Eichner, T. | bioRxiv Science

bioRxiv2025

M5 positive allosteric modulation alleviates parkinsonian motor deficits

Parkinsons disease is a neurodegenerative movement disorder which is characterized by cardinal motor symptoms of tremor at rest, rigidity, bradykineasia, and postural instability. Underlying these cardinal motor symptoms is thought to be death and dysfunction of nigrostriatal dopamine neurons, and the gold-standard treatment of Parkinsons disease is dopamine replacement therapy with the dopamine precursor L-DOPA. While efficacious, L-DOPA does not treat all motor symptoms and can have serious treatment-related side effects called L-DOPA induced dyskinesias, indicating an immense need for new targets to modulate dopaminergic function for anti-parkinsonian efficacy. One such potential target is the M5 muscarinic acetylcholine receptor, which has a unique expression profile where it is selectively expressed in midbrain dopaminergic neurons and their terminals in the striatum, and previous studies have indicated that M5 can modulate dopamine release and patterning of firing of dopamine neurons. Given this unique expression profile and function of M5, this receptor has an untested potential to modulate parkinsonian motor phenotypes. To test the potential for M5 to modulate Parkinsonian-like motor deficits and dyskinesia, we employed the unilateral 6-OHDA lesioned mouse model to create a hemi-parkinsonian state. Using multiple behavioral assays, including the cylinder test, forepaw adjusting steps assay, and in the Erasmus ladder, in conjunction with prototypical M5 pharmacological tool compounds, we investigated the ability of M5 to modulate parkinsonian motor deficits. Additionally, we tested the ability of M5 to modulate established L-DOPA induced dyskinesia or cause dyskinesia on its own. Overall, we found that M5 PAM alleviates forepaw asymmetry, bradykinesia, and spatial aspects of gait in the Erasmus ladder. Excitingly, M5 PAM does not cause robust dyskinesia, does not affect already established L-DOPA-induced dyskinesia, and does not affect L-DOPA motor efficacy. Taken together with previous findings, the current study suggests that M5 receptors are an exciting novel therapeutic strategy for ameliorating parkinsonian motor deficits even in late-stage models of severe PD without lessening L-DOPAs motor benefit and without affecting existing symptoms of L-DOPA-induced dyskinesia.

neuroscience↗

bioRxiv2023

Holistic in silico developability assessment of novel classes of small proteins using publicly available sequence-based predictors

The development of novel therapeutic proteins is a lengthy and costly process, with an average attrition rate of 91% (Thomas et al., 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., 2023). 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(R) 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.

biochemistry↗