acetyl-prolyl-histidyl-seryl-cysteinyl-asparaginamide and Prostatic-Neoplasms

Activated alpha5beta1 integrin occurs specifically on tumor cells and on endothelial cells of tumor-associated vasculature, and plays a key role in invasion and metastasis. The PHSCN peptide (Ac-PHSCN-NH(2)) preferentially binds activated alpha5beta1, to block invasion in vitro, and inhibit growth, metastasis and tumor recurrence in preclinical models of prostate cancer. In Phase I clinical trial, systemic Ac-PHSCN-NH(2) monotherapy was well tolerated, and metastatic disease progression was prevented for 4-14 months in one-third of treated patients. We have developed a significantly more potent derivative, the PHSCN-polylysine dendrimer (Ac-PHSCNGGK-MAP). Using in vitro invasion assays with naturally serum-free basement membranes, we observed that the PHSCN dendrimer was 130- to 1900-fold more potent than the PHSCN peptide at blocking alpha5beta1-mediated invasion by DU 145 and PC-3 human prostate cancer cells, whether invasion was induced by serum, or by the Ac-PHSRN-NH(2) peptide, under serum-free conditions. The PHSCN dendrimer was also approximately 800 times more effective than PHSCN peptide at preventing DU 145 and PC-3 extravasation in the lungs of athymic mice. Chou-Talalay analysis suggested that inhibition of both invasion in vitro and extravasation in vivo by the PHSCN dendrimer are highly synergistic. We found that many extravasated DU 145 and PC-3 cells go onto develop into metastatic colonies, and that a single pretreatment with the PHSCN dendrimer was 100-fold more affective than the PHSCN peptide at reducing lung colony formation. Since many patients newly diagnosed with prostate cancer already have locally advanced or metastatic disease, the availability of a well-tolerated, nontoxic systemic therapy, like the PHSCN dendrimer, which prevents metastatic progression by inhibiting invasion, could be very beneficial.

Topics: Angiogenesis Inhibitors; Animals; Dendrimers; Disease Progression; Humans; Inhibitory Concentration 50; Integrin alpha5beta1; Lung Neoplasms; Male; Mice; Mice, Nude; Neoplasm Invasiveness; Oligopeptides; Prostatic Neoplasms; Tumor Cells, Cultured

Copolymers of N-(2-hydroxypropyl)methacrylamide (HPMA) are prototypic and well-characterized polymeric drug carriers that are being broadly implemented in the delivery of anticancer therapeutics. To better predict the in vivo potential of the copolymers and to describe the biodistributional consequences of functionalization, 13 physicochemically different HPMA copolymers were synthesized, varying in molecular weight and in the nature and amount of functional groups introduced. Upon radiolabeling, the copolymers were injected i.v., and their circulation kinetics, tissue distribution and tumor accumulation were monitored in rats bearing subcutaneous Dunning AT1 tumors. It was found that increasing the average molecular weight of HPMA copolymers resulted in prolonged circulation times and in increased tumor concentrations. Conjugation of carboxyl and hydrazide groups, as well as introduction of spacer, drug and peptide moieties reduced the long-circulating properties of the copolymers and as a result, lower levels were found in tumors and in all organs other than kidney. Interestingly, however, in spite of the reduced (absolute) tumor concentrations, hardly any reduction in the relative levels localizing to tumors was found. Tumor-to-organ ratios were comparable to unmodified control for the majority of chemically modified copolymers, indicating that functionalization does not necessarily affect the tumor targeting ability of the copolymers and suggesting that HPMA copolymer-based drug delivery systems may prove to be attractive tools for more effectively treating various forms of advanced solid malignancy.

Topics: Animals; Antineoplastic Agents; Cell Line, Tumor; Doxorubicin; Drug Carriers; Male; Methacrylates; Molecular Weight; Neoplasm Transplantation; Oligopeptides; Polymethacrylic Acids; Prostatic Neoplasms; Rats; Tissue Distribution

Using naturally serum-free SU-ECM basement membranes as invasion substrates showed that plasma fibronectin was necessary to stimulate invasion by DU 145 human and metastatic MATLyLu (MLL) rat prostate carcinoma cells. This activity mapped to the PHSRN sequence, which induced invasion through alpha5beta1 integrin. PHSCN, a competitive inhibitor, blocked both PHSRN- and serum-induced invasion. Acetylated, amidated PHSCN (Ac-PHSCN-NH2) was 30-fold more potent; however, Ac-HSPNC-NH2 was inactive. Rats receiving injections s.c. with 100,000 MLL cells were treated systemically by i.v. injection three times weekly with 1 mg of either Ac-PHSCN-NH2 or Ac-HSPNC-NH2 beginning 24 h later, three times weekly with 1 mg of Ac-PHSCN-NH2 beginning only after surgery to remove large (2 cm) MLL tumors, or were left untreated. MLL tumors grew rapidly in Ac-HSPNC-NH2-treated and in untreated rats. MLL tumor growth in rats treated with Ac-PHSCN-NH2 beginning 1 day after MLL cell injection was reduced by 99.9% during the first 16 days of treatment, although subsequent tumor growth occurred. MLL tumor cryosections immunostained with anti-PECAM-1 showed that Ac-PHSCN-NH2 inhibited neovascularization by 12-fold during this time. Whether initiated after MLL cell injection or only after MLL tumor removal, Ac-PHSCN-NH2 treatment reduced the numbers of MLL lung colonies and micrometastases by 40- to >100-fold, whereas Ac-HSPNC-NH2 was inactive. Thus, Ac-PHSCN-NH2 may be a potent antitumorigenic and antimetastatic agent for postsurgical use prior to extensive metastasis.

Topics: Amino Acid Sequence; Animals; Antineoplastic Agents; Basement Membrane; Cell Survival; Chemotaxis; Fibronectins; Humans; Lung Neoplasms; Male; Neoplasm Invasiveness; Neoplasm Metastasis; Oligopeptides; Prostatic Neoplasms; Rats; Receptors, Fibronectin; Structure-Activity Relationship; Tumor Cells, Cultured