Doxorubicin conjugated with a trastuzumab epitope and an MMP-2 sensitive peptide linker for the treatment of HER2-positive breast cancer

Abstract

HER2-positive breast cancer correlates with more aggressive tumor growth, a poorer prognosis and reduced overall survival. Currently, trastuzumab (Herceptin), which is an anti-HER2 antibody, is one of the key drugs. There is evidence indicating that conjugation of trastuzumab with chemotherapy drugs, such as doxorubicin (DOX), for multiple targets could be more effective. However, incomplete penetration into tumors has been noted for those conjugates. Compared to an antibody, peptides may represent an attractive alternative. For HER2, a similar potency has been observed for a 12-amino-acid anti-HER2 peptide mimetic YCDGFYACYMDV-NH2 (AHNP, disulfide-bridged) and full-length trastuzumab. Thus, a peptide, GPLGLAGDDYCDGFYACYMDV-NH2, which consists of AHNP and an MMP-2 cleavable linker GPLGLAGDD, was first designed, followed by conjugation with DOX via a glycine residue at the N-terminus to form a novel DOX-peptide conjugate MAHNP-DOX. Using HER2-positive human breast cancer cells BT474 and SKBR3 as in vitro model systems and nude mice with BT474 xenografts as an in vivo model, this conjugate was comprehensively characterized, and its efficacy was evaluated and compared with that of free DOX. As a result, MAHNP-DOX demonstrated a much lower in vitro IC50, and its in vivo extent of inhibition in mice was more evident. During this process, enzymatic cleavage of MAHNP-DOX is critical for its activation and cellular uptake. In addition, a synergistic response was observed after the combination of DOX and AHNP. This effect was probably due to the involvement of AHNP in the PI3K–AKT signaling pathway, which can be largely activated by DOX and leads to anti-apoptotic signals.

African Vegetables (Clerodendrum volubile Leaf and Irvingia gabonensis Seed Extracts) Effectively Mitigate Trastuzumab-Induced Cardiotoxicity in Wistar Rats

“…Trastuzumab use as a key treatment therapy for advanced HER2-positive breast carcinoma has also been reported to have yielded unequivocal improvements in the clinical treatment outcome of this disease [ 3 ]. Clinically, trastuzumab is either used alone or in combination with other cytotoxic agents especially with the anthracycline doxorubicin usually in a pegylated form although it is reported to be most effective in its combination form [ 4 ] since DOX enters its target cells by simple diffusion, intercalates into DNA, and inhibits topoisomerase II to hinder and completely stall DNA replication [ 5 ]. However, wide-scale clinical use of trastuzumab-based therapies has been significantly limited by its adverse cardiac dysfunctions and dilated cardiomyopathy-related congestive heart failures, which have been reported to occur in up to 27% of HER2-positive metastatic breast cancer patients on its combination therapy with doxorubicin [ 2 ].…”

Antibody–Drug Conjugates: The Last Decade

“…Smaller conjugate formats [64,68,69] have been explored to remedy these drawbacks, in particular peptides [70], single domain antibody fragments (sdAb or VHH) [71], single chain variable fragments (scFv) [72], antigen binding fragments (Fab) [73] or small immunoproteins (SIP) in the form of scFv dimerized using a CHε4 domain [74,75]. Surprisingly, only a few examples of drugs effectively vectorized with these new antibody formats have been published: Janda's team has described several scFv-duocarmycin conjugates with a DAR of 1 or 2, targeting laminin-binding integrin α 3 β 1 , with an EC 50 between 2.7 and 180.8 nM on pancreatic carcinoma cells SW1990 [76].…”

The Product of Matrix Metalloproteinase Cleavage of Doxorubicin Conjugate for Anticancer Drug Delivery: Calorimetric, Spectroscopic, and Molecular Dynamics Studies on Peptide–Doxorubicin Binding to DNA

“…In the first step, 1 mL of HBSS was added to 1 g of collagenase IV (205 u/mg) and mixed to complete dissolution. Activation of collagenase IV was performed using 1 mM APMA in 0.15 M NaOH for 1.5 h at 37 • C [24,43]. The octapeptide stock solution was added to activated 1 mM collagenase IV and incubated at 37 • C. The aliquots were removed and analyzed by HPLC, between 2 and 12 h. The HPLC Dionex Ultimate 3000 (Waltham, MA, USA) was equipped with a bioZen Intact XB-C8 column (3.6 µm, 2.1 × 150 mm; Phenomenex, Aschaffenburg, Germany).…”

“…MMPs are overexpressed in newly formed tumor tissues, thereby promoting angiogenesis, and are involved in cancer progression; MMP-2 and MMP-9, in particular, play a critical role in this process [22,23]. Therefore, MMPs could constitute a promising strategy for targeted therapeutic agent delivery via an enzyme-triggered release mechanism [24]. According to the literature, there are a series of substrate peptide sequences degradable in the presence of MMPs, e.g., Gly-Pro-Leu-Gly-Ile-Ala-Gly-Gln, Pro-Val-Gly-Leu-Ile-Gly, or Pro-Leu-Gly-Val-Arg, where cleavage occurs between leucine (Leu) and glycine (Gly) [25][26][27].…”