Cenderitide: structural requirements for the creation of a novel dual particulate guanylyl cyclase receptor agonist with renal-enhancingin vivoandex vivoactions

Abstract

Aim Cenderitide is a novel dual natriuretic peptide (NP) receptor chimeric peptide activator, which targets the particulate guanylyl cyclase B (pGC-B) receptor and pGC-A unlike native NPs. Cenderitide was engineered to retain the anti-fibrotic properties of CNP/pGC-B with renal enhancing actions facilitated by fusion to the carboxyl-terminus of Dendroaspis NP (DNP), a pGC-A agonist, to CNP. Here, we address significance of the DNP carboxyl-terminus in dual pGC receptor activation and actions of cenderitide compared to CNP on renal function and cGMP in vivo and ex vivo in normal canines. Methods and Results In vitro, only cenderitide and not CNP or three CNP-based variants was a potent dual pGC-A/pGC-B activator of cGMP production (from 5 to 237 pmol/ml) in HEK293 cells overexpressing human pGC-A while in pGC-B overexpressing cells cenderitide increased cGMP production (from 4 to 321 pmol/ml) while the three CNP-based variants were weak agonists. Based upon our finding that the DNP carboxyl-terminus is a key structural requirement for dual pGC-A/pGC-B activation, we defined in vivo the renal enhancing actions of cenderitide compared to CNP. Cenderitide increased urinary cGMP excretion (from 989 to 5977 pmol/ml), net generation of renal cGMP (821 to 4124 pmol/min), natriuresis (12 to 242 ueq/min) and GFR (37 to 51 ml/min) while CNP did not. We then demonstrated the transformation of CNP ex vivo into a renal cGMP activating peptide which increased cGMP in freshly isolated glomeruli 8 fold greater than CNP. Conclusion The current study establishes that dual pGC-A and pGC-B activation with CNP requires the specific carboxyl-terminus of DNP. In normal canines in vivo and in glomeruli ex vivo, the carboxyl-terminus of DNP transforms CNP into a natriuretic and GFR enhancing peptide. Future studies of cenderitide are warranted in cardiorenal disease states to explore its efficacy in overall cardiorenal homeostasis.

Natriuretic peptide based therapeutics for heart failure: Cenderitide: A novel first-in-class designer natriuretic peptide

“…4), which needs confirmation in a larger trial. These renal actions especially related to GFR are consistent with pGC-A action, as supported by previous studies in isolated glomeruli [38].…”

“…The importance of the unique structure of Cenderitide was recently reported by, Lee et al [38] Specifically, Lee and co-workers addressed the hypothesis that the 15-AA carboxyl-terminus of DNP, which is fused to CNP, uniquely facilitates dual pGC-A and pGC-B activation produced by Cenderitide. The actions of Cenderitide on cGMP activation in vitro in HEK293 cells selectively overexpressing human pGC-A or pGC-B were compared to native CNP and three variants of Cenderitide with altered carboxyl-termini.…”

Snake Venom Peptides: Tools of Biodiscovery

“…In vivo studies revealed that CD-NP, like ANP, BNP, and DNP (but unlike CNP) possesses renal-enhancing actions through pGC-A/cGMP activation. CD-NP was under clinical trials to treat cardiorenal syndrome [ 41 ]. However, Capricor Beverly Hills, California, recently completed an additional study with patients having stable heart failure in combination with moderate renal impairment.…”

Structural insight into hormone recognition and transmembrane signaling by the atrial natriuretic peptide receptor

“…Compared with ANP, BNP, and CNP, DNP has elongated N-and C-termini and is more stable against neutral endopeptidase 38 , which is the primary inactivator of natriuretic peptides. Based on the enhanced stability and high potency of DNP, chimeras of DNP and other natriuretic peptides have been created and assessed in clinical trials for heart failure 39,40 . Thus, understanding the difference between the binding of ANP and DNP to ANPR is essential to future drug discovery and research for heart failure therapies.…”