Simon L. Rössler

Covalent peptide-encoded libraries are assembled through a Heck coupling method allowing modification of resin-bound peptides and enabling affinity-based selection of small molecules bearing structurally diversified acrylamide warheads.

Protein-protein interactions (PPIs) are intriguing targets in drug discovery and development. Peptides are well suited to target PPIs, which typically present with large surface areas lacking distinct features and deep binding pockets. To improve binding interactions with these topologies and advance the development of PPI-focused therapeutics, potential ligands can be equipped with electrophilic groups to enable binding through covalent mechanisms of action. We report a strategy termed electrophile scanning to identify reactivity hotspots in a known peptide ligand and demonstrate its application in a model PPI. Cysteine mutants of a known ligand are used to install protein-reactive modifiers via a palladium oxidative addition complex (Pd-OAC). Reactivity hotspots are revealed by cross-linking reactions with the target protein under physiological conditions. In a model PPI with the 9-mer peptide antigen VL9 and major histocompatibility complex (MHC) class I protein HLA-E, we identify two reactivity hotspots that afford up to 87% conversion to the protein-peptide conjugate within 4 h. The reactions are specific to the target protein in vitro and dependent on the peptide sequence. Moreover, the cross-linked peptide successfully inhibits molecular recognition of HLA-E by CD94-NKG2A possibly due to structural changes enacted at the PPI interface. The results illustrate the potential application of electrophile scanning as a tool for rapid discovery and development of covalent peptide binders.

Molecular vaccines comprising antigen peptides and inflammatory cues make up a class of therapeutics that promote immunity against cancer and pathogenic diseases but often exhibit limited efficacy. Here, we engineered an antigen peptide delivery system to enhance vaccine efficacy by targeting dendritic cells and mediating cytosolic delivery. The delivery system consists of the nontoxic anthrax protein, protective antigen (PA), and a single-chain variable fragment (scFv) that recognizes the XCR1 receptor on dendritic cells (DCs). Combining these proteins enabled selective delivery of the N-terminus of lethal factor (LF_N) into XCR1-positive cross-presenting DCs. Incorporating immunogenic epitope sequences into LF_N showed selective protein translocation in vitro and enhanced the priming of antigen-specific T cells in vivo. Administering DC-targeted constructs with tumor antigens (Trp1/gp100) into mice bearing aggressive B16-F10 melanomas improved mouse outcomes when compared to free antigen, including suppressed tumor growth up to 58% at 16 days post tumor induction (P < 0.0001) and increased survival (P = 0.03). These studies demonstrate that harnessing DC-targeting anthrax proteins for cytosolic antigen delivery significantly enhances the immunogenicity and antitumor efficacy of cancer vaccines.

Encoding small-molecule information in DNA has been leveraged to accelerate the discovery of ligands for therapeutic targets such as proteins. However, oligonucleotide-based encoding is hampered by inherent limitations of information stability and density. In this study, we establish abiotic peptides for next-generation information storage and apply them for the encoding of diverse small-molecule synthesis. The chemical stability of the peptide-based tag allows the use of palladium-mediated reactions to efficiently synthesize peptide-encoded libraries (PELs) with broad chemical diversity and high purity. We demonstrate the successful de novo discovery of small-molecule protein ligands from PELs by affinity selection against carbonic anhydrase IX and the oncogenic protein targets BRD4(1) and MDM2. Collectively, this work establishes abiotic peptides as carriers of information for the encoding of small-molecule synthesis, leveraged herein for the discovery of protein ligands.

In this Review, we highlight recent advances in the understanding and design of N‐functionalized pyridinium scaffolds as redox‐active, single‐electron, functional group transfer reagents. We provide a selection of representative methods that demonstrate reactivity and fundamental advances in this emerging field. The reactivity of these reagents can be divided into two divergent pathways: homolytic fragmentation to liberate the N‐bound substituent as the corresponding radical or an alternative heterolytic fragmentation that liberates an N‐centered pyridinium radical. A short description of the elementary steps involved in fragmentation induced by single‐electron transfer is also critically discussed to guide readers towards fundamental processes thought to occur under these conditions.

In diesem Aufsatz sind die jüngsten Fortschritte beim Verständnis und Design von N‐funktionalisierten Pyridiniumsalzen als redoxaktivierte Einelektronentransfer‐Reagenzien zur Übertragung funktioneller Gruppen zusammengefasst. Anhand einer Auswahl an repräsentativen Methoden werden Reaktivität und grundlegende Fortschritte in diesem sich rasch entwickelnden Bereich diskutiert. Diese Reagenzien weisen zwei grundsätzlich divergierende Reaktionspfade auf: homolytische Fragmentierung zur Freisetzung des N‐gebundenen Substituenten als entsprechendes Radikal oder eine alternative, heterolytische Fragmentierung zur Freisetzung eines radikalischen Pyridiniumkations. Die elementaren Schritte der durch Einelektronentransfer induzierten Fragmentierung werden kritisch diskutiert.

The catalytic, asymmetric synthesis of complex molecules has been a core focus of our research program for some time because developments in the area can have an immediate impact on the identification of novel strategies for the synthesis of value-added molecules. In concert with this central interest, we have emphasized the design of ligand scaffolds as a tactic to discover and develop novel chemistry and overcome well-recognized synthetic challenges. Based on our group’s work on chiral pool-derived diolefin ligands, we designed and implemented a class of hybrid (phosphoramidite,olefin) ligands, which combines the properties of both phosphoramidite and olefin motifs to impact, fine-tune, and even override the inherent reactivity of the metal center. Specifically, we have utilized these unique modifying ligands to address several recognized limitations in the field of iridium-catalyzed, asymmetric allylic substitution. The methods we have documented typically employ branched, unprotected allylic alcohols as substrates and obviate the need for rigorous exclusion of air and moisture. Following Takeuchi’s seminal report demonstrating the high aptitude of Ir(I)-phosphite catalysts for branch-selective allylic substitution, concerted efforts from numerous research laboratories have led to a broadening of the synthetic utility of this reaction class. The first section of this Account outlines the process leading to our discovery of an unprecedented (phosphoramidite,olefin) ligand and its validation in the first iridium-catalyzed amination of branched, unprotected allylic alcohols. This section continues with our work involving heteroatom-based nucleophiles within inter- and intramolecular etherification, thioetherification and spiroketalization processes. The second section highlights the use of readily available carbon nucleophiles possessing sp, sp², and sp³ hybridization in a series of enantioselective carbon–carbon bond-forming reactions. We describe how alkylzinc, allylsilane, and several classes of organotrifluoroborate nucleophiles can be coupled enantioselectively to enable construction of several key motifs including 1,5-dienes, 1,4-dienes, and 1,4-enynes. Since the unique electronic and steric properties of this class of ligands renders the (η³-allyl)-Ir(III) intermediate highly electrophilic, even weak nucleophiles such as alkyl olefins can be used. We also show that more nucleophilic alkene motifs such as enamines and in situ generated ketene acetals smoothly participate in substitution reactions with allylic alcohols to yield valuable piperidines and γ,δ-unsaturated esters, respectively. The concept of stereodivergent dual catalysis, which synergistically combines chiral amine catalysis with iridium catalysis to furnish α-allylated aldehydes containing two independently controllable stereocenters is then discussed. This process has enabled the independent, stereoselective synthesis of all four possible product stereoisomers from a single set of starting materials, and was highlighted in the stereodivergent synthesis of Δ⁹-tetrahydrocannabinol. This Account concludes with an overview of our organometallic mechanistic studies regarding relevant intermediates within the catalytic cycle of this class of allylic substitution. These studies have allowed us to better understand the origin of the unique characteristics exhibited by this catalyst in comparison to related systems.

Morpholine ketene aminal is employed in iridium‐catalyzed asymmetric allylic alkylation reactions as a surrogate for amide enolates to prepare γ,δ‐unsaturated β‐substituted morpholine amides. Kinetic resolution or, alternatively, stereospecific substitution affords the corresponding products in high enantiomeric excess. The utility of the products generated by this method has been showcased by their further elaboration into amines, ketones, or acyl silanes. A putative catalytic intermediate (η³‐allyl)iridium(III) with achiral P,Olefin‐ligand was synthetized and characterized for the first time.

Morpholine ketene aminal is employed in iridium‐catalyzed asymmetric allylic alkylation reactions as a surrogate for amide enolates to prepare γ,δ‐unsaturated β‐substituted morpholine amides. Kinetic resolution or, alternatively, stereospecific substitution affords the corresponding products in high enantiomeric excess. The utility of the products generated by this method has been showcased by their further elaboration into amines, ketones, or acyl silanes. A putative catalytic intermediate (η³‐allyl)iridium(III) with achiral P,Olefin‐ligand was synthetized and characterized for the first time.

Electron‐transfer photocatalysis provides access to the elusive and unprecedented N‐pyridyl radical cation from selected N‐substituted pyridinium reagents. The resulting C(sp²)−H functionalization of (hetero)arenes furnishes versatile intermediates for the development of valuable aminated aryl scaffolds. Mechanistic studies that include the first spectroscopic evidence of a spin‐trapped N‐pyridyl radical adduct implicate SET‐triggered, pseudo‐mesolytic cleavage of the N−X pyridinium reagents mediated by visible light.