23rd and 24th February 2022
Liverpool, UK
Agenda
Conference Day 1 - 23rd February
| Time | Topic | Speaker |
|---|---|---|
| 9.00 - 9.30 | Registration | |
| 9.30 - 9.55 | “Further advancements in peptide synthesis and purification” | Dr Brunello Nardone |
| 10.00 - 10.30 | "Microwave-Assisted Solid-Phase Peptide Synthesis, A Unique Tool for Peptide Research" | Prof Fernando Albericio |
| Coffee Break | ||
| 11.00 - 11.25 | "Large Scale Solid Phase Peptide Synthesis (SPPS) at Elevated Temperatures: Advances, Process Development, and Considerations" | Dr Giorgio Marini |
| 11.30 - 12.00 | "Fluorinated peptides in Medicinal and Biological Chemistry" | Dr Christopher R Coxon |
| 12.00 - 12.30 | Poster snapshot session | |
| 12.30 - 13.30 | Lunch and Poster Session | |
| 13.30 - 14.00 | "Bioportides: Applications with Spermatozoa and Stem Cells." | Prof John Howl |
| 14.00 - 14.30 | "Peptide-Based Therapeutics: Opportunities and Challenges" | Dr Christian Behn |
| 14.30 - 15.00 | Coffee Break | |
| 15.00 -16.00 | Practical sessions | In the lab |
| 17.00 - 18.30 | Bus tour - historic sights of Liverpool | Hope Street Hotel |
| 18.30 | Conference Dinner | Liverpool - Albert Docks |
Conference Day 2 - 24th February
| Time | Topic | Speaker |
|---|---|---|
| 9.30 - 9.45 | Welcome Day 2 | |
| 9.45 - 10.15 | "Flip & Tag: Studying Protein with Chemistry. Flip: Enantiomeric Miniprotein Synthesis. Tag: Chemo-enzymatic Protein Bioconjugation." | Dr Louis Luk |
| 10.15 - 10.45 | "Radiolabelling of peptides for Positron Emission Tomography" | Dr Sergio Dall'Angelo |
| 10.45 - 11.15 | Coffee Break | |
| 11.15 - 11.45 | "Design, synthesis, and biological evaluations of Teixobactins to combat multidrug-resistant bacterial pathogens" | Dr Ishwar Singh |
| 11.45 - 12.00 | Poster prize awards and final remarks | Dr Brunello Nardone |
| 12.00 - 13.00 | Lunch | |
| Close |
Academic Guest Speakers
Microwave-Assisted Solid-Phase Peptide Synthesis, A Unique Tool for Peptide Research
Fernando Albericio,1,2 Beatriz G. de la Torre1
1University of KwaZulu-Natal, Durban 4001, South Africa; 2University of Barcelona, 08028-Barcelona, Spain
During the last five years (2016-2020), 18 drugs containing peptides have been approved by the US FDA. In addition, several hundred are in clinical phases or advanced preclinical studies. The development of new synthetic strategies has facilitated this explosion in the world of peptides. In this regard, the development of new resins, coupling reagents, protecting groups, and, more importantly, reliable and robust peptide synthesizers have been vital for the consolidation of the peptide drug market.
In the field of automatic synthesizers, the last breakthrough occurred when at the beginning of this century, CEM launched the new concept of Microwave-Assisted SPPS. Our group and others have demonstrated that both couplings and Fmoc-removal are better performed in the microwave mode compared with classical synthesis or even with conventional heating. More important, this better performance in the two key reactions is not accompanied by any significant side-reaction.
This presentation will discuss our last results on the use of Microwave synthesis for the preparation of intriguing peptides and/or for developing new synthetic strategies: (i) the preparation of staple peptides via late-stage C(sp2)-H Pd activation using the microwave; (ii) synthesis of N-alkyl amino acid containing peptides; (iii) development of new and more efficient coupling reagents; and (iv) microwave-based Green Solid-Phase Peptide Synthesis (GSPPS).
In the GSPPS, the CEM Liberty Blue Microwave technology by itself could be considered green in terms of solvent consumption and time-saving and due to the excellent quality of the crude peptides, which enormously facilitates the purification with the consequent increase of yield and reduction of chromatography solvents.
Professor John Howl
University of Wolverhampton
Biography
Bioportides: Applications with Spermatozoa and Stem Cells.
Molecular Pharmacology Group, Research Institute in Healthcare Science, University of Wolverhampton, UK
Intrinsically bioactive cell penetrating peptides (CPPs), or bioportides, access discrete intracellular compartments to modulate protein-protein interactions. Collaborative studies with the University of Aveiro have identified bioportides which regulate the activity of a novel isoform of Protein Phosphatase 1 (PPP1) differentially expressed in spermatozoa. These perturb the interactions of PPP1 with key regulatory proteins that are cell-type specific to rapidly inhibit the motility of human sperm. As progressive motility is a key prognostic indicator of male fertility, our current objective is to further develop STOPSPERM bioportides as a potential contraceptive option.
The triplobastic bilateral planarian Schmiditea mediterranea, a free living platyhelminth, is a model organism routinely employed for studies of developmental cell biology, regeneration, embryology and stem cell function.
S. mediterranea is also a convenient platform to assess the entry of CPPs and bioportides into a three-dimensional and relatively complex tissue. As totipotent stem cells, planarian neoblasts are a viable model for the development of bioportides which regulate stem cell function. Several transcription factors that control cellular differentiation and eye-regeneration in planaria are a source of highly efficient polycationic CPPs. Moreover, proteomimetic biportides which mimic the helical EYA domain of Eyes Absent (EYA) proteins inhibit head regeneration in decapitated planaria.
Fluorinated peptides in Medicinal and Biological Chemistry
Dr Christopher R Coxon
Senior Lecturer in Medicinal Chemistry
Director of Pepmotec Ltd
EaStChem School of Chemistry,
University of Edinburgh
Joseph Black Building,
The King’s Buildings
EH9 3FJ, UK
Peptides and proteins are becoming increasingly valuable as medicines, diagnostic agents and as tools for biomedical sciences. Much of this has been underpinned by the emergence of new methods for the manipulation and augmentation of native biomolecules. Perfluoroaromatic reagents are perhaps one of the most useful tools with which to modify peptides and proteins, due principally to their nucleophilic substitution chemistry, high electron deficiency and the ability for their reactivity to be tuned towards specific nucleophiles. The introduction of fluorine atoms or fluorinated groups is common in small molecule pharmaceuticals and agrochemicals, as a means of fine-tuning the physicochemical and pharmacokinetic properties. However, examples of this in peptides are very rare.
In this talk, we will discuss the optimisation and uses of fluoroaromatic reagents for applications in chemoselective ‘tagging’, stapling and bioconjugation of peptides, as well as tuning of ‘drug-like’ properties in the optimisation of anti-migraine peptides. A major interest is in the use of fluorinated ‘reporters’ for 19F NMR with which to measure rates of peptide nanoparticle metabolism and to probe the role of prolines in protein folding and dynamics. We will also consider possible future applications of these reagents in biological chemistry.
Design, synthesis, and biological evaluations of Teixobactins to combat multidrug-resistant bacterial pathogens
Anish Parmar, 1 Sanjit Das, Enas Newire, 1, 1 Ishwar Singh 1*
1 Antimicrobial Pharmacodynamics and Therapeutics, Institute of Systems, Molecular & Integrative Biology, University of Liverpool, Liverpool, UK;
* Corresponding author: i.singh@liverpool.ac.uk
One of the biggest challenges faced by medical science is how to treat infections caused by bacteria that have become resistant to antibiotics. It is predicted that by 2050, an additional 10 million people will succumb to drug-resistant infections each year. The development of new antibiotics, which are effective where other drugs fail, is, therefore, a crucial area of study. New classes of effective antibiotics are urgently needed to address the global health challenges associated with resistant bacterial infections and save lives.
The recently discovered natural compound, teixobactin kills a broad range of MDR bacterial pathogens such as methicillin-resistant Staphylococcus aureus (MRSA), Enterococcus spp. (vancomycin resistant enterococci, VRE) and Mycobacterium tuberculosis without detectable resistance.1 Importantly, teixobactin is an attractive scaffold for antibiotic development as bacterial resistance is less likely to develop as it can target a range of highly conserved critical non protein components of bacterial cell walls such as lipid II and lipid III.2
The focus of our work is to develop viable simplified teixobactin analogues with desirable drug like properties such potent antibacterial activity, good safety and low cost. These are important requirements to realise the therapeutic potential of teixobactins.3 We have developed simplified designs of teixobactins by replacing the key bottleneck enduracididine with simpler and economical building blocks such as leucine. The simplified teixobactin analogues showed identical or superior potency against superbugs such as MRSA to natural texiobactin.4 Synthetic teixobactins have the potential to save thousands of lives lost to resistant bacterial infections every year. In this presentation, I will share our efforts to develop simplified teixobactins.
Flip & Tag: Studying Protein with Chemistry
Flip: Enantiomeric Miniprotein Synthesis
Tag: Chemo-enzymatic Protein Bioconjugation
Louis Luk, Cardiff University
In this presentation, Louis will present two active branches of his research, Flip and Tag:
Flip: Enantiomeric Miniprotein Synthesis. At the current state of the art, polypeptides entirely composed of D-amino acids must be chemically synthesised. Though their preparation can be challenging, the availability of protein enantiomers is valuable for research. In this presentation, we will report the chemical synthesis of the bacteriocins aureocin A53 and lacticin Q as well as mechanistic insights gained from the racemic protein crystallography analysis. As a result of the current pandemic, we have also designed non-proteolytic D-miniprotein binders through synthesising enantiomers of cytokine storm targets.
Tag: Chemo-enzymatic Protein Bioconjugation. A chemo-enzymatic labelling system that exploits the substrate promiscuity of the plant transpeptidase AEP and the facile chemical reaction between N-terminal cysteine and 2-formyl phenylboronic acid (FPBA) was developed. This work enables protein labelling at the terminus of choice for protein of different sequences with reduced equivalent of labelling agents.
Radiolabelling of peptides for Positron Emission Tomography
Positron Emission Tomography (PET) is an imaging modality widely used in clinical diagnosis as well as in drug development. PET radiopharmaceuticals are bioactive molecules that are labelled with a positron-emitting radionuclide such as fluorine-18, gallium-68, or copper-64. The essential component of a PET tracer is the targeting moiety, which is designed to have high affinity and specificity towards a biological target associated with a specific disease. Initially, the targeting entities were developed as biologically active small molecules, for example the most widely used PET tracer is [18F]fluorodeoxyglucose ([18F]FDG). However, in recent years there has been a shift in the use of biologics, such as peptides, proteins, antibodies, and antibody fragments as targeting moieties.
In this talk, we will discuss different labelling strategies to prepare radiopharmaceuticals having peptides as targeting moiety. We will explore the use of readily available fluorine-18 containing small molecules (prosthetic groups) to perform a bioconjugation via a linker. We will also consider the potential of enzymatic methodologies and examine new procedures based on silicon-, boron- and aluminium-18F bond. Eventually, we will consider the use of chelators to trap radiometals like Gallium-68 or Copper-64.
CEM Speakers
Large Scale Solid Phase Peptide Synthesis (SPPS) at Elevated Temperatures: Advances, Process Development, and Considerations
Method development for further advancing the efficiency of SPPS is of the utmost importance. Microwave irradiation provides simplified optimization, higher peptide purity, and an overall “greener” process. Compared to conventional heating methods, microwave irradiation provides rapid and direct energy exchange with the reagents.
Rapid scale-up for clinical trials and peptide production has been accomplished using similar technology. Crude purity from R&D to production scale is preserved if not improved and unwanted side reactions such as epimerization and aspartimide formation are easily controlled. The result, easier purification and reduced labor cost.
More recently, the usage of a large microwave cavity (up to fifteen-liter capability) allows us the possibility to scale up laboratory conditions in order to make bigger amount of peptides still taking advantages of the benefits of microwave technology. Cycle times at the production scale range from 30 – 60 min with the capability to produce up to 1 KG crude peptide in a single batch with considerably shorter synthesis time compared to the conventional methods at room temperature.
Peptide-Based Therapeutics: Opportunities and Challenges
Peptides have tremendous chemical and biological diversity and represents a very attractive and interesting class of pharmaceutical compounds, molecularly poised between small molecules and proteins, yet biochemically and therapeutically distinct from both.
The peptide chemistry era started in 1901, when Emil Fisher reported the preparation of the first dipeptide, glycin-glycin. The field of peptide therapeutics began in 1922 with the discovery and the first medical use of insulin — extracted from animal pancreases — which revolutionized the treatment of type 1 diabetes. Four decades passed before synthetically produced peptide hormones, namely oxytocin and vasopressin, entered the clinic and initiated the field of peptide drug development.
Soon, the importance of peptides as biological mediators were recognized.
Their role as mediators of key biological functions and their unique intrinsic properties makes them particularly attractive therapeutic agents: peptides show high biological activity associated with low toxicity, high specificity and these characteristics include little unspecific binding to molecular structures other than the desired target, and less accumulation in tissues reducing risks of complications due to intermediate metabolites. In addition, peptides offer valuable chemical and biological diversity compared to small molecules, for which intellectual property is still widely used.
The peptide therapeutics market is providing new commercial opportunities to biotechnology and pharmaceutical industries. Recent advances in drug delivery have re-focused attention on peptides. Today over 80 peptide drugs are approved in the United States and other major markets, for a wide range of diseases, including diabetes, cancer, osteoporosis, multiple sclerosis, HIV infection and chronic pain.
Peptide arrays derived from protein sequences provide the basic tools to elucidate interactions between a protein and a ligand. Scans of overlapping peptides signify that the entire protein sequence is synthesised as short, linear, overlapping peptides that are tested for ligand binding. The use of synthetic peptide libraries on membrane supports with high local peptide concentrations at each “spot” is becoming extensively popular for protein–ligand interaction studies. With the MultiPep the SPOT synthesis option allows for the synthesis of up to 2400 peptides in a batch for high-throughput screening applications. The peptides are chemically synthesized; they are easier to produce and handle relative to proteins. Peptide synthesis is also essential to incorporate modified or non-natural amino acid with 100% specificity.
Further advancements in peptide synthesis and purification
Peptides are recognized for being highly selective and efficacious and, at the same time, relatively safe and well tolerated. With several high revenue peptide drugs on the market the demand for a highly robust synthetic method is crucial.5 In 2018 CEM launched the Liberty Prime technology, officially removing the “bottle-neck” in solid-phase peptide synthesis.6
In 2021 CEM released the Prodigy Prep-HPLC system, which is complementary to the CEM range of microwave peptide synthesisers. In this presentation the green microwave synthesis and purification of neo-antigens and long peptides is discussed.
