Activation of the receptor tyrosine kinase MET is associated with poor clinical outcome in certain cancers. To target MET more effectively, we developed an antagonistic antibody mixture, Sym015, consisting of two humanized mAbs directed against nonoverlapping epitopes of MET. We screened a large panel of well-annotated human cancer cell lines and identified a subset with highly elevated MET expression. In particular, cell lines of lung cancer and gastric cancer origin demonstrated high MET expression and activation, and Sym015 triggered degradation of MET and significantly inhibited growth of these cell lines. Next, we tested Sym015 in patient- and cell line-derived xenograft models with high MET expression and/or exon 14 skipping alterations, and in models harboring amplification as a mechanism of resistance to EGFR-targeting agents. Sym015 effectively inhibited tumor growth in all these models and was superior to an analogue of emibetuzumab, a monoclonal IgG4 antibody against MET currently in clinical development. Sym015 also induced antibody-dependent cellular cytotoxicity (ADCC) , suggesting that secondary effector functions contribute to the efficacy of Sym015.Retrospectively, all responsive, high MET-expressing models were scored as highly-amplified by hybridization, pointing to amplification as a predictive biomarker for efficacy. Preclinical toxicology studies in monkeys showed that Sym015 was well tolerated, with a pharmacokinetic profile supporting administration of Sym015 every second or third week in humans. The preclinical efficacy and safety data provide a clear rationale for the ongoing clinical studies of Sym015 in patients with -amplified tumors..

SD and MD of NNC0195-0092 administered to healthy Japanese and non-Asian male subjects were well tolerated at all doses. The present trial suggests that NNC0195-0092 has the potential for an efficacious, well-tolerated, once-weekly GH treatment.

New insulin analogues with a longer duration of action and a flatter pharmacodynamic profile are developed to improve convenience and safety for diabetic patients. During the nonclinical development of such analogues, safety studies must be conducted in nondiabetic rats, which consequently are rendered chronically hypoglycemic. A rat comparator model using human insulin would be valuable, as it would enable differentiation between effects related to either persistent insulin-induced hypoglycemia (IIH) or a new analogue per se. Such a model could alleviate the need for an in-study-comparator and thereby reduce the number of animals used during development. Thus, the aims of the present study were i) to develop a preclinical animal model of persistent hypoglycemia in rats using human insulin infusion for four weeks and ii) to investigate histopathological changes in sciatic nerves and quadriceps femoris muscle tissue, as little is known about the response to persistent hypoglycemia in these tissues. Histopathologic changes in insulin-infused animals included axonal degeneration and myofibre degeneration. To our knowledge, this is the first study to show that persistent IIH provokes peripheral nerve and skeletal myofiber degeneration within the same animals. This suggests that the model can serve as a nonclinical comparator model during development of long-acting insulin analogues.

The aim of this study was to develop a structure-property model for membrane partitioning of oligopeptides using statistical design methods and multivariate data analysis. A set of 20 tetrapeptides with optional N-methylations at residues 2 and 4 was designed by a D-optimal design procedure. After synthesis and purification, the membrane partitioning abilities of the peptides were tested in two chromatographic systems with phospholipids as the stationary phase: immobilized artificial membrane chromatography (IAM) and immobilized liposome chromatography (ILC). The relationship between these measures and three different sets of calculated descriptors was analyzed by partial least-squares projection to latent structures (PLS). The descriptors used were the molecular surface area, Molsurf parameters, and Volsurf parameters. All three models were of good statistical quality and supported that a large hydrogen-bonding potential and the presence of a negative charge impair membrane partitioning, whereas hydrophobic parameters promote partitioning. The findings are in accordance with what has been found for absorption of known drugs and have implications for the design of peptide-like drugs with good oral bioavailability.

NN1177 is a glucagon/glucagon-like peptide 1 receptor coagonist investigated for chronic weight management and treatment of nonalcoholic steatohepatitis. Here, we show concentration-dependent downregulation of cytochrome P450 (P450) enzymes using freshly isolated human hepatocytes treated with this linear 29-amino acid peptide. Notably, reductions in CYP3A4 mRNA expression (57.2%-71.7%) and activity (18.5%-51.5%) were observed with a clinically relevant concentration of 100 nM NN1177. CYP1A2 and CYP2B6 were also affected but to a lesser extent. Physiologic-based pharmacokinetic modeling simulated effects on CYP3A4 and CYP1A2 probe substrates (midazolam and caffeine, respectively) and revealed potential safety concerns related to drug-drug interactions (DDIs). To investigate the clinical relevance of observed in vitro P450 downregulation, a phase 1 clinical cocktail study was initiated to assess the DDI potential. The study enrolled 45 study participants (body mass index 23.0-29.9 kg/m 2 ) to receive a DDI cocktail (midazolam, caffeine, omeprazole, dextromethorphan, and S-warfarin/vitamin K) alone and following steady-state NN1177 exposure. The analysis of pharmacokinetic profiles for the cocktail drugs showed no significant effect from the coadministration of NN1177 on AUC 0-inf for midazolam or S-warfarin. Omeprazole, caffeine, and dextromethorphan generally displayed decreases in AUC 0-inf and C max following NN1177 coadministration. Thus, the in vitro observations were not reflected in the clinic. These findings highlight remaining challenges associated with standard in vitro systems used to predict DDIs for peptide-based drugs as well as the complexity of DDI trial design for these modalities. Overall, there is an urgent need for better preclinical models to assess potential drug-drug interaction risks associated with therapeutic peptides during drug development. SIGNIFICANCE STATEMENTThis study highlights significant challenges associated with assessing drug-drug interaction risks for therapeutic peptides using in vitro systems since potential concerns identified by standard assays did not translate to the clinical setting. Further research is required to guide investigators involved in peptide-based drug development towards better preclinical models in order to more accurately evaluate potential drug-drug interactions.