作者: Felderer, Karin ; Silva, Sandra D. ; Mayer, Jan-Peter A. ; Kumar, Raman ; Santos, Sofia T. ; Batalha, Maria B ; Eichner, Timo ; Santos, Sofia T ; Kaufmann, Hitto ; Mayer, Jan-Peter A ; Pais, Daniel A M ; Pais, Daniel A. M. ; Silva, Sandra D ; Batalha, Maria B.

The development of novel therapeutic proteins is a lengthy and costly process, with an average attrition rate of 91% (Thomas et al. Clinical Development Success Rates and Contributing Factors 2011-2020, 2021). To increase the probability of success and ensure robust drug supply beyond approval, it is essential to assess the developability profile of new potential drug candidates as early and broadly as possible in development (Jain et al. MAbs, 2023. https://doi.org/10.1016/j.copbio.2011.06.002 ). Predicting these properties in silico is expected to be the next leap in innovation as it would enable significantly reduced development timelines combined with broader screens at lower costs. However, developing predictive algorithms typically requires substantial datasets generated under very defined conditions, a limiting factor especially for new classes of therapeutic proteins that hold immense clinical promise. Here we describe a strategy for assessing the developability of a novel class of small therapeutic Anticalin® proteins using machine learning in conjunction with a knowledge-driven approach. The knowledge-driven approach considers developability attributes such as aggregation propensity, charge variants, immunogenicity, specificity, thermal stability, hydrophobicity, and potential post-translational modifications, to calculate a holistic developability score. Based on sequence-derived descriptors as input parameters we established novel statistical models designed to predict the developability scores for Anticalin proteins. The best models yielded low root mean square errors across the entire dataset and were further validated by removing input data from individual screening campaigns and predicting developability scores for those drug candidates. The adoption of the described workflow will enable significantly streamlined preclinical development of Anticalin drug candidates and could potentially be applied to other therapeutic protein scaffolds.

2021-12-01·BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY

Selection of an artificial paraquat-specific binding protein from a ribosome display library based on a lipocalin scaffold.

Article

作者: Sun, Zhiyong ; Liu, Li ; Ning, Baoan ; Jiang, Yongqiang ; Yao, Sun ; Li, Hua ; Liu, Jianqing

In this study, an anticalin that could specifically bind paraquat (PQ), a quaternary nitrogen herbicide, as a new set of engineered receptor protein with antibody-like properties was generated to detect PQ concentration. To this end, a native and random library was constructed and engineered to allow in vitro transcription and translation using an Escherichia coli lysate system. Meanwhile, a PQ derivative that carries an active aliphatic carboxylate group at the end of an aliphatic spacer arm was synthesized. Then, this compound was coupled covalently to the carrier protein bovine serum albumin/ovalbumin and amino-functionalized paramagnetic beads. Alternating selection in solution and immobilization in microtiter wells were used to pan mRNA-ribosome-antibody complexes. After several rounds of ribosome display, three variants were selected from a random library of the bilin-binding protein. The variants that could bind complex PQ with high affinity and exhibit IC₅₀ values as low as 14.039 ± 0.970 ng/mL were identified. Moreover, the limits of detection reached 0.083 ± 0.011 ng/mL. Our data suggest that the generation of anticalins may provide a promising alternative to recombinant antibody fragments to create a stable receptor protein against hapten with bioanalytical relevance.

2021-04-01·EXPERT OPINION ON BIOLOGICAL THERAPY

Anticalin® proteins: from bench to bedside.

Review

作者: Ilyukhina, Elena ; Deuschle, Friedrich-Christian ; Skerra, Arne

INTRODUCTION:

Anticalin proteins are engineered versions of lipocalins that constitute a novel class of clinical-stage biopharmaceuticals. The lipocalins exhibit a central β-barrel with eight antiparallel β-strands and an α-helix attached to its side. Four structurally variable loops at the open end of the β-barrel form a pronounced binding pocket, which can be reshaped to generate specificities toward diverse disease-relevant molecular targets.

AREAS COVERED:

This article reviews the current status of Anticalin engineering, from the basic principles to the development of Anticalins with high target affinity and specificity via combinatorial protein design and directed evolution, including examples of Anticalin-based drug candidates under preclinical and clinical development.

EXPERT OPINION:

Combinatorial gene libraries together with powerful molecular selection techniques have enabled the expansion of the natural ligand specificities of lipocalins from small molecules to peptides and proteins. This biomolecular concept has been validated by structural analyses of a series of Anticalin•target complexes. Promising Anticalin lead candidates have reached different preclinical and clinical development stages in the areas of (immuno)oncology, metabolic, and respiratory diseases, as antidotes to treat intoxications and as novel antibiotics. Thus, Anticalins offer an alternative to antibodies with promising and potentially superior features as next-generation biologics.

项与 Anticalin H1GA 相关的新闻（医药）

2024-09-04

·生物谷

Nat Commun：靶向作用β淀粉样蛋白分子或有望帮助开发新型阿尔兹海默病预防性疗法

本文研究结果表明β淀粉样蛋白单体的清除或在阿尔兹海默病早期阶段修复神经元功能方面发挥着至关重要的作用。突触毒性β-淀粉样蛋白（Aβ）寡聚体所介导的过度活跃（hyperactivity）是阿尔兹海默病中最早出现的神经元功能障碍之一。近日，一篇发表在国际杂志Nature Communications上题为“β-amyloid monomer scavenging by an anticalin protein prevents neuronal hyperactivity in mouse models of Alzheimer’s Disease”的研究报告中，来自慕尼黑工业大学等机构的科学家们通过研究开发了一种非常有前途的预防性治疗性策略，文章中，研究人员能专门针对β淀粉样蛋白生物分子，其能诱发大脑中神经细胞的过度活跃，而这是早期脑部疾病的典型特征。研究者Benedikt Zott博士表示，这项研究中，我们成功开发并利用一种蛋白质药物来抑制有害分子所产生的影响效应。通过对实验室小鼠进行研究后，我们发现，神经元的功能异常甚至能被修复，因此我们希望所调查的这种蛋白质（称之为β淀粉样蛋白结合anticalin，H1GA）能帮助在早期阶段减缓严重神经变性疾病的进展。据研究人员推测，目前全球大约有5500万人患有痴呆症，其中大多数患者为阿尔兹海默病，而且每年仍有大约1000万例病例被诊断出来，目前并没有药物能对抗这种疾病的基本机制，只有患者出现的智力表现下降等症状能被治疗。研究者Zott强调道，我们距离开发能用于人类机体的疗法的开发还有很长一段路要走，但如今动物实验结果让我们甚至欣慰，在疾病早期阶段完全抑制神经元过度活跃的效果尤为显著。靶向作用β淀粉样蛋白分子或有望帮助开发新型阿尔兹海默病预防性疗法图片来源：Nature Communications (2024). DOI:10.1038/s41467-024-50153-y 文章中，研究人员通过蛋白质设计获得了anticalin H1GA，随后利用遗传修饰的工程化大肠杆菌来大量产生这种蛋白质，紧接着他们将有效成分注射到小鼠大脑的海马体区域，在可测量的行为方面，此前过度活跃的脑细胞或许就不再能与健康的神经元细胞区分开来了。目前研究人员并不清楚这种效应是否也能在实验室之外的人类患者中被观察到，然而无论如何，目前他们正在开发一种更有效的活性成分来进行给药。早在2016年，被认为具有类似效果的活性物质索拉珠单抗（solanezumab）就在大规模的临床试验中被证明是失败的，这或许能利用其不同的分子结构来进行解释。此外，研究人员还在试验中直接将新型活性成分与索拉珠单抗进行比较，结果发现，H1GA能表现出明显正向影响效应。综上，本文研究结果表明β淀粉样蛋白单体的清除或在阿尔兹海默病早期阶段修复神经元功能方面发挥着至关重要的作用。（生物谷Bioon.com）参考文献： Zott, B., Nästle, L., Grienberger, C. et al. β-amyloid monomer scavenging by an anticalin protein prevents neuronal hyperactivity in mouse models of Alzheimer’s Disease. Nat Commun 15, 5819 (2024). doi：10.1038/s41467-024-50153-y