Shandong Boan Biotechnology Co., Ltd.

Charge heterogeneity is a critical quality attribute in biopharmaceutical development, necessitating thorough characterization to ensure product quality, efficacy, and stability. Dulaglutide, a glucagon-like peptide-1 (GLP-1) receptor agonist consists of two GLP-1 analogs fused to a human IgG4-Fc fragment. Being an acidic Fc-fusion protein, investigating charge variants of dulaglutide poses significant challenges due to its complex charge heterogeneity, molecular instability, and technical constraints. A dual-mode semi-preparative anion-exchange chromatography method was developed to fractionate charge variants, providing insights into structural differences in charge heterogeneity and their potential impact on biological functions. This method offers high purity, low cost, large preparation volume, and wide instrument availability, while preserving protein structural integrity. Subsequent comprehensive characterizations with respect to charge heterogeneity, sialic acid, post-translational modifications, size heterogeneity, related substances and biological activities were conducted to enhance understanding of modifications related to microheterogeneity such as phosphorylation, sialylation, aggregation, truncation, deamidation, or oxidation, and to establish critical structure-activity relationship. Surprisingly, the aggregates in basic variants are primarily bound through non-covalent interactions, whereas covalently linked aggregates are present in acidic variants. Charge variants slightly affected biological activity potentially due to the presence of aggregates. The comparative study of the innovator product Trulicity® and biosimilar candidate revealed that minor differences in acidic variants were possibly attributed to variations in phosphorylation and sialylation. This study elucidated the structural origins and functional implications of dulaglutide's charge heterogeneity, offering promising and comprehensive solutions for fractionation and detailed characterization of charge heterogeneity in complex proteins.

Denosumab (Xgeva®) is a standard treatment for the prevention of skeletal-related events (SREs) in patients with bone metastases (BM). This trial was designed to assess the equivalence of LY01011 to denosumab in terms of efficacy and safety.

Eligible patients with BM from solid tumors were randomized at a 1:1 ratio to receive 120 mg of LY01011 or 120 mg of denosumab subcutaneously every four weeks during a 12-week double-blind treatment period, and then all enrolled patients continued to receive LY01011 until week 53. The primary endpoint was the natural logarithm of change of the urinary N-terminal crosslinked telopeptide of type I collagen level normalized to the urine creatinine level (uNTX/uCr) at week 13 from baseline. Other endpoints included the uNTX/uCr ratio, serum bone-specific alkaline phosphatase level alteration, status of anti-drug antibodies and neutralizing antibodies, adverse events and SREs.

850 eligible patients were randomized into the LY01011 group (n = 424) or the denosumab group (n = 426). The least-squares means (SEs) of the natural logarithms of the changes in the uNTX/uCr ratios at week 13 from baseline were -1.810 (0.0404) in the LY01011 group and -1.791 (0.0406) in the denosumab group. The LSM difference [90 % CI] between two arms was -0.019 [-0.110, 0.073] within the equivalence margins (-0.135, 0.135) and met the predetermined primary endpoint. The AEs, ADAs and the PK data showed no statistically significant difference.

This study demonstrated the equivalent efficacy and safety of LY01011 to denosumab in patients with BM.

A review.Organic small-mol. fluorescent probes play a pivotal role in fluorescence-based sensing applications, owing to their exceptional responsiveness to diverse stimuli.These probes can be classified into two categories based on their emission characteristics: those that emit strongly in isolated states and those that exhibit robust emission in densely packed states.While the former grapples with self-quenching issues, a phenomenon known as aggregation-caused quenching (ACQ), the latter triumphs over this challenge through aggregation-induced emission (AIE) properties.Nevertheless, both ACQ and AIE fluorophores are constrained to single emission states, necessitating the development of dual-state emission (DSE) fluorophores capable of emitting strongly in both isolated and aggregated states.DSE materials, unlike traditional fluorescent dyes, offer versatility across different phases, spanning from dilute solutions to solid states.They have demonstrated significant utility in biosensing and chemosensing, empowering the detection of various substances with exceptional sensitivity, specificity, and adaptivity.However, despite the remarkable progress achieved, a comprehensive understanding of the underlying mechanisms and the resolution of practical challenges remain ongoing endeavors.Future pursuits in mol. engineering, novel material development, and broader application exploration hold the key to fully harnessing the vast potential of DSE materials across diverse fields.