Biophysical platforms have become established and versatile tools in drug discovery, especially effective during lead generation. Whilst they rely on a single readout, the variety of assay configurations available can significantly affect both the information depth and the speed of data provision, facilitating timely decision-making in drug projects. This will be illustrated through two specific examples: expediting hit validation from DNA-encoded libraries and investigating ternary-complex formation induced by PROTACs.

The availability of small molecule leads is an important aspect of small molecule drug discovery. This presentation describes the origin of small molecule leads across the two research organisations Roche and Genentech. Different lead-finding approaches such as HTS, focused or fragment screening, DNA-encoded library technology, in-licensing, and structure-based design are included. The translation of the lead series to identify in vivo tool compounds or development candidates will be discussed.

The discovery efforts using our RAPID (Reactive Affinity Probe Interaction Discovery) chemoproteomics platform will be described that led to small molecules that inhibited transporter SLC6A19 and demonstrated in vivo activity in the Pahenu2 model. This work led to the identification of JNT-517, a potential first-in-class clinical candidate for the treatment of phenylketonuria which has demonstrated positive POC in a Ph1b clinical trial.

Pyruvate dehydrogenase kinases (PDHKs) act as a key regulator in glycolysis and oxidative phosphorylation. PDHK inhibition presents a promising therapeutic target for diabetes, cancers, autoimmune, and inflammatory diseases. This talk highlights the PDHK hit identification campaign utilising iterative screening/FBDD approaches. Following that, the structural-based hit optimisation process to achieve PDHK1/2 selectivity and cellular potency will be elaborated.

This presentation will disclose several examples where X-ray crystallographic fragment screening has been used to identify novel allosteric binding sites across different protein classes. We will discuss computational methodologies used to characterise the binding pockets, the application of multiple biophysical approaches to measure affinity, and the structure-guided optimisation work undertaken to develop hits into chemical probes which were used to elucidate the functional relevance of these sites.

X-ray crystallography molecular replacement (MR) is a highly versatile tool for the detailed characterization of lead compound and binding modes. Application to drug discovery depends on (i) the availability of a similar protein structure, and (ii) well-diffracting crystals of the ligand-protein complex. While (i) is usually not a limitation, (ii) obtaining well-diffracting crystals may be difficult. I present how to determine structure of a protein-ligand complex by liquid-state NMR.

I will present innovations we’ve implemented to obtain an average hit rate of 20% for diverse protein targets using HZB’s synchrotron BESSY II for crystallographic fragment screening. We support academic and industrial users with a specialized fragment library, a crystal harvesting tool, high-throughput beamlines, a web-based data management tool and a computational workflow to perform a first optimization round. A few case studies will also be presented.

The drug discovery landscape is changing, with an increased focus on rapid hit-finding, new therapeutic modalities, and pursuit of increasingly challenging novel targets. While these targets present innovative ways to treat disease, their lack of binding sites or inert nature presents challenges for hit-finding. We will share experiences from implementing affinity screening approaches, such as ASMS, to enable rapid hit discovery and facilitate the use of these challenging targets.

The design of covalent drugs targeting Lys, His, or Tyr is gaining significant traction. In our experience, aryl-fluorosulfates represent suitable, stable electrophiles to react with the side chains of these residues. I will discuss strategies and opportunities to design covalent ligands targeting Lys, His, or Tyr. I will report on both ligand-first structure-based design and fragment-flurosulfates screening strategies, and I will present on our successful implementations of both approaches.

I will show recent results of "PureDEL" technology, a new solid phase-based DEL synthesis approach for the construction of pure DELs of more sets of diversity elements. Further, enzyme-assisted DEL synthesis ("EnzyDEL") and macrocyclic dual-display DELs will be presented.

Using a co-factor directed screening strategy and DNA-encoded libraries, a class of MTA-cooperative PRMT5 inhibitors was identified. Structural studies show that the hit series occupies the arginine substrate pocket of MTA-bound PRMT5, while simultaneously exhibiting a hydrophobic interaction to MTA. Further optimisation led to lead compounds, which potently and selectively inhibit PRMT5 in MTAP-deleted cells and show in vivo efficacy in an MTAP-deleted cancer cell model.