Abstract 4349330: Bispecific Activation of GC-A and GLP-1 Receptors: A Novel Peptide Therapeutic for Cardiometabolic Diseases

Introduction: Diabetes mellitus (DM) is strongly associated with metabolic and cardiorenal derangements. Natriuretic peptides (NPs), and glucagon-like peptide 1 receptor agonists (GLP-1 RA) have well-established cardiometabolic and cardiorenal protective effects. We developed a novel bispecific biologic, CRRL 101, which agonizes guanylyl cyclase A (GCA) NP receptors and GLP-1 receptors. In this study, we sought to evaluate the in vitro effects and in vivo impact of CRRL 101 in a DM rat model. Methods: In vitro experiments evaluated the impact of CRRL 101 treatment of GCA receptor transfected human embryonic kidney (HEK) cells and INS-1 rat pancreatic beta cells. In vivo experiments were conducted by inducing DM in Wistar rats (male, 6-8-week-old) by intraperitoneal Streptozotocin (STZ) injection (65 mg/kg). One week after STZ injection, DM was confirmed (fasting blood glucose > 350 mg/dL). Rats were then divided into 3 groups: saline control (n=6), 1 pmol/kg/min CRRL-101 (“1pmol” n= 4), and 10 pmol/kg/min CRRL-101 (“10 pmol” n=5). Treatment was infused for 120 min after which rat blood and urine were collected. Data were analyzed using GraphPad Prism 10.0.0. Results: In vitro, there was a marked increase in cyclic GMP (cGMP) levels (pmol/mL) in HEK treated with 10 -6 M CRRL 101 (mean difference ± SEM:1.603 ± 0.29 p = 0.0048). In INS-1 cells treated with CRRL 101 at 10 -6 M in a 20 mM glucose medium, cAMP levels (pmol/mL) markedly increased (16.7 ± 1.47 vs 23.19 ± 1.51 p=0.0036) and insulin levels (ng/mL) doubled (4.4 ng/mL ± 0.44 vs 9.5 ng/mL ± 0.34 in 20 mM glucose p < 0.0001). Similar increases were observed at lower doses (10 -7 M, 10 -8 M) of CRRL 101. In vivo , after 120 minutes of treatment, mean plasma cGMP levels were 6.5x greater in both groups (mean difference± SEM) (1pmol: 38.89 ± 7.93, p = 0.016; 10pmol: 39.40 ± 4.84 p=0.001). Plasma insulin (ng/mL) more than doubled with both doses (1pmol: 0.39 ± 0.148, p = 0.039; 10 pmol: 0.198 ± 0.051, p = 0.004) with no significant increase with placebo ( 0.03 ± 0.08, p = 0.67). Rate of natriuresis (mEq/min) more than doubled with peptide administration vs placebo (1 pmol: 0.36 ± 0.093 p = 0.014; 10 pmol: 0.38 ± 0.13, p = 0.034). Conclusions: Our novel bispecific biologic, CRRL 101, activates GC-A, increasing plasma cGMP and inducing natriuresis, and stimulates GLP-1R, raising cAMP and plasma insulin. Chronic administration studies to eludicate its therapeutic efficacy are warranted.