Oxime bond-linked daunorubicin-GnRH-III bioconjugates exert antitumor activity in castration-resistant prostate cancer cells via the type I GnRH receptor

Abstract

It is well established that gonadotropin-releasing hormone receptors (GnRH-R) are expressed in different types of cancers, including castration-resistant prostate cancer (CRPC) and mediate the antiproliferative effect of GnRH analogs. Thus, these compounds are employed as targeting moieties to selectively deliver chemotherapeutic agents to cancer cells. GnRH-III, the decapeptide isolated from the sea lamprey brain, has lower potency than GnRH in stimulating gonadotropin secretion, but it exerts antiproliferative effects on many tumors expressing the GnRH-R. GnRH-III-based peptides are considered promising targeting moieties for the preparation of anticancer drug delivery systems. These studies were aimed at i) evaluating the antitumor activity of two cytotoxic oxime bond-linked daunorubicin (Dau)-GnRH-III derivative bioconjugates (Dau-GnRH-III, in which daunorubicin was coupled to the 8Lys in the native form of GnRH-III, and Dau-[4Lys(Ac)]-GnRH-III, in which daunorubicin was attached to the 8Lys of a GnRH-III derivative where 4Ser was replaced by an acetylated lysine) on CRPC cells; and ii) to elucidate the involvement of the classical GnRH-R (type I GnRH-R) in this antitumor activity. Our results demonstrated that both Dau-GnRH-III and Dau-[4Lys(Ac)]-GnRH-III were rapidly internalized into DU145 prostate cancer cells and exerted a significant cytostatic effect. Both bioconjugates increased the levels of the active form of caspase-3, indicating the involvement of apoptosis in their antitumor activity. The antiproliferative effect of both Dau-GnRH-III and Dau-[4Lys(Ac)]-GnRH-III was counteracted by the simultaneous treatment of the cells with Antide, an antagonist of the GnRH-R. Moreover, after silencing the type I GnRH-R the antitumor activity of both bioconjugates was completely abolished. These data demonstrate that in CRPC cells, daunorubicin-GnRH-III derivative bioconjugates: i) inhibit tumor cell proliferation, by triggering the apoptosis process; ii) exert their antitumor effect through the activation of the type I GnRH-R expressed on these cells. Cytotoxic-GnRH-III derivative may represent promising targeted chemotherapeutics for the treatment of CRPC patients.

Comparative cell biological study of in vitro antitumor and antimetastatic activity on melanoma cells of GnRH-III-containing conjugates modified with short-chain fatty acids

“…A similar enhanced antitumor activity was also detected for conjugates modified with acylated 4 Lys in the HT-29 human colon [ 19 ], LNCaP [ 17 ] and DU145 [ 37 ] human prostate cancer cell lines compared to the conjugate containing native GnRH-III sequence. Despite the increased activity of conjugates substituted with acylated 4 Lys, they displayed their dose-dependent cytotoxic activity at higher concentrations comparing to the free Dau.…”

GnRH and GnRH receptors in the pathophysiology of the human female reproductive system

“…In humans, as reported for GnRH-II, also GnRH-III seems to exert its effects through the activation of the classical form of GnRHRs (Montagnani Marelli et al, 2015); however, GnRH-III seems to exert a significant anti-tumour effect on human cancer cells expressing the GnRHR, while causing very low activity on gonadotrophin secretion in mammals (Kovacs et al, 2002).…”

Dissecting the Hormonal Signaling Landscape in Castration-Resistant Prostate Cancer

“…In our laboratory, we investigated the antitumor activity of bioconjugates consisting of daunorubicin linked to the GnRH-III isoform in CRPC cells. We found that these compounds are rapidly internalized and exert a significant antitumor activity in these cells [ 292 ]. Further studies are needed to confirm the efficacy and the lack of toxicity of GnRH-based cytotoxic bioconjugates in PCa.…”