Dual Constant Domain-Fab: A novel strategy to improve half-life and potency of a Met therapeutic antibody.
The kinase receptor encoded by the Met oncogene is a sensible target for cancer therapy. The chimeric monovalent Fab fragment of the DN30 monoclonal antibody (MvDN30) has an odd mechanism of action, based on cell surface removal of Met via activation of specific plasma membrane proteases. However, the short half-life of the Fab, due to its low molecular weight, is a severe limitation for the deployment in therapy. This issue was addressed by increasing the Fab molecular weight above the glomerular filtration threshold through the duplication of the constant domains, in tandem (DCD-1) or reciprocally swapped (DCD-2). The two newly engineered molecules showed biochemical properties comparable to the original MvDN30 in vitro, acting as full Met antagonists, impairing Met phosphorylation and activation of downstream signaling pathways. As a consequence, Met-mediated biological responses were inhibited, including anchorage-dependent and -independent cell growth. In vivo DCD-1 and DCD-2 showed a pharmacokinetic profile significantly improved over the original MvDN30, doubling the circulating half-life and reducing the clearance. In pre-clinical models of cancer, generated by injection of tumor cells or implant of patient-derived samples, systemic administration of the engineered molecules inhibited the growth of Met-addicted tumors.
Candiolo Cancer Institute, FPO-IRCCS, Str Prov 142, 10060 Candiolo, Italy; University of Turin, Department of Oncology, Str Prov 142, 10060 Candiolo, Italy.
Simona
S
Cignetto
Candiolo Cancer Institute, FPO-IRCCS, Str Prov 142, 10060 Candiolo, Italy; University of Turin, Department of Oncology, Str Prov 142, 10060 Candiolo, Italy.
Chiara
C
Modica
Candiolo Cancer Institute, FPO-IRCCS, Str Prov 142, 10060 Candiolo, Italy.
Cristina
C
Chiriaco
Candiolo Cancer Institute, FPO-IRCCS, Str Prov 142, 10060 Candiolo, Italy.
Lara
L
Fontani
University of Turin, Department of Science and Drug Technology, Via P. Giuria 9, 10125 Turin, Italy.
Paola
P
Milla
Candiolo Cancer Institute, FPO-IRCCS, Str Prov 142, 10060 Candiolo, Italy; University of Turin, Department of Oncology, Str Prov 142, 10060 Candiolo, Italy.
Paolo
P
Michieli
Candiolo Cancer Institute, FPO-IRCCS, Str Prov 142, 10060 Candiolo, Italy; University of Turin, Department of Oncology, Str Prov 142, 10060 Candiolo, Italy. Electronic address: pcomoglio@gmail.com.
Paolo M
PM
Comoglio
Candiolo Cancer Institute, FPO-IRCCS, Str Prov 142, 10060 Candiolo, Italy; University of Turin, Department of Oncology, Str Prov 142, 10060 Candiolo, Italy. Electronic address: elisa.vigna@ircc.it.
Elisa
E
Vigna
D000072283:A549 Cells; D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D000970:Antineoplastic Agents / Q000097:blood / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D003106:Colon / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D005260:Female; D057809:HEK293 Cells; D006207:Half-Life; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000097:blood / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D010766:Phosphorylation / Q000187:drug effects; D000072417:Protein Domains; D015202:Protein Engineering; D019859:Proto-Oncogene Proteins c-met / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects
D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D007140:Immunoglobulin Fab Fragments; D019859:Proto-Oncogene Proteins c-met
Antibody; Cancer targeted therapy; Fab; Half-life; Met; Protein engineering