectively. The in vitro cytotoxicity of the conjugate was evaluated toward A2780 sensitive and A2780/AD resistant human ovarian carcinoma cells. An HPMA copolymer, containing ADR bound via a tetrapeptide (GFLG) sequence susceptible to cleavage catalyzed by lysosomal enzymes (P-GFLG-ADR), was used as control. The IC₅₀ doses seemed to indicate that the total hydrolysis of P-aconityl-ADR in prelysosomal and lysosomal compartments proceeded faster than the release of ADR from P-GFLG-ADR catalyzed by lysomal cysteine proteinases. Both HPMA copolymer-ADR conjugates appeared to overcome the ATP-driven P-glycoprotein efflux pump expressed in A2780/AD cells.

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