GDF15 is a circulating polypeptide associated with cellular stress, and recently linked to metabolic adaptation. GDF15 has a half-life of approximately 3 hours in and acts at the GFRAL receptor selectively expressed in the area postrema. To characterize the effects of sustained GFRAL agonism on food intake (FI) and body weight (BW), we developed a half-life extended analog of GDF15 (Compound H; CpdH) suitable for reduced dosing frequency and tested its effects in obese cynomolgus monkeys. Animals were treated once weekly for 12 weeks with 0.048, 0.16, or 1.6 mg/Kg of CpdH or with 0.02 mg/Kg of the long-acting GLP-1 analog dulaglutide as a positive control. FI was measured daily and BW was measured biweekly. Mechanism-based longitudinal exposure-response (E-R) modeling was performed to characterize the effects of CpdH and dulaglutide on FI and BW. The integrated novel model accounts for both acute, exposure-dependent effects of treatments to reduce FI and the compensatory changes in energy expenditure (EE) and FI that occur over time in response to weight loss. CpdH had approximately linear, dose-proportional pharmacokinetics with a half-life of ≈8 days and treatment with CpdH led to dose- and exposure-dependent reductions in FI and BW. The 1.6 mg/Kg CpdH dose reduced mean FI by 57.5% at 1 week and provided sustained FI reductions of 31.5% from weeks 9-12, leading to a peak reduction in BW of 16±5%. Dulaglutide had more modest effects on FI (reductions between 15-40%) and peak BW loss was 3.8±4.0%. Longitudinal modeling of both the FI and BW profiles suggested reductions in BW observed with both CpdH and dulaglutide were fully explained by the exposure-dependent reductions in FI without any increase in EE. Upon verification of pharmacokinetic/pharmacodynamic relationship established in monkeys and human for dulaglutide, we predicted that CpdH could reach double digit BW loss in human. In summary, treatment with a long-acting GDF15 analog led to sustained dose- and exposure-dependent reductions in food intake in a monkey model of obesity and holds potential for effective clinical obesity pharmacotherapy. Significance Statement GDF15 activation of GFRAL receptors in the hindbrain controls food intake and body weight. Here we describe the effect and durability of a circulating half-life extended analog of GDF15 (Compound H) on food intake and body weight loss in a spontaneously obese cynomolgus monkey model. Inclusion of a translational treatment arm with a weight loss agent, dulaglutide, permitted pharmacokinetic/pharmacodynamic modeling and comparison of both GDF15 and GLP-1 based weight loss mechanisms, and development of an allometric scaling based mathematical model to estimate the efficacy of Compound H in human obese subjects.

The long circulating half-life and inherently bivalent architecture of IgGs provide an ideal vehicle for presenting otherwise short-lived G-protein-coupled receptor agonists in a format that enables avidity-driven enhancement of potency. Here, we describe the site-specific conjugation of a dual agonist peptide (an oxyntomodulin variant engineered for potency and in vivo stability) to the complementarity-determining regions (CDRs) of an immunologically silent IgG4. A cysteine-containing heavy chain CDR3 variant was identified that provided clean conjugation to a bromoacetylated peptide without interference from any of the endogenous mAb cysteine residues. The resulting mAb-peptide homodimer has high potency at both target receptors (glucagon receptor, GCGR, and glucagon-like peptide 1 receptor, GLP-1R) driven by an increase in receptor avidity provided by the spatially defined presentation of the peptides. Interestingly, the avidity effects are different at the two target receptors. A single dose of the long-acting peptide conjugate robustly inhibited food intake and decreased body weight in insulin resistant diet-induced obese mice, in addition to ameliorating glucose intolerance. Inhibition of food intake and decrease in body weight was also seen in overweight cynomolgus monkeys. The weight loss resulting from dosing with the bivalently conjugated dual agonist was significantly greater than for the monomeric analog, clearly demonstrating translation of the measured in vitro avidity to in vivo pharmacology.

Obesity is associated with increased risk of diabetes and cardiovascular disease. Evidence is accumulating that significant WL (>10%) may reduce this risk. GDF15 is a secreted circulating polypeptide associated with energy balance. The receptor for GDF15, Gfral, is expressed in the area postrema and mediates the inhibitory effects of GDF15 on food intake. Compound H (CpdH) is a GDF15 agonist on a half-life extension platform, suitable for once-weekly dosing. We sought to test the durability of the food intake suppression and body weight loss with once weekly dosing of CpdH in the spontaneously obese cynomolgus non-human primate (NHP) model. As a translational control we included dulaglutide (dula), a long-acting GLP-1 agonist, to enable efficacy modeling and validation of the animal model. PK/PD simulations derived from single-dose lean and obese NHP studies were used for dose selection; CpdH or dula was administered SC once weekly in a biologic naïve spontaneously obese cynomolgus (population BMI 46.0±1.0Kg/m2 with 15.1±1.5% total body fat (n=32) at baseline). Chronic treatment of animals with CpdH at 1 and 10nmol/kg once weekly reduced food intake in a dose related manner relative to vehicle. A commensurate, sustained reduction in body weight (up to 16.2±4.9%) was observed at steady state exposures. The injection of dulaglutide control caused rapid, transient reduction in food intake and a 3.8% reduction in vehicle corrected body weight over 12 weeks. A dose-response relationship for CpdH was formally demonstrated. In conclusion, CpdH is a novel agent for the treatment of human obesity and type 2 diabetes. Unless otherwise noted, all abstracts presented at ENDO are embargoed until the date and time of presentation. For oral presentations, the abstracts are embargoed until the session begins. Abstracts presented at a news conference are embargoed until the date and time of the news conference. The Endocrine Society reserves the right to lift the embargo on specific abstracts that are selected for promotion prior to or during ENDO.

The discovery of a novel series of N-arylpyrroles as agonists of GPR120 (FFAR4) is discussed. One lead compound is a potent GPR120 agonist, has good selectivity for related receptor GPR40 (FFAR1), has acceptable PK properties, and is active in 2 models of Type 2 Diabetes in mice.

Compound 12 is a GPR40 agonist that realizes the full magnitude of efficacy possible via GPR40 receptor agonism. In vitro and in vivo studies demonstrated superior glucose lowering by 12 compared to fasiglifam (TAK-875), in a glucose dependent manner. The enhanced efficacy observed with the full agonist 12 was associated with both direct and indirect stimulation of insulin secretion.

Adiponectin (ADN) is a crucial mediator of insulin sensitivity and vascular health. The discoveries of the adiponectin receptors AdipoR1 and AdipoR2 (R1 and R2) prompted efforts to identify ADN-mimetics. R1 and R2 are especially difficult drug targets due to the complex higher-order structures of ADN as well as challenges in creating receptor knock-out cell lines. We approached R1 and R2 as possible targets beginning with a rigorous validation of the ligand and receptors. ADN preparations from multiple internal and external sources were characterized in physicochemical, cell binding and in vitro assays (pAMPK/pACC and ceramidase). It is important to note that ADN from only one source passed this evaluation. This ADN triggered in vitro activities associated with this protein, including stimulation of phosphorylation of ACC (EC50 = 9.3 ug/ml), calcium influx (EC50 = 2.2 ug/ml), glucose uptake, fatty acid oxidation and ceramidase activity. With a biologically active ADN preparation identified, we strove to understand the interactions of ADN with R1 and R2. However, both R1 and R2 are ubiquitously expressed, complicating the in vitro confirmation of receptor-dependent functional activity. We selected HAP1 cells, an ADN-responsive haploid CML derived line well-suited for gene deletion via CRISPR-Cas9 technology, to generate R1, R2 and R1/R2 deficient cells. Surprisingly, the stimulatory activity of adiponectin in pACC and ceramidase assays in HAP1 cells deficient in R1, R2 or both R1 and R2 were indistinguishable from those in wild type HAP1 cells. In addition, transient overexpression of R1 in HAP1 cells or pretreatment with antibodies to block R1 did not alter their pACC responses to ADN. These data indicate R1 and R2 are not responsible for all the in vitro biological activity attributed to ADN and suggest that there must be other yet unidentified receptors for ADN. Disclosure J.W. Gunnet: Employee; Self; Janssen Research & Development. W. Li: None. Y. Wang: None. J.A. Chavez: None. M. Husovsky: None. R. Meng: None. W. Lang: None. J.R. Jeyaseelan: Employee; Self; Janssen Pharmaceuticals, Inc.. G.W. Caldwell: None. S. Edavettal: Employee; Self; Janssen Research & Development.

A novel series of 5-membered heterocycle-containing phenylpropanoic acid derivatives was discovered as potent GPR120 agonists with low clearance, high oral bioavailability and in vivo antidiabetic activity in rodents.

We have discovered a novel series of isothiazole-based phenylpropanoic acids as GPR120 agonists. Extensive structure-activity relationship studies led to the discovery of a potent GPR120 agonist 4x, which displayed good EC50 values in both calcium and β-arrestin assays. It also presented good pharmaceutical properties and a favorable PK profile. Moreover, it demonstrated in vivo antidiabetic activity in C57BL/6 DIO mice. Studies in WT and knockout DIO mice showed that it improved glucose handling during an OGTT via GPR120. Overall, 4x possessed promising antidiabetic effect and good safety profile to be a development candidate.

Newcastle disease virus (NDV) is an important pathogen hazardous to poultry industry, and the pathogenicity of NDV strains varies with different virulence. Chicken Embryo Fibroblasts can serve as a laboratory host systems for NDV, and cytopathic effects (CPE) has some relationship to the strain’s virulence. In order to explore the rhythm of cytokine expression in CEF following velogenic or lentogenic strain infection, NDV virulent strain F48E9 and vaccine strain Lasota were used to infect CEF separately, and cells were collected at 0, 4, 8, 12, 24, 36 hours post-infection (h.p.i.). The total RNA was isolated and real-time quantitative RT-PCR was used to detect the expression of interleukin (IL)-15 and IL-18 gene. The results showed that the expression of both IL-15 and IL-18 gene of F48E9 and Lasota groups were significantly lower than the control group at 4 h, 8 h, 12 h, 24 h.p.i.. But at 36 h.p.i., the expression level of both IL-15 and IL-18 gene of F48E9 group increased rapidly to the level higher than that of control group, while in the Lasota group, the picture was quite different, the expression level of both IL-15 and IL-18 are still lower than that of the control group. The expression change of other cytokines showed no clear regularity in F48E9 or Lasota group. Expression of IL-15 and IL-18 may play an important role in mediation of NDV pathogenesis in CEF.

The env gene of subgroup J avian leukosis virus SCMS1 strain isolated from Sichuan area was amplified by PCR.Sequencing results indicated that there were 6 base pair deletion in gp85 gene.A 248 bp deletion in the nucleotides region corresponding to 6,995 bp to 7,242 bp of the HPRS-103 strain,including the entire domains of rTM.These mutations could play a meaningful role for virus pathogenicity variations.