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Development of a novel prodrug of paclitaxel that is cleaved by prostate-specific antigen: an in vitro and in vivo evaluation study

 

Bakheet Elsadek, Ralph Graeser, Norbert Esser, Cynthia Schäfer-Obodozie,

Khalid Abu Ajaj, Clemens Unger, André Warnecke, Tahia Saleem, Nagla El-Melegy, Hafez Madkor, Felix Kratz

 

 

European Journal of Cancer, 2010, 46 (18), 3434–3444

 

Keywords: paclitaxel; prodrug; human serum albumin; prostate-specific antigen; orthotopic animal model, in vivo bioluminescence; prostate cancer.

 

Abstract:

In developed countries, prostate cancer is the third most common cause of death from cancer in men. Unfortunately, while accumulating clinical data have suggested that taxanes may prolong the survival in a subset of men with prostate carcinoma, the dose and duration of administration of these drugs are limited by their significant systemic toxicities due to a lack of tumor selectivity. In an attempt to improve both the water solubility and tumor targeting properties of paclitaxel (Taxol®), we set out to develop a water soluble paclitaxel prodrug that is activated specifically by prostate-specific antigen (PSA) which is almost exclusively expressed in prostate tissue and prostate carcinoma making it an ideal molecular target for prodrugs strategies. Using albumin as drug carrier, we describe a novel albumin-binding prodrug of paclitaxel, EMC-Arg-Ser-Ser-Tyr-Tyr-Ser-Leu-PABC-paclitaxel [EMC: ε-maleimidocaproyl; PABC: p-aminobenzyloxycarbonyl] that was synthesized by reacting EMC-Arg-Ser-Ser-Tyr-Tyr-Ser-OH with H-Leu-PABC-paclitaxel. This prodrug was water soluble and was bound to endogenous and exogenous albumin. Moreover, incubation studies with PSA demonstrated that the albumin-bound form of the prodrug was cleaved rapidly at the P1–P1' scissile bond releasing the paclitaxel dipeptide H-Ser-Leu-PABC-paclitaxel. Last but not least, due to the incorporation of a PABC self-eliminating linker, this dipeptide was rapidly degraded to liberate paclitaxel as a final cleavage product within a few hours in prostate tumor tissue homogenates. Of note was that the albumin-bound form of the prodrug was approximately three-fold more active in killing PSA-positive LNCaP cells than paclitaxel. In addition, orientating toxicity studies in mice showed that the maximum tolerated dose of the novel paclitaxel prodrug was twice that of conventional paclitaxel. When tested in vivo in an orthotopic mouse model of human prostate cancer using luciferase-transduced LNCaP LLN cells, both paclitaxel and the new paclitaxel prodrug showed distinct antitumor efficacy on the primary tumor and metastases that was significantly better than the effect of doxorubicin that was used as a comparison and showed no antitumor efficacy. The new paclitaxel prodrug (3 × 24 mg paclitaxel equivalents) showed comparable antitumor activity on the primary tumor to paclitaxel at its maximum tolerated dose (3 × 12 mg/kg), redciced circulating PSA levels and demonstrated   a better antitumour effect on lung metastases but not on bone metastases.

 

 

 

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