Background Programmed cell death 1 (PD-1) is an inhibitory immune checkpoint expressed on activatedT cells. Upon the formation of T cell receptor (TCR)-pMHC complexes, concomitant PD-1 ligation to its ligands programmed death-ligand 1 (PD-L1) or programmed death-ligand 2 (PD-L2) downregulates TCR signaling and effector function. Here we describe the preclinical characterization of Sintilimab, a fully human IgG4 antibody that potently blocks PD-1 interactions with PD-L1 and PD-L2. Methods The binding affinity and blockade function were detected by using surface plasmon resonance (SPR), Enzyme-linked immunosorbent assay (ELISA) and flow cytometry. The biology function properties were measured with luciferase assay and mixed lymphocyte reaction assay. In vivo anti-tumor function and preclinical pharmacokinetic (PK) were identified with human PD-1 transgenic mice and non-human primates separately. Results Sintilimab can specifically and strongly bind to human PD-1 (hPD-1) and cynomolgus PD-1 and the affinity of Sintilimab to human PD-1 was measured at 0.3 nm via surface SPR, and displayed slow dissociation kinetics. Sintilimab can block the interaction of PD-1 to PD-L1 and PD-L2 and induce high secretion levels of interferon (IFN)-γ and interleukin (IL)-2 in primary T cell assays. In humanized hPD-1 knock-in mouse models, Sintilimab showed potent anti-tumor activity and increased tumor-infiltrating CD8/CD4 T cell and CD8/ Treg ratios. Preclinical experimentation in non-human primates following a single intravenous infusion of Sintilimab at 1, 6 and 30 mg/kg presented with no signs of drug-related toxicity, and showed typical PK characteristics of an IgG antibody. Conclusions Sintilimab has desirable preclinical attributes that supports its clinical development for cancer treatment.

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Maintaining pancreatic β-cell mass and function is essential for normal insulin production and glucose homeostasis. Regenerating islet-derived 2 (Reg2, Reg II, human ortholog Reg1B) gene is normally expressed in pancreatic acinar cells and is significantly induced in response to diabetes, pancreatitis, and high-fat diet (HFD) and during pancreatic regeneration. To evaluate the role of endogenous Reg2 production in normal β-cell function, we characterized Reg2 gene-deficient (Reg2-/-) mice under normal conditions and when subjected to several pathological challenges. At a young age, Reg2 gene deficiency caused no obvious change in normal islet morphology or glucose tolerance. There was no change in the severity of streptozotocin-induced diabetes or caerulein-induced acute pancreatitis in the Reg2-/- mice, indicating that the increased Reg2 expression under those conditions was not essential to protect the islet or acinar cells. However, 13- to 14-month-old Reg2-/- mice developed glucose intolerance associated with significantly decreased islet β-cell ratio and serum insulin level. Similarly, after young mice were fed an HFD for 19 weeks, diminished islet mass expansion and serum insulin level were observed in Reg2-/- vs wild-type mice. This was associated with a decline in the rate of individual β-cell proliferation measured by Ki67 labeling. In both conditions, the β-cells were smaller in gene-deficient vs wild-type mice. Our results indicate that normal expression of Reg2 gene is required for appropriate compensations in pancreatic islet proliferation and expansion in response to obesity and aging.

High concentration (5 and 10 μmol/L) of berberine could induce the apoptosis of MIN6 cells through cytochrome C/Apaf-1/caspase-3 and apoptosis inducing factor (AIF) pathway.