Intraneoplastic polymer-based delivery of cyclophosphamide for intratumoral bioconversion by a replicating oncolytic viral vector

rRp450 is an oncolytic herpesvirus that expresses the CYP2B1 cDNA, responsi ble for bioconverting cyclophosphamide (CPA) into the active metabolites 4- hydroxyCPA/aldophosphamide (AP). However, formal proof of prodrug activatio n is lacking. We report that activation of CPA in cells infected with rRp45 0 generates a time-dependent increase of diffusible 4-hydroxyCPA/AP. For in vivo applications, a CPA-impregnated polymer was implanted into human tumo r xenografts inoculated with rRp450. The area under the curve for 4-hydroxy CPA/AP was 806 mug/g of tumor tissue/h when CPA was administered via intran eoplastic polymer and 3 mug/g of tumor tissue/h when CPA was administered i .p. Therefore, (a) a lytic virus expressing a "suicide" gene can activate a prodrug; and (b) within rRp450-infected tumor, more prolonged and higher c oncentrations of activated metabolites are generated by intraneoplastic com pared with systemic administration of prodrug.