27Th Annual GP2A Medicinal Chemistry Conference

pharmaceuticals

Meeting Report 27th Annual GP2A Medicinal Chemistry Conference

Shailesh N. Mistry 1,* , Pascal Marchand 2,* and Barrie Kellam 1 1 School of Pharmacy, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK; [email protected] 2 Université de Nantes, Cibles et Médicaments des Infections et du Cancer, IICiMed, EA 1155, F-44000 Nantes, France * Correspondence: [email protected] (S.N.M.); [email protected] (P.M.); Tel.: +44-115-8467983 (S.N.M.); +33-253-009-155 (P.M.) Received: 30 November 2019; Accepted: 2 December 2019; Published: 6 December 2019

Abstract: The 27th annual GP2A (Groupement des Pharmacochimistes de l0Arc Atlantique/Group of Medicinal Chemists in the Atlantic Arc) conference took place from 21 to 23 August 2019, at the East Midlands Conference Centre (University Park, Nottingham, United Kingdom) and was hosted by the Division of Biomolecular Science and Medicinal Chemistry (BSMC), within the School of Pharmacy at the University of Nottingham. The event brought together an international delegation of researchers with interests in medicinal chemistry and interfacing disciplines. In addition, a pre-conference workshop provided an opportunity for younger researchers to develop their theoretical knowledge in quantitative pharmacology. Abstracts of presentations by the 14 invited speakers and 6 young researchers, in addition to 41 posters, are included in this report.

Keywords: medicinal chemistry; drug design; chemical biology; ligand kinetics

1. Aim and Scope of the Meeting

The GP2A (Groupement des Pharmacochimistes de l0Arc Atlantique/Group of Medicinal Chemists in the Atlantic Arc) network is made up of researchers in the ﬁeld of medicinal chemistry working in universities and research institutes across Europe. It was founded in 1992 with the aim of bringing together researchers to exchange ideas and expertise and facilitate a friendly collaborative networking environment. Historically, it has included members from France, Spain, Portugal, Ireland and the United Kingdom (the “Atlantic Arc”). More recently, it has expanded both geographically and in terms of research ﬁelds, now including researchers from areas that range from physical and pharmaceutical chemistry to molecular pharmacology. The annual GP2A conference host city alternates between France and another country and brings together established scientists and those earlier in their career. In addition, the network facilitates short visits between laboratories. A consistent theme of the annual conference is to provide young researchers an opportunity to present their work to an international audience through either poster presentation or by sharing the stage with invited speakers. The 2019 conference was held at the East Midlands Conference Centre and hosted by the Division of Biomolecular Science and Medicinal Chemistry (BSMC), within the School of Pharmacy at the University of Nottingham (UoN). Ahead of the workshop, young researchers were invited to attend a pre-conference workshop, led by Prof. Steven J. Charlton (UoN/Excellerate Bioscience) and Dr Elizabeth Rosethorne (UoN). The meeting opened and closed with two internationally recognised plenary speakers, Prof. Peter Gmeiner (Friedrich-Alexander University, Erlangen-Nürnberg, Germany) and Prof. Rob Leurs (Vrije-Universiteit, Amsterdam, The Netherlands). In addition, there were a further 12 invited speaker talks, 6 young researcher talks and 41 posters, selected from over 50 submitted abstracts.

Pharmaceuticals 2019, 12, 179; doi:10.3390/ph12040179 www.mdpi.com/journal/pharmaceuticals Pharmaceuticals 2019, 12, 179 2 of 47

Reflective of the multidisciplinary nature of medicinal chemistry research, the meeting included a broad range of topics of interest. These included infectious and neurodegenerative diseases, chemoprevention of cancer, approaches to target identification, hit identification and optimisation for drug candidates, natural products, inflammatory diseases/pain and the application of structural cheminformatics.

2. Pre-Conference Workshop on Quantitative Pharmacology Comparing the pharmacological characteristics of compounds across different systems (e.g., different assays and/or cell types) can be a complex task. More difficult still is the extrapolation of these in vitro data to in vivo systems through PK/PD modelling. In order to make these tasks as accurate as possible, it is critical to define quantitative, system independent measures of compound action. This workshop will introduce the concept of quantitative molecular pharmacology and detail the experimental and analytical methods used to determine key system-independent pharmacological descriptors of both drug binding and functional efficacy. It will culminate in a team-based exercise where you will put the knowledge and skills gained from the morning0s lectures into practice. You will play the role of the pharmacologist in a (fictitious) pharmaceutical company0s due diligence team, tasked with identifying the best compound to in-license for a respiratory indication. These deals can be very expensive—will you be confident enough in your analysis to make a recommendation to senior management ... ?

3. Opening Plenary Lecture

Selective Ligands Based on GPCR Structures Peter Gmeiner Department of Chemistry and Pharmacy, Medicinal Chemistry, Emil Fischer Center, Friedrich-Alexander University, Nikolaus-Fiebiger- Str. 10, 91058 Erlangen, Germany; [email protected] GPCR crystal structures may leverage an effective development of novel drug candidates because they can be used for structure-based in silico docking screens, giving access to new chemotypes and, as a consequence, to new biological profiles (Shoichet, B.K., Kobilka, B.K. Trends Pharmacol. Sci. 2012, 33, 268–272). Furthermore, they can guide lead modification and optimization programs by providing insights into crucial ligand–receptor interactions and the bioactive ligand conformation. Both strategies can be performed on the basis of either the crystal structure of a given GPCR or from a homology model of a structurally highly similar congener. Structures of different activity states of GPCRs allow us to identify molecular interactions discriminating between inverse agonists, antagonists and agonists. These fundamental results also contribute to the rational discovery of drugs selectively binding to particular conformational states. We aim to leverage GPCR structures to discover new chemotypes and bioisosteres with beneficial physical and biological properties and use these to probe and control signalling pathways. Thus, we develop highly specific agonists and antagonists for Class A GPCRs for structural and functional investigations and for structure–function relationship studies. The final goal is the development of novel drug candidates. The seminar will show very recent developments in the design, synthesis and biological investigation of orthosteric and allosteric GPCR ligands with unprecedented activity profiles. Pharmaceuticals 2019, 12, 179 3 of 47 Pharmaceuticals 2019, 12, x FOR PEER REVIEW 3 of 47

Structure-guided development of highly specific GPCR ligands.

4. Closing Plenary Lecture

It isis AllAll AboutAbout TimeTime…...... Rob LeursLeurs Department of of Medicinal Medicinal Chemistry, Chemistry, Vrije Vrije Universiteit Universiteit Amsterdam, Amsterdam, 1081 HZ 1081 Amsterdam, HZ Amsterdam, The TheNetherlands; Netherlands; e-mail: [email protected] [email protected] In light of of the the recent recent interest interest in in drug drug binding binding kinetics, kinetics, we we have have investigated investigated the thekinetics kinetics of the of thebinding binding interactions interactions between between the classical the classical antihist antihistaminesamines and the and H1R the within H1R within the framework the framework of the ofInnovative the Innovative Medicines Medicines Initiative-sponsored Initiative-sponsored program program “Kinetics “Kinetics for Drug forDrug Discovery Discovery (K4DD)”. (K4DD)”. We Wedeveloped developed new new methodologies methodologies to tomeasure measure and and st studyudy GPCR-ligand GPCR-ligand kinetics kinetics and and investigated the relationship betweenbetween thethe drug structure and the drug drug-receptor-receptor binding kinetics (i.e., structure kinetics relationship, SKR).SKR). A A better better understanding understanding of thisof this relationship relationship should should provide provide a handle a handle for the rationalfor the drugrational optimization drug optimization of the kinetic of the binding kinetic profilebinding of profile antihistamines. of antihistamines. Large didifferencesfferences inin H1RH1R bindingbinding kineticskinetics werewere observedobserved between didifferentfferent antihistaminesantihistamines that werewere notnot alwaysalways reflectedreflected byby thethe strengthstrength ofof thethe bindingbinding interactioninteraction (a(affinity).ffinity). It was additionallyadditionally shown that didifferencesfferences in thethe bindingbinding kinetics of antihistamines at the H1R aaffectffect their mode of antagonism of the GPCR, independent of their bindingbinding aaffinity.ffinity. For ligands binding the H1R, several sub-structures werewere linked linked to to an an increase increase in thein the receptor receptor residence residence time. time. Moreover, Moreover, several several of the newerof the generationnewer generation antihistamines antihistamines (e.g., levocetirizine, (e.g., levocetirizine, olopatadine, olopatadine, rupatadine) rupatadine) contained onecontained or multiple one ofor thesemultiple chemical of these structures, chemical explainingstructures, explaining their long residencetheir long time residence at the time H1R at and the most H1R likelyand most also likely their therapeuticalso their therapeutic success. success. This, together with evidence presented in the scientificscientific literature, suggests that kinetic analyses of drug-receptordrug-receptor binding have added valuevalue forfor thethe developmentdevelopment ofof newnew drugs.drugs. Focus on a better understanding ofof the molecular interactions thatthat areare formed between the drug and receptor over time and thethe relationship relationship with with ligand ligand binding binding rate constants rate constants are therefore are therefore a step forward a step in theforward development in the ofdevelopment effective GPCR of effective ligands. GPCR ligands.

5. Keynote Lectures

5.1. The Design and Development of Keap1-Nrf2 InteractionInteraction Inhibitors and Their Biological ActivityActivity (KL1)(KL1)

Nikolaous Georgakopoulos,Georgakopoulos, Jemma Jemma Gatli Gatliffﬀ and and Geo Geoffﬀ Wells Wells UCL School of Pharmacy, University College London, London WC1N 1AX, United UK; [email protected] Kingdom; e-mail: [email protected] The therapeutic potential of Nrf2-inducing molecules spans several disease states including chronic neurodegenerative diseases, inflammatory conditions and possible roles in cancer

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The therapeutic potential of Nrf2-inducing molecules spans several disease states including chronic neurodegenerative diseases, inflammatory conditions and possible roles in cancer chemoprevention. Conversely, targeted use of Nrf2 inhibitors may have applications in cancer treatment regimens (Cuadrado, A., et al. Nat. Rev. Drug Disc. 2019, 18, 295–317). Our research has been directed towards identifying both reversible and irreversible inhibitors of the Keap1-Nrf2 protein–protein interaction. Reactive inducers have been developed from a previous class of multi-targeted redox homeostasis modulators. On the other hand, reversible inhibitors of Keap1 are the product of structure-based design utilising peptide and small molecule leads. We have identified high affinity sulphonamide ligands for Keap1 (IC50 < 1 nM) and compounds with non-standard binding modes utilising X-ray crystallography studies. In our analysis of the biological activity of these compounds, we characterised effects on mitochondrial function and quality control processes, and autophagy that differ between reversible and irreversible Nrf2 inducers. Differences in mitochondrial turnover, ROS and biogenesis appear to distinguish the two types of Nrf2 inducer across several distinct chemotypes. The activity of these small molecules can contribute to understanding the biological activities of different classes of Nrf2 inducers, their therapeutic utility and potential off-target effects and toxicity.

5.2. Novel Antikinetoplastid Nitroaromatics Bioactivated by Type 1 NTRs (KL2) P. Verhaeghe Faculté des Sciences Pharmaceutiques, Université Paul Sabatier and Laboratoire de Chimie de Coordination, UPR CNRS 8241, 205 route de Narbonne, 31077 Toulouse cedex, France; [email protected] Kinetoplastids are a group of flagellated parasites including Leishmania ssp. and Trypanosoma ssp. that are responsible for several neglected tropical diseases (NTD): visceral leishmaniasis (VL), human African trypanosomiasis (HAT) and Chagas disease (CD), globally responsible for about 30,000 annual deaths, according to the WHO. There are few drugs on the market against these parasitic diseases that affect people living in developing countries. Moreover, most of these drugs present severe side effects and are not orally available. In this worrying context, a new drug called Fexinidazole, a 5-nitroimidazole derivative developed by Sanofi and DNDi, was approved in 2018 by the EMA against HAT and is being evaluated in phase II against CD. Nevertheless, there is still no new chemical entity that clinically studied against VL. Working on nitroaromatic derivatives displaying anti-infective potential, our group identified two novel antileishmanial pharmacophores in 8-nitroquinolin-(1H)-one and 3-nitroimidazo [1, 2-a] pyridine series (Paloque, L., et al. Eur. J. Med. Chem. 2012, 54, 75–86; Castera-Ducros, C., et al. Bioorg. Med. Chem. 2013, 21, 7155–7164; Kieffer, C., et al. Eur. J. Med. Chem. 2015, 92, 282–294; Kieffer, C., et al. Bioorg. Med. Chem. 2015, 23, 2377–2386). By synthesizing more than 200 derivatives, electrochemistry-guided pharmacomodulation studies progressively led to several potent and selective in vitro hit-compounds presenting IC50 values ranging 10 from 10 µM to nM against L. donovani (pro. & ama.), L. infantum (axe. ama.), T. b. brucei (trypo.) and T cruzi (epi.), low cytotoxicities on the human HepG2 cell line and high selectivity indices (from 10 to 400). We then demonstrated that, like for fexinidazole, these nitroaromatic molecules are selectively bioactivated by type 1 nitroreductases, probably leading to cytotoxic electrophilic reduction metabolites such as nitroso and hydroxylamine derivatives. In both series, hit-compounds were not genotoxic in the comet assay and even the mutagenicity Ames test was negative for several imidazopyridine derivatives. The determination of some preliminary in vitro pharmacokinetic parameters highlighted good microsomal stability (T1/2 > 40 min), high albumin binding (98%–99%) and blood–brain barrier diffusion (BBB PAMPA) in quinolinone series, whereas some metabolic issues needed additional work in imidazopyridine series to reach optimized hit-molecules that are now able to undergo in vivo evaluations on mouse models (Pedron, J., et al. Eur. J. Med. Chem. 2018, 155, 135–152; Pedron, J., et al. Pharmaceuticals 2019, 12, 179 5 of 47

ChemMedChem 2018, 13, 2217–2228; Fersing, C., et al. Eur. J. Med. Chem. 2018, 157, 115–126; PharmaceuticalsFersing, C., et2019 al., 12ACS, x FOR Med. PEER Chem. REVIEW Lett. 2019, 10, 34–39). 5 of 47

E° = -0.4 to -0.6 V R8 R6 R3 N R N 2 N O R6 NO NO2 H 2 E°=-0.6to-0.7V

Selective antiparasitic molecules af ter bioactivation by type 1 nitroreductases

5.3. Small Small Molecule Molecule Drugs Drugs and Where to Find Them Them—Approaches—Approaches to Hit Finding, New and Old (KL3) Prof.Michael Michael J. Waring J. Waring Chemistry, School of Natural and Environmental Sciences, Newcastle University, University, Bedson Building, Newcastle upon Tyne NE1 7RU, UK;United [email protected] Kingdom; e-mail: [email protected] The technologies available to the medicinal chemist for finding finding hit molecules for proteins of interest have expanded rapidly in recent years. Fr Fromom extensive use of high-throughput screening to now well-established fragment-based fragment-based methods methods to to emerging emerging technologies technologies (Murray, (Murray, C.W., C.W., Rees, Rees, D.C. D.C. Nature Chem. 20092009,, 1,, 187–192)187–192) suchsuch asas DNA-encodedDNA-encoded chemicalchemical librarieslibraries (Favalli, N., et al. FEBSFEBS Lett. Lett. 2018, 592,, 2168 2168–2180),–2180), the the range range of of complementary complementary techniques techniques is is enabling enabling more more rapid rapid drug-discovery, drug-discovery, including approaches to more challengingchallenging targets.targets. The The central central premises premises to to all these approaches remain—how to select protein targets that are am amenableenable to small molecule modulation in a manner compatible withwith ADMETADMET properties properties and and how how to designto des screeningign screening libraries libraries that generate that generate the best the quality best qualityhits for hits optimisation. for optimisation. The talk will describe past optimisationoptimisation programmes demonstrating where where the hits came from and why these enabled the progression of the project. It will then outline how some of the identified identified concepts might bebe appliedapplied toto futurefuture hit hit finding finding and and some some new new approaches approaches we we are are developing, developing, such such as asFraglite Fraglite screening, screening, which which uses uses minimal minimal halogenated halogenated fragments fragments that express that express hydrogen hydrogen bonding bonding doublet doubletmotifs to motifs assess to druggability assess druggability and inform and future inform hit future finding hit (Wood,finding D.J., (Wood, et al. D.J.,J. Med. et al. Chem. J. Med.2019 Chem., 62, 20193741–3752)., 62, 3741–3752).

5.4. Development Development of of Small Small Molecule Molecule Glial Cell Line-D Line-Derivederived Neurotrophic Factor (GDNF) Family Ligand Mimetics for the Treatment ofof NeurodegenerativeNeurodegenerative DiseasesDiseases andand PainPain (KL4)(KL4) Yulia A.A. Sidorova,Sidorova, Arun Arun Mahato Mahato and and Mart Mart Saarma Saarma

InstituteInstituteofBiotechnology,HiLIFE,UniversityofHelsinki, of Biotechnology, HiLIFE, University of FIN-00014Helsinki,Helsinki, FIN-00014 Finland; Helsinki, [email protected] Finland; e-mail: [email protected] factors are secretory proteins that by binding to their cognate receptors trigger intracellularNeurotrophic signalling factors promoting are secretory neuron proteins survival, that neurite by binding outgrowth to andtheir regulate cognate neuronal receptors plasticity. trigger intracellularGlial cell line-derived signalling neurotrophic promoting neuron factor (GDNF) survival, family neurite consist outgrowth of four factorsand regulate GDNF, neurturinneuronal plasticity.(NRTN), artemin Glial cell (ARTN) line-derived and persephin neurotrophic (PSPN) fact thator (GDNF) first bind family to the consist GPI-anchored of four GDNFfactors receptorGDNF, neurturinfamily alpha (NRTN), (GFRa) artemin and then (ARTN) activate and the persephin transmembrane (PSPN) tyrosinethat first kinase bind to RET. the GDNFGPI-anchored family ligandsGDNF receptor(GFLs) have family neurotrophic alpha (GFRa) effects and on sensory,then activate dopamine, the transmembrane sympathetic, parasympathetic, tyrosine kinase enteric RET. neurons GDNF familyand motoneurons. ligands (GFLs) A recently have discovered neurotrophic novel membereffects ofon the sensory, GFL family, dopamine, a protein calledsympathetic, GDF15, parasympathetic,signals via the GFRAL-RET enteric neurons pathway and motoneurons. and regulates A bodyrecently weight discovered in many novel diseases member (Saarma, of the GFL M., family,Goldman, a protein A. Nature called2017 GDF15,, 550, 195–197). signals GDNFvia the and GFRAL-RET NRTN have pathway been tested and inregulates phase II body clinical weight trials onin manyParkinson diseases0s disease (Saarma, patients M., showing Goldman, modest A. Nature effects 2017 that,are 550 in, 195–197). principle promisingGDNF and although NRTN statisticallyhave been testedinsignificant. in phase ARTN II clinical has been trials tested on Parkinson in phase I′s clinical disease trials patients for neuropathic showing modest pain. Toeffects understand that are the in principle promising although statistically insignificant. ARTN has been tested in phase I clinical trials for neuropathic pain. To understand the molecular mechanisms of ligand receptor interactions, we solved the GDNF-GFRa1 complex structure at 2.35 Å resolution, identified amino acid residues involved in this interaction and residues possibly interacting with RET (Parkash, V., et al. J. Biol.

Pharmaceuticals 2019, 12, 179 6 of 47 molecular mechanisms of ligand receptor interactions, we solved the GDNF-GFRa1 complex structure at 2.35 Å resolution, identified amino acid residues involved in this interaction and residues possibly interacting with RET (Parkash, V., et al. J. Biol. Chem. 2008, 283, 35164–35172). Results of structural studies together with the notion that GFL proteins have poor pharmacological properties, encouraging us to develop small molecules that act similarly to GFLs. The major drawback hindering clinical use of GFLs is inability of these proteins to cross tissue barriers and limited tissue distribution, which restricts their clinical efficacy. We developed medium- and high-throughput methods to identify agonists of GFL receptors and screened a chemical library of 18,000 drug-like compounds. We identified a molecule named BT13 that activated GFL receptor RET and its down-stream signalling cascades in immortalized cells, as well as in sensory and dopamine neurons. In cultured dorsal root ganglion neurons BT13 activated Akt, Erk and Src intracellular signalling cascades and, in the ARTN-like fashion, stimulated elongation and arborisation of neurites of peptidergic neurons (Sidorova, Y.A., et al. Front. Pharmacol. 2017, 8, 365). In rodent spinal nerve ligation (SNL)-induced neuropathic pain model BT13, with equal efficacy with ARTN protein, the surgery-induced mechanical hypersensitivity and normalized the expression of multiple neuronal markers were decreased. BT13 penetrated through the blood–brain barrier, supported the survival of cultured dopamine neurons at 100 nM, alleviated motor symptoms of PD in a rat 6-OHDA model and showed a trend to increase the density of DA fibres in striatum. Using medicinal chemistry and different screening methods, we developed next generation GFL mimetics with improved efficacy, solubility and stability of the compounds. They are currently being tested in animal models of neuropathic pain and Parkinson0s disease. Novel GFL mimetics can be hopefully used also for therapeutic intervention in obesity and in palliative cancer care, as RET activation by GDF15 regulates body weight.

5.5. The Holy Grail of Opioid Analgesia? (KL5) Gerta Cami-Kobeci 1, Mei-Chuan Ko 2, John R Traynor 3 and S M Husbands 1 1 Department of Pharmacy and Pharmacology, University of Bath, Bath BA2 7AY, UK 2 Department of Physiology and Pharmacology, Wake Forest School of Medicine, Winston-Salem, NC, USA 3 Department of Pharmacology, University of Michigan, Ann Arbor, USA; [email protected] The opioid epidemic has rapidly evolved into a medical crisis, with recent figures suggesting 11.5 million people in the United States had misused prescription opioids and >17,000 deaths had been caused by commonly prescribed opioids in 2016 alone. A long sought-after but elusive solution has been the development of a strong but safe opioid analgesic—The Holy Grail of opioid research. Our approach has been to develop analgesics, such as BU08028 (Ding, H., et al. PNAS, 2016, 113, E5511–E5518), with partial agonist activity at both mu and nociceptin/orphanin FQ peptide receptors, as evidence suggests a synergistic interaction between these receptors for analgesia. BU08028 proved to be a potent, long-acting analgesic in primates and did not cause respiratory dependence or reduce respiration. Having defined a pharmacophore for mixed mu and nociception receptor binding, we were able to design a further series of mixed ligands based on the oxymorphone scaffold. BU10038 was identified from within this series as having an almost identical receptor profile to BU08028 and has now been evaluated in a battery of assays to define its analgesic and side effect profile (Kiguchi, N., et al. Brit. J. Anaesthesia, 2019, 122, e146–e156). Following systemic administration in non-human primates, BU10038 1 (0.001–0.01 mg kg− ) dose-dependently produced long-lasting antinociceptive and antihypersensitive effects. Unlike the standard analgesic oxycodone, BU10038 lacked reinforcing effects (i.e., had little or no abuse liability), and compromised the physiological functions of primates including respiration, cardiovascular activities and body temperature at antinociceptive doses and at a 10–30-fold higher dose 1 (0.01–0.1 mg kg− ). Unlike morphine, BU10038 did not cause the development of physical dependence and tolerance after repeated and chronic administration. This work strongly supports the development Pharmaceuticals 2019, 12, 179 7 of 47 of bifunctional MOP/NOP agonists as improved analgesics and an alternative solution for the ongoing Pharmaceuticalsprescription 2019 opioid, 12, x crisis. FOR PEER REVIEW 7 of 47

Docking poses of the analogues in the mu opioid peptide receptor

Funding from the National Institute on Drug Abuse and Orexigen Therapeutics. Funding from the National Institute on Drug Abuse and Orexigen Therapeutics. 5.6. Design of Matrix Metalloproteinase Inhibitors for Inflammatory Bowel Disease (KL6) 5.6. Design of Matrix Metalloproteinase Inhibitors for Inflammatory Bowel Disease (KL6) Maria Pigott, Jun Wang, Shane O′Sullivan, Carlos Medina and John F Gilmer Maria Pigott, Jun Wang, Shane O0Sullivan, Carlos Medina and John F Gilmer School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, D2, Ireland; e-mail: School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, D2, Ireland; [email protected] [email protected] Matrix metalloproteinases (MMPs) is an umbrella term for a large family of widely distributed Matrix metalloproteinases (MMPs) is an umbrella term for a large family of widely distributed enzymes that influence diverse biochemical processes relevant to normal physiology and to disease. enzymes that influence diverse biochemical processes relevant to normal physiology and to disease. MMPs are interesting drug targets but they are challenging because of their distribution, range of MMPs are interesting drug targets but they are challenging because of their distribution, range of effects and similarity. The so-called gelatinase B enzyme, also known as MMP-9, is a key player effects and similarity. The so-called gelatinase B enzyme, also known as MMP-9, is a key player in in the progression of inflammatory bowel disease (IBD) and a marker of inflammation and disease the progression of inflammatory bowel disease (IBD) and a marker of inflammation and disease progression (O Sullivan, S., et al. Mediat. Inflamm. 2015, 2015, 1–18). This talk will describe the biology progression (O0′Sullivan, S., et al. Mediat. Inflamm. 2015, 2015, 1–18). This talk will describe the biology of MMP-9 in IBD and provide an overview of our attempts to design inhibitors that selectively affect of MMP-9 in IBD and provide an overview of our attempts to design inhibitors that selectively affect its function. These efforts relate to three broad strategies: (i) attempts at increasing MMP-9 selectivity its function. These efforts relate to three broad strategies: (i) attempts at increasing MMP-9 selectivity in the pyrimidinetrione class through conventional medicinal chemistry approaches (Wang, J., et al. in the pyrimidinetrione class through conventional medicinal chemistry approaches (Wang, J., et al. Bioorg. Med. Chem. 2011, 19, 16, 4985–4999); (ii) hybrid tactics using nitric oxide mimetic functionality Bioorg. Med. Chem. 2011, 19, 16, 4985–4999); (ii) hybrid tactics using nitric oxide mimetic functionality to influence MMP transcription (O Sullivan, S., et al. Br. J. Pharmacol. 2017, 174, 512–524; Wang. J.; to influence MMP transcription (O′0Sullivan, S., et al. Br. J. Pharmacol. 2017, 174, 512–524; Wang. J.; O Sullivan, S., et al. J. Med. Chem. 2012, 55, 2154–2162); (iii) quasi pharmaceutical strategies to confine O′0Sullivan, S., et al. J. Med. Chem. 2012, 55, 2154–2162); (iii) quasi pharmaceutical strategies to confine broadly unselective pyrimidinetrione inhibitors to disease tissue through manipulation of chemical broadly unselective pyrimidinetrione inhibitors to disease tissue through manipulation of chemical properties. The outcomes of these attempts and lessons learned will be related through data from properties. The outcomes of these attempts and lessons learned will be related through data from in in vitro/in vivo models of ulcerative colitis. vitro/in vivo models of ulcerative colitis. 5.7. Computational Medicinal Chemistry Approaches for GPCR Structure-Based Drug Discovery (KL7) 5.7. Computational Medicinal Chemistry Approaches for GPCR Structure-Based Drug Discovery (KL7) Chris De Graaf Chris De Graaf Sosei Heptares; [email protected] Sosei Heptares; e-mail: [email protected] Novel crystal structures of GPCR-ligand complexes solved at Sosei Heptares and elsewhere continueNovel to crystal reveal structures a diversity of of GPCR-ligand potential ligand comp bindinglexes solved sites. at Emerging Sosei Heptares sets of and GPCR elsewhere crystal continuestructures to of reveal multiple a diversity diverse ligands of potential bound ligand to closely binding related sites. receptors Emerging furthermore sets of finallyGPCR enable crystal a structuresprotein structure-based of multiple diverse view ofligands how di boundfferent to ligands closely bind related this receptors major drug furthermore target class. finally enable a proteinThis structure-based presentation will view address of how several different important ligands bind repercussions this major and drug learnings target class. from the analysis of GPCRThis presentation structures for will ligand address design several that importan should bet repercussions transferable and and learnings relevant forfrom many the analysis targets, ofboth GPCR GPCRs structures and enzymes, for ligand including: design that should be transferable and relevant for many targets, both GPCRs and enzymes, including: • Caveats in using pharmacophore-based similarity principles for modeling receptor ligand complexes different ligand chemotypes. • The important roles of lipophilic hot spots and water networks as drivers of GPCR druggability, ligand binding and selectivity.

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Caveats in using pharmacophore-based similarity principles for modeling receptor ligand • complexes different ligand chemotypes. The important roles of lipophilic hot spots and water networks as drivers of GPCR druggability, • ligand binding and selectivity. Binding mode diversity of (chemically similar) ligands across the structural GPCRome. • This presentation will show how the breakthroughs in GPCR structural biology can be complemented by computational and experimental studies for a more accurate description and prediction of molecular and structural determinants of ligand-receptor binding affinity, kinetics, potency and selectivity. Integrated cheminformatics workflows will be described that combine structural, pharmacological and chemical data to explore receptor–ligand interaction space and steer structure-based virtual ligand screening. Novel cheminformatics-driven molecule design approaches will be discussed, combining retrosynthetic analysis, library enumeration approaches (e.g., matched molecular pairs analysis) and recurrent neural network-driven molecule generation. We will discuss how the accumulated GPCR structural chemogenomics data can be used to construct customized structure-based medicinal chemistry toolboxes for hit optimization and library design. Orthogonal physics-based (Molecular Dynamics, e.g., Free Energy Perturbation FEP+, WaterMap from Schrödinger) and empirical (e.g., GRID and WaterFLAP from Molecular Discovery) structure-based drug design methods will be presented to target lipophilic hotspots, water networks and cryptic ligand binding pockets for a variety of GPCR subfamilies.

5.8. Phenotypic Activity Driven Target Identification and Discovery of Drug Candidates (KL8) Paul Wyatt Drug Discovery Unit, Wellcome Centre for Anti-Infectives Research, University of Dundee, School of Life Sciences, Dundee DD1 5EH, UK; [email protected] There are considerable opportunities for drug discovery centres based in universities to generate benefit to patients, provided they do not compete with or merely replicate BioPharma. The centres have to develop activities complementary to, or in partnership with the BioPharma industry. These include partially de-risking novel drug discovery targets, or tackling neglected and orphan diseases. The Drug Discovery Unit seeks to achieve this by combining a globally rare university-based, fully integrated drug discovery group capable of delivering new drug candidates with a world-class life sciences research environment and multiple partners to develop the drug discovery approaches to major global diseases. The talk describes how we are delivering much needed clinical candidates, for example, malaria and visceral leishmaniasis (two) by taking a phenotypic approach to hit identification and optimisation, using mode of action studies to identify novel drug targets and to manage a portfolio of phenotypically derived projects.

5.9. Resin Acid-Driven Innovations in Antimicrobial Research (KL9) Vânia M. Moreira 1,2 1 Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, United Kingdom, 161 Cathedral Street, G4 0RE Glasgow; [email protected] 2 Drug Research Program, Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, Finland, Viikinkaari 5 E, P.O. Box 56, FI-00014 University of Helsinki Humans have employed resins for esthetic, ceremonial or therapeutic uses for millennia (Klemens, F., Dieter, G. “Resins, Natural”. Ullmann0s Encyclopedia of Industrial Chemistry, 2000). Gum rosin, in particular, has an annual worldwide production of more than 1 million tons and is used as an ingredient for inks, varnishes, adhesives, cosmetics, medicines and chewing gums. The ecological role of resins is protective, that is, when trees are cut, the volatiles in the resin evaporate leaving Pharmaceuticals 2019, 12, 179 9 of 47 the solid portion that contains “resin acids”, which shelter them from external stress and invaders, promoting healing. Our research has revealed that these “resin acids”, belonging to the class of abietane-type diterpenoids, have antimicrobial and anti-biofilm activity towards gram-positive bacteria and can be exploited for human use (Fallarero, A., et al. Int. J. Mol. Sci. 2013, 14, 12054–12072). Through chemistry, we have created a library of over 50 new chemically pure “resin acids” derivatives, which includes, to the best of our knowledge, the most potent agents reported within their class so far (Manner, S., et al. Eur. J. Med. Chem. 2015, 102, 68–79; Moreira, V.M., et al. WO 2016/051013 A1, 7th April 2016). It is noteworthy that this effect is accompanied by a good tolerability profile and low potential for spreading resistance. Moreover, combination of these compounds with nanocellulose, an advanced biopolymer with excellent mechanical properties and biocompatibility, resulted in innovative and cost-effective functional surfaces, bearing a broad spectrum of action against bacteria, high biocompatibility and no cross-resistance with drug-resistant strains (Hassan, G., et al. ACS Sustainable Chem. Eng. 2019, 7, 5002–5009). This presentation will detail the outcomes of our work with this class of compounds and further illustrate how natural products and their derivatives foster innovation in antimicrobial research. Acknowledgements: The Finnish Funding Agency for Technology and Innovation (project 1297/31/2016).

5.11. Drug-Like Properties and Ligand Efficiency-Guided Optimisation (KL11) Paul Leeson Paul Leeson Consulting Ltd. and School of Pharmacy,University of Nottingham, UK; [email protected] It has been acknowledged that significant attrition and delay in clinical drug development pipelines has been caused by poor compound properties such as inadequate pharmacokinetics and poor solubility. Generic compound quality guidelines, such as the rule of five, may not always be helpful because differing drug binding sites on biological targets can strongly influence the allowable physicochemical property ranges of their ligands. The application of ligand efficiency measures, which quantify in vitro activities per unit of a physical property such as size or lipophilicity, provides an Pharmaceuticals 2019, 12, x FOR PEER REVIEW 9 of 47 includes, to the best of our knowledge, the most potent agents reported within their class so far (Manner, S., et al. Eur. J. Med. Chem. 2015, 102, 68–79; Moreira, V.M., et al. WO 2016/051013 A1, 7th April 2016). It is noteworthy that this effect is accompanied by a good tolerability profile and low potential for spreading resistance. Moreover, combination of these compounds with nanocellulose, an advanced biopolymer with excellent mechanical properties and biocompatibility, resulted in innovative and cost-effective functional surfaces, bearing a broad spectrum of action against bacteria, high biocompatibility and no cross-resistance with drug-resistant strains (Hassan, G., et al. ACS Sustainable Chem. Eng. 2019, 7, 5002–5009). This presentation will detail the outcomes of our work with this class of compounds and further illustrate how natural products and their derivatives foster innovation in antimicrobial research. Acknowledgements: The Finnish Funding Agency for Technology and Innovation (project 1297/31/2016).

5.10. Glycoconjugates Disrupting Bacterial and Fungal Adhesion Mechanisms (KL10) Sébastien G. Gouin University of Nantes, CEISAM, Chimie Et Interdisciplinarité, Synthèse, Analyse, Modélisation, UMR CNRS 6230, UFR des Sciences et des Techniques, 2, rue de la Houssinière, BP 92208, 44322 NANTES Cedex 3, France; e-mail: [email protected] Increasing resistance to antibiotics or antifungals is a serious health problem, which is worsening with the constant identification of strains resilient to commonly available chemotherapeutic agents. Among the therapeutic alternatives developed at the academic level, the anti-adhesive strategy has Pharmaceuticalsseen a growing2019, interest12, 179 in the last 25 years (Cecioni, S., et al. Chem. Rev. 2015, 115, 525–561).10 ofThe 47 concept is to disrupt the lectin-mediated adhesion of the pathogen to eukaryotic cells. This therapeutic approach should be less prone to the development of resistances and selection pressures, improved perspective of compound quality over physicochemical properties alone. Thus, candidate as the pathogens are not killed during the decolonisation process. drugs and approved drugs are frequently amongst the most ligand efficient for their specific target, Our group is interested in developping antagonists of carbohydrate-binding and processing on the basis of measures using both size (ligand efficiency, LE) and lipophilicity (ligand lipophilic proteins (adhesins and glycosidases) to disrupt the attachment of pathogenic bacteria and fungi to efficiency, LLE). This is seen even with drugs having extreme physicochemical properties, such as host cells. We will discuss the design of mono- and multivalent glycoconjugates and their in vitro those breaking the rule of five, for example, the new raft of drugs for treatment of the hepatitis C and in vivo antiadhesif potential. A specific focus will be made on antagonists of adhesins (Figure 1) virus. The desirable optimisation destination in the property efficiency space of a novel target can be (Brument, S., et al. J. Med. Chem. 2013, 56, 5395–5406; Brissonnet, Y., et al. J. Am. Chem. Soc. 2013, 135, anticipated once enough screening data are available, usually in the hit-to-lead phase. In conjunction 18427–18435; Brissonnet, Y., et al. Chem. Eur. J. 2019, 25, 2358–2365; Alvarez Dorta, D., et al. with the application of multiparameter ADMET optimisation, the approach can help to refine drug ChemBioChem 2016, 17, 936–952) or invasive aspergillosis (Lehot, V., et al. Chem. Eur. J. 2018, 24, 19243– design options, prioritise synthesis and reduce the numbers of compounds made (Young, R.J., et al. 19249). J. Med. Chem. 2018, 61, 6421–6467).

Figure 1. Illustration of the antiadhesive concept. Oral administration of Escherichia coli antiadhesives Figure 1. Illustration of the antiadhesive concept. Oral administration of Escherichia coli antiadhesives in Crohn s disease mouse model was shown to lower the bacterial load in the gut and to reduce the in Crohn0′s disease mouse model was shown to lower the bacterial load in the gut and to reduce the inﬂammatory syndromes. AIEC: adherent-invasive E. coli. inflammatory syndromes. AIEC: adherent-invasive E. coli. 5.12. Natural Products and Analogues as Potential Therapeutic Agents (KL12) 5.11. Drug-Like Properties and Ligand Efficiency-Guided Optimisation (KL11) Diego Cortes 1, María Jesús Sanz 1,2 and Nuria Cabedo 2

1 Department of Pharmacology, University of Valencia, Burjassot, Spain, 46100 2 Biomedical Research Institute INCLIVA, 46010 Valencia, Spain; [email protected] Natural products have been the most valuable inspiration for the synthesis of novel therapeutic agents. Among them, heterocyclic moieties are prevalent in pharmacologically relevant drugs (Cortes, D. Bioactive Secondary Metabolites: The Drugs that Nature provides. 2018. Cortes D. (Ed). ISBN-10: 171877768X; ISBN-13: 978–1718777682. CreateSpace). In the course of our research on synthesis of bioactive heterocycles, we synthetized novel isoquinolines as dopaminergic ligands, including tetrahydroisoquinolines carbamates (Parravicini, O., et al. J. Mol. Model. 2017, 23, 273), hexahydrocyclopenta-IQ (HCPIQ)(Figure2) (Parraga, J., et al. Eur. J. Med. Chem. 2016, 122, 27–42), aporphines (Moreno, L., et al. Bioorg. Med. Chem. Lett. 2013, 23, 4824–4827), phenanthrenes (Moreno, L, et al. Bioorg. Med. Chem. Lett. 2013, 23, 4824–4827) and protoberberines (Párraga, J., et al. Eur. J. Med. Chem. 2013, 68, 150–166). Dopaminergic ligands can bind to subfamily D1-like (D1,D5) and D2-like (D2-D4) dopamine receptors (DR) to “restore” the dopaminergic pathway. From a therapeutic point of view, DR are the main target for antipsychotic (neuroleptics) and antiparkinsonian drugs. Recently, we demonstrated by binding experiments and functional assays that some of these compounds displayed aﬃnity for DR with high selectivity for D2-like DR, behaving as dopaminergic agonists. Molecular modelling studies have established the main molecular interactions that stabilize the diﬀerent ligand-receptor complexes (Parravicini, O., et al. J. Mol. Model. 2017, 23, 273; Parraga, J., et al. Eur. J. Med. Chem. 2016, 122, 27–42). Pharmaceuticals 2019, 12, x FOR PEER REVIEW 10 of 47

Paul Leeson Paul Leeson Consulting Ltd. and School of Pharmacy, University of Nottingham, United Kingdom; e-mail: [email protected] It has been acknowledged that significant attrition and delay in clinical drug development pipelines has been caused by poor compound properties such as inadequate pharmacokinetics and poor solubility. Generic compound quality guidelines, such as the rule of five, may not always be helpful because differing drug binding sites on biological targets can strongly influence the allowable physicochemical property ranges of their ligands. The application of ligand efficiency measures, which quantify in vitro activities per unit of a physical property such as size or lipophilicity, provides an improved perspective of compound quality over physicochemical properties alone. Thus, candidate drugs and approved drugs are frequently amongst the most ligand efficient for their specific target, on the basis of measures using both size (ligand efficiency, LE) and lipophilicity (ligand lipophilic efficiency, LLE). This is seen even with drugs having extreme physicochemical properties, such as those breaking the rule of five, for example, the new raft of drugs for treatment of the hepatitis C virus. The desirable optimisation destination in the property efficiency space of a novel target can be anticipated once enough screening data are available, usually in the hit-to-lead phase. In conjunction with the application of multiparameter ADMET optimisation, the approach can help to refine drug design options, prioritise synthesis and reduce the numbers of compounds made (Young, R.J., et al. J. Med. Chem. 2018, 61, 6421–6467).

5.12. Natural Products and Analogues as Potential Therapeutic Agents (KL12)

Diego Cortes 1, María Jesús Sanz 1,2, and Nuria Cabedo 2

1 Department of Pharmacology, University of Valencia, Burjassot, Spain, 46100. 2 Biomedical Research Institute INCLIVA, 46010 Valencia, Spain; e-mail: [email protected] Natural products have been the most valuable inspiration for the synthesis of novel therapeutic agents. Among them, heterocyclic moieties are prevalent in pharmacologically relevant drugs (Cortes, D. Bioactive Secondary Metabolites: The Drugs that Nature provides. 2018. Cortes D. (Ed). ISBN-10: 171877768X; ISBN-13: 978–1718777682. CreateSpace). In the course of our research on synthesis of bioactive heterocycles, we synthetized novel isoquinolines as dopaminergic ligands, including tetrahydroisoquinolines carbamates (Parravicini, O., et al. J. Mol. Model. 2017, 23, 273), hexahydrocyclopenta-IQ (HCPIQ)(Figure 2) (Parraga, J., et al. Eur. J. Med. Chem. 2016, 122, 27–42), aporphines (Moreno, L., et al. Bioorg. Med. Chem. Lett. 2013, 23, 4824–4827), phenanthrenes (Moreno, L, et al. Bioorg. Med. Chem. Lett. 2013, 23, 4824–4827) and protoberberines (Párraga, J., et al. Eur. J. Med. Chem. 2013, 68, 150–166). Dopaminergic ligands can bind to subfamily D1-like (D1, D5) and D2-like (D2-D4) dopamine receptors (DR) to “restore” the dopaminergic pathway. From a therapeutic point of view, DR are the main target for antipsychotic (neuroleptics) and antiparkinsonian drugs. Recently, we demonstrated by binding experiments and functional assays that some of these compounds displayed affinity for DR with high selectivity for D2-like DR, behaving as dopaminergic agonists. Molecular modelling studies have established the main molecular interactions that stabilize the different ligand-receptor complexes (Parravicini, O., et al. J. Mol. Model. 2017, 23, 273; Parraga, J., Pharmaceuticalset al. Eur. J. Med.2019, 12Chem, 179. 2016, 122, 27–42). 11 of 47

Figure 2. Molecular graph of the molecular interactions between hexahydrocyclopenta-IQ (HCPIQ) 3e and hydrophobic residues: Val115, Ile183, Ile184, Trp386, Phe389, Phe390, His393, Thr412 and Tyr416 of

D2 dopamine receptors (DR).

In recent years, considerable attention has been paid on melatonergic (MT) receptor agonists given their application for treating insomnia and depression. We synthetized novel hexahydroindenopyridines (HHIP), which were able to bind MT1 and/or MT2 receptors. These HHIP emerged as promising leads for the development of new melatoninergic agents (Parraga, J., et al. Eur. J. Med. Chem. 2014, 30, 700–709). This work was funded by grants SAF2017–89714-R, SAF2014–57845R, CP15/00150 and PI18/01450 from Ministerio de Ciencia, Innovación, Instituto de Salud Carlos III and co-funded by European Union (ERDF/ ESF) “Investing in your future”.

6. Young Researcher Communications

6.1. Pyrroloquinolones as Novel Dual-Targeting and Dual-Stage Oral Efficacious Antimalarials (YRC01) Gustavo da Silva 1,2, Lis Coelho 3, Diana Fontinha 4, Vera L. M. Silva 2, Artur M. S. Silva 2 and Rui Moreira 1 1 iMed.ULisboa, Faculdade de Farmácia da Universidade de Lisboa, Lisboa, Portugal, 1649–003 2 Department QOPNA, Departamento de Química da Universidade de Aveiro, 3810–193 Aveiro, Portugal 3 Centro da Malária e Doenças Tropicais, IHMT, Universidade Nova de Lisboa, 1349–008 Lisboa, Portugal 4 Instituto de Medicina Molecular, Faculdade de Medicina Universidade de Lisboa, 1649–028 Lisboa, Portugal; galfdsilva@ff.ulisboa.pt In this communication, we report the results from a medicinal chemistry programme focused on a pyrroloquinolone (PQ) series targeting the Plasmodium falciparum mETC (mitochondrial electron transport chain). This previously unknown series 1 was tested against blood- and liver-stage malaria parasites and our hit compound revealed excellent dual-stage inhibitory activity. The plausible structure–activity relationships and the inhibition of mETC components (cytochrome bc1, dihydroorotate dehydrogenase) will be discussed in this communication. In addition, our hit compound interfered with calcium homeostasis in transgenic malaria parasites. The interactions between the lead compounds and target were confirmed by docking. Our hit compound also revealed a good selectivity index, in vivo efficacy and favourable pharmacokinetic properties (Figure3). This work was supported by Fundação para a Ciência e Tecnologia (FCT), FEDER and PT2020 through projects UID/DTP/04138/2013 and UID/QUI/00062/2013. We also thank FCT for the fellowships SFRH/BPD/108807/2015 (V.L.M.S.) and SFRH/BD/103412/2014 (G.S.). Pharmaceuticals 2019, 12, 179 12 of 47 Pharmaceuticals 2019, 12, x FOR PEER REVIEW 12 of 47

FigureFigure 3.3. StructureStructure andand relevantrelevant resultsresultsof ofour ourlead lead compounds. compounds.

6.2. Molecular Docking, Semisynthesis and Biological Evaluation of Garcinoic Acid Amides as 5-Lipoxygenase This work was supported by Fundação para a Ciência e Tecnologia (FCT), FEDER and PT2020 Inhibitors (YRC02) through projects UID/DTP/04138/2013 and UID/QUI/00062/2013. We also thank FCT for the Chaufellowships Phi Dinh SFRH/BPD/108807/20151, Alexia Villea, Anais Barb (V.L.M.S.)é 1, Thuy and Linh SFRH/BD/103412/2014 To 1, Konstantin Neukirch (G.S.).2, Guillaume Viault 1, Veronika Temml 3, Andreas Koeberle 2, Daniela Schuster 4, Oliver Werz 2, Pascal Richomme 1, Jean-Jacques6.2. Molecular Helesbeux Docking, Semisynthesis1 and Denis Sandéraphin Biolog1ical Evaluation of Garcinoic Acid Amides as 5- Lipoxygenase Inhibitors (YRC02) 1 SONAS, EA921, UNIV Angers, SFR QUASAV, Faculty of Health Sciences, Department of Pharmacy, 16Chau bd Daviers, Phi Dinh 49045 1,*, Alexia Angers Ville Cedexa, Anais 01, France Barbé 1, Thuy Linh To 1, Konstantin Neukirch 2, Guillaume 2ViaultChair of1, PharmaceuticalVeronika Temml/Medicinal 3, Andreas Chemistry, Koeberle Institute 2, Daniela of Pharmacy, Schuster Friedrich-Schiller-University 4, Oliver Werz 2, Pascal Jena,Richomme 07743 Jena, 1, Jean-Jacques Germany Helesbeux 1 and Denis Séraphin 1 3 1 Institute SONAS, of PharmacyEA921, UNIV/Pharmacognosy Angers, SFR QUASAV, University Faculty of Innsbruck, of Health Sciences, 6020 Innsbruck, Department Austria of Pharmacy, 16 bd 4 DepartmentDaviers, 49045 of Pharmaceutical Angers Cedex 01, and France. Medicinal Chemistry, Paracelsus Medical University Salzburg, 50202 Salzburg, Chair of Pharmaceutical/Medicinal Austria; [email protected] Chemistry, Institute of Pharmacy, Friedrich-Schiller-University Jena, 07743 Jena, Germany. Prostaglandins and leukotriens (LTs), produced from arachidonic acid by cyclo-oxygenase and 3 Institute of Pharmacy/Pharmacognosy University of Innsbruck, 6020 Innsbruck, Austria. 5-lipoxygenase (5-LOX) pathways, respectively, are key mediators in inflammation (Funk, C. D. 4 Department of Pharmaceutical and Medicinal Chemistry, Paracelsus Medical University Salzburg, 5020 ScienceSalzburg,2001, 294 Austria;, 1871–1875). e-mail: [email protected] Our recent results showed an anti-inflammatory activity of ω-oxidized tocotrienols by inhibiting 5-LOX (Pein, H., et al. Nat. Commun. 2018, 9, 3834). Amongst these compounds,Prostaglandinsδ-garcinoic and acid leukotriens (δ-GA) was (LTs), the produced most active from one. arachidonic It inhibited acid LTs formationby cyclo-oxygenase in 5-LOX andand polymorphonuclear5-lipoxygenase (5-LOX) leukocyte pathways, assays respectively, with IC at are 0.04 key0.02 mediatorsµM and in 0.35 inflammation0.08 µM, (Funk, respectively. C. D. 50 ± ± MolecularScience 2001 docking, 294, 1871–1875). with an in-house Our recent dataset results was performedshowed an andanti-inflammatory identified new allostericactivity of binding ω-oxidized site fortocotrienols these compounds. by inhibiting 5-LOX (Pein, H., et al. Nat. Commun. 2018, 9, 3834). Amongst these compounds,In order toδ-garcinoic further understand acid (δ-GA) the was mode the of most action active and one. to improve It inhibited the 5-LOX LTs formation inhibitory in activity 5-LOX ofandδ-GA polymorphonuclear or its pharmacological leukocyte profile, assays the with rational IC50 at design 0.04 ± 0.02 of a µM series and of 0.35 amides ± 0.08 followed µM, respectively. by their semisynthesisMolecular docking was carriedwith an out.in-house Here, dataset we present was pe arformed dataset ofand novel identifiedδ-GA-based new allosteric anti-inflammatory binding site agentsfor these with compounds promising. in vitro and in cellulo anti-inflammatory activity.

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 13 of 47 Pharmaceuticals 2019, 12, 179 13 of 47

Docking pose of a GA amide into an allosteric binding site of 5-lipoxygenase (5-LOX).

In order to further understand the mode of action and to improve the 5-LOX inhibitory activity 6.3. Polypharmacological Compounds in Alzheimer0s Disease: Innovative 5-HT4R Agonist with Promising in Celluloof δ-GA Antioxidant or its pharmacological Properties (YRC03) profile, the rational design of a series of amides followed by their semisynthesis was carried out. Here, we present a dataset of novel δ-GA-based anti-inflammatory 1 2 3 1 1 1 Carolineagents with Lanthier promising, Irene in vitro liparulo and ,in Hugo cellulo Payan anti-inflammatory,Cédric Lecoutey activity., Marc Since , Audrey Davis , Sylvie Claeysen 3, Maria-Laura Bolognesi 2, Patrick Dallemagne 1 and Christophe Rochais 1 16.3.Normandie Polypharmacological Univ, UNICAEN, Compounds CERMN, in Alzheimer 14000′ Caen,s Disease: France Innovative 5-HT4R Agonist with Promising in 2CelluloDepartment Antioxidant of Pharmaceutical Properties (YRC03) Sciences, University of Bologna, Via Belmeloro 6, 40126 Bologna, Italy 3 CarolineIGF, Univ. Lanthier Montpellier, 1, Irene CNRS, liparulo INSERM, 2, Hugo Montpellier, Payan 3, Cédric France; Lecoutey [email protected] 1, Marc Since 1, Audrey Davis 1, SylvieIn a Claeysen world where 3, Maria-Laura life expectancy Bolognesi is increasing, 2, Patrick Alzheimer Dallemagne disease 1 and (AD)Christophe is the mainRochais cause 1 of dementia,1 Normandie and a ffUniv,ects UNICAEN, approximatively CERMN, 17% 14000 of people Caen, France. who are older than 75 years in France. This is a progressive2 Department neurodegenerative of Pharmaceutical disorder Sciences, characterized University of byBologna, memory Via lossBelmeloro and cognitive 6, 40126 Bologna, decline. Italy. Despite the3 fact IGF, that Univ. the Montpellier, physiopathology CNRS, INSERM, of AD is Montpellier, not entirely France; known e-mail: at the [email protected] current time, some molecular causes were found such as the ß-amyloid peptides aggregation, tau-dependent neurofibrillary tangles, In a world where life expectancy is increasing, Alzheimer disease (AD) is the main cause of as well as oxidative stress and neuroinflammation. Currently, treatments available for patients are dementia, and affects approximatively 17% of people who are older than 75 years in France. This is a mainly acetylcholine esterase (AChE) inhibitors, which only have symptomatic benefits and do not progressive neurodegenerative disorder characterized by memory loss and cognitive decline. Despite cure AD. Thus, there is still a strong medical need in the AD population. the fact that the physiopathology of AD is not entirely known at the current time, some molecular In this context, the concept of multi-target directed ligands (MTDLs) was applied to design a drug causes were found such as the ß-amyloid peptides aggregation, tau-dependent neurofibrillary with several therapeutic targets. The envisaged MTDL (targeted structure—Figure4) should be able tangles, as well as oxidative stress and neuroinflammation. Currently, treatments available for to limit the development of ß-amyloid plaques obtained by the aggregation of ß-amyloid peptides patients are mainly acetylcholine esterase (AChE) inhibitors, which only have symptomatic benefits (Aß).and do Indeed, not cure our AD. compounds Thus, there are is still designed a strong to promotemedical need the cleavage in the AD of population. amyloid protein precursor α α (APP)In by this -secretasecontext, the activation concept inof ordermulti-target to produce directed a neuroprotective ligands (MTDLs) and was soluble applied peptide to design sAPP a. Thisdrug iswith the several role of thetherapeutic 5HT4R agonists,targets. The which envisage are alreadyd MTDL studied (targeted in thestructure—Figure CERMN (Centre 4) should d’Etudes be etable de to Recherche limit the sur development le Médicament of deß-amyloid Normandie) plaques in other obtained MTDL by projects the aggregation and led to theof ß-amyloid discovery J. Med. Chem 2015 ofpeptides donecopride (Aß). Indeed, (blue part—Figure our compounds1) (Rochais, are designed C., et al. to promote the., cleavage, 58, 3172–3187).of amyloid protein On the J. Med. Chem otherprecursor hand, (APP) it appears by α -secretase that oxidative activation stress in plays order a centralto prod roleuce ina neuroprotective AD (Rosini, M., and et al. soluble peptide., 2014 sAPP,α 57,. This 2821–2831). is the role Adding of the antioxidant5HT4R agonists, moiety which such are as polyphenol,already studied lipoic in andthe CERMN ferulic acid (Centre (red part—Figured'Etudes et de1) couldRecherche trap free sur radicals le Médicament or reactive de oxygen Normandie) species in (ROS) other and MTDL also haveprojects a neuroprotective and led to the ediscoveryffect. This of aspect donecopride has been (blue widely part studied—Figure in Prof.1) (Rochais, Maria-Laura C., et al. Bolognesi J. Med. Chem0s laboratory., 2015, 58, over 3172–3187). the years J. Med. Chem (OnBolognesi, the other M.-L., hand, et it al. appears that oxidative., 2009, stress 52, 7883–7886). plays a central To that role end, in AD di ff(Rosini,erent compounds M., et al. J. Med. will beChem designed., 2014, and57, 2821–2831). synthesized, Adding with both antioxidant the expertise moiety of CERMNsuch as polyphenol, and Prof. Maria-Laura lipoic and ferulic Bolognesi, acid in(red order part—Figure to evaluate 1) their couldin vitrotrap /infree vivo radicalsproperties or reactive regarding oxygen their species agonist (ROS) activity and on 5-HTalso 4haveR and a antioxidantneuroprotective properties. effect. The This development aspect has and been promising widelyin studied vitro/in celluloin Prof. results Maria-Laura of the chloroaniline Bolognesi0′ss moietylaboratory line over will bethe described years (Bolognesi, in this presentation. M.-L., et al. J. Med. Chem., 2009, 52, 7883–7886). To that end, different compounds will be designed and synthesized, with both the expertise of CERMN and Prof. Maria-Laura Bolognesi, in order to evaluate their in vitro/in vivo properties regarding their agonist

Pharmaceuticals 2019, 12, 179 14 of 47

Figure 4. Targeted structure, with 5-HT4R agonist moiety in blue and antioxidant moieties in red.

6.4. Reactivity of Pazopanib and Sunitinib Aldehyde Metabolites toward Proteins: Potential Involvement in Hepatotoxicity and Drug–Drug Interactions (YRC04) Marie-Noëlle Paludetto 1, Jean-Luc Stigliani 2, Anne Robert 2, Vania Bernardes-Génisson 2, Etienne Chatelut 1, Florent Puisset 1 and Cécile Arellano 1 1 Centre de Recherches en Cancérologie de Toulouse (CRCT), University of Toulouse, 31037 Toulouse, France 2 Laboratoire de Chimie de Coordination du CNRS (LCC-CNRS), University of Toulouse, 31077 Toulouse, France; [email protected] Pazopanib and sunitinib are tyrosine kinase inhibitors (TKI) approved for the treatment of various cancers. Their hepatotoxicity has been highlighted by the black boxed warnings issued by the FDA in their drug labels. These TKI are mainly metabolized by hepatic cytochromes P450 (CYP), and previous in vitro studies have suggested that they are CYP3A4 time-dependant inhibitors (Kenny, J. R., et al. Pharm. Res. 2012, 29, 1960–1976; Filppula, A. M., et al. Drug Metab. Dispos. 2014, 42, 1202–1209). New reactive metabolites (RM) with aldehyde structures have been discovered during pazopanib and sunitinib metabolism (Paludetto, M-N., et al. J. Med. Chem. 2018, 61, 7849–7860). Aldehydes are hard electrophiles that have a high reactivity potential toward proteins and that are thought to be responsible for CYP inactivation, drug–drug interactions (DDI) and hepatotoxicity (Kalgutkar, A. S. Chem. Res. Toxicol. 2017, 30, 220–238). The reactivity of pazopanib and sunitinib aldehyde RM toward different hepatic (human liver microsomes (HLM), recombinant CYP3A4) and plasma (albumin) proteins was investigated by LC-MS/MS to better understand how these RM could be involved in liver toxicity and DDI. Because of the presence of a free amino group on its side chain, lysine could be a good target for reactive electrophiles. Reactions between aldehydes and amines affording imine derivatives are known to be reversible in aqueous media. Thus, adduct formation with these different proteins was examined after reductive amination of imine derivatives by NaBH3CN in vitro and after proteolytic digestion. Preliminary studies were carried out with free lysine and aldehyde RM generated beforehand by biomimetic oxidation reactions catalysed by metalloporphyrins. Lysine adduct formation was then studied with aldehyde RM generated in situ in HLM and recombinant CYP3A4 incubations. Aldehyde-lysine adducts on both CYP3A4 and plasma proteins were detected for the first time by LC-MS for pazopanib and sunitinib and characterized by the related [M+H]+ ions and MS/MS fragmentation patterns. In addition, the formation of aldehyde RM in vivo was investigated in plasma samples from patients treated, separately, with pazopanib or sunitinib. These aldehyde RM have been unambiguously detected in patient plasma samples during drug monitoring. Thus, it could be hypothesized that the coupling of pazopanib or sunitinib aldehydes with lysine residues of CYP3A4 and hepatic proteins may be involved both in hepatotoxicity and DDI.

6.5. Click Chemistry Tools to Unravel New Targets in Malaria (YRC05) Jorge Grilo 1,2, Lara Fidalgo 1, Diana Fontinha 2, Beatriz Anacleto 1, Charlie Saunders 3, Edward W. Tate 3, Rui Moreira 1, Miguel Prudêncio 2 and Ana S. Ressurreição 1 Pharmaceuticals 2019, 12, x FOR PEER REVIEW 15 of 47

Pharmaceuticals6.5. Click Chemistry2019, 12, 179Tools to Unravel New Targets in Malaria (YRC05) 15 of 47 Jorge Grilo 1,2, Lara Fidalgo 1, Diana Fontinha 2, Beatriz Anacleto 1, Charlie Saunders 3, Edward W. 3 1 2 1 1TateResearch , Rui Institute Moreira for , Miguel Medicines—iMed. Prudêncio ULisboa, and Ana Faculdade S. Ressurreição de Farm ácia da Universidade de Lisboa, 1649–0031 Research Lisbon, Institute Portugal for Medicines—iMed. ULisboa, Faculdade de Farmácia da Universidade de Lisboa, 2 iMM—Instituto1649–003 Lisbon, de MedicinaPortugal. Molecular, Faculdade de Medicina da Universidade de Lisboa, 1649–028 Lisbon,2 iMM—Instituto Portugal de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Lisbon, 3 DepartmentPortugal, 1649–028. of Chemistry, Imperial College London White City Campus, London W12 0BZ, UK; 3 jhfgrilo@ Departmentff.ulisboa.pt of Chemistry, Imperial College London White City Campus, London W12 0BZ, United Kingdom; e-mail: [email protected] Despite the best efforts and investment from the last decade, malaria s death toll still amounts to Despite the best efforts and investment from the last decade, malaria0 ′s death toll still amounts nearly half a million people a year, and no reduction of the diagnosed cases was observed in the last to nearly half a million people a year, and no reduction of the diagnosed cases was observed in the two years. Furthermore, parasite resistance is already reported for every drug class in clinical use. last two years. Furthermore, parasite resistance is already reported for every drug class in clinical These makes this protozoan infection a major public health concern mainly in the tropical regions of use. These makes this protozoan infection a major public health concern mainly in the tropical regions the globe (WHO. World Malaria Report. 2018). To overcome these hurdles, it is critical to advance new of the globe (WHO. World Malaria Report. 2018). To overcome these hurdles, it is critical to advance drugs with different mechanisms of action. new drugs with different mechanisms of action. With that in mind, we have already disclosed Torin2, known as an ATP-competitive mTOR kinase With that in mind, we have already disclosed Torin2, known as an ATP-competitive mTOR inhibitor (Liu, Q., et al. J. Med. Chem. 2011, 54, 1473), as a portent antimalarial lead compound that kinase inhibitor (Liu, Q., et al. J. Med. Chem. 2011, 54, 1473), as a portent antimalarial lead compound possesses in vivo activity in mice against the blood and liver stages. Compound activity was shown to that possesses in vivo activity in mice against the blood and liver stages. Compound activity was be independent of the host mTOR pathway and does not involve the mechanisms already knows for shown to be independent of the host mTOR pathway and does not involve the mechanisms already drugs in clinic (Hanson, K. K., et al. Proc. Natl. Acad. Sci. U.S.A. 2013, 110, E2838). knows for drugs in clinic (Hanson, K. K., et al. Proc. Natl. Acad. Sci. U.S.A. 2013, 110, E2838). Here, we report the development of Torin2 photoaffinity-based probes to deconvolute the Here, we report the development of Torin2 photoaffinity-based probes to deconvolute the molecular target(s) of this compound in the parasite. This was achieved through the modification of molecular target(s) of this compound in the parasite. This was achieved through the modification of the core scaffold with a photocrosslinker and a handle suitable for ‘click chemistry’. Furthermore, we the core scaffold with a photocrosslinker and a handle suitable for ‘click chemistry’. Furthermore, we disclose the application of these chemical tools in a mass spectrometry-based proteome profiling of disclose the application of these chemical tools in a mass spectrometry-based proteome profiling of P. falciparum lysates and live cultures of the disease blood stage (Scheme1). The results thus obtained P. falciparum lysates and live cultures of the disease blood stage (Scheme 1). The results thus obtained are fundamental optimization of Torin2-based compounds as a new alternative to current antimalarials, are fundamental optimization of Torin2-based compounds as a new alternative to current a key breakthrough to address the existing therapeutic gap. antimalarials, a key breakthrough to address the existing therapeutic gap.

Scheme 1. (A) Torin2 hit compound; (B) structure of the photo-aﬃnity labelling probe I, derived Scheme 1. (A) Torin2 hit compound; (B) structure of the photo-affinity labelling probe I, derived from from Torin2 by the introduction of a diazirine photoreactive moiety (in red) and a terminal alkyne Torin2 by the introduction of a diazirine photoreactive moiety (in red) and a terminal alkyne handle handle for CuAAC chemistry (in green) and (C) schematic overview of the methodology applied for for CuAAC chemistry (in green) and (C) schematic overview of the methodology applied for Plasmodium falciparum cell-based proteome proﬁling. In red: Diazirine before (star) and after (circle) Plasmodium falciparum cell-based proteome profiling. In red: Diazirine before (star) and after (circle) irradiation. In pink: TAMRA and/or Biotin azide capture reagents. irradiation. In pink: TAMRA and/or Biotin azide capture reagents. Acknowledgements: FCT-Portugal, (PTDC/QEQ-MED/7097/2014)(PD/BD/128260/2016) (IF/01034Acknowledgements:/2014). FCT-Portugal, (PTDC/QEQ- MED/7097/2014)(PD/BD/128260/2016)(IF/01034/2014). 6.6. The Development of a 6-Chloro-3-(Thiazole)Methyl-Quinazolin-4(3H)-One Series as Inhibitors of PqsR, the6.6. Transcriptional The Development Regulator of a 6-Chloro-3-(Thiazole)Methyl of the Pqs Quorum Sensing-Quinazolin-4(3H)-One System in Pseudomonas Series aeruginosa. as Inhibitors (YRC06) of PqsR, the Transcriptional Regulator of the Pqs Quorum Sensing System in Pseudomonas aeruginosa. (YRC06) Scott Grossman 1, Fadi Soukarieh 2, Paul Williams 2, Miguel Cámara 2 and Michael J. Stocks 1 Scott Grossman 1, Fadi Soukarieh 2, Paul Williams 2, Miguel Cámara 2 and Michael J. Stocks 1 1 School of Pharmacy, Nottingham NG7 2RD, UK 21 School School of Life of Pharmacy, Sciences, National Nottingh Biofilmsam NG7 Innovation2RD, United Centre, Kingdom. Centre for Biomolecular Sciences, University of2 Nottingham, School of Life Nottingham, Sciences, National Nottinghamshire Biofilms Innovation NG7 2RD, Centre, UK; Centre [email protected] for Biomolecular Sciences, University of Nottingham, Nottingham, Nottinghamshire NG7 2RD, United Kingdom; e-mail: [email protected] the rise in antimicrobial resistance continues to reduce treatment options for a variety of pathogens at an alarming rate, the need for alternate antimicrobial agents grows ever more urgent. Pharmaceuticals 2019, 12, x FOR PEER REVIEW 16 of 47

PharmaceuticalsAs the 2019rise, in12, 179antimicrobial resistance continues to reduce treatment options for a variety16 of of 47 pathogens at an alarming rate, the need for alternate antimicrobial agents grows ever more urgent. One such approach gaining ever more interest is the attenuation of virulence through the silencing One such approach gaining ever more interest is the attenuation of virulence through the silencing of quorum sensing, a form of community-based cell-to-cell communication. The opportunistic of quorum sensing, a form of community-based cell-to-cell communication. The opportunistic pathogen Pseudomonas aeruginosa is the greatest contributor to morbidity in cystic fibrosis patients, pathogen Pseudomonas aeruginosa is the greatest contributor to morbidity in cystic ﬁbrosis patients, and causes chronic and persistent infections in immunocompromised patients, particularly in a and causes chronic and persistent infections in immunocompromised patients, particularly in a hospital environment. hospital environment. Previous studies have evidenced the attenuation of virulence through the suppression of its Previous studies have evidenced the attenuation of virulence through the suppression of its alkylquinolone-based quorum sensing system, Pseudomonas quinolone system (pqs) (Ilangovan, A., et alkylquinolone-based quorum sensing system, Pseudomonas quinolone system (pqs)(Ilangovan, A., et al. al. PLoS Pathog. 2013, 9, e1003508). Herein, we report the synthesis and biological evaluation of a PLoS Pathog. 2013, 9, e1003508). Herein, we report the synthesis and biological evaluation of a novel series of 6-chloro-3-(thiazole)methyl-quinazolin-4(3H)-one compounds (Figure 1) through the novel series of 6-chloro-3-(thiazole)methyl-quinazolin-4(3H)-one compounds (Figure1) through the determination of their biological activity using both P. aeruginosa PAO1-L mCTX:PpqsA-lux and PA14 determination of their biological activity using both P. aeruginosa PAO1-L mCTX:PpqsA-lux and PA14 mCTX:PpqsA-lux bioreporter strains. We report structural evaluation of their binding mode through mCTX:PpqsA-lux bioreporter strains. We report structural evaluation of their binding mode through generating protein–ligand crystal structures of this class of novel inhibitors in the PqsR ligand generating protein–ligand crystal structures of this class of novel inhibitors in the PqsR ligand binding binding domain. The structure–activity relationship (SAR) study verified the binding mode to be domain. The structure–activity relationship (SAR) study veriﬁed the binding mode to be comparable comparable with that of the natural agonists HHQ and PQS, whilst providing lead pqs inhibitors with with that of the natural agonists HHQ and PQS, whilst providing lead pqs inhibitors with IC50s in the IC50s in the 100 nM range. 100 nM range.

(left) The structures of cognate ligands HHQ and Pseudomonas quinolone system (PQS); (right) the 6-chloro-3-ithiazole)methyl-quinazolin-4(3H)-one scaffold of PqsR inhibitors detailed herein.

7. Posters

7.1. Design and Synthesis of New Integrated Stress Response Inhibitor (ISRIB) Analogues TargetingTargeting eIF2BeIF2B as Potentialas Potential Candidates Candidates for for Enhancing Enhancing Protein Protein Synthesis Synthesis Rate Rate (P01) (P01)

Akeel AboAbo AlardAlard1,2 1,2, David RonRon 33,, AnaAna Crespillo-CresadoCrespillo-Cresado3 ,3, Alisa Alisa Zyryanova Zyryanova3 ,3, Christophe Christophe Fromont Fromont1, Dave1,Dave Barrett Barrett1 1,, Catherine Catherine Oteri Oteri 11, ,Pavel Pavel Gershkovich Gershkovich 1, Jong1, Jong Bong Bong Lee Lee 1, Barrie1, Barrie Kellam Kellam 1 and1 Peterand PeterFischer Fischer 1 1 1 Center Center for for Biomolecular Biomolecular Science, Science, University of of Nottingham, Nottingham, Nottingham Nottingham NG7 NG7 2RD, 2RD, United UK Kingdom. 2 Department Department of of Pharmaceutical Pharmaceutical Chemistry, Chemistry, FacultyFaculty of of Pharmacy, Pharmacy, University University of ofKufa, Kufa, Najaf, Najaf Iraq, 54001, 54001. Iraq 3 Cambridge Cambridge Institute Institute for for Medical Medical Research,Research, UniversityUniversity of of Cambridge, Cambridge, Cambridge Cambridge CB2 CB2 0XY, 0XY, United UK; [email protected]; e-mail: [email protected] Neurodegenerative diseases represent the major cause of dementia, which is predominantly a disease of the elderly, andand isis characterised by a decline in cognitive functions and memory. All these diseases share a similar aetiology, that is, the productionproduction and accumulation of misfolded proteins in the brain. This This results results in in increasing increasing the the activity activity of of the the protein protein kinase kinase RNA- RNA-likelike endoplasmic endoplasmic reticulum reticulum kinase PERKPERK/eIF2/eIF2α-P pathway of the unfolded protein response (UPR), which in turn leads to the inhibition of proteinprotein synthesis.synthesis. According According to to a a recent study the small-molecule integrated stress response inhibitor (ISRIB) reduces the level of eIF2eIF2α-P byby activatingactivating eukaryoticeukaryotic initiation factor 2B (eIF2B), a guanineguanine nucleotidenucleotide exchange factor (GEF) for eIF2α, resulting in enhancementenhancement of proteinprotein synthesis based on in vivo study in the cell-based luciferase reporter assay (Sidrauski, (Sidrauski, C., C., et et al. Elife.

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 17 of 48

inhibition of protein synthesis. According to a recent study the small-molecule integrated stress Pharmaceuticals 2019, 12, 179 17 of 47 response inhibitor (ISRIB) reduces the level of eIF2α-P by activating eukaryotic initiation factor 2B (eIF2B), a guanine nucleotide exchange factor (GEF) for eIF2α, resulting in enhancement of protein 2013synthesis, 2, e00498; based on Halliday, in vivo study M., et in al. theCell cell death-based dis. luciferase2015, 6 reporter(3), e1672.). assay Herein,(Sidrauski, ISRIB C., et analogues al. Elife. were2013, 2 designed, e00498; H andalliday, synthesised, M., et al. focusingCell death on dis. modifications 2015, 6 (3), e1672.). of the Herein, central ISRIB core cyclohexylanalogues were ring, Odesigned-arylglycolamide and synthesised, side chains focusing and terminal on modifications aromatic rings of of the the prototype central core ISRIB cyclohexyl molecule (Figure ring, 5O).- Synthesisarylglycolamide of truncated side chains ISRIB and analogues terminal revealedaromatic thatrings even of the subtle prototype modifications ISRIB molecule have an (Figure obvious 5). impactSynthesis on of activity. truncated Central ISRIB core analogues ring was revealed also explored, that even and itsubtle was found modifications that its replacement have an obvious with 1,4-diaminopiperazineimpact on activity. Central and core 1,4-diaminopiperdine ring was also explored, retained and high it was activity. found Furthermore,that its replacement to assist with in observe1,4-diaminopiperazine ISRIB0s interaction and with 1,4-diaminopi eIF2B, weperdine designed retained and synthesised high activity. fluorescently Furthermore, label to derivative, assist in consideringobserve ISRIB the′s interaction most active with compound, eIF2B, we with designed a modified and synthesised aryl ether portionfluorescently that was label then derivative used to, elaborateconsidering the the fluorescently most active label compound, compound. with The a modified derivatized aryl compound ether portion contained that was a flexible then used linker to terminatingelaborate the in fluorescently a 5/6-carboxyfluorescein label compound moiety. The (Zyryanova,derivatized compound A. F., et al. containedScience 2018 a flexible, 359 (6383),linker 1533–1536).terminating in a 5/6-carboxyfluorescein moiety (Zyryanova, A. F., et al. Science 2018, 359 (6383), 1533– 1536).

(A) (B)

Figure 5.5. (A) General overview of int integratedegrated stress response inhibitor (ISRIB) structural modificationmodification investigated inin thisthis study,study, ((BB)) bindingbinding modemode ofof ISRIBISRIB inin thethe activeactive sitesite ofof eIF2B.eIF2B. 7.2. Novel Synthesis of 3-Fluoro and 3,3-Difluoro Substituted -Lactams: Evaluation as Potential Anticancer 7.2. Novel Synthesis of 3-Fluoro and 3,3-Difluoro Substitutedβ β-Lactams: Evaluation as Potential Agents (P02) Anticancer Agents (P02) Azizah Malebari 1,2 and Mary J. Meegan 2 Azizah Malebari 1,2 and Mary J. Meegan 2 1 Department of Pharmaceutical Chemistry, College of Pharmacy, King Abdulaziz University, 1 Department of Pharmaceutical Chemistry, College of Pharmacy, King Abdulaziz University, Jeddah, KSA. Jeddah,2 School KSA of Pharmacy and Pharmaceutical Sciences, Trinity Biomedical Sciences Institute, 152–160 Pearse 2 SchoolStreet, of Trinity Pharmacy College and Dublin, Pharmaceutical Dublin 2, Ireland; Sciences, e-mail: Trinity [email protected] Biomedical Sciences Institute, 152–160 Pearse Street, Trinity College Dublin, Dublin 2, Ireland; [email protected] Combretastatin A-4 (CA-4), a natural product stilbene, is a potent microtubule-disrupting agent that bindsCombretastatin at the colchicine A-4 (CA-4),-binding a natural site of producttubulin. stilbene,The design, is a synthesis potent microtubule-disrupting and biochemical evaluation agent thatof a series binds atof theanalogues colchicine-binding of the microtubule site of tubulin.-destabilising The design, agent CA synthesis-4 is described. and biochemical The monocyclic evaluation β- oflactam a series CA of-4 analogues ofcontaining the microtubule-destabilising halogen substituents agentat the CA-4C-3 position is described. of β-lactam The monocyclic ring were βsynthesized-lactam CA-4 usin analoguesg the Staudinger containing reaction halogen (Greene, substituents T.F.; at et the al. C-3J. Med. position Chem. of β, -lactam 2016, 59 ring, 90– were113; synthesizedMalebari, A.M., using et al. the Eur. Staudinger J. Med. Chem. reaction 2017 (Greene,, 130, 261 T.F.;–285). et Previous al. J. Med. investigations Chem., 2016 described, 59, 90–113; two Malebari,approaches A.M., for the et al. constructionEur. J. Med. of Chem. 3-fluoro2017-β,-130lactams, 261–285). using Previousthe ketene investigations-imine condensation described or twothe approachesenolate-imine for condensation the construction method of 3-fluoro- (Araki, βK.,-lactams et al. Tetrahedron using the ketene-imine Lett. 1991, 32 condensation, 5461–5464). orIn the enolate-iminepresent work, condensationthe synthesis of method 3-fluoro (Araki, and 3,3 K.,-difluoro et al. Tetrahedron substituted Lett. β-lactams1991, 32 was, 5461–5464). developed Ineasily the presentby a convenient work, the microwave synthesis ofassiste 3-fluorod Reformatsky and 3,3-difluoro reaction substituted using ethylβ-lactams bromofluoroacetate was developed and easilyethyl bybromodifluoroacetate a convenient microwave, respectively assisted (Scheme Reformatsky 2). To reaction the best using to our ethyl knowledge, bromofluoroacetate this is the first and report ethyl bromodifluoroacetate,of this new synthetic respectively approach for (Scheme 3-fluoro2). To and the 3,3 best-difluoro to our knowledge, β-lactams as this CA is-4 the analogues. first report The of thisreaction new syntheticwas successful approach with for short 3-fluoro reaction and 3,3-difluoro time comparedβ-lactams to the as conventional CA-4 analogues. Staudinger The reaction reaction, was successfulwith moderate with yield short and reaction few timesteps compared required, toand the it conventionalwas demonstrated Staudinger to be reaction,a convenient with and moderate facile yieldmethod and for few the steps preparation required, of anda variety it was of demonstrated 3-fluoro and 3,3 to- bedifluoro a convenient β-lactams. and The facile 3-fluoro method β-lactams for the preparation of a variety of 3-fluoro and 3,3-difluoro β-lactams. The 3-fluoro β-lactams (1–8) and 3,3-difluoro β-lactams (9–16) in this series contained the 3,4,5-trimethoxyphenyl ring A, (required Pharmaceuticals 2019, 12, 179 18 of 47 for potent activity of CA-4), together with various ring B substituents. The structure of β-lactams 6 and 14 was confirmed by X-ray crystallography. Preliminary cell viability studies demonstrated the antiproliferative effects of these novel compounds, with an IC50 value of 0.153 µM for 6 in MCF-7 humanPharmaceuticals breast 2019 cancer, 12, x cellFOR line.PEER REVIEW 18 of 47

Scheme 2.2. Synthesis of 3-fluoro3-fluoro and 3,3-difluoro3,3-difluoro ββ-lactams by thethe microwavemicrowave assistedassisted ReformatskyReformatsky reaction. Reagents Reagents and and conditions: conditions: (a (a) )Zn Zn dust, dust, (CH (CH3)3SiCl,)3SiCl, 40 40°C,◦C, 2 min, 2 min, microwave; microwave; C6H C6,H 1006, 100 °C,◦ 30C, 30min, min, microwave; microwave; products products obtained obtained as asa ra acemic racemic mixture, mixture, one one enantiomer enantiomer represented. represented. 7.3. The Influence of Fluorination on Aliphatic Lipophilicity (P03) 7.3. The Influence of Fluorination on Aliphatic Lipophilicity (P03) Bruno Linclau Bruno Linclau School of Chemistry, University of Southampton, Southampton SO171BJ, UK; [email protected] School of Chemistry, University of Southampton, Southampton SO171BJ, United Kingdom; e-mail: [email protected] of bioactive compounds is commonly employed in the drug optimisation process, given its potential to modulate a range of properties, and its ability to prevent or slow down undesired Fluorination of bioactive compounds is commonly employed in the drug optimisation process, processes such as oxidation and/or acid-catalysed degradation. In this context, aliphatic fluorination given its potential to modulate a range of properties, and its ability to prevent or slow down (as opposed to aromatic fluorination) is receiving increasing interest. Our group is interested in undesired processes such as oxidation and/or acid-catalysed degradation. In this context, aliphatic investigating how fluorination affects molecular conformation (Bogdan, E., et al. Chem. Eur. J. fluorination (as opposed to aromatic fluorination) is receiving increasing interest. Our group is 2015, 21, 11462–11474) and lipophilicity ((a) Linclau, B., et al. Angew. Chem. Int. Ed. 2016, 55, 674–678 interested in investigating how fluorination affects molecular conformation (Bogdan, E., et al. Chem. (VIP); (b) Effries, B., et al. J. Med. Chem. 2018, 61, 10602–10618), as well as hydrogen bond properties of Eur. J. 2015, 21, 11462–11474) and lipophilicity ((a) Linclau, B., et al. Angew. Chem. Int. Ed. 2016, 55, adjacent functional groups including chiral aliphatic structures and carbohydrates ((a) Graton, J., et al. 674–678 (VIP); (b) Effries, B., et al. J. Med. Chem. 2018, 61, 10602–10618), as well as hydrogen bond Angew. Chem. Int. Ed. 2012, 51, 6176–6180 (VIP). (b) Graton, J., et al. Chem. Eur. J. 2017, 23, 2811–2819) properties of adjacent functional groups including chiral aliphatic structures and carbohydrates ((a) and how this impacts on binding affinity ((a) van Straaten, K.E., et al. J. Am. Chem. Soc. 2015, 137, Graton, J., et al. Angew. Chem. Int. Ed. 2012, 51, 6176–6180 (VIP). (b) Graton, J., et al. Chem. Eur. J. 2017, 1230–1244; (b) N Go, I., et al. S. Chem. Eur. J. 2014, 20, 106–112). 23, 2811–2819) and0 how this impacts on binding affinity ((a) van Straaten, K.E., et al. J. Am. Chem. Soc. The introduction of fluorine on aliphatic (sp3) carbon often leads to a reduction in lipophilicity, 2015, 137, 1230–1244; (b) N′Go, I., et al. S. Chem. Eur. J. 2014, 20, 106–112). and is the result of a number of effects including hydrophobic surface and dipole introduction, as well The introduction of fluorine on aliphatic (sp3) carbon often leads to a reduction in lipophilicity, as reduction of polarizability. The importance of these opposing effects depends also on a number and is the result of a number of effects including hydrophobic surface and dipole introduction, as of factors such as absolute lipophilicity and conformational effects, the latter of which can, in turn, well as reduction of polarizability. The importance of these opposing effects depends also on a be very different in water and octanol. number of factors such as absolute lipophilicity and conformational effects, the latter of which can, We developed a convenient 19F NMR-based method for the logP-determination of fluorinated in turn, be very different in water and octanol. compounds, thus not relying on UV-activity, and used this methodology for an extensive study of We developed a convenient 19F NMR-based method for the logP-determination of fluorinated aliphatic lipophilicity based on an alkanol scaffold. compounds, thus not relying on UV-activity, and used this methodology for an extensive study of This presentation will give an overview of these, and other, unpublished findings, and will involve aliphatic lipophilicity based on an alkanol scaffold. an extensive list of aliphatic fluorination motifs. This presentation will give an overview of these, and other, unpublished findings, and will 7.4.involve Exploring an extensive the Impact list ofof Polymer aliphatic Size fluorination on the Biological motifs. Activity of PEG-Haloperidol Conjugates (P04)

Az7.4. AlddienExploring Natfji, the Impact Camilla of Polymer Pegoraro, Size Kim on the Watson, Biological Helen Activity Osborn of PEG- and FrancescaHaloperidol Greco Conjugates (P04)

SchoolAz Alddien of Pharmacy, Natfji, Camilla University Pegoraro of Reading, , Kim WhiteknightsWatson, Helen PO Osborn BOX 224 and Reading Francesca RG6 Greco 6AD Berkshire, UK; [email protected] School of Pharmacy, University of Reading, Whiteknights PO BOX 224 Reading RG6 6AD Berkshire, United Kingdom;Haloperidol e-mail: (HA)[email protected] is a well-known antagonist of D2 receptors, clinically used as an antipsychotic. Moreover,Haloperidol HA has (HA) been is shown a well-known to be a ligand antagonist of diﬀ erentof D2 receptors, types of receptors clinically (centrally used as andan antipsychotic. peripherally) Moreover, HA has been shown to be a ligand of different types of receptors (centrally and peripherally) including σ receptors, which have a role in the growth and development of different types of solid tumours (van Waarde, A., et al., Biochim. Biophys. Acta Biomembr. 2015, 1848, 2703–2714). However, HA, as a small molecule, has the ability to permeate across the blood–brain barrier (BBB) and, consequently, produces central nervous system (CNS) (extrapyramidal) side effects, which limits its peripheral clinical applications. We have previously suggested that conjugating HA to a non-biodegradable polymer through a biologically stable linker could minimise or even prevent its

Pharmaceuticals 2019, 12, 179 19 of 47 including σ receptors, which have a role in the growth and development of different types of solid tumours (van Waarde, A., et al., Biochim. Biophys. Acta Biomembr. 2015, 1848, 2703–2714). However, HA, as a small molecule, has the ability to permeate across the blood–brain barrier (BBB) and, consequently, produces central nervous system (CNS) (extrapyramidal) side effects, which limits its peripheral clinical applications.Pharmaceuticals 2019 We, 12 have, x FOR previously PEER REVIEW suggested that conjugating HA to a non-biodegradable polymer 19 of 47 through a biologically stable linker could minimise or even prevent its diffusion though the BBB while diffusion though the BBB while retaining its biological efficacy (Figure 6). Therefore, conjugation retaining its biological efficacy (Figure6). Therefore, conjugation between PEG, a non-biodegradable between PEG, a non-biodegradable polymer, and HA through a carbamate linkage (biologically polymer, and HA through a carbamate linkage (biologically stable) was developed for the first time by stable) was developed for the first time by our group (Heath, F., et al. Polym. Ch. 2016, 7204–7210). In our group (Heath, F., et al. Polym. Ch. 2016, 7204–7210). In silico and in vitro studies revealed that the silico and in vitro studies revealed that the conjugated HA is extremely unlikely to cross the BBB and conjugated HA is extremely unlikely to cross the BBB and that it maintains binding activity towards that it maintains binding activity towards D2 receptors, but at low level. In the present study, we D2 receptors, but at low level. In the present study, we evaluated the impact of polymer size on the evaluated the impact of polymer size on the biological activity of PEG-HA on σ receptors in vitro. In biological activity of PEG-HA on σ receptors in vitro. In addition to this, molecular docking studies, addition to this, molecular docking studies, in σ and D2 receptors, were performed to understand the in σ and D2 receptors, were performed to understand the effect of conjugation on the binding of HA to effect of conjugation on the binding of HA to these receptors. these receptors.

Figure 6. Comparison of the peripheral administration of free haloperidol (HA) and PEG-HA conjugate. Figure 6. Comparison of the peripheral administration of free haloperidol (HA) and PEG-HA PEG-HA conjugates, using different sizesconjugate. of PEG (6000, 2000 Da), were synthetized and characterised. A two-step synthesis was carried out: first, HA was modified to have an amine handlePEG-HA that allows conjugates, conjugation; using second, different the HA-aminesizes of PEG handle (6000, was 2000 conjugated Da), were to NHS-activated synthetized PEG.and characterised. A two-step synthesis was carried out: first, HA was modified to have an amine handle In vitro cytotoxicity studies, via σ receptors, were performed on MCF-7. The IC50 values of free HA andthat PEG-HAallows conjugation; conjugates weresecond, determined the HA-amine using the handle MTT assay.was conjugated In parallel, to molecular NHS-activated docking PEG. studies In werevitro conducted.cytotoxicity Thestudies, results via revealed σ receptors, that conjugatingwere performed HA toon PEG MCF-7. affected The itsIC50 binding values toof thefree receptors. HA and PEG-HA conjugates were determined using the MTT assay. In parallel, molecular docking studies In the case of D2 receptors, the position of PEG-HA inside the pocket was different compared to free HAwere while conducted. retaining The the results salt bridge revealed interaction that conjugating with Asp114, HA which to PEG has been affected indicated its binding to be essential to the forreceptors. the biological In the activity.case of D These2 receptors, findings the might position justify of the PEG-HA reduced inside biological the pocket activity was observed different for PEG-HAcomparedin to vitro free. RegardingHA while theretainingσ receptors, the salt initial bridge results interaction indicated with that Asp114, conjugating which HA has to been PEG influencedindicated to its be binding essential to for these the receptors.biological activity. These findings might justify the reduced biological activity observed for PEG-HA in vitro. Regarding the σ receptors, initial results indicated that 7.5.conjugating Design, SynthesisHA to PEG and influenced Evaluation its of Novelbinding PqsR to Inhibitorsthese receptors. as Adjuvant Therapy to Treat Pseudomonas aeruginosa Infections (P05) Alaa7.5. Design, Mashabi Synthesis1, Fadi and Soukarieh Evaluation2, of William Novel PqsR Richardson Inhibitors1 ,as Barrie Adjuvant Kellam Therapy1, Miguel to Treat C Pseudomonasámara 2 and Michaelaeruginosa Stocks Infections1 (P05) 1 2 1 1 2 1AlaaSchool Mashabi of Pharmacy, , Fadi NottinghamSoukarieh NG7, William 2RD, Richardson UK , Barrie Kellam , Miguel Cámara and 1 2MichaelSchool Stocks of Life Sciences, National Biofilms Innovation Centre, Centre for Biomolecular Sciences,1 School University of Pharmacy, of Nottingh Nottingham,am NG7 Nottingham 2RD, United NG7 Kingdom. 2RD, UK; [email protected], [email protected] School of Life Sciences, National Biofilms Innovation Centre, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom; e-mail: [email protected], [email protected] sensing (QS) is a cell-to-cell communication mechanism used by bacterial populations to control virulence traits and resistance mechanisms (Williams, P., et al. Philos. Trans R SocLond.Abstract B Biol.Quorum Sci. sensing2007, 362 (QS), 1119–1134). is a cell-to-cellP. aeruginosa communicationhas several mechanism QS systems, used one by ofbacterial which, thepopulationsPseudomonas to controlquinolone virulence system traits (pqs and) uses resistance alkyl-quinolone mechanisms (AQ)-derived (Williams, QSP., et molecules. al. Philos. Trans PqsR isR theSocLond. key regulatory B Biol. Sci. protein 2007, 362, responsible 1119–1134). for the P. activationaeruginosa ofhas the severalpqs system QS systems, upon interaction one of which, with the Pseudomonas quinolone system (pqs) uses alkyl-quinolone (AQ)-derived QS molecules. PqsR is the key regulatory protein responsible for the activation of the pqs system upon interaction with the P. aeruginosa ligands (2-heptyl-3-hydroxy-4(1H)-quinolone (PQS) and 2-heptyl-4-hydroxyquinoline (HHQ)) to drive the upregulation of AQ production (feedback activation) and virulence factors biosynthesis (Ilangovan, A., et al. PLoS Pathog. 2013, 9(7), e1003508). It has been shown that the inhibition of the PqsR receptor attenuates bacterial pathogenicity, without affecting bacterial growth and viability. Our aim was to design, synthesise and evaluate a series of new PqsR inhibitors using a quinazolinone-derived molecule (SEN016, IC50 3.2 µM) that was found during a high-throughput

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 20 of 48

Pharmaceuticals 2019, 12, 179 20 of 47 the Pseudomonas quinolone system (pqs) uses alkyl-quinolone (AQ)-derived QS molecules. PqsR is the key regulatory protein responsible for the activation of the pqs system upon interaction with the P. aeruginosaP. aeruginosa ligandsligands (2 (2-heptyl-3-hydroxy-4(1H)-quinolone-heptyl-3-hydroxy-4(1H)-quinolone (PQS) (PQS) and and 2-heptyl 2-heptyl-4-hydroxyquinoline-4-hydroxyquinoline (HHQ))(HHQ)) to to drive drive the the upregulation upregulation of of AQ AQ production production (feedback (feedback activation) activation) and and virulence virulence factors factors biosynthesisbiosynthesis (Ilangovan, (Ilangovan, A., A.,et al. et al.PLoSPLoS Pathog. Pathog. 20132013, 9(7),, 9(7), e1003508). e1003508). It has It has been been shown shown that that the the inhibitioninhibition of the of the PqsR PqsR receptor receptor attenuates attenuates bacterial bacterial pathogenicity, pathogenicity, without without affecting affecting bacterial bacterial growth growth andand viability. viability. Our Our aim aim was was to design, to design, synthesise synthesise and and evaluate evaluate a series a series of new of new PqsR PqsR inhibitors inhibitors using using a 50 quinazolinonea quinazolinone-derived-derived molecule molecule (SEN016, (SEN016, IC IC3.250 µM)3.2 µ thatM)that was was found found during during a high a high-throughput-throughput screeningscreening (HTS) (HTS) (Figure (Figure 7)7. ). Replacement Replacement of of the the quinazolinone quinazolinone ring ring by by benzimidazole benzimidazole (SEN089) (SEN089) 50 enhancedenhanced the the activity activity (IC (IC0.06750 0.067 µM).µM). However, However, further further lead lead optimisation optimisation was was required required to enhance to enhance its its biological biological and and physiochemical physiochemical profile. profile. The The co- co-crystalcrystal structure structure of of SEN089 SEN089 with with PqsR PqsRLBDLBD waswas determineddetermined and and used used to guide to guide the the lead lead optimisation optimisation strategy. strategy. Herein Herein,, we we report report further further SAR SAR analysis analysis 1 2 3 aroundaround modifications modifications (R (R, R1,R and2 and R ) Rof3 )SEN089 of SEN089,, which which resulted resulted in a in series a series of novel of novel PqsR PqsR inhibitors inhibitors withwith enhanced enhanced biological biological activity activity and and improved improved physiochemical physiochemical properties. properties.

(a) (b)

FigureFigure 7. ( 7.a)( a Improvement) Improvement in in the the activity activity from SEN016 SEN016 (hit) (hit) to to SEN089 SEN089 (lead) (lead) and and the the structure–activity structure–

activityrelationship relationship (SAR) (SAR) study study of SEN089 of SEN089 based based on the on modiﬁcations the modifications of R 1of,R R2 1and, R2 and R3.( Rb3). The(b) The co-crystal co- crystalstructure structure of SEN089 of SEN089 complexed complexed with with PqsR PqsRCBD domain.CBD domain.

7.67.6.. A Novel A Novel in inVivo Vivo Anti Anti-Amnesic-Amnesic Agent Agent,, Specially Specially Designed Designed to to Express Express Both Both Acetylcholinesterase Acetylcholinesterase (AChE) Inhibitory, Serotonergic Subtype 4 Receptor (5-HT4R) Agonist and Serotonergic Subtype 6 Receptor (5-HT6R) (AChE) Inhibitory, Serotonergic Subtype 4 Receptor (5-HT4R) Agonist and Serotonergic Subtype 6 Inverse Agonist Activities, with a Potential Interest against Alzheimer0s Disease (P06) Receptor (5-HT6R) Inverse Agonist Activities, with a Potential Interest against Alzheimer′s 1,2 1 1 1 3 DiseaseBérénice (P06) Hatat , Samir Yahiaoui ,Cédric Lecoutey , Audrey Davis , Thomas Freret , Michel Boulouard 3, Sylvie Claeysen 2, Christophe Rochais 1 and Patrick Dallemagne 1 Bérénice Hatat 1,2, Samir Yahiaoui 1, Cédric Lecoutey 1, Audrey Davis 1, Thomas Freret 3, Michel 1 BoulouardNormandie 3, Sylvie Universit Claeysené, UNICAEN, 2, Christophe Centre Rochais d0Etudes 1 and et de Patrick Recherche Dallemagne sur le Médicament 1 de Normandie (CERMN), Caen, France 1 Normandie Université, UNICAEN, Centre d′Etudes et de Recherche sur le Médicament de Normandie 2 IGF, University of Montpellier, CNRS, INSERM, Montpellier, France 3 (CERMN), Caen, France. 2 NormandieIGF, University Universit of Montpellier,é, UNICAEN, CNRS, INSERM, INSERM, Montpellier, U1075, GIPFrance. CYCERON, COMETE, Caen, France; [email protected] 3 Normandie Université, UNICAEN, INSERM, U1075, GIP CYCERON, COMETE, Caen, France; e-mail: cedric.lecoutey@uThis work describesnicaen.fr the conception and synthesis in vitro and in vivo biological evaluation of novel multi-target directed ligands (MTDL) able to both activate 5-HT4 receptors, block 5-HT6 receptors and This work describes the conception and synthesis in vitro and in vivo biological evaluation of inhibit acetylcholinesterase activity (AChE), in order to exert a synergistic anti-amnesic eﬀect, potentially novel multi-target directed ligands (MTDL) able to both activate 5-HT4 receptors, block 5-HT6 useful in the treatment of Alzheimer0s disease (AD). Indeed, both activation of 5-HT4 and blockage

Pharmaceuticals 2019, 12, 179 21 of 47

of 5-HT6 receptors led to an enhanced acetylcholine release, suggesting that it could lead to efficient restoration of the cholinergic neurotransmission deficit observed in AD. Furthermore, 5-HT4 receptor agonists are able to promote the non-amyloidogenic cleavage of the amyloid precursor protein (APP) and to favour the production of the neurotrophic protein sAPPα. Finally, we identified a pleiotropic compound, [1-(4-amino-5-chloro-2-methoxyphenyl)-3-(1-(3-methylbenzyl)piperidin-4-yl)propan-1-one fumaric acid salt, which displayed in vivo an anti-amnesic effect in a model of scopolamine-induced deficitPharmaceuticals of working 2019, 12 memory, x FOR PEER at aREVIEW dose of 0.3 mg/kg. 21 of 47

Summary of the programme

This work was supported by funding from the Fondation Vaincre Alzheimer (#FR-15072) and This work was supported by funding from the Fondation Vaincre Alzheimer (#FR-15072) and the the Fondation Plan Alzheimer (AAP2015 Project TRIAD 016). Fondation Plan Alzheimer (AAP2015 Project TRIAD 016). 7.7. Exploring Brain–Immune Interaction with Hyper-Polarised Xenon-129 Magnetic Resonance Imaging 7.7. Exploring Brain–Immune Interaction with Hyper-Polarised Xenon-129 Magnetic Resonance Imaging(P07) (P07)

1 1 2 3 1,4,5 ClClémentément VigierVigier1 ,, EmmanuelleEmmanuelle DubostDubost1 ,, DenisDenis Vivien Vivien2 ,, Christian Christian Fernandez Fernandez3 ,, Thomas Thomas Cailly Cailly1,4,5 1 and Frederic FabisFabis1

11 Normandie Univ, UNICAEN, Centre d′Etudes et de Recherche sur le Médicament de Normandie Normandie Univ, UNICAEN, Centre d0Etudes et de Recherche sur le Médicament de Normandie (CERMN), 14000 Caen, France. (CERMN), 14000 Caen, France 2 Physiopathology and Imaging of Neurological Disorders (PhIND), 14000 Caen, France. 2 Physiopathology and Imaging of Neurological Disorders (PhIND), 14000 Caen, France 3 Laboratoire Catalyse et Spectrochimie (LCS), 14000 Caen, France. 3 4 Laboratoire Normandie Catalyse Univ, UNICAEN, et Spectrochimie IMOGERE, (LCS), 14000 14000 Caen, Caen, France. France 4 5 Normandie Department Univ, of UNICAEN, Nuclear IMOGERE,Medicine, CHU 14000 Caen,Cotê Francede Nacre, 14000 Caen, France; e-mail: 5 [email protected] of Nuclear Medicine, CHU Côte de Nacre, 14000 Caen, France; [email protected] InflammationInflammation isis aa hallmark of most neurological disordersdisorders andand thethe ability to detect, quantify and monitor the inflammatoryinflammatory response ofof the central nervous system (CNS) couldcould have large implication for bothboth diagnosisdiagnosis andand therapeutic therapeutic response response prediction prediction (V. (V. Wee Wee Yong, Yong,Neuroscientist Neuroscientist. 2010. 2010; 16:; 16: 408–20). 408– Among20). Among the di thefferent different diagnostic diagnost modalitiesic modalities suitable tosuitable detect neuro-inflammation,to detect neuro-inflammation, plasmatic biomarkers plasmatic canbiomarkers be used, can but nobe used, reliable but and no specific reliable plasmatic and specific biomarker plasmatic of neuro-inflammation biomarker of neuro-inflammation has been identified has tobeen date. identified Recently, to molecular date. Recently, imaging molecular of neuro-inflammation imaging of neuro-inflammation has been developed—the has beenin vivo developed—detection ofthe a in protein, vivo detection the P-selectin, of a protein, which is the over-expressed P-selectin, which at the is luminal over-expressed surface of at endothelial the luminal cells surface during of neuro-inflammationendothelial cells during was neuro-inflammation performed using micro-sizedwas performed particles using micro-sized of iron oxide particles (MPIO). of iron However, oxide this(MPIO). biosensor However, cannot this be biosensor used in humans cannot be because used in of humans its toxicity. because of its toxicity. Hyper-polarised xenon-129 (HP-129Xe) has recently emerged as a promisingpromising biocompatiblebiocompatible contrast agent to improve sensitivity of MRI, successfully used to acquire images of the humanhuman pulmonary system (Liburn, D. M. L., et al. J. Magn. Reson. 2013, 7, 173) and brain (Swanson, S. D., et al. Magn. Reson. Med. 1997, 38, 695). However, this gas is not specific of a biological target, and therefore it has to be vectorised using a molecular host to be a valuable biosensor. Among them, cryptophanes showed very good xenon encapsulation properties, leading to a number of in vitro studies using HP-129Xe cryptophane-based biosensors reported in the literature since the 2000s (Wang, Y., et al. Acc. Chem. Res. 2016, 49, 2179–2187). Here, we propose the design and the synthesis of a new biocompatible MRI biosensor composed of a cryptophane core able to encapsulate xenon, and a well-characterized P-selectin antibody able to selectively bind endothelial P-selectin with high affinity. The two parts will be connected with a linker using copper-free click chemistry to afford the desire biosensor (Figure 8).

Pharmaceuticals 2019, 12, 179 22 of 47 pulmonary system (Liburn, D. M. L., et al. J. Magn. Reson. 2013, 7, 173) and brain (Swanson, S. D., et al. Magn. Reson. Med. 1997, 38, 695). However, this gas is not specific of a biological target, and therefore it has to be vectorised using a molecular host to be a valuable biosensor. Among them, cryptophanes showed very good xenon encapsulation properties, leading to a number of in vitro studies using HP-129Xe cryptophane-based biosensors reported in the literature since the 2000s (Wang, Y., et al. Acc. Chem. Res. 2016, 49, 2179–2187). Here, we propose the design and the synthesis of a new biocompatible MRI biosensor composed of aPharmaceuticals cryptophane 2019 core, 12, ablex FOR to PEER encapsulate REVIEW xenon, and a well-characterized P-selectin antibody able 22 to of 47 selectively bind endothelial P-selectin with high affinity. The two parts will be connected with a linker using copper-freeIn this presentation, click chemistry we will to focus afford on the the desire conception biosensor and (Figure the synthesis8). of the biosensor.

Figure 8. Representation of the biosensor. Figure 8. Representation of the biosensor. In this presentation, we will focus on the conception and the synthesis of the biosensor. 7.8. Multivalent Inhibitors of Sialidases (P08)

7.8.Coralie Multivalent Assailly Inhibitors 1, Yoan of SialidasesBrissonnet (P08) 1, Amélie Saumonneau 2, Charles Tellier 2, Franck Daligault 2, CoralieCyrille Assailly Grandjean1, Yoan 2, David Brissonnet Deniaud1, Am 1 andélie Sébastien Saumonneau Gouin2, Charles 1 Tellier 2, Franck Daligault 2, 2 1 1 Cyrille1 Grandjean CEISAM, UMR, David CNRS Deniaud 6230, UFR desand Sciences Sébastien et des Gouin Techniques, 2 rue de la Houssinière, BP 92208, 44322 1 CEISAM,Nantes UMR Cedex CNRS 3. 6230, UFR des Sciences et des Techniques, 2 rue de la Houssinière, BP 92208, 2 UFIP, UFR des Sciences et des Techniques, 2 rue de la Houssinière, BP92208, 44322 Nantes Cedex 3; e-mail: 44322 Nantes Cedex 3 [email protected] 2 UFIP, UFR des Sciences et des Techniques, 2 rue de la Houssinière, BP92208, 44322 Nantes Cedex 3; [email protected] recent years much effort has been devoted to the design of potent and selective glycosidase inhibitors. However, potential candidates often lack of glycosidase selectivity, and non-specific In recent years much effort has been devoted to the design of potent and selective glycosidase inhibitions may lead to severe side-effects. Limiting selectivity issues is a challenge unmet with the inhibitors. However, potential candidates often lack of glycosidase selectivity, and non-specific first generation of inhibitors. inhibitions may lead to severe side-effects. Limiting selectivity issues is a challenge unmet with the Sialidases are glycosidases involved in many physiological and pathological functions. Many first generation of inhibitors. viruses, bacteria and parasites produce this enzyme to cleave sialic acid residues from host cells, Sialidases are glycosidases involved in many physiological and pathological functions. Many unmasking membrane receptors for adhesion and invasion (Newstead, S. L.; et al. J. of Biol. Chem. viruses, bacteria and parasites produce this enzyme to cleave sialic acid residues from host cells, 2008, 283 (14), 9080–9088). There are several classes of sialidases, some display a unique catalytic site unmasking membrane receptors for adhesion and invasion (Newstead, S. L.; et al. J. of Biol. Chem. (CAT) to cleave the sialosides, whereas others have an additional lectinic site called CBM 2008, 283 (14), 9080–9088). There are several classes of sialidases, some display a unique catalytic (carbohydrate-binding module), which is an anchoring point for glycans, thus increasing the catalytic site (CAT) to cleave the sialosides, whereas others have an additional lectinic site called CBM efficiency of the enzyme (Thobhani, S., et al. J. Am. Chem. Soc. 2003, 125 (24), 7154–7155). These (carbohydrate-binding module), which is an anchoring point for glycans, thus increasing the catalytic sialidases with CAT and CBM are virulence factors common to many protozoal and pathogenic efficiency of the enzyme (Thobhani, S., et al. J. Am. Chem. Soc. 2003, 125 (24), 7154–7155). bacteria, and are potential therapeutic targets. Furthermore, human sialidases involved in important These sialidases with CAT and CBM are virulence factors common to many protozoal and pathogenic physiological functions do not express CBMs. bacteria, and are potential therapeutic targets. Furthermore, human sialidases involved in important In this work, we developed an alternative approach to the “lock and key concept” to achieve physiological functions do not express CBMs. high affinities and selectivities for sialidases (Brissonnet, Y., et al. Chem. Eur. J. 2019, 25 (9), 2358–2365). In this work, we developed an alternative approach to the “lock and key concept” to achieve high Multivalent thiosialosides were designed to bind to both the CAT and the CBM of pathogenic affinities and selectivities for sialidases (Brissonnet, Y., et al. Chem. Eur. J. 2019, 25 (9), 2358–2365). sialidases. Multivalent thiosialosides were designed to bind to both the CAT and the CBM of pathogenic sialidases. Sialidases from Vibrio cholerae, Trypanosoma cruzi and Streptococcus pneumoniae were produced with and without the CBM to determine the importance of dual targeting and the validity of the multivalent concept (Figure 9).

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Sialidases from Vibrio cholerae, Trypanosoma cruzi and Streptococcus pneumoniae were produced with and without the CBM to determine the importance of dual targeting and the validity of the multivalent conceptPharmaceuticals (Figure 20199,). 12, x FOR PEER REVIEW 23 of 47

Figure 9. Inhibition by multivalent compound.

Inhibition tests of a polyvalent thiosialoside against the representative bacterial, parasitic and Inhibition tests of a polyvalent thiosialoside against the representative bacterial, parasitic and fungal sialidases showed inhibitory activity up to the nanomolar range with a strong synergistic fungal sialidases showed inhibitory activity up to the nanomolar range with a strong synergistic effect. These results extend the multivalent concept to this important class of enzymes for which effect. These results extend the multivalent concept to this important class of enzymes for which transition-state inhibitors failed to reach the submicromolar level. transition-state inhibitors failed to reach the submicromolar level. 7.9. Discovery of Highly Selective PI3K δ Inhibitors for Treatment of Blood Cancers and Autoimmune Diseases7.9. Discovery (P09) of Highly Selective PI3K δ Inhibitors for Treatment of Blood Cancers and Autoimmune Diseases (P09) Anas Buzrieda 1, Simon Macdonald 2, Shailesh Mistry 1 and Michael J. Stocks 1 Anas Buzrieda 1, Simon Macdonald 2, Shailesh Mistry 1 and Michael J. Stocks 1 1 Department of Pharmacy, University of Nottingham, Nottingham NG7 2QL, UK 1 2 GSK, Department Gunnels Woodof Pharmacy, Rd, SG1 University 2NY Stevenage, of Nottingham, Hertfordshire; Nottingham, [email protected] United Kingdom, NG7 2QL. 2 GSK, Gunnels Wood Rd, SG1 2NY Stevenage, Hertfordshire; e-mail: [email protected] Phosphatidylinositol-3-kinases (PI3Ks) are lipid kinases that regulate a plethora of cellular Phosphatidylinositol-3-kinases (PI3Ks) are lipid kinases that regulate a plethora of cellular processes including proliferation, survival and motility. PI3Ks have been divided into three classes. processes including proliferation, survival and motility. PI3Ks have been divided into three classes. Of these, the most characterized is the class I enzymes, which are activated by cell surface receptors to Of these, the most characterized is the class I enzymes, which are activated by cell surface receptors generate phosphatidylinositol-3,4,5-trisphosphate (PIP3). Class I PI3Ks are further divided into four to generate phosphatidylinositol-3,4,5-trisphosphate (PIP3). Class I PI3Ks are further divided into isoforms (α, β, γ and δ), the former two isoforms are ubiquitously expressed, whereas PI3K γ and δ four isoforms (α, β, γ and δ), the former two isoforms are ubiquitously expressed, whereas PI3K γ are found mainly in the haematopoietic system (Schwehm, C., et al. J. Med. Chem. 2017, 60, 1534–1554). and δ are found mainly in the haematopoietic system (Schwehm, C., et al. J. Med. Chem. 2017, 60, 1534– PI3K δ has been identified as an attractive drug target, where it plays a key role in the pathogenesis of 1554). PI3K δ has been identified as an attractive drug target, where it plays a key role in the blood cancer (lymphoma, leukaemia) and autoimmune disease, such as rheumatoid arthritis. Selective pathogenesis of blood cancer (lymphoma, leukaemia) and autoimmune disease, such as rheumatoid inhibition of PI3K δ is desired to avoid the side effects associated with the inhibition of PI3k α and β, arthritis. Selective inhibition of PI3K δ is desired to avoid the side effects associated with the which play a major role in insulin signalling and platelet aggregation. Idelalisib is the first approved inhibition of PI3k α and β, which play a major role in insulin signalling and platelet aggregation. selective PI3K δ inhibitor for the treatment of chronic lymphocytic leukaemia (CLL) and follicular Idelalisib is the first approved selective PI3K δ inhibitor for the treatment of chronic lymphocytic lymphoma (FL). However, idelalisib has demonstrated serious side effects involving hepatic toxicity, leukaemia (CLL) and follicular lymphoma (FL). However, idelalisib has demonstrated serious side colitis, serious diarrhoea and pneumonitis (Somoza, J., et al. J. Biol. Chem. 2015, 290, 8439–8446). effects involving hepatic toxicity, colitis, serious diarrhoea and pneumonitis (Somoza, J., et al. J.Biol. Starting from a non-selective PI3K inhibitor “pictilisib”, we identified a series of potent indole-based Chem. 2015, 290, 8439–8446). Starting from a non-selective PI3K inhibitor “pictilisib”, we identified a inhibitors of PI3K δ with good selectivity versus other PI3K isoforms—the high δ selectivity of these series of potent indole-based inhibitors of PI3K δ with good selectivity versus other PI3K isoforms— compounds could be attributed to favourable interaction with “tryptophan shelf”. The “tryptophan the high δ selectivity of these compounds could be attributed to favourable interaction with shelf” defines the face of Trp760, that can be accessed only in PI3K δ due to movement of Thr750. “tryptophan shelf”. The “tryptophan shelf” defines the face of Trp760, that can be accessed only in On the other hand, in the other isoforms, the face of the tryptophan is “blocked” by basic residues that PI3K δ due to movement of Thr750. On the other hand, in the other isoforms, the face of the can form strong -cation interactions with Trp760 (Figure 10). tryptophan is “blocked” by basic residues that can form strong -cation interactions with Trp760 (Figure 10).

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Figure 10. Comparison of the tryptophan region of crystal structure of PI3K δ, α, β and γ. Figure 10. Comparison of the tryptophan region of crystal structure of PI3K δ, α, β and γ. 7.10. Development of Novel Antimicrobial Biopolymers (P10) 7.10. Development of Novel Antimicrobial Biopolymers (P10) Declan Mullen 1, Atiya Sarmin 2, John Connelly 2, Louise Young 1, John A. Parkinson 3 and DeclanVânia M. Mullen Moreira 1, Atiya1,4 Sarmin 2, John Connelly 2, Louise Young 1, John A. Parkinson 3 and Vânia M. Moreira 1,4 1 Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral 1Street, Strathclyde Glasgow Institute G4 0RE, of UK; Pharmacy [email protected] and Biomedical Sciences, University of Strathclyde, United Kingdom, 2 Centre161 Cathedral for Cell BiologyStreet, G4 and 0RE CutaneousGlasgow; e-mail: Research, declan.mulle [email protected] and the London School of Medicine and 2 Dentistry, Centre Queen for Cell Mary Biology University and Cutaneous of London Research, Barts and the London School of Medicine and Dentistry, Queen Mary University of London. 3 Department of Pure & Applied Chemistry, University of Strathclyde, 295 Cathedral Street, Glasgow 3 Department of Pure & Applied Chemistry, University of Strathclyde, United Kingdom, 295 Cathedral G1 1XL,Street, UK Glasgow G1 1XL. 4 4 Drug Drug Research Research Program, Program, Division Division of of Pharmaceutical Pharmaceutical Chemistry Chemistry andand Technology,Technology, FacultyFaculty of Pharmacy, UniversityUniversity of Helsinki, of Helsinki, Finland, Finland, Viikinkaari Viikinkaari 55 E,E, P.O. Box 56, 56, FI-00014 FI-00014 University University of Helsinki. of Helsinki The declinedecline inin discovery discovery of of novel novel antimicrobial antimicrob compoundsial compounds over over the last the two last decades two decades has become has becomea worldwide a worldwide problem problem (Spellberg, (Spellberg, B., et al. B.,Clin. et al. Infect. Clin. Dis.Infect., 2004 Dis., 38(9), 2004,, 1279–1286). 38(9), 1279–1286). With the With ability the abilityof microorganisms of microorganisms to evolve to and evolve adapt and to surviveadapt to exposure survive to exposure antibiotics, to thatantibiotics, is, the emergence that is, the of emergenceantimicrobial of resistanceantimicrobial (AMR), resistance human (AMR), health human is severely health threatened is severely (O0 Neill,threatened J.; 2014 (O(Accessed:′Neill, J.; 2014 May (Accessed:2019)). For May instance, 2019)). AMR For instance, hampers AMR the treatment hampers of the post-surgery treatment of infections post-surgery caused infections by bacteria caused such by as bacteriaStaphylococcus such aureusas Staphylococcus, and severely aureus complicates, and severely the healing complicates of infected the chronic healing wounds. of infected chronic wounds.Previous work in our group showed that amino acid-functionalised dehydroabietic acid derivatives are capablePrevious of killingwork in planktonic our group bacteria showed and that limit amino their ability acid-functionalised to form biofilms dehydroabietic (a. Manner, S., etacid al. derivativesEur. J. Med. are Chem. capable2015 ,of102 killing, 68–79; planktonic b. Helfenstein, bacteria A., and et al. limitBioorg. their Med. ability Chem to, form2017, 25biofilms, 132–137; (a. Manner,c. Fallarero, S., et A., al. et Eur. al. Int. J. Med. J. M Chem. ol. Sci. 20152013, 102, 14,, 68–79; 12054–12072). b. Helfenstein, This is A., particularly et al. Bioorg. relevant, Med. Chem as biofilms’, 2017, 25contribution, 132–137; c. to Fallarero, AMR are A., highly et al. Int. resilient J. M ol. to Sci. traditional 2013, 14, antimicrobials 12054–12072). This (a. Manner, is particularly S., et al.relevant,Eur. J. asMed. biofilms’ Chem. contribution2015, 102, to 68–79; AMR b.are Helfenstein,highly resilient A., to et traditional al. Bioorg. antimicrobials Med. Chem ,(a.2017 Manner,, 25, 132–137;S., et al. Eur.c. Fallarero, J. Med. Chem. A., et al.2015Int., 102 J., Mol. 68–79; Sci. b. Helfenstein,2013, 14, 12054–12072). A., et al. Bioorg. The Med. group Chem has, also2017, shown 25, 132–137; that the c. Fallarero,functionalisation A., et al. of nanocelluloseInt. J. Mol. Sci. with 2013 the, abietane14, 12054–12072). dehydroabieylamine The group resultedhas also in shown innovative that andthe functionalisationcost-efficient eco-friendly of nanocellulose surfaces with with the antimicrobial abietane dehydroabieylamine properties and good resulted biocompatibility in innovative (Hassan, and cost-efficientG, et al. ACS eco-friendly Sust. Chem. Eng.surfaces, 2019 with, 7, 5002–5009). antimicrobial As properties the primary and structural good biocompatibility component of (Hassan, the plant G,cell et wall, al. ACS cellulose Sust. Chem. is the Eng. most, 2019 common, 7, 5002–5009). organic compound As the primary on the structural planet. Itcomponent can be harvested of the plant and cellprocessed wall, cellulose from many is the diff erentmost renewablecommon organic plant sources compound such ason woodthe planet. and cotton It can (a. be Schultz, harvested C., etand al. processedACS Sustainable from many Chem. different Eng., 2018 renewable, 67, 8317–8324; plant sour b. Lin,ces N.,such et as al. woodMolecules and, 2018cotton, 23(10) (a. Schultz,, 2684–2708). C., et al.This ACS combination Sustainable ofChem. renewability, Eng., 2018, low 67, 8317–8324; cost, strong b. structural Lin, N., et integrityal. Molecules and, 2018 its innate, 23(10) non-cytotoxic, 2684–2708). Thisproperties combination make it of an renewability, ideal basis for low development cost, strong of structural new antimicrobial integrity biomaterials.and its innate non-cytotoxic propertiesThe goal make of ourit an current ideal basis work for is development to build from of this new knowledge antimicrobial and chemicallybiomaterials. modify medically relevantThe biopolymersgoal of our current with our work in-house is to build library from of antimicrobialthis knowledge abietanes and chemically to produce modify materials medically with relevantinnate antibacterial biopolymers properties with our andin-house explore library their of potential antimicrobial applications abietanes in wound-healing. to produce materials with innate antibacterial properties and explore their potential applications in wound-healing.

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 25 of 47 Pharmaceuticals 2019, 12, 179 25 of 47 Acknowledgements: Tenovus Scotland, The Engineering and Physical Sciences Research Council (EPSRC) Acknowledgements: Tenovus Scotland, The Engineering and Physical Sciences Research Council (EPSRC) 7.11. Stories from Staudinger: Synthesis of Chiral Beta-Lactams (P11)

7.11.Niamh Stories O′Boyle, from Staudinger:Mary Meegan Synthesis and Eavan of Chiral McLoughlin Beta-Lactams (P11)

Niamh1 School O0Boyle, of Pharmacy Mary Meeganand Pharmaceut and Eavanical Sciences, McLoughlin Trinity College, University of Dublin, Dublin, Ireland; e- mail: [email protected] 1 School of Pharmacy and Pharmaceutical Sciences, Trinity College, University of Dublin, Dublin, Ireland;Combretastatin [email protected] A-4 (CA-4) is the most potent antimitotic agent of the combretastatin A series, a group of diaryl stilbenes isolated from the wood of the South African tree Combretum caffrum (Watt, Combretastatin A-4 (CA-4) is the most potent antimitotic agent of the combretastatin A series, J. M., et al. The Medicinal and Poisonous Plants of Southern and Eastern Africa. E. & S. Livingstone Ltd.: a group of diaryl stilbenes isolated from the wood of the South African tree Combretum caﬀrum Edinburgh and London, United Kingdom, 1962). It has significant anticancer activity through (Watt, J. M., et al. The Medicinal and Poisonous Plants of Southern and Eastern Africa. E. & S. Livingstone inhibition of tubulin polymerization and microtubule assembly (Pettit, G. R., et al. Experientia 1988, Ltd.: Edinburgh and London, United Kingdom, 1962). It has signiﬁcant anticancer activity through 45). While cis stilbene structures demonstrate superior biological activity, the corresponding trans inhibition of tubulin polymerization and microtubule assembly (Pettit, G. R., et al. Experientia derivatives are inherently more stable. Isomerization of cis CA-4 to the trans form is observed both 1988, 45). While cis stilbene structures demonstrate superior biological activity, the corresponding during storage and in vivo during metabolism, which dramatically reduces anti-tumour activity trans derivatives are inherently more stable. Isomerization of cis CA-4 to the trans form is observed (Ohsumi, K., et al. Bioorg. Med. Chem. Lett. 1998, 8, 3153–3158). Our group previously employed the both during storage and in vivo during metabolism, which dramatically reduces anti-tumour activity Staudinger reaction to synthesize novel 3-hydroxy-1,4-diaryl-2-azetidinones. The problem of CA-4′s (Ohsumi, K., et al. Bioorg. Med. Chem. Lett. 1998, 8, 3153–3158). Our group previously employed cis–trans isomerization has been overcome via chemical manipulation of CA-4′s alkene bridge, the Staudinger reaction to synthesize novel 3-hydroxy-1,4-diaryl-2-azetidinones. The problem of utilizing a β-lactam ring to induce cis restriction. A number of analogues (Figure 11) have shown CA-4 s cis–trans isomerization has been overcome via chemical manipulation of CA-4 s alkene bridge, potent0 nanomolar antiproliferative activity in MCF-7 and HL-60 cells with enhanced0 activity relative utilizing a β-lactam ring to induce cis restriction. A number of analogues (Figure 11) have shown to CA-4 (O′Boyle, N. M., et al. J. Med. Chem. 2010, 53, 8569; Azizah, M., et al. Eur. J. Med. Chem. 2017, potent nanomolar antiproliferative activity in MCF-7 and HL-60 cells with enhanced activity relative to 130, 261–285). Typical Staudinger reactions form mixtures of cis and trans isomers depending on CA-4 (O Boyle, N. M., et al. J. Med. Chem. 2010, 53, 8569; Azizah, M., et al. Eur. J. Med. Chem. 2017, 130, reaction0 conditions employed, and additionally, at the 3-hydroxy position′s chiral center, racemic 261–285). Typical Staudinger reactions form mixtures of cis and trans isomers depending on reaction mixtures of R&S enantiomers. Trans isomers of 3-substituted-2-azetidinones have been shown to be conditions employed, and additionally, at the 3-hydroxy position s chiral center, racemic mixtures up to 50 times more potent than the corresponding cis derivatives0 emphasizing the requirement to of R&S enantiomers. Trans isomers of 3-substituted-2-azetidinones have been shown to be up to optimize the Staudinger approach to minimize yields of the undesirable cis isomer. Levo- and dextro- 50 times more potent than the corresponding cis derivatives emphasizing the requirement to optimize rotatory enantiomers hold potential to display lesser or greater biological activity relative to one the Staudinger approach to minimize yields of the undesirable cis isomer. Levo- and dextro-rotatory another. enantiomers hold potential to display lesser or greater biological activity relative to one another.

Figure 11. Structures of discussed compounds Figure 11. Structures of discussed compounds Our current work aims to (1) improve the available yield for chiral resolution by determiningOur current work the aims necessary to (1) improve conditions the availa to achieveble yield stereoselectivefor chiral resolution synthesis by determining of trans 3-hydroxy-1,4-diaryl-2-azetidinonesthe necessary conditions to achieve in the stereoselective Staudinger reaction synthesis and (2)of providetrans 3-hydroxy-1,4-diaryl-2- puriﬁcation of racemic mixturesazetidinones using inN the-(tert-butoxycarbonyl)-L-Proline Staudinger reaction and (2) provide as a chiral purification resolving of agent racemic to aﬀ mixturesord optically using pure N-

Pharmaceuticals 2019, 12, 179 26 of 47 enantiomers for further biological evaluation. Trans 3-hydroxy β-lactams have been separated from cis derivatives using chromatographic puriﬁcation. We have since optimized the Staudinger reaction to return relative yields of 95% relative ratio of trans/cis isomers, as indicated by integration of protons at position 3 and 4 of the β-lactam on 1H-NMR. Diastereomeric resolution using ﬂash column chromatography followed by hydrolysis of chiral resolving agents successfully yielded enantiomers of EMCL001 and 2 (Figure 11). Preliminary biochemical data for the enantiomers in breast cancer cells will be reported.

7.12. Promising Biologically Active Peptides of Hirudo medicinalis (P12) Ekaterina Grafskaia 1,2, Kirill Nadezhdin 2,3, Irina Talyzina 3,4, Nadezhda Polina 1, Oleg Podgorny 1,5, Pavel Bobrovsky 1, Ivan Latsis 1, Valentin Manuvera 1,2 and Vassili Lazarev 1,2 1 Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, 119435 Moscow, Russia 2 Moscow Institute of Physics and Technology (State University), 141701 Dolgoprudny, Russia 3 M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia 4 Federal State Budget Educational Institution of Higher Education M.V.Lomonosov Moscow State University (Lomonosov MSU or MSU), 119991 Moscow, Russia 5 Koltzov Institute of Developmental Biology of Russian Academy of Sciences, 119991 Moscow, Russia; [email protected] Our previous research aimed at complete genome sequence of medicinal leech Hirudo medicinalis, allowing for in silico implementation of the searching algorithms to identify new biologically active peptides, such as thrombolytic, anticoagulant and antibacterial agents. We applied computational algorithms to the medicinal leech Hirudo medicinalis genome assembly and identified homologs of serine proteinase inhibitors, promising anticoagulant proteins and candidate antimicrobial peptides (AMPs). Identified AMPs were chemically synthesised and assayed antimicrobial activity against three bacterial species (Escherichia coli, Bacillus subtilis and Chlamydia thrachomatis), cytotoxic and hemolytic activities and determined their secondary structures by NMR spectroscopy. Eight peptides exhibited antimicrobial activity, and two peptides, 3967 and 536–1, reduced the survival of E. coli, B. subtilis and C. thrachomatis cells through the disruption of cellular integrity at a concentration of 10 µM. Almost all the examined AMPs exhibited low-to-moderate haemolytic activity with no effects on the viability of cultured eukaryotic cells. Moreover, it was shown that three peptides, 536_2, 12530 and 3967, adopted α-helical conformation in membrane-mimetic DPC micelles. Overall, our experimental data verify the utility of the developed computational algorithms for the discovery of potential biologically active peptides. Using developed computational algorithm, we identified two promising medicinal leech AMPs, 3967 and 536–1, which are able to be considered in perspective as therapeutic agents. This work was supported by the Russian Science Foundation (project №17–75–20099).

7.13. Subtype-Selective Fluorescent Ligands as Pharmacological Tools for the Human Adenosine hA2A AR Receptor (P13) Eleonora Comeo 1,3, Nicholas D. Kindon 1,3, Mark Soave 2,3, Leigh A. Stoddart 2,3, Stephen J. Hill 2,3 and Barrie Kellam 1,3 1 Division of Biomolecular Sciences and Medicinal Chemistry, School of Pharmacy, University of Nottingham, NG7 2RD, UK 2 Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, NG7 2UH, UK 3 Centre of Membrane Proteins and Receptors (COMPARE) University of Birmingham and Nottingham, UK; [email protected] Pharmaceuticals 2019, 12, 179 27 of 47

The last decades have witnessed a significant growth and development of fluorescence-based techniques as a means to facilitate understanding of the signalling and dynamics of drug targets such as G protein-coupled receptors (GPCRs) in their native cellular environment (Vernall, A. J, et al. Br. J. Pharmacol. 2014, 171 (5), 1073–1084; Stoddart, L. A., et al. Sci. Rep. 2018, 8 (1), 1–19). Among the class A GPCRs, the human adenosine hA2AAR receptor represents an attractive drug target, which has been the subject of intensive medicinal chemistry research during the last 40 years due to the broad tissue distribution and implication in modulating a variety of biological processes (Ruiz, M., et al. J. Med. Chem. 2014, 57, 3623–3650). The hA2AAR mainly signals through the stimulatory GPCR and, upon receptor activation, adenyl cyclase is stimulated, leading to Cyclic adenosine monophosphate cAMP accumulation. The therapeutic potential of targeting the hA2AAR has been broadly investigated, leading to the development of an extensive number of synthetic agonists and antagonists for the treatment of several diseases including cardiovascular, inflammatory and CNS disorders. More recently, targeting the hA2AAR has generated a great interest in cancer immunotherapy. The tumour micro-environment features high concentrations of immunosuppressive adenosine that, upon A2AAR engagement, silences the immune response in the host, leading to tumour growth and proliferation. Indeed, A2AAR antagonism is being investigated as a therapeutic approach to defeat the immune system evasion by the tumour (Leone, R. D., et al. J. Immunother. Cancer 2018, 6 (1), 1–9). In this regard, understanding the complexity of the adenosinergic signalling is an essential prerequisite to address and develop more efficacious therapies. This can be facilitated by exploiting fluorescent probes selectively targeting the hA2AAR both in vitro and in vivo (Vernall, A. J., et al. iScience 2018, 6, 280–288). Herein, we report the design synthesis and pharmacological evaluation of two generations of subtype-selective fluorescently-labelled hA2AAR antagonists based on preladenant. Of particular interest for in vivo studies was the second generation of ligands, which features water soluble probes and, therefore, might display better stability in plasma and improved detection after IV-administration. Molecular modelling analysis has corroborated previously reported SAR studies indicating the optimal position for fluorophore attachment. The newly synthesised fluorescent probes retained functional activity and binding affinity at the hA2AAR, displaying a selectivity of >1000 nM over the other receptor subtypes. The effectiveness of the probes as pharmacological tools was also investigated in high resolution confocal imaging studies, revealing specific co-localisation with SNAP-hA2AAR at the cell membrane, which was surmounted by incubating the cells with high concentration of unlabelled ligand. In conclusion, given their excellent pharmacological and photochemical properties, the reported novel fluorescent ligands embody valuable tools to study the signalling and dynamics of the hA2AAR, not only in vitro but also in vivo, providing a robust and specific platform for further drug discovery investigations.

7.14. Synthesis and Pharmacological Evaluation of 5,6,7,8-Tetrahydroimidazo[1–a]Pyrazine-Based Thiosemicarbazones against Trypanosoma cruzi (P14) Marlene S. de Araújo Neta 1,2, Felipe N. Coutinho 1,2, Antônio R. de Faria 2, Carine Picot 1, Fabrice Pagniez 1, Patrice Le Pape 1, Teresinha G. da Silva 3 and Pascal Marchand 1 1 Université de Nantes, Cibles et médicaments des infections et du cancer, IICiMed, EA 1155, F-44000 Nantes, France 2 Department of Pharmaceutical Sciences, University of Pernambuco, Recife-PE 50740–520, Brazil 3 Department of Antibiotics, University Federal of Pernambuco, Recife-PE 50740–520, Brazil; [email protected] Neglected tropical diseases (NTDs) aﬀect mainly underdeveloped or developing countries located in Africa, Asia and Latin America. Worldwide, there are more than one billion people aﬀected by these diseases (World Health Organization, Neglected Tropical Diseases. Access: 26 May 2017. Information site: http://www.who.int/neglected_diseases/diseases/en/). These include Chagas disease, also known as American trypanosomiasis, a potentially life-threatening illness caused by the protozoan parasite Pharmaceuticals 2019, 12, 179 28 of 47

TrypanosomaPharmaceuticals cruzi 2019(T., 12 cruzi, x FOR). PEER Diﬃ cultyREVIEW of treatment due to the amount of side eﬀects of the drugs 28 of 47 currently used, the resistance of the parasite and late detection are concerns to address. Therefore, the discoveryaddress. Therefore, of new substances the discovery for the of treatmentnew substances of Chagas for the disease treatment is extremely of Chagas necessary. disease Thus, is extremely we developednecessary. a medicinal Thus, we chemistrydeveloped program a medicinal dealing chemistr withy the program synthesis dealing of 5,6,7,8-tetrahydroimidazo with the synthesis of 5,6,7,8- [1, 2-tetrahydroimidazoa] pyrazine-based thiosemicarbazones[1, 2-a] pyrazine-based as antiparasitic thiosemicarbazones agents (Moreno-Rodr as antiparasiticíguez, A.,agents et al. Eur.(Moreno- J. Med.Rodríguez, Chem. 2014 A.,, 87 et, al. 23–29; Eur. Leite,J. Med. F., Chem. et al. 2014Eur.J., 87 Med., 23–29; Chem. Leite,2016 F., ,et123 al., Eur.J. 639–648; Med. Marchand, Chem. 2016 P.,, 123 et al., 639– Eur.J.648; Med. Marchand, Chem. 2015 P.,, et103 al., 381–395.)Eur.J. Med. (Figure Chem. 122015). , 103, 381–395.) (Figure 12).

N N 2 S 1 N N R 3 NH 2 H NHR

Figure 12. General structure of target compounds. Figure 12. General structure of target compounds. The first developments and biological results of this novel series of molecules will be discussed. TheThe authors first developments would like to acknowledgeand biological Capes results/Cofecub of this novel Program series France-Brazil of molecules Mewill 865–15 be discussed. for financialThe support. authors would like to acknowledge Capes/Cofecub Program France-Brazil Me 865–15 for financial support. 7.15. Synthesis and Biological Evaluation of Inhibitors Targeting Signal Transducers and Activators of Transcription 5 (STAT5) Proteins in Myeloid Leukemias (P15) 7.15. Synthesis and Biological Evaluation of Inhibitors Targeting Signal Transducers and Activators of MarionTranscription Polomski 5 (STAT5)1, Marie Proteins Brachet-Botineau in Myeloid Leukemias2, Ludovic(P15) Juen 1, Fabrice Gouilleux 2, Marie-Claude Viaud-Massuard 1 and Gildas Prié 1 Marion Polomski 1, Marie Brachet-Botineau 2, Ludovic Juen 1, Fabrice Gouilleux 2, Marie-Claude 1 TeamViaud-Massuard IMT, GICC EA 1 7501,and Gildas University Prié of1 Tours, Labex SYNORG, 37200 Tours, France 2 Team LNOx, GICC ERL 7001 CNRS, University of Tours, 37032 Tours, France; [email protected] 1 Team IMT, GICC EA 7501, University of Tours, Labex SYNORG, 37200 Tours, France. 2 Myeloid Team leukemiasLNOx, GICC are ERL myeloproliferative 7001 CNRS, University diseases of To thaturs, a37032ffect Tours, hematopoietic France; e-mail: stem gildas.prie@univ- cells (HSC), and are dividedtours.fr in two types acute (AML) and chronic (CML) according to a fast or slower cell growth. CMLMyeloid is mainly leukemias due to the are t(9,22) myeloproliferative genomic translocation-derived diseases that affect BCR-ABL hematopoietic fusion oncogene stem cells coding (HSC), for theand tyrosine are divided kinase in Bcr-Abl, two types which acute activates (AML) the and transcription chronic (CML) factors according STAT5 (signal to a transducersfast or slower and cell activatorsgrowth. of transcription 5). The latter plays a crucial role in the initiation and maintenance of CML and mediatesCML resistanceis mainly todue Bcr-Abl to the kinase t(9,22) inhibitors genomic suchtranslocation-derived as imatinib mesylate BCR-ABL (IM, Glivec) fusion (Hoelbl oncogene et al.codingEMBO for Mol. the Med. tyrosine2010 , kinase2, 98–110; Bcr-Abl, Kavalerchik, which etactivates al. J. Clin. the Oncol. transcription2008, 26 ,factors 2911–2915). STAT5 AML (signal resultstransducers mainly from and internal activators tandem of transcription duplication (Itd)5). The mutations latter plays in the a juxtamembrane crucial role in regionthe initiation or point and mutationmaintenance in fms like of CML tyrosine and kinasemediates 3, FLT3. resistance This oncoproteinto Bcr-Abl kinase FLT3-Itd inhibitors (internal such tandem as imatinib duplication) mesylate has(IM, a tyrosine Glivec) kinase (Hoelbl activity, et al.which EMBO activates Mol. Med. STAT5 2010 (Birkenkamp,, 2, 98–110; Kavalerchik, et al. Leukemia et al.2001 J. Clin., 15, Oncol. 1923–1931). 2008, 26, 2911–2915).As a result, inhibitingAML results STAT5 mainly would contributefrom internal to reduce tandem the survivalduplication of CML (Itd) and mutations AML cells andin the moreoverjuxtamembrane tackle their region potential or point chemoresistance. mutation in fms like tyrosine kinase 3, FLT3. This oncoprotein FLT3- ItdA first(internal structure–activity tandem duplication) relationship has a study tyrosine allowed kinase us activity, to identify which one activates compound, STAT517f (Figure (Birkenkamp, 13), whichet al. inhibited Leukemia the 2001 growth, 15, of1923–1931). AML and CML cell lines as well as phosphorylation and transcriptional activity ofAs STAT5.a result, These inhibiting results STAT5 suggest would that contribute17f might to be reduce a new the lead survival molecule of CML targeting and STAT5AML cells signallingand moreover in myeloid tackle leukemias their potential (Juen, et chemoresistance. al. J. Med. Chem. 2017, 60, 6119–6136). Thanks to these results, synthesis of 17f new analogues with modulation around the tetrahydroquinoleine (THQ) ring were undertaken and their biological evaluation was carried out on model CML (KU812, K562) and AML (MV-4–11) cell lines. Among these new results, one compound, with the 4-pyridinyl in position 5 on the THQ ring, showed slightly better results than 17f on all cell lines. This outcome will guide further modulation work on the THQ ring with others nitrogen heterocycles (pyridazine, triazine, etc.).

Figure 13 Lead 17f, new analogs and hit compound.

A first structure–activity relationship study allowed us to identify one compound, 17f (Figure 13), which inhibited the growth of AML and CML cell lines as well as phosphorylation and

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 28 of 47

address. Therefore, the discovery of new substances for the treatment of Chagas disease is extremely necessary. Thus, we developed a medicinal chemistry program dealing with the synthesis of 5,6,7,8- tetrahydroimidazo [1, 2-a] pyrazine-based thiosemicarbazones as antiparasitic agents (Moreno- Rodríguez, A., et al. Eur. J. Med. Chem. 2014, 87, 23–29; Leite, F., et al. Eur.J. Med. Chem. 2016, 123, 639– 648; Marchand, P., et al. Eur.J. Med. Chem. 2015, 103, 381–395.) (Figure 12).

N N 2 S 1 N N R 3 NH 2 H NHR

Figure 12. General structure of target compounds.

The first developments and biological results of this novel series of molecules will be discussed. The authors would like to acknowledge Capes/Cofecub Program France-Brazil Me 865–15 for financial support.

7.15. Synthesis and Biological Evaluation of Inhibitors Targeting Signal Transducers and Activators of Transcription 5 (STAT5) Proteins in Myeloid Leukemias (P15)

Marion Polomski 1, Marie Brachet-Botineau 2, Ludovic Juen 1, Fabrice Gouilleux 2, Marie-Claude Viaud-Massuard 1 and Gildas Prié 1

1 Team IMT, GICC EA 7501, University of Tours, Labex SYNORG, 37200 Tours, France. 2 Team LNOx, GICC ERL 7001 CNRS, University of Tours, 37032 Tours, France; e-mail: gildas.prie@univ- tours.fr Myeloid leukemias are myeloproliferative diseases that affect hematopoietic stem cells (HSC), and are divided in two types acute (AML) and chronic (CML) according to a fast or slower cell growth. CML is mainly due to the t(9,22) genomic translocation-derived BCR-ABL fusion oncogene coding for the tyrosine kinase Bcr-Abl, which activates the transcription factors STAT5 (signal transducers and activators of transcription 5). The latter plays a crucial role in the initiation and maintenance of CML and mediates resistance to Bcr-Abl kinase inhibitors such as imatinib mesylate (IM, Glivec) (Hoelbl et al. EMBO Mol. Med. 2010, 2, 98–110; Kavalerchik, et al. J. Clin. Oncol. 2008, 26, 2911–2915). AML results mainly from internal tandem duplication (Itd) mutations in the juxtamembrane region or point mutation in fms like tyrosine kinase 3, FLT3. This oncoprotein FLT3- Itd (internal tandem duplication) has a tyrosine kinase activity, which activates STAT5 (Birkenkamp, et al. Leukemia 2001, 15, 1923–1931). PharmaceuticalsAs a result,2019, 12 inhibiting, 179 STAT5 would contribute to reduce the survival of CML and AML cells 29 of 47 and moreover tackle their potential chemoresistance.

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 29 of 47 Figure 13 13. LeadLead 17f17f, new, new analogs analogs and andhit compound. hit compound. 7.16.transcriptional From Phloridzin activity towards of STAT5. Semisynthetic These Bioactiveresults suggest Dihydrochalcones that 17f (P16)might be a new lead molecule targetingA first STAT5 structure–activity signalling inrelationship myeloid stleukemiasudy allowed (Juen, us to et identify al. J. Med.one compound, Chem. 2017 17f, 60(Figure, 6119–6136). 1 2 1 2 3 2 1 A.Thanks13), Cala which Peraltato inhibitedthese, F. results, Mayr the growth , G.synthesis Viaultof AML ,of V.and Temml17f CM newL cell, D. analogueslines Schuster as well with, H.as phosphorylation Stuppnermodulation, D. around Sandéraphin the, 1 1 P.tetrahydroquinoleineRichomme and J-J. (THQ) Hélesbeux ring were undertaken and their biological evaluation was carried out on 1modelSONAS, CML EA921, (KU812, UNIV K562) Angers, and AML SFR QUASAV, (MV-4–11) Faculty cell lines. of Health Among Sciences, these new Department results, one of compound, Pharmacy, 16with bd the Daviers, 4-pyridinyl 49045 Angersin position Cedex 5 on 01, the France THQ ring, showed slightly better results than 17f on all cell 2lines.Institute This of outcome Pharmacy will/Pharmacognosy guide further modulation and Center forwork Molecular on the THQ Biosciences ring with Innsbruck others (CMBI),nitrogen Universityheterocycles of (pyridazine, Innsbruck, 6020 triazine, Innsbruck, etc.). Austria 3 Department of Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy, Paracelsus Medical University7.16. From Salzburg,Phloridzin 5020towards Salzburg, Semisynthetic Austria; Bioactive [email protected] Dihydrochalcones (P16) 1 2 1 2 3 2 1 A. CalaPlant Peralta secondary , F. Mayr metabolism , G. Viault produces , V. aTemml large number , D. Schuster of specialized , H. Stuppner compounds , D. (overSéraphin 300,000) , P. 1 1 includingRichomme terpenoids, and J-J. Hélesbeux alkaloids and (poly)phenols. Often referred to as “natural products” (NPs), many1 SONAS, of these EA921, compounds UNIV Angers, are used SFR for QUASAV, medicinal, Faculty cosmetical of Health and Sciences,/or nutraceutical Department purposes.of Pharmacy, Apple 16 bd trees (Daviers,Malus x 49045 domestica AngersBrokh.) Cedex are01, widelyFrance. cultivated all around the world, and apple is the first fruit to2 be produced Institute of inPharmacy/Pharmacognosy Europe. In France, the “Randé gionCenter des for Pays Molecular de la Loire”Biosciences appears Innsbruck as the (CMBI), second University national producerof Innsbruck, of apples, 6020 with Innsbruck, apple trees Austria. being the main cultivated surface in this area. Apple fruits/trees 3 Department of Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy, Paracelsus Medical are characterized with a cell specific accumulation—in different tissues—of varying levels of bioactive University Salzburg, 5020 Salzburg, Austria; e-mail: [email protected] polyphenols including proanthocyanidins, anthocyanins, flavonols and dihydrochalcones (DHCs) (FigurePlant 14). secondary metabolism produces a large number of specialized compounds (over 300,000) including terpenoids, alkaloids and (poly)phenols. Often referred to as “natural products” (NPs), many of these compounds areTable used 1. forGeneral medicinal, structure cosmetical of the compounds. and/or nutraceutical purposes. Apple trees (Malus x domestica Brokh.) are widely cultivated all around the world, and apple is the first fruit Compound R1 R2 R3 to be produced in Europe. In France, the “Région des Pays de la Loire” appears as the second national producer of apples, with applePhloretin trees being (Pt) the main OH cultivated OH surface H in this area. Apple fruits/trees are characterized with a cell specificPhlorizin accumulation—in OH OGludifferent Htissues—of varying levels of 3-Hydroxyphloretin bioactive polyphenols including proanthocyanidins,OH anthocyanins, OH flavonol OH s and dihydrochalcones (HPt) (DHCs). Trilobatin OGlu OH H Sieboldin OGlu OH OH

FigureFigure 14. 14.Structures Structures of naturalof natural dihydrochalcones dihydrochalcones (DHCs) (DHC isolateds) isolated fromapple fromtree apple leaves, tree and leaves, example and ofexample semisynthetic of semisynthetic modifications modifications (Table1). (Table 1). To date, around 250 DHCs are known, either as glycosylated derivatives or C-alkylated compounds Compound R1 R2 R3 (C. Rivière in Studies in Natural Products Chemistry, 2016, 51, 253–281). Amongst the latter, some Phloretin (Pt) OH OH H C-benzylated compounds have demonstrated significant pharmacological effects (anti-inflammatory, Phlorizin OH OGlu H 3-Hydroxyphloretin (HPt) OH OH OH Trilobatin OGlu OH H Sieboldin OGlu OH OH

Table 1. General structure of the compounds.

To date, around 250 DHCs are known, either as glycosylated derivatives or C-alkylated compounds (C. Rivière in Studies in Natural Products Chemistry, 2016, 51, 253–281). Amongst the latter, some C-benzylated compounds have demonstrated significant pharmacological effects (anti- inflammatory, anti-tumoral) (Somsrisa, J., et al. Molecules 2013, 18, 6898–6907; Prawat, U., et al. Planta

Pharmaceuticals 2019, 12, 179 30 of 47 Pharmaceuticals 2019, 12, x FOR PEER REVIEW 30 of 47

Med.anti-tumoral) 2013, 79, 83–86). (Somsrisa, Therefore J., et al. thisMolecules series2013 of ,compou18, 6898–6907;nds is Prawat,of high U.,interest et al. Plantabecause Med. of 2013their, 79, pharmacological83–86). Therefore potential. this series The of main compounds goal of is this of high project interest is to because (semi-)synthesize of their pharmacological original DHCs, potential. in particular,The main C-benzylated goal of this derivatives, project is to starting (semi-)synthesize from phloridzin original or DHCs,sieboldin in extracted particular, fromC-benzylated locally producedderivatives, apple starting tree leaves from phloridzinas renewable or sieboldin sources. extracted The semi-synthetic from locally analogues produced applewill then tree leavesbe submittedas renewable to biological sources. assays The semi-syntheticto evaluate their analogues anti-tumoural will thenpotential. be submitted to biological assays to evaluate their anti-tumoural potential.

7.17. Synthesis and Pharmacological Study of Alprenolol Analogues at the Secondary Conformation of the β1- 7.17. Synthesis and Pharmacological Study of Alprenolol Analogues at the Secondary Conformation of the Adrenoceptor (P17) β1-Adrenoceptor (P17) Emanuel P. Sousa a,b, Peter J. Scammells c, Jillian G. Baker b and Shailesh N. Mistry a Emanuel P. Sousa 1,2, Peter J. Scammells 3, Jillian G. Baker 2 and Shailesh N. Mistry 1 1 School of Pharmacy, Centre for Biomolecular Sciences, University of Nottingham, Nottingham, United 1 School of Pharmacy, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 Kingdom, NG7 2RD. 2 2RD, Cell UKSignalling, School of Life Sciences, Queen′s Medical Centre, University of Nottingham, Nottingham, 2 UnitedCell Signalling, Kingdom, NG7 School 2UH. of Life Sciences, Queen0s Medical Centre, University of Nottingham, 3 Nottingham Medicinal Chemistry, NG7 2UH, Monash UK Institute of Pharmaceutical Sciences, Monash University, 3052 Victoria, 3 MedicinalAustralia; e-mail: Chemistry, [email protected] Monash Institute of Pharmaceutical Sciences, Monash University, 3052 Victoria, Australia; [email protected] The β1-adrenergic receptor (β1-AR) exists in at least two distinct agonist conformations: (1) a primaryThe conformationβ1-adrenergic where receptor responses (β1-AR) are existsreadily in inhibited at least two by distinctantagonists agonist and conformations: (2) a secondary (1) a conformationprimary conformation of which the where precise responses nature areis un readilyknown, inhibited where byagonist antagonists responses and are (2) relatively a secondary resistantconformation to antagonism of which (Kaumann, the precise A.J., nature et al. is Pharmacol. unknown, Ther. where 2008 agonist, 118, responses 303–336). are relatively resistant toEndogenous antagonism (Kaumann,catecholamines A.J., (e.g., et al. adrenaline)Pharmacol. Ther.and conventional2008, 118, 303–336). agonists (e.g., isoprenaline and cimaterol)Endogenous mediate a catecholamines response mainly (e.g., through adrenaline) the andprimary conventional conformation. agonists Other (e.g., isoprenalineligands (e.g., and CGP12177cimaterol) 1) mediate a response mainlythrough through the secondary the primary conformation, conformation. even Other though ligands the concentration (e.g., CGP12177 required1) mediate to stimulate a response this through response the is secondary considerably conformation, greater than even the though concentration the concentration required requiredto bind to thestimulate primary conformation this response (Pak, is considerably M.D., et al. greater J Recept than Signal the Transduct concentration Res., 1996 required, 16, 1–23). to bind Alprenolol the primary (2) conformationstimulates a response (Pak, M.D., through et al. J. both Recept conformations Signal Transduct and Res it., also1996 ,requires16, 1–23). higher Alprenolol concentrations (2) stimulates to a activateresponse the throughsecondary both conformation conformations (Baker, and it J.G., also et requires al. Mol. higher Pharmacol concentrations, 2003, 63, to1312–1321). activate the To secondary date, no conformationligand has been (Baker, reported J.G., etthat al. bindsMol. the Pharmacol second,ary2003 conformation, 63, 1312–1321). with To higher date, affinity no ligand than has the been primaryreported conformation. that bindsthe secondary conformation with higher affinity than the primary conformation. AlprenololAlprenolol bis bisanalogueanalogue 3 has3 has been been previously previously identified identified as as the the first first sub-micr sub-micromolaromolar compound compound to to have similar aaffinityffinity forfor bothboth conformationsconformations (Sousa, (Sousa, E.P, E.P, et et al. al.2018 2018, Poster, Poster presented presented at EFMC-ISMCat EFMC- ISMC2018). 2018). In order In order to identify to identify the molecular the molecula descriptorsr descriptors responsible responsible for the interaction for the interaction and activation and of activationthe secondary of the conformationsecondary conformation of the β1-AR, of several the β1 analogues-AR, several were analogues synthesised were and synthesised pharmacologically and pharmacologicallycharacterised through characterised competitive through radioligand competitive binding radioligand assays andbinding cAMP assays reporter and cAMP gene (CRE-SPAP) reporter genefunctional (CRE-SPAP) assays. functional In this assays. communication, In this communi wecation, report we the report affinity the ofaffinity these of analogues these analogues for both for conformationsboth conformations of the ofβ 1the-AR. β1-AR.

Representative compounds

7.18.7.18. X-ray X-ray Diffraction Diﬀraction Investigation Investigation of Aqueous of Aqueous Microgels Microgels for forDrug Drug Delivery Delivery (P18) (P18)

1 2,3 2,3 2,3,4 AndreyAndrey V. V.Dolgopolov Dolgopolov , 1Kseniia, Kseniia N. N. Grafskaia Grafskaia 2,3, Denis, Denis V. V. Anokhin Anokhin 2,3, Dimitri, Dimitri A. A. Ivanov Ivanov 2,3,4, , 1 1 1 AndrijAndrij Pich Pich , Xiaomin1, Xiaomin Zhu Zhu and1 and Martin Martin Möller Möller 1 1 1 DWI—Leibniz-Institute DWI—Leibniz-Institute for for Interactive Interactive Materials Materials e.V. e.V. and and Institute Institute for for Technical Technical and and Macromolecular Macromolecular Chemistry of RWTH Aachen University, D-52056 Aachen, Germany. Chemistry of RWTH Aachen University, D-52056 Aachen, Germany 2 Moscow Institute of Physics and Technology (State University), 141700 Moscow, Russia. 3 Institute of Problems of Chemical Physics of RAS, 142432 Chernogolovka, Russia.

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Pharmaceuticals2 Moscow Institute 2019, 12, ofx FOR Physics PEER andREVIEW Technology (State University), 141700 Moscow, Russia 31 of 47 3 Institute of Problems of Chemical Physics of RAS, 142432 Chernogolovka, Russia 4 4 Institut Institutde de ScienceScience dede Matériaux Matériaux de de Mulhouse Mulhouse (IS2M-CNRS), (IS2M-CNRS), F-68057 F-68057 Mulhouse, Mulhouse, France; France;e-mail: [email protected]@phystech.edu Aqueous microgels based on poly(N-vinylcaprolactam) with reversable temperature-induced Aqueous microgels based on poly(N-vinylcaprolactam) with reversable temperature-induced volume transition are promising “smart” materials for various applications, in particular for drug volume transition are promising “smart” materials for various applications, in particular for drug delivery (Saunders, B. R., et al. Adv. Colloid. Interfac. 2009, 147–48, 251–262). Here, the aqueous delivery (Saunders, B. R., et al. Adv. Colloid. Interfac. 2009, 147–48, 251–262). Here, the aqueous microgels are modified via acid–base interaction by ligand molecules. They are wedge-shaped microgels are modified via acid–base interaction by ligand molecules. They are wedge-shaped amphiphilic sulfonic acid molecules with an azobenzene grou and alkyl chains of different lengths. amphiphilic sulfonic acid molecules with an azobenzene grou and alkyl chains of different lengths. Modified microgels remain colloidally stabile in water and show different responses to the change of Modified microgels remain colloidally stabile in water and show different responses to the change of temperature and pH within the physiologic condition (Grafskaia, K. N., et al. Chem. Commun. 2017, temperature and pH within the physiologic condition (Grafskaia, K. N., et al. Chem. Commun. 2017, 53, 53, 13217). The hydrophobicity of the microgel interior increases with the increase of the modification 13217). The hydrophobicity of the microgel interior increases with the increase of the modification degree. Hardening of hydrophobic nanodomains and a decrease of the hydrodynamic diameter for degree. Hardening of hydrophobic nanodomains and a decrease of the hydrodynamic diameter sample complexed with C8AzoSO3H at the highest neutralization degrees can be explained by for sample complexed with C8AzoSO3H at the highest neutralization degrees can be explained by formation of the alkyl sub-lattice (Figure 15). formation of the alkyl sub-lattice (Figure 15).

Figure 15. GIWAXS curves of PVCL/AAEM/VIm/(C8AzoSO3H) microgels with DN = 25 (red), 50 (green) Figure 15. GIWAXS curves of PVCL/AAEM/VIm/(C8AzoSO3H) microgels with DN = 25 (red), and 100% (black) (left); GIWAXS pattern of PVCL/AAEM/VIm/(C8AzoSO3H) microgels with DN = 25 50 (green) and 100% (black) (left); GIWAXS pattern of PVCL/AAEM/VIm/(C8AzoSO3H) (right). microgels with DN = 25 (right). The characterisation of local structure of microgels was performed by GIWAXS technique. In wide-angleThe characterisation region, a narrowof local st reﬂectionructure of corresponding microgels was to performed d-spacing by of GIWAXS 4.82 Å can technique. be clearly In wide-angleseen. One region, can assume a narrow that reflection this peak corresponding originated to from d-spacing the linear of 4.82 alkyl Å can chain be clearly packing seen. of One the wedge-shapedcan assume that mesogens this peak (Zhu, originated X. M., from et al. thePhys. linear Chem.alkyl chain Chem. packing Phys. of2010 the, wedge-shaped 12, 1444–1452; mesogensGrafskaia, (Zhu, K. N., X. et M., al. etChem. al. Phys. Commun. Chem. Chem.2017, 53,Phys 13217).. 2010, 12, Formation 1444–1452; of alkylGrafskaia, sub-lattice K. N., canet al. explain Chem. Commun.hardening 2017 of hydrophobic, 53, 13217). nanodomainsFormation of alkyl and decrease sub-latti ofce hydrodynamiccan explain hardening diameter of for hydrophobic the sample nanodomains and decrease of hydrodynamic diameter for the sample PVCL/AAEM/VIm/(C8AzoSO3H)100%. PVCL/AAEM/VIm/(C8AzoSO3H)100%. 7.19. DYRK2 Inhibition—A Potential Targeted Therapy for TNBC (P19) 7.19. DYRK2 Inhibition—A Potential Targeted Therapy for TNBC (P19) Nicholas C. O. Tomkinson 1, Simon Mackay 2, Laura M. Bain 1, Laureano de la Vega 3 and NicholasJoanne Edwards C. O. Tomkinson4 1, Simon Mackay 2, Laura M. Bain 1, Laureano de la Vega 3 and Joanne Edwards 4 1 Department of Pure and Applied Chemistry, University of Strathclyde, Glasgow G1 1XL, Scotland 1 2 Strathclyde Department Institute of Pureof and Pharmacy Applied Chemistry, and Biomedical University Sciences, of Strathclyde, University G1 1XL of Strathclyde, Glasgow, Scotland. Glasgow G1 2 1XL,Strathclyde Scotland Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, G1 1XL. 3 School of Medicine, University of Dundee, DD1 9SY Dundee, Scotland. 3 School of Medicine, University of Dundee, DD1 9SY Dundee, Scotland 4 Institute of Cancer Sciences, University of Glasgow, G61 1QH Glasgow, Scotland; e-mail: [email protected] Triple negative breast cancer (TNBC) is the most aggressive form of breast cancer. It accounts for approximately 20% of all breast cancers and has an increased rate of relapse compared to non- TNBCs. This aggressive nature is due to its lack of receptors for targeted therapies and thus results

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4 Institute of Cancer Sciences, University of Glasgow, G61 1QH Glasgow, Scotland; [email protected] Triple negative breast cancer (TNBC) is the most aggressive form of breast cancer. It accounts for Pharmaceuticalsapproximately 2019 20%, 12, x of FOR all PEER breast REVIEW cancers and has an increased rate of relapse compared to non-TNBCs. 32 of 47 This aggressive nature is due to its lack of receptors for targeted therapies and thus results in inchemotherapy chemotherapy being being the the onlyonly optionoption for treatment treatment (Gonçalves (Gonçalves Jr, Jr, H., H., et etal. al.Clin.Clin. Med. Med. In. Onc. In. 2018, Onc. 122018, 1–10)., 12, This 1–10). reveals This revealsthe need the to need investigate to investigate other means other meansfor targeting for targeting and treating and treating TNBC, TNBC,more specifically,more speciﬁcally, through through the use the of use small of small molecule molecule inhibitors. inhibitors. We We discovered discovered that that dual dual specificity speciﬁcity tyrosine(Y)tyrosine(Y) regulated regulated kinase-2 kinase-2 (DYRK2) (DYRK2) stabilises stabilises heat heat shock shock factor-1 factor-1 (HSF1), (HSF1), the the master master regulator regulator of proteotoxicof proteotoxic stress stress pathways pathways (Rashmi, (Rashmi, K. C., K. C.,et al. et Cell al. CellStress Stress and andChaperones Chaperones, 2017, 2017, 22, 22751–766)., 751–766). In addition,In addition, we weshowed showed that that low low DYRK2 DYRK2 expression expression correlates correlates with with an an increased increased survival survival rate rate in in TNBC TNBC patientspatients (Figure (Figure 16A).16A).

(A) (B)

FigureFigure 16. 16. (A(A) )Levels Levels of of dual dual specificity speciﬁcity tyrosine(Y) regulatedregulated kinase-2kinase-2 (DYRK2) (DYRK2) expression expression correlate correlate to totriple triple negative negative breast breast cancer cancer (TNBC) (TNBC) survival survival rate. rate. (B ()B Model) Model of of LB35 LB35 in in DYRK2 DYRK2 active active site. site.

ToTo date, date, there there are are no no selective selective small small molecule molecule inhibitors inhibitors for for DYRK2. DYRK2. With With the the aid aid of of molecular molecular modelling,modelling, we we developed developed an an SAR SAR profile profile of of DYRK2 DYRK2 inhibitors, inhibitors, with with our our most most potent potent inhibitor inhibitor being being LB35LB35 Ki Ki 19 19 nM nM (Figure (Figure 16B).16B). These These inhibitors inhibitors reveal revealeded an an impressive impressive selectivity selectivity profile, profile, whereby whereby a a structurallystructurally similar similar compound compound showed showed <30%<30% inhibition inhibition against against a apanel panel of of 40 40 structurally structurally related related kinases.kinases. Within Within this this poster, poster, we we aim aim to to describe describe ou ourr ongoing ongoing efforts efforts to to provide provide an an inhibitor inhibitor for for DYRK2 DYRK2 withwith increased increased potency potency and and examine examine the the DMPK DMPK chal challengeslenges associated associated with with this this inhibitor inhibitor series.

7.20.7.20. Synthesis Synthesis of of Conformation Conformationallyally Restricted Restricted Adrenaline Adrenaline An Analogsalogs Selectively Selectively Activating Activating ββ22-AR-AR with with Substantial Subtype Selectivity (P20) Substantial Subtype Selectivity (P20) L. Maul 1, H. Hübner 1, X. Liu 2, J. Shonberg 1, A. Stößel 1, M. Stanek 1, D. Weikert 1, B.K. Kobilka 2,3 L. Maul 1, H. Hübner 1, X. Liu 2, J. Shonberg 1, A. Stößel 1, M. Stanek 1, D. Weikert 1, B.K. Kobilka and P. Gmeiner 1 2,3 and P. Gmeiner 1 1 1 Department Department of of Chemistry Chemistry and Pharmacy, Pharmacy, Friedrich-Alexander Friedrich-Alexander University, University, Erlangen, Erlangen, Germany. Germany 2 2 Beijing Beijing Advanced Advanced Innovation Innovation Center Center for for Structural Structural Biology, Biology, Tsinghua Tsinghua Univ University,ersity, Beijing, Beijing, China. China 3 3 Department Department ofof MolecularMolecular and and Cellular Cellular Physiology, Physiology, Stanford Stanford University, University, California, California, USA; email: USA; [email protected]@fau.de

TheThe loss ofof aa ligand ligand0s′ entropys entropy upon upon receptor receptor binding binding can becan reduced be reduced by rigidification by rigidification of its chemical of its chemicalstructure, structure, often leading often to leading enhanced to potency, enhanced depending potency, on depending whether the on bioactive whether conformation the bioactive is conformationmatched or not. is matched Conformational or not. restrictionConformational can also restriction induce selectivity can also forinduce receptor selectivity subtypes, for asreceptor slightly subtypes,different shapesas slightly of orthosteric different binding shapes pockets of orth mayosteric require binding a diff erentpockets conformational may require adaption a different of the conformational adaption of the ligand. Taking advantage of those effects can lead to the development of novel molecular probes for scientific research and medications with improved pharmacodynamic or pharmacokinetic properties (Ring, et al. Nature. 2013, 502, 575–579). Using the pharmacologically important target β2-AR, we aimed at synthetically constraining highly flexible agonists of the catecholamine type into their bioactive conformation. Thus, we synthesized a set of eight different isomers of ethylene bridged isoprenaline, with one of them (“super-iso”) being clearly superior regarding affinity, potency and selectivity for β2 over β1. Super-

Pharmaceuticals 2019, 12, 179 33 of 47 ligand. Taking advantage of those effects can lead to the development of novel molecular probes for scientific research and medications with improved pharmacodynamic or pharmacokinetic properties (Ring, et al. Nature. 2013, 502, 575–579). Using the pharmacologically important target β2-AR, we aimed at synthetically constraining highly Pharmaceuticals 2019, 12, x FOR PEER REVIEW 33 of 47 flexible agonists of the catecholamine type into their bioactive conformation. Thus, we synthesized a set of eight different isomers of ethylene bridged isoprenaline, with one of them (“super-iso”) being epi, super-norepi, etc., refer to compounds with the same constitution and stereochemical clearly superior regarding affinity, potency and selectivity for β over β . Super-epi, super-norepi, etc., configuration (Figure 17). 2 1 refer to compounds with the same constitution and stereochemical configuration (Figure 17).

Figure 17. Illustration of the ß2-AR active state crystal structures in complex with either epinephrine or Figure 17. Illustration of the ß2-AR active state crystal structures in complex with either epinephrine super-epi, which were shown to occupy virtually the same binding pocket in the receptor. or super-epi, which were shown to occupy virtually the same binding pocket in the receptor. Structural insights into the receptor state and the binding poses of super-epi and super-iso were Structural insights into the receptor state and the binding poses of super-epi and super-iso were obtained by co-crystal structures of β2-AR bound to Nb6B9. obtained by co-crystal structures of β2-AR bound to Nb6B9. The poster will also give information about efficient chemical synthesis, biological investigations The poster will also give information about efficient chemical synthesis, biological investigations and structure–activity relationships (SAR) of the conformationally restricted catecholamines. and structure–activity relationships (SAR) of the conformationally restricted catecholamines. 7.21. Synthesis of Lung Tissue Retentive Prodrugs (P21) 7.21. Synthesis of Lung Tissue Retentive Prodrugs (P21) J. Ayre 1, D. Speed 1, R. Weaver 2, G. Vitulli 3, J. Morrell 3, C. Bosquillon 1, J. M. Redmond 1 and M. J.J. Stocks Ayre 1 1, D. Speed 1, R. Weaver 2, G. Vitulli 3, J. Morrell 3, C. Bosquillon 1, J. M. Redmond 1 and M. J. Stocks 1 1 School of Pharmacy, University of Nottingham, Nottingham, NG7 2RD 1 2 XenoGesis, School BioCity, of Pharmacy, Pennyfoot University Street, of Nottingham Nottingham, NG1Nottingham, 1GF NG7 2RD. 2 XenoGesis, BioCity, Pennyfoot Street, Nottingham NG1 1GF. 3 DMPK, GSK, Gunnels Wood Rd, Stevenage, Hertfordshire SG1 2NY 3 DMPK, GSK, Gunnels Wood Rd, Stevenage, Hertfordshire SG1 2NY. 4 Chemical Biology, GSK, Gunnels Wood Rd, Stevenage, Hertfordshire SG1 2NY; 4 Chemical Biology, GSK, Gunnels Wood Rd, Stevenage, Hertfordshire SG1 2NY; emails: [email protected],[email protected], [email protected]*, [email protected]*, [email protected] [email protected] The pulmonaryThe pulmonary drug deliverydrug delivery route possesses route possesses significant significant benefits overbenefits other over administrative other administrative routes, includingroutes, targeted including delivery targeted to the delivery site of action,to the site by-pass of action, of first by-pass pass metabolism of first pass in metabolism the liver and in fewer the liver sideand effects fewer owing side to effects a reduced owing drug to a dosage. reduced However, drug dosage. rapid However, drug elimination rapid drug from elimination lung tissue from in to lung the hepatictissue in vein to the and hepatic the pan vein antagonistic and the naturepan antagonist of receptoric nature antagonists of receptor often leadsantagonists to increased often sideleads to effects.increased By exploiting side effects. the highly By exploiting lung retentive the highly nature lu ofng dibasic retentive compounds, nature of it dibasic is possible compounds, to reduce it is the ratepossible of elimination to reduce fromthe rate the lungsof elimination and thus from reduce the side lungs eff ects.and Thisthus workreduce describes side effects. the designThis work and synthesisdescribes ofthe a rangedesign of and dibasic synthesis prodrugs of thata range have of the dibasic potential prodrugs to be lysosomally that have trappedthe potential within to be lunglysosomally tissue and cleave trapped slowly within at alung desired tissue pH and to ecleaveffectively slowly deliver at a andesired active pH drug to ateffectively a slower deliver more an consistentactive rate.drug at a slower more consistent rate. 7.22. Design and Synthesis of Novel Flavone Derivatives as Anti-Cancer Agents (P22) 7.22. Design and Synthesis of Novel Flavone Derivatives as Anti-Cancer Agents (P22) Mai Khater, Francesca Grecoa and Helen M.I. Osborna Mai Khater,a Francesca Grecoa and Helen M.I. Osborna

Schoola of School Pharmacy, of UniversityPharmacy, ofUniversity Reading, Readingof Reading, RG6 Reading 6AD, UK; RG6 [email protected] 6AD, United Kingdom; e-mail: [email protected] Breast cancer is the most common cancer in women, resulting in 2.09 million cases worldwide (WHO Fact Sheets https://www.who.int/en/news-room/fact-sheets/detail/cancer (12 Sepetmber 2018). Flavones are one of the most active classes of flavonoids exhibiting anti-tumour activities. Flavopiridol, for example, has reached phase-II clinical trials as an anti-cancer agent against leukaemia, lymphoma and solid tumours. Selenium and ruthenium atoms have demonstrated promising anti-proliferative activities when incorporated into small molecular ligands (Olla S., et al. Bioorg. Med. Chem. Lett. 2009, 19, 1512–1516). Replacement of flavonoids′ oxo-carbonyl with a seleno-

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Breast cancer is the most common cancer in women, resulting in 2.09 million cases worldwide (WHO Fact Sheets https://www.who.int/en/news-room/fact-sheets/detail/cancer (12 Sepetmber 2018). Flavones are one of the most active classes of ﬂavonoids exhibiting anti-tumour activities. Flavopiridol, for example, has reached phase-II clinical trials as an anti-cancer agent against leukaemia, lymphoma

Pharmaceuticalsand solid tumours. 2019, 12, x Selenium FOR PEER andREVIEW ruthenium atoms have demonstrated promising anti-proliferative 34 of 47 activities when incorporated into small molecular ligands (Olla S., et al. Bioorg. Med. Chem. Lett. carbonyl2009, 19, 1512–1516).can result inReplacement up to a 25-fold of ﬂavonoidsdecrease in0 oxo-carbonylIC50 values on with breast a seleno-carbonyl cancer cell lines can (Martins resultin I. upL., etto al. a 25-fold J. Med. decrease Chem. 2015 in IC, 5850, 4250–4256)values on breast (Scheme cancer 2). cellOn the lines other (Martins hand, I. coordination L., et al. J. Med. with Chem. metal2015 atoms, 58, such4250–4256) as ruthenium (Scheme 3is). reported On the other to enhance hand, coordination flavonoids′ withwater metal solubility atoms up such to as10-fold, ruthenium which is reportedcan lead to better enhance clinical ﬂavonoids applications.0 water solubility up to 10-fold, which can lead to better clinical applications.

Scheme 2. Synthesis 3. Synthesis of flavone derivatives. of ﬂavone (I) Benzoyl derivatives. chloride halide, (I) 1,8-DiazaBicyclo[5.4.0]Undec- Benzoyl chloride halide, 7-ene DBU, pyridine, 75 °C, 1 h; (II) pyridine, KOH, 50–5 °C, 2 h; (III) glacial acetic acid, 1% H2SO4, 1,8-DiazaBicyclo[5.4.0]Undec-7-ene DBU, pyridine, 75 ◦C, 1 h; (II) pyridine, KOH, 50–5 ◦C, 90–110 °C, 1 h; (IV) dry toluene, Lawesson′s reagent, 110 °C, 4 h; (V) acetonitrile, Woollin′s reagent, 2 h; (III) glacial acetic acid, 1% H2SO4, 90–110 ◦C, 1 h; (IV) dry toluene, Lawesson0s reagent, 110 ◦C, microwave,4 h; (V) acetonitrile, 175 W, 150 Woollin °C, 50 smi reagent,n; (VI) MeOH, microwave, NaOMe, 175 W,MLn. 150 ◦C, 5 min; (VI) MeOH, NaOMe, MLn.

We aim to develop a new series of flavones flavones for for a applicationpplication against against breast breast cancer. cancer. Two Two previously previously identifiedidentified flavone flavone leads (1 and 2 inin Schemescheme 32 fromfrom whichwhich newnew seleniumselenium andand rutheniumruthenium derivativesderivatives can bebe synthesized). synthesized). The The synthesis synthesis provides provides molecular molecular frameworks, frameworks, following afollowing Baker–Venkataraman a Baker– Venkataramanrearrangement pathway..rearrangement This will pathway.. be followed This by willin vitro be followedcytotoxicity by andin vitro anti-angiogenic cytotoxicity assessments and anti- angiogenicin addition assessments to probing the in possibleaddition mechanisms to probing the of actionpossible of themechanisms most active of compounds.action of the most active compounds. 7.23. Imidazo [1, 2-a] Pyrazines Displaying In Vivo Antileishmanial Activity (P23) 7.23.Marc-Antoine Imidazo [1, Bazin 2-a] Pyrazi1, Sandrinenes Displaying Cojean 2 ,In Fabrice Vivo Antileishmanial Pagniez 1, Marie-Ren Activityée (P23) Nourrisson 1, Carine Picot 1, 2 2 3 4 4 Marc-AntoineChristian Cav éBazin, Guillaume 1, Sandrine Bernadat Cojean ,2, St Fabriceéphane Pagniez Bach , 1 Najma, Marie-Renée Rachidi Nourrisson, Olivier Leclercq 1, Carine, 2 1 1 PicotPhilippe 1, Christian Loiseau Cavé, Patrice 2, Guillaume Le Pape Bernadatand Pascal 2, MarchandStéphane Bach 3, Najma Rachidi 4, Olivier Leclercq 4 2 1 1 1, UniversitPhilippe éLoiseaude Nantes, , Patrice Cibles Le et Pape médicaments and Pascal des infectionsMarchand et du cancer, IICiMed, EA 1155, F-44000 1Nantes, Université France de Nantes, Cibles et médicaments des infections et du cancer, IICiMed, EA 1155, F-44000 Nantes, 2 UniversitFrance. é Paris-Sud, Université Paris-Saclay, Chimiothérapie Antiparasitaire, BIOmolécules: 2Conception, Université Isolement Paris-Sud, et Université Synthèse—BioCIS Paris-Saclay, UMR Chim 8076iothérapie CNRS, Antiparasitaire, F-92296 Châtenay-Malabry, BIOmolécules: Conception, France 3 SorbonneIsolement Universit et Synthèse—BioCISés, UPMC Paris UMR 06, 8076 CNRS CN USR3151RS, F-92296 “Protein Châtenay-Malabry, Phosphorylation France. and Human Disease” 3 group, Sorbonne Station Universités, Biologique, UPMC F-29680 Paris Rosco 06, ffCNRS, France USR3151 “Protein Phosphorylation and Human Disease” group, Station Biologique, F-29680 Roscoff, France. 4 Institut Pasteur and INSERM U1201, Unité de Parasitologie Moléculaire et Signalisation, F-75015 4 Institut Pasteur and INSERM U1201, Unité de Parasitologie Moléculaire et Signalisation, F-75015 Paris, Paris,France; France; e-mail: [email protected] [email protected] According to a recent report from the World Health Organization (WHO), leishmaniases— visceral leishmaniasis (VL), cutaneous leishmaniasis (CL) and mucocutaneous leishmaniasis (MCL) —collectively affect 12 million people in 98 countries, 350 million more are at risk of infection and 40,000 deaths are attributed to leishmaniases each year. Currently, there are no effective vaccines and a number of drugs are used in the treatment of these parasitic infections: pentavalent antimonials, amphotericin B, miltefosine, pentamidine, paromomycin and sitamaquine (Sangshetti, J.N., et al. RSC Adv. 2015, 5, 32376–32415). Unfortunately, most of them cause side effects and high toxicities, and an inevitable resistance has developed in recent times in Leishmania parasites. Consequently, there is an

Pharmaceuticals 2019, 12, 179 35 of 47

According to a recent report from the World Health Organization (WHO), leishmaniases— visceral leishmaniasis (VL), cutaneous leishmaniasis (CL) and mucocutaneous leishmaniasis (MCL) —collectively affect 12 million people in 98 countries, 350 million more are at risk of infection and 40,000 deaths are attributed to leishmaniases each year. Currently, there are no effective vaccines and a number of drugs are used in the treatment of these parasitic infections: pentavalent antimonials, amphotericin B, miltefosine, pentamidine, paromomycin and sitamaquine (Sangshetti, J.N., et al. RSCPharmaceuticals Adv. 2015 2019, 5,, 12 32376–32415)., x FOR PEER REVIEW Unfortunately, most of them cause side effects and high toxicities, 35 of 47 and an inevitable resistance has developed in recent times in Leishmania parasites. Consequently, thereurgent is anneed urgent to speed need toup speed the development up the development of a new of ageneration new generation of more of effective more eff ectiveand safe and safeantileishmanials. antileishmanials. InIn the the course course of ourof ongoingour ongoing synthetic synthetic and screening and screening programs programs for the obtention for the obtention of new biologically of new activebiologically imidazo active [1, 2- aimidazo] azines, we[1, decided2-a] azines, to develop we decided bioisostere to develop analogues bioisostere of the previously analogues described of the 2,3-diarylimidazo[1,2-previously described a2,3-diarylimidazo[1,2-]pyridines as antileishmaniala]pyridines agents as antileishmanial (Marhadour, S., agents et al. (Marhadour,Eur. J. Med. Chem.S., et 2012al. Eur., 58, J. 543–556). Med. Chem. 2012, 58, 543–556). TwoTwo promising promising analogues analogues were were highlightedhighlighted asas hithit compoundscompounds (Figure 18),18), exhibiting exhibiting very very good good inin vitro vitro and and in in vivo vivo activity activity associated associated withwith highhigh therapeutictherapeutic index, especially on on amastigote amastigote stage stage of of thethe parasite parasite (Marchand, (Marchand, P., P., et et al. al.Eur. Eur. J.J. Med.Med. Chem.Chem. 2015, 103, 381–395). 381–395).

H3C CH3 O N amastigotes IC = 0.8-12.8 μM (CL, VL) N N 50 NH amastigotes IC = 0.2-5.8 μM (CL, VL) N μ 50 F IC50 MRC-5 = 42.0 M IC MRC-5 = 93.2 μM N CK1 IC = 0.99 μM N N 50 Lm 50 F LmCK1 IC > 10 μM CK1 IC = 0.32 μM N 50 Hs 50

I N Figure 18. Structures of hit compounds. Figure 18. Structures of hit compounds. The mechanism of action involved in the antiparasitic properties will be discussed, as 4-pyridyl The mechanism of action involved in the antiparasitic properties will be discussed, as 4-pyridyl moiety at the C-3 position of the heterocyclic core was associated with L. major casein kinase 1 moiety at the C-3 position of the heterocyclic core was associated with L. major casein kinase 1 (LmCK1) inhibition, validated as a potential molecular target for antileishmanial drug development (LmCK1) inhibition, validated as a potential molecular target for antileishmanial drug development (Durieu, E., et al. Antimicrob. Agents Chemother. 2016, 60, 2822–2833). (Durieu, E., et al. Antimicrob. Agents Chemother. 2016, 60, 2822–2833). 7.24. Inhibiting X-Linked Inhibitor of Apoptosis Protein (XIAP), Design of New Non-Peptidic Foldamers of SMAC7.24. Inhibiting (P24) X-Linked Inhibitor of Apoptosis Protein (XIAP), Design of New Non-Peptidic Foldamers of SMAC (P24) Meziane Yahia-Ouahmed, Martin Giret, Kévin Antraygues, Marie Jouanne, Charline Kieﬀer and JanaMeziane Sopkova-de Yahia-Ouahmed, Oliveira Santos Martin Giret, Kévin Antraygues, Marie Jouanne, Charline Kieffer and Jana Sopkova-de Oliveira Santos Normandie Univ, UNICAEN, EA 4258 CERMN—FR CNRS INC3M, Caen, France Normandie Univ, UNICAEN, EA 4258 CERMN—FR CNRS INC3M, Caen, France The X-linked inhibitor of apoptosis protein (XIAP) binds to and inhibits caspases 3, 7 and 9, which The X-linked inhibitor of apoptosis protein (XIAP) binds to and inhibits caspases 3, 7 and 9, are some of the enzymes responsible for cell death, and therefore stopping apoptosis. XIAP is naturally which are some of the enzymes responsible for cell death, and therefore stopping apoptosis. XIAP is regulated (inhibited) by the mitochondrial protein SMAC/DIABLO. However, XIAP is overexpressed naturally regulated (inhibited) by the mitochondrial protein SMAC/DIABLO. However, XIAP is in tumorous cells (particularly in ovarian cancer) making them resist cell death. This is why XIAP overexpressed in tumorous cells (particularly in ovarian cancer) making them resist cell death. This makes a suitable drug target. is why XIAP makes a suitable drug target. The project aims at designing new non-peptidic inhibitors of XIAP in order to promote apoptotic The project aims at designing new non-peptidic inhibitors of XIAP in order to promote apoptotic cell death in ovarian tumours. This could be done by “mimicking” the AVPI tetra-peptide of SMAC cell death in ovarian tumours. This could be done by “mimicking” the AVPI tetra-peptide of SMAC (β strand) that is used by the latter to bind to the BIR3 domain of XIAP. (β strand) that is used by the latter to bind to the BIR3 domain of XIAP. First, a pharmacophore model was built on the basis of two ligands of XIAP-BIR3 found in the First, a pharmacophore model was built on the basis of two ligands of XIAP-BIR3 found in the literature. Then, a series of compounds were designed. The binding of these compounds to the active literature. Then, a series of compounds were designed. The binding of these compounds to the active site was studied by molecular docking. Molecular dynamics simulations were then performed to site was studied by molecular docking. Molecular dynamics simulations were then performed to assessassess the the stability stability of of the the ligands ligands inin thethe activeactive site.site. The best compounds were were synthetized synthetized and and tested tested in-vitroin-vitro by by ﬂuorescence fluorescence polarisationpolarisation assayassay (FPA).(FPA). TheThe mostmost promisingpromising candidates were were selected selected in in orderorder to to carry carry out out new new pharmacomodulations. pharmacomodulations.

7.25. Discovery of Novel pqs Inhibitors for Pseudomonas aeruginosa Infections (P25)

Rui-Ling Liu 1, Fadi Soukarieh 2, William Richardson 1, Shailesh N. Mistry 1, Miguel Cámara 2 and Michael J. Stocks 1

1 School of Pharmacy, Nottinghamshire NG7 2RD, United Kingdom. 2 School of Life Sciences, National Biofilms Innovation Centre, Centre for Biomolecular Sciences, University of Nottingham, Nottingham, Nottinghamshire NG7 2RD, United Kingdom; e-mail: [email protected]

Pharmaceuticals 2019, 12, 179 36 of 47

7.25. Discovery of Novel pqs Inhibitors for Pseudomonas aeruginosa Infections (P25) Rui-Ling Liu 1, Fadi Soukarieh 2, William Richardson 1, Shailesh N. Mistry 1, Miguel Cámara 2 and Michael J. Stocks 1 1 School of Pharmacy, Nottinghamshire NG7 2RD, UK 2PharmaceuticalsSchool of Life 2019,Sciences, 12, x FOR PEER National REVIEW Biofilms Innovation Centre, Centre for Biomolecular Sciences, 36 of 47 University of Nottingham, Nottingham, Nottinghamshire NG7 2RD, UK; [email protected] Current antibiotic treatments for P. aeruginosa infections have proven ineffective due to the increasingCurrent rate antibiotic of resistance treatments to them. for InhibitorsP. aeruginosa of theinfections pqs quorum have sensing proven signalling, ineffective key due for to thethe increasingcontrol of virulence, rate of resistance have been to them.regarded Inhibitors as a promising of the pqs alternativequorum sensing(Melissa, signalling, S., et al. PLoS. key forPathog the. control2014, 10, of e1004321). virulence, have been regarded as a promising alternative (Melissa, S., et al. PLoS. Pathog. 2014,In10 ,this e1004321). work, we report the discovery, design, synthesis and biological evaluation of a series of novelIn inhibitors this work, of we the report PqsR theregulatory discovery, protein, design, key synthesis for the activation and biological of the evaluation pqs system, of resulting a series ofin novelthe identification inhibitors of of the compound PqsR regulatory 2. The initial protein, hit keycompound for the activation1 (IC50 = 0.98 of theµM)pqs wassystem, obtained resulting through in thea virtual identification screening of performed compound on 2. ThePqsR initial ligand-binding hit compound domain 1 (IC using50 = 0.98 an in-houseµM) was compound obtained through library a(Figure virtual 19a). screening Exploration performed of compound on PqsR ligand-binding 1 gave the first domain generation using of an compounds in-house compound evaluated library using (Figureboth PAO1-L 19a). ExplorationmCTX: PpqsA- oflux compound and PA14 mCTX: 1 gave P thepqsA first-lux P. generation aeruginosa of pqs compounds bioreporter evaluated strains. Further using bothstructure PAO1-L activity mCTX: exploration, PpqsA-lux and to improve PA14 mCTX: potenc PpqsAy -andlux P.physiochemical aeruginosa pqs bioreporter properties, strains.gave a Furthersecond structuregeneration activity of compounds exploration, with toimproved improve inhibitory potency andactivity. physiochemical Notably, the properties,introduction gave of an a electron second generationdeficient tail of group compounds gave compound with improved 2 (IC inhibitory50 = 0.25 µM), activity. one of Notably, the most the potent introduction known ofpqs an inhibitors electron deficientto date. The tail crystallization group gave compound of compound 2 (IC50 2 =in0.25 PqsRµ M),ligand one binding of the most domain potent was known obtainedpqs inhibitors(Figure 19b) to date.and detailed The crystallization SAR studies of of compound this series 2 of in compounds PqsR ligand will binding be discussed domain was in the obtained poster. (Figure 19b) and detailed SAR studies of this series of compounds will be discussed in the poster.

(a) (b)

FigureFigure 19.19. General structurestructure ofof compoundscompounds andand PqsR.PqsR.

7.26.7.26. In Silico Screening andand DevelopmentDevelopment ofof SmallSmall MoleculeMolecule NeurotrophinNeurotrophin ReceptorReceptor LigandsLigands (P26)(P26)

MirjanaMirjana Antonijevic, Antonijevic, Ronan Ronan Bureau, Bureau, Patrick Patrick Dallemagne Dallemagne and and Christophe Christophe Rochais Rochais

CentreCentre d d0′EtudesEtudes et et de de Recherche Recherche sur surle Médicament le Médicament de Normandie de Normandie (CERMN), (CERMN), University University of Caen of Caen (UNICAEN),(UNICAEN), 1400014000 Caen, Caen, France; France; e-mail [email protected]: [email protected] Neurodegenerative diseases (ND), such as Alzheimer′s disease, Parkinson′s disease, multiple Neurodegenerative diseases (ND), such as Alzheimer0s disease, Parkinson0s disease, multiple sclerosissclerosis andand motormotor neuronneuron disease,disease, areare onon thethe riserise worldwide.worldwide. Currently, therethere isis nono curecure forfor anyany NDND andand mostmost of of the the available available drugs drugs fail tofail tackle to tackle ND pathogenesis. ND pathogenesis. Numerous Numerous studies have studies been have published been aboutpublished the role about of the neurotrophinthe role of receptorsthe neurotrophin in the pathogenesis receptors of several in the neurodegenerative pathogenesis of conditions several (Simmons,neurodegenerative D.A., et al. conditionsJ. Neurosci. (Simmons,2013, 33(48) D.A.,, 18712–27). et al. J. There Neurosci. are two 2013 types, 33(48) of neurotrophin, 18712–27). There receptors: are atwo non-enzymatic, types of neurotrophin trans-membrane receptors: protein a of non-enzy the tumourmatic, necrosis trans-membrane receptor (TNFR) protein family—p75, of the andtumour the tyrosinenecrosis kinasereceptor receptors (TNFR) (Trks)family—p75, A, B and and C. Trkthe receptorstyrosine kinase are activated, receptors speciﬁcally, (Trks) A, andB and with C. high Trk areceptorsﬃnity, by are nerve activated, growth factorspecifically, (NGF) and (TrkA), with brainderived high affinity, growth by nerve factor growth and neurotrophin factor (NGF) 4/5 (TrkA), (BDNF brainderived growth factor and neurotrophin 4/5 (BDNF and NT-4/5) (TrkB), and neurotrophin 3 (NT3) (TrkC), whereas p75 is activated by all mentioned neurotrophins (Meldolesi, J., et al. Pharmacol. Res. 2017, 121, 129–137). The implication of the TrkB receptor in the pathogenesis of several neurodegenerative conditions (Alzheimer′s, Parkinson′s, and Huntington′s disease) point to the therapeutic potential of TrkB agonists (Nomura, T., et al. J. Neurosci. 2017, 37(47), 11298–11310; Zheng, H., et al. J. Metab. Brain Dis. 2018, 33, 1961).

Pharmaceuticals 2019, 12, 179 37 of 47 and NT-4/5) (TrkB), and neurotrophin 3 (NT3) (TrkC), whereas p75 is activated by all mentioned neurotrophins (Meldolesi, J., et al. Pharmacol. Res. 2017, 121, 129–137). The implication of the TrkB receptor in the pathogenesis of several neurodegenerative conditions (Alzheimer0s, Parkinson0s, and Huntington0s disease) point to the therapeutic potential of TrkB agonists (Nomura, T., et al. J. Neurosci. 2017, 37(47), 11298–11310; Zheng, H., et al. J. Metab. Brain Dis. 2018, 33, 1961). After conducting in silico studies on the crystal structure of the TrkB receptor in order to determine the most important interactions between TrkB and its natural ligand—neurotrophin-4/5 (NT-4/5) (Figure 20a)—we performed in silico screening of the unique CERMN (Centre d’Etudes et de Recherche sur le Médicament de Normandie) library of compounds. The screening was done on the basis of the pharmacophore hypothesis (Figure 20b) developed from interactions of TrkB and its synthetic ligand—LM22A-4 (Massa, S.M., et al. J. Clin. Invest. 2010, 120, 1774–1785). On the basis of the main structural features of the LM22A-4, we designed a small dataset of the new potential TrkB agonists and conductedPharmaceuticals the 2019 synthesis, 12, x FOR of thePEER compounds REVIEW with best in silico results. 37 of 47

FigureFigure 20. 20.(a )(a Representation) Representation of of main main interactions interactions between betwee Asp298n Asp298 (tyrosine (tyrosine kinase kinase receptor receptor B: B: TrkB) TrkB) —green—green and Arg11and Arg11 (neurotrophin (neurotrophin (NT)-4/5)—blue; (NT)-4/5)—blue; (b) representation (b) representation of the developed of pharmacophorethe developed hypothesispharmacophore with its hypothesis features. with its features. This project received funding from the European Union s Horizon 2020 framework programme After conducting in silico studies on the crystal structure0 of the TrkB receptor in order to for research and innovation under grant agreement no. 765704. determine the most important interactions between TrkB and its natural ligand—neurotrophin-4/5 7.27.(NT-4/5) Isolation (Figure and Structure 20a)—we Elucidation performed of Aspergillus in silico scre Metabolitesening of Inducedthe unique with EpigeneticCERMN (Centre Modulators d'Etudes (P27) et de Recherche sur le Médicament de Normandie) library of compounds. The screening was done on 1,2 1 Mohammedthe basis of Aldholmi the pharmacophoreand A. Ganesan hypothesis (Figure 20b) developed from interactions of TrkB and its 1 Schoolsynthetic of Pharmacy,ligand—LM22A-4 University (Massa, of East S.M., Anglia, et al. Norwich J. Clin. Invest. Research 2010 Park,, 120 Norwich, 1774–1785). NR4 On 7TJ, the UK basis of 2 Naturalthe main Products structural and features Alternative of the Medicine, LM22A-4, College we designed of Clinical a small Pharmacy, dataset Imam of theAbdulrahman new potential BinTrkB Faisalagonists University, and conducted Dammam the 31441, synthesis Saudi of Arabia; the compounds [email protected] with best in silico results. This project received funding from the European Union′s Horizon 2020 framework programme Aspergillus for research andspecies innovation are rich under source grant of agreement bioactive metabolites.no. 765704. However, most natural product biosynthetic pathways are silent under laboratory conditions. The epigenetic processes, including the covalent7.27. Isolation modifications and Structure of DNA Elucidation and amino of acidsAsperg onillus histones, Metabolites have beenInduced revealed with Epigenetic to be extensively Modulators used by(P27) organisms to regulate the expression of genes involved in natural product biosynthesis (Felsenfeld, G., et al. Nature 2003, 421(6921), 448–453; Cichewicz, R. H. et al. Nat. Prod. Rep. 2010, 27(1), 11–22). 1,2 1 Therefore,Mohammed it is possibleAldholmi to influenceand A. Ganesan these processes during microbial fermentation by the addition of chemical1 School epigenetic of Pharmacy, modulators University (Cichewicz, of East R.Anglia, H, et al.NorwichNat. Prod.Research Rep .Park,2010 ,Norwich27(1), 11–22; NR4 Williams,7TJ, United R.B., etKingdom. al. Org. Biomol. Chem. 2008, 6(11), 1895–1897). 2 Five NaturalAspergillus Productsstrains and Alternative were cultivated Medicine, in theCollege presence of Clinical of Histone Pharmacy, Deacetylase Imam Abdulrahman Activity (HDAC) Bin Faisal inhibitorsUniversity, (e.g., vorinostat)Dammam 31441, or DNASaudi Arab methyltransferaseia; e-mail: [email protected] (DNMT) inhibitors (e.g., 5-azacytidine). MorphologicalAspergillus changes species were are monitoredrich source for of 7 bioactive days before metabolites. extraction withHowever, ethyl acetatemost natural or methanol. product Thebiosynthetic extracts were pathways analysed are bysilent HPLC under and laboratory LC-MS, and conditions. the metabolic The epigenetic profiles wereprocesses, compared including using the chromatogramcovalent modifications overlay and of volcanoDNA and plots. amino acids on histones, have been revealed to be extensively usedThe by overlayered organisms chromatograms to regulate the and expression volcano plotsof genes presented involved significant in natural diff erencesproduct between biosynthesis the cultures(Felsenfeld, treated G., with et al. epigenetic Nature 2003 modifiers, 421(6921), and control 448–453; cultures. Cichewicz, Two inducedR. H. et al. bioactive Nat. Prod. metabolites Rep. 2010 were, 27(1), 11–22). Therefore, it is possible to influence these processes during microbial fermentation by the addition of chemical epigenetic modulators (Cichewicz, R. H, et al. Nat. Prod. Rep. 2010, 27(1), 11–22; Williams, R.B., et al. Org. Biomol. Chem. 2008, 6(11), 1895–1897). Five Aspergillus strains were cultivated in the presence of Histone Deacetylase Activity (HDAC) inhibitors (e.g., vorinostat) or DNA methyltransferase (DNMT) inhibitors (e.g., 5-azacytidine). Morphological changes were monitored for 7 days before extraction with ethyl acetate or methanol. The extracts were analysed by HPLC and LC-MS, and the metabolic profiles were compared using chromatogram overlay and volcano plots. The overlayered chromatograms and volcano plots presented significant differences between the cultures treated with epigenetic modifiers and control cultures. Two induced bioactive metabolites were isolated from Aspergillus westerdijkiae and Aspergillus calidoustus, and their structures were elucidated using mass spectrometry and NMR. I would like to thank Imam Abdulrahman Bin Faisal University for their financial support.

7.28. Pharmacomodulation of Antiplasmodial Trichloromethylated-Heterocycles Using Ring Variation Strategy (P28)

Pharmaceuticals 2019, 12, 179 38 of 47 Pharmaceuticals 2019, 12, x FOR PEER REVIEW 38 of 47 isolatedDyhia Amrane from Aspergillus 1, Nicolas westerdijkiae Primas 1, Sébastienand Aspergillus Hutter 2 calidoustus, Nadine Azas, and 2 their, Pierre structures Verhaeghe were 3 and elucidated Patrice usingVanelle mass 1 spectrometry and NMR. I would like to thank Imam Abdulrahman Bin Faisal University for their financial support. 1 Aix-Marseille Univ, CNRS, ICR UMR 7273, PCR, Faculté de Pharmacie, 13385 Marseille. 2 Aix-Marseille Univ, IHU Méditerranée Infection, UMR VITROME, 13005 Marseille. 7.28.3 Pharmacomodulation of Antiplasmodial Trichloromethylated-Heterocycles Using Ring Variation Strategy Université (P28) Paul Sabatier, CNRS UPR 8241, LCC, 31077 Toulouse; e-mail: [email protected] Malaria is still the leading cause of death among parasitic infections worldwide (WHO, World Dyhia Amrane 1, Nicolas Primas 1,Sébastien Hutter 2, Nadine Azas 2, Pierre Verhaeghe 3 and Malaria report, 2018, http://www.who.int/malaria/publications/world-malaria-report- Patrice Vanelle 1 2018/report/en/). The emergence and the expansion of Plasmodium strains resistant to the artemisinin- 1 basedAix-Marseille combination Univ, therapies CNRS, ICR (ACTs) UMR are 7273, now PCR, threatening Faculté dethe Pharmacie, efficacy of 13385malaria Marseille treatment, notably 2 inAix-Marseille the Greater Mekong Univ, IHU sub-region. Méditerran Therefore,ée Infection, newUMR molecules VITROME, displaying 13005 the Marseille original mode of action 3 areUniversit urgentlyé Paul required. Sabatier, Aiming CNRS UPRat developing 8241, LCC, ne 31077w antiplasmodials, Toulouse; [email protected] our laboratory previously describedMalaria the is synthesis still the leading and the cause biological of death activities among parasiticof a library infections of 2-trichloromethylquinazoline worldwide (WHO, World Malariaderivatives, report, which2018 highlighted, http://www.who.int a hit molecule/malaria 1 (IC/publications50 = 0.4 µM,/ world-malaria-report-2018CC50 = 16 µM) (Cominetti, /M.report D. D.,/en et/). Theal. Org. emergence Biomol. andChem. the 2016 expansion, 14, 10161–10164; of Plasmodium Verhaeghe,strains resistant P., et al. to Bioorg. the artemisinin-based Med. Chem. 2009 combination, 17, 4313– therapies4322)(Figure (ACTs) 21). are Moreover, now threatening a scaffold-hopping the efficacy of malariastrategy treatment, showed notablythat the in replacement the Greater Mekong of the sub-region.quinazoline Therefore,moiety by a new quinoxaline molecules one displaying improved the the original cytotoxicity mode profile of action (Desroches. are urgently J., et required.al. Eur J. AimingMed. Chem. at developing 2017, 125, new68–86). antiplasmodials, Thus, we synthetized our laboratory a new previouslyseries of 2-trichloromethylquinoxaline described the synthesis and theanalogues. biological The activities in vitro of biological a library ofeval 2-trichloromethylquinazolineuations against the multi-resistant derivatives, K1 P. which falciparum highlighted strain highlighted two new hit molecules substituted by an electron-withdrawing group in para-position. a hit molecule 1 (IC50 = 0.4 µM, CC50 = 16 µM) (Cominetti, M. D. D., et al. Org. Biomol. Chem. 2016Accordingly, 14, 10161–10164; with theseVerhaeghe, results, we P., prepared et al. Bioorg. new 2-trichloromethylquinoxaline Med. Chem. 2009, 17, 4313–4322) analogues (Figure bearing 21). Moreover,other electron-withdrawing a scaffold-hopping groups strategy such showed as -CF3 that, -SF the5 or replacement-OCF3. of the quinazoline moiety by a quinoxalineIn parallel, one to improved complete the the cytotoxicity global SAR profilestudy us (Desroches.ing the scaffold J., et hopping al. Eur J. approach, Med. Chem. we2017 replaced, 125, 68–86).the quinoxaline Thus, we ring synthetized with a phthalazine a new series one. of 2-trichloromethylquinoxalineThe importance for the antiplasmodial analogues. activity The in of vitro the biologicalphenyl ring evaluations contained againstin the bicyclic the multi-resistant quinazoline K1andP. quinoxaline falciparum strain scaffolds highlighted was also two studied new hitby moleculesstructural simplification, substituted by leading an electron-withdrawing to the related pyrimidine group in andpara pyrazine-position. analogues. Accordingly The with synthesis these results,details of we the prepared new analogues new 2-trichloromethylquinoxaline of hit 1 in various series and analogues the biological bearing otherresults electron-withdrawing will be described in the poster. groups such as -CF3, -SF5 or -OCF3.

Figure 21. Ring variation strategy adopted in order to explore SARs. Figure 21. Ring variation strategy adopted in order to explore SARs. In parallel, to complete the global SAR study using the scaffold hopping approach, we replaced the This work was supported by the ANR NINTARMAL project, grant ANR-17-CE11–0017. quinoxaline ring with a phthalazine one. The importance for the antiplasmodial activity of the phenyl ring contained in the bicyclic quinazoline and quinoxaline scaffolds was also studied by structural 7.29. Optimisation of Peptide Linker-Based Fluorescent Ligands for Histamine H1 Receptor (P29) simplification, leading to the related pyrimidine and pyrazine analogues. The synthesis details of the newZhi Yuan analogues Kok 1 of, Shailesh hit 1 in various N. Mistry series 1, Stephen and the biologicalJ. Hill 2 and results Barrie will Kellam be described 1 in the poster. 1 This School work of Pharmacy, was supported University by of the No ANRttingham, NINTARMAL NG7 2RD, United project, Kingdom. grant ANR-17-CE11–0017. 2 School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, United Kingdom; e-mail: 7.29. Optimisation of Peptide Linker-Based Fluorescent Ligands for Histamine H1 Receptor (P29) [email protected] Zhi Yuan Kok 1, Shailesh N. Mistry 1, Stephen J. Hill 2 and Barrie Kellam 1 The histamine H1 receptor (H1R) is a class A GPCR and is widely expressed throughout the 1humanSchool body. of Pharmacy, H1R-dependent University signalling of Nottingham, is responsible NG7 2RD,for allergic UK reactions, plays an important role 2inSchool immunomodulation of Life Sciences, University and, as ofrecent Nottingham, studies Nottingham have suggested, NG7 2UH, is UK;involved [email protected] in mediating cell proliferation and thus cancer progression (Panula, P., et al. Pharmacol Rev. 2015, 67 (3), 601–655). High affinity H1R fluorescent ligands with optimum physicochemical properties would serve as a valuable

Pharmaceuticals 2019, 12, 179 39 of 47

The histamine H1 receptor (H1R) is a class A GPCR and is widely expressed throughout the human body. H1R-dependent signalling is responsible for allergic reactions, plays an important role in immunomodulation and, as recent studies have suggested, is involved in mediating cell proliferation and thus cancer progression (Panula, P., et al. Pharmacol Rev. 2015, 67 (3), 601–655). High affinity H1R fluorescent ligands with optimum physicochemical properties would serve as a valuable tool to probe in further detail H1R cancer pharmacology and drug discovery (McGrath, J. C., et al. The Adrenergic Receptors, Springer: 2006; pp 151–172; Vernall, A. J., et al. British J. Pharmacol. 2014, 171 (5), 1073–1084). Fluorescent ligands comprise a receptor binding element (either an orthostere or allostere) linker connect to an appropriate fluorophore. Whilst modulating the overall physicochemical properties of the fluorescent ligand, a peptide-based linker could engage in receptor binding through its side chain functional groups, potentially improving binding affinity of the overall conjugate and providing information on SAR along the receptor exit tunnel. The crystal structure of H1R in complex with the antagonist doxepin was reported in 2011, providing detailed information of the orthosteric binding site and revealing an additional binding site lined by the basic amino acids of Lys179, Lys191 and His450, which could be targeted to improve ligand affinity (Shimamura, T., et al. Nature 2011, 475 (7354), 65–70). We envisaged optimising the peptide linker for a previously reported H1R fluorescent ligand (Stoddart, L. A., et al. Scientific Reports 2018, 8 (1), 1572) with pKD of 8.1 by sequential screening for amino acids along the peptide chain in order to identify those that could provide the greatest improvement in binding affinity. By incorporating these amino acids into the linker, we synthesised H1R fluorescent ligands with pKD0s ranging from 7.8 to 8.4. Molecular docking of these ligands into an H1R crystal structure suggested the existence of small binding pockets along the receptor exit tunnel that were exploited by the functionalised peptide linkers. Further experimentation with fluorescent ligands of varying fluorophores suggested that the attachment of the boron-dipyrromethene BODIPY630/650 fluorophore was the main driving force in binding affinity in H1R fluorescent ligands. Despite not being able to significantly improve binding affinities of H1R fluorescent ligands, optimisation of the peptide linker provided information on SAR along the receptor exit tunnel.

7.30. First Design of MT5-MMP Inhibitors, a Challenging New Target for Alzheimer0s Disease (P30) Pauline Zipfel 1, Julien Lalut 1, Camille Denis 1, Ronan Bureau 1, Peggy Suzanne 1, Audrey Davis 1, Aurélie Malzert-Fréon 1, Kevin Baranger 2, Michel Khrestchatisky 2, Santiago Rivera 2, Christophe Rochais 1 and Patrick Dallemagne 1

1 Centre d0Etudes et de Recherche sur le Médicament de Normandie (CERMN), Normandie Univ, UNICAEN, 14000 Caen, France 2 Institute of Neuropathophysiology (INP), UMR7051, CNRS, Aix Marseille Université, France; [email protected] In 2018, the number of people living with dementia in the world was estimated at 50 million, and Alzheimer0s disease (AD) is the most common form of dementia. AD is a neurodegenerative and incurable brain disorder; only treatments for symptoms are available at this time. Because of the heavy economic and societal impacts, there is an urgent need to find new treatments that target the molecular causes of neuronal cell death. Two abnormal structures in the brain called β-amyloid (Aβ)-containing plaques and neurofibrillary tangles are considered as two of the main features of AD. In this context, several studies support the hypothesis that alterations in the processing of amyloid precursor protein (APP) contributes to AD pathogenesis, resulting in the accumulation of β-amyloid peptides (Aβ) and other proteolytic products. Thus, current research focuses on the enzymes involved in APP cleavage such as α-, β- and γ-secretases. However, recent studies have revealed the existence of another physiological APP processing pathway, mediated by a novel AD-related enzyme, membrane-type 5-matrix metalloproteinase (MT5-MMP), which can process APP and promote Aβ accumulation, as well as the inflammatory process in AD transgenic mice (Baranger, K., et al. Cell. Mol. Life Sci. Pharmaceuticals 2019, 12, 179 40 of 47

2016, 73(1), 217–236); Baranger, K. et al. J. Neuroinflammation 2016, 13(1), 167–170); Baranger, K., et al., Front. Mol. Neurosci. 2017, 9, 1–17). Moreover, MT5-MMP can cleave APP upstream from the β-secretase cleavage site (the so called η-cleavage site) (Ahmad, M. et al., J. Biochem. 2006, 139(3), 517–526) and release an N-terminally elongated Aβ fragment (Aη-α), which appears to be synaptotoxic (Willem, M., et al. Nature 2015, 526(7573), 443–447). We aim to design and synthesize first selective MT5-MMP inhibitors through an interdisciplinary approach including molecular modelling, medicinal chemistryPharmaceuticals and 2019 biology., 12, x FOR Starting PEER REVIEW from a cyclopentathiophene derivative identified by in silico screening, 40 of 47 we are currently investigating the pharmacomodulations on this hit compound to gain in affinity and identified by in silico screening, we are currently investigating the pharmacomodulations on this hit selectivity for MT5-MMP. compound to gain in affinity and selectivity for MT5-MMP.

Drug Discovery strategy

7.31. Design, Synthesis and EvaluationEvaluation of Quinolines for Drug-Resistant Malaria (P31) Rachael MagwazaMagwaza1, Buthaina1, Buthaina Hussein Hussein1, Muna 1, Muna Abubaker Abubaker2, Sally 2 Freeman, Sally Freeman1 and Niroshini 1 and NirmalanNiroshini2 Nirmalan 2 1 Department of Pharmacy and Optometry, University of Manchester, Manchester M13 9PL, UK 21 Department Department of life of sciences, Pharmacy University and Optometry, of Salford, Universi Salford M5ty of 4WT,UK; Manchester, [email protected] Manchester M13 9PL, United Kingdom. 2 Malaria Department is anofinfectious life sciences, disease University caused of by Salf theord, parasite Salford of theM5 genus4WT, PlasmodiumUnited Kingdom;with humane-mail: [email protected] caused by species falciparum , malaria, knowlesi, vivax and ovale (De Koning-ward, T. F., et al. Nat. Publ. Gr. 2016, 14, 494–507). P. falciparum is associated with the most severe form of Malaria is an infectious disease caused by the parasite of the genus Plasmodium with human malaria, responsible for 400,000 deaths/year, especially in Africa. A number of antimalarial drugs infection caused by species≈ falciparum, malaria, knowlesi, vivax and ovale (De Koning-ward, T. F., et al. are currently used to treat malaria. However, P. falciparum has developed resistance to all currently Nat. Publ. Gr. 2016, 14, 494–507). P. falciparum is associated with the most severe form of malaria, used medicines, hence, there is a great need for the development of new drugs. Here, we report responsible for ≈400,000 deaths/year, especially in Africa. A number of antimalarial drugs are novel quinolines as antimalarial leads. The quinolines were originally designed to be inhibitors of currently used to treat malaria. However, P. falciparum has developed resistance to all currently used NRH—quinone oxidoreductase 2 (NQO2), a potential therapeutic target in cancer chemotherapy medicines, hence, there is a great need for the development of new drugs. Here, we report novel (Alnabulsi, S., et al. Bioorg. Med. Chem. Lett. 2018; 9, 1–6). P. falciparum contains a functionally quinolines as antimalarial leads. The quinolines were originally designed to be inhibitors of NRH— similar type II NADH/dehydrogenase known as Pf NDH2; therefore, these compounds were tested quinone oxidoreductase 2 (NQO2), a potential therapeutic target in cancer chemotherapy (Alnabulsi, against Plasmodium falciparum as antimalarial leads. The quinolines were identiﬁed as a potent in vitro S., et al. Bioorg. Med. Chem. Lett. 2018; 9, 1–6). P. falciparum contains a functionally similar type II inhibitor of the multi-drug-resistant strain K1 of P. falciparum (with EC ranging from 0.5–20 µM). NADH/dehydrogenase known as PfNDH2; therefore, these compounds50 were tested against The EC speed assays carried out to verify onset of antimalarial activity showed that the compounds Plasmodium50 falciparum as antimalarial leads. The quinolines were identified as a potent in vitro were fast-acting, as the EC50 values were achieved at 24 h. Cell cytotoxicity assays showed that the inhibitor of the multi-drug-resistant strain K1 of P. falciparum (with EC50 ranging from 0.5–20 µM). compounds were very toxic towards HepG2 hepatic cancer cell lines seeded at 4000 cells/well, with low The EC50 speed assays carried out to verify onset of antimalarial activity showed that the compounds selectivity indices (SI 0.6–2.7). The compounds were screened further against MDBK bovine kidney were fast-acting, as the EC50 values were achieved at 24 h. Cell cytotoxicity assays showed that the cells, which showed improved selectivity indices (SI 1.6 > 10). To achieve further dose reduction and compounds were very toxic towards HepG2 hepatic cancer cell lines seeded at 4000 cells/well, with to minimize toxicity, Calcusyn-based drug interactivity assays are ongoing. Further studies are in low selectivity indices (SI 0.6–2.7). The compounds were screened further against MDBK bovine kidney cells, which showed improved selectivity indices (SI 1.6 > 10). To achieve further dose reduction and to minimize toxicity, Calcusyn-based drug interactivity assays are ongoing. Further studies are in progress to characterize stage specificity, mechanism of action, cytotoxicity profiles and drug interaction with existing antimalarial drugs.

7.32. Design, Synthesis and Biological Evaluation of New Fluorinated Indazole Derivates as Potential New 5-HT4R PET Radiotracers (P32)

Reynald Mangeant 1, Silvia Stiebing 1, Thomas Cailly 1,2, Audrey Davis 1, Frederic Fabis 1 and Valerie Collot 1

Pharmaceuticals 2019, 12, 179 41 of 47 Pharmaceuticals 2019, 12, x FOR PEER REVIEW 41 of 47 progress1 Normandie to characterize univ, UNICAEN, stage specificity, CERMN, mechanism 14000 Caen, of France. action, cytotoxicity profiles and drug interaction with2 existing Department antimalarial of Nuclear drugs. Medecine, CHU Côte de Nacre, 14000 Caen, France; e-mail: [email protected] 7.32. Design, Synthesis and Biological Evaluation of New Fluorinated Indazole Derivates as Potential New 5-HT4SinceR PET its Radiotracers discovery (P32) in 1988, the serotonin 4 receptor subtype (5-HT4R) has emerged as a promising target for drug discovery and development, resulting from their implications in learning, Reynald Mangeant 1, Silvia Stiebing 1, Thomas Cailly 1,2, Audrey Davis 1, Frederic Fabis 1 and cognition, memory processes and many neuropsychiatric disorders such as Alzheimer′s disease, Valerie Collot 1 anxiety, depression or anorexia nervosa (Bockaert, J., et al. Neuropharmacology 2008, 55 (6), 922–931). 1 Thus,Normandie discovery univ, of UNICAEN, active 5-HT CERMN,4R agonists 14000 and Caen, antagonists France remains a continuing interest in clinical 2 research.Department To ofthis Nuclear end, positron Medecine, emission CHU Cô tomographyte de Nacre, 14000 (PET) Caen, (Marner, France; L., [email protected] et al. Neuroimage 2010, 50 (3), 855–861; Caillé, F., et al. Bioorg. Med. Chem. Lett. 2013, 23 (23), 6243–6247) coupled with effective Since its discovery in 1988, the serotonin 4 receptor subtype (5-HT4R) has emerged as a promising targetradioligands for drug constitutes discovery a and valuable development, tool, both resulting in clinical from studies their and implications drug discovery in learning, programmes. cognition, On the basis of previous works in CERMN (Lam, B. V., et al. Chem. Eur. J. 2016, 22 (13), 4440–4446), we memory processes and many neuropsychiatric disorders such as Alzheimer0s disease, anxiety, depressionaimed to develop or anorexia new nervosafluorinated (Bockaert, indazole J., derivates et al. Neuropharmacology as potential brain2008 5-HT, 554R (6) PET, 922–931). tracers (Figure Thus, 22). discovery of active 5-HT4R agonists and antagonists remains a continuing interest in clinical research. To thisThis end, synthesis positron was emission carried tomography out in three (PET)essential (Marner, steps. L.,A convergent et al. Neuroimage synthesis2010 pathway, 50 (3), 855–861;to obtain Caillcoldé ,ligands F., et al. (CL)Bioorg. was Med. established. Chem. Lett. A methodology2013, 23 (23) ,allowing 6243–6247) selective coupled functionalization with effective radioligands at position 3 constitutesleading to apolyfunctional valuable tool, bothindazoles in clinical in a studies few steps and drugwas developed. discovery programmes. New pharmacomodulations On the basis of previousstudies were works realized in CERMN in order (Lam, to B. increase V., et al. receptorChem. Eur. affinity, J. 2016 decrease, 22 (13), lipophilicity 4440–4446), weand aimed increase to metabolic resistance. develop new fluorinated indazole derivates as potential brain 5-HT4R PET tracers (Figure 22).

Figure 22. Essential steps to obtain radiotracers. CL: cold ligands, PET: positron emission tomography. Figure 22. Essential steps to obtain radiotracers. CL: cold ligands, PET: positron emission This synthesis was carried out in three essentialtomography. steps. A convergent synthesis pathway to obtain cold ligands (CL) was established. A methodology allowing selective functionalization at position 3 leadingMore to polyfunctionalthan 20 compounds indazoles with na innomolar a few steps affinity was were developed. synthesized. New One pharmacomodulations of them, MR35806, is studiescurrently were radiolabeled realized inwith order fluorine to increase 18 to obtain receptor our first affinity, brain decrease 5-HT4R PET lipophilicity tracer. This and compound increase metabolicwill be tested resistance. in vivo on mouse brain. MoreThe authors than 20 are compounds grateful for with the nanomolarfinancial support affinity by were regional synthesized. council of One Normandy, of them, FEDER MR35806, and isCrunch currently Network. radiolabeled with fluorine 18 to obtain our first brain 5-HT4R PET tracer. This compound will be tested in vivo on mouse brain. 7.33.The Acetoxystachybotrydial authors are grateful Acetate, for the a financial Natural Comp supportound by Isolated regional from council Stachybotrys of Normandy, Chartarum FEDER is a and CrunchPotent Inhibitor Network. of Human Protein Casein Kinase 2 (CK2) (P33) Samer Haidar 1, Franziska M Jürgens 2, Dagmar Aichele 1, Annika Jagels 2, Hans-Ulrich Humpf 2 7.33. Acetoxystachybotrydial Acetate, a Natural Compound Isolated from Stachybotrys Chartarum is a Potent and Joachim Jose 1 Inhibitor of Human Protein Casein Kinase 2 (CK2) (P33) 1 Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, und 2Institut für Samer Haidar 1, Franziska M Jürgens 2, Dagmar Aichele 1, Annika Jagels 2, Hans-Ulrich Humpf 2 and Lebensmittelchemie,1 Westfälische Wilhelms-Universität Münster, 48149 Münster, Germany. Joachim2 Institute Jose of Food Chemistry, Westfälische Wilhelms-Universität Münster, 48149 Münster, Germany; e-mail: [email protected]

Pharmaceuticals 2019, 12, 179 42 of 47

1 Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, und 2Institut für Lebensmittelchemie, Westfälische Wilhelms-Universität Münster, 48149 Münster, Germany Pharmaceuticals 2019, 12, x FOR PEER REVIEW 42 of 47 2 Institute of Food Chemistry, Westfälische Wilhelms-Universität Münster, 48149 Münster, Germany; [email protected] protein kinase CK2 is an emerging target for drug design. Overexpression of CK2 is closelyHuman related proteinto many kinase types CK2 of human is an emerging cancer. It target has been for drug shown design. that elevated Overexpression levels of of CK2 CK2 protect is closely tumourrelated cells to many from typesapoptosis of human (Faust cancer. M, Montenarh It has been M. shownCell Tissue that Res elevated 2000, levels301, 329–340). of CK2protect Inhibition tumour of CK2cells activity from apoptosiscan lead tumour (Faust M, cells Montenarh into apoptosis, M. Cell whereas Tissue Res viability2000, 301of healthy, 329–340). cells Inhibition is not affected of CK2 (Ruzzene,activity can M., lead et al. tumour Biochim. cells Biophys. into apoptosis, Acta 2010 whereas, 1804, 499–504). viability ofUp healthy until now, cells isone not ATP affected competitive (Ruzzene, inhibitorM., et al. ofBiochim. CK2, silmitasertib, Biophys. Acta is 2010in phase-II, 1804, 499–504).clinical trials Up until(Gowda, now, C., one et ATPal. Curr. competitive Pharm. Des. inhibitor 2017, of 23CK2,, 95–107). silmitasertib, Here, iswe in phase-IIreport clinicalon the trials screening (Gowda, C.,of etnatural al. Curr. compounds Pharm. Des. 2017isolated, 23, 95–107).from StachybotryschartarumHere, we report on the(Gratz, screening A., et al. of Electrophoresis natural compounds 2010, 31 isolated, 634–640) from. TwelveStachybotryschartarum phenylspirodrimanes(Gratz, andA., three et al. triphenylphenolesElectrophoresis 2010 were, 31, 634–640).investigated Twelve on inhibitory phenylspirodrimanes activity towards and threeCK2 using triphenylphenoles a capillary electrophoresis-basedwere investigated on assay inhibitory (Jagels, activity A., towardset al. Mycotoxin CK2 using aRes. capillary 2018, electrophoresis-based34, 179–185). Triphenyl assay phenolestachybotrychromene(Jagels, A., et al. Mycotoxin Res.C, 2018phenylsp, 34, 179–185).irodrimanes Triphenyl stachybotrydial phenolestachybotrychromene acetate and C, acetoxystachybotrydial acetate with IC50 values of 0.3 µM, 0.7 µM and 1.9 µM, respectively, were phenylspirodrimanes stachybotrydial acetate and acetoxystachybotrydial acetate with IC50 values of identified0.3 µM, 0.7as µpotentM and CK2 1.9 µinhibitorsM, respectively, (Figure were 23). identifiedThe effect asof potentthese compounds CK2 inhibitors on (Figurethe proliferation 23). The e ffofect breastof these cancer compounds cells MCF-7 on the was proliferation determined of breastusing canceran EdU cells (5-ethynyl-2’-deoxyuridine) MCF-7 was determined using assay. an For EdU comparison,(5-ethynyl-2’-deoxyuridine) viability of breast assay. cancer For cells comparison, is shown below. viability of breast cancer cells is shown below.

Acetoxystachybotrydial acetate Stachybotrychromene C

FigureFigure 23. 23. 2D 2Drepresentation representation of ofthe theinteraction interaction between between the the ATP ATP binding binding site site of ofCK2 CK2 with with acetoxystachybotrydialacetoxystachybotrydial acetate acetate and and stachybotrychromene stachybotrychromene C. C.

CancerCancer cells cells MCF-7 MCF-7 as aswell well as aslung lung cancer cancer cell cellss A427 A427 and and epidermal epidermal cancer cancer cells cells A A431 431 were were testedtested using using MTT MTT assay. assay. In particular, In particular, acetoxystachybot acetoxystachybotrydialrydial acetate acetate turned turned out to be out the to most be the active most compound.active compound. After treatment After treatment of MCF-7 of cells MCF-7 with cells 1 µM with for 124µ Mh, cell for 24proliferation h, cell proliferation was blocked was almost blocked completelyalmost completely (99%), whereas (99%), whereascell viability cell viability was decreased was decreased only onlyby by63%. 63%. Here, Here, we we describe describe phenlyspirodrimanesphenlyspirodrimanes as asa new a new class class of of CK2 CK2 inhibitors, inhibitors, among among them them acetoxystachybotrydial acetoxystachybotrydial acetate acetate whichwhich has has been been proven proven to to be be the the most most potent potent representative representative of of this this series. series.

7.34.7.34. Synthesis Synthesis and and Biological Biological Eval Evaluationuation of ofNew New Indolo Indolo [2, [2, 3-b] 3-b] Qu Quinolineinoline Derivatives Derivatives as asPotential Potential Antimalarial Candidates (P34) Antimalarial Candidates (P34) Ludovic Doudet 1, Ingrid Allart-Simon 1, Alexandre Maciuk 2, Janos Sapi 1 and Stéphane Gérard 1 Ludovic Doudet 1, Ingrid Allart-Simon 1, Alexandre Maciuk 2, Janos Sapi 1 and Stéphane Gérard 1 1 Université de Reims Champagne-Ardenne, Institut de Chimie Moléculaire de Reims (ICMR), UMR 1 Université de Reims Champagne-Ardenne, Institut de Chimie Moléculaire de Reims (ICMR), UMR CNRS CNRS7312, 7312, UFR UFRPharmacie, Pharmacie, 51 rue 51Cogn rueacq-jay, Cognacq-jay, 51096 Reims, 51096 France. Reims, France 2 2 Universit Universitéé dede ParisParis Sud,Sud, BioCIS UMR UMR CNRS CNRS 8076, 8076, UFR UFR Pharmacie, Pharmacie, 92296 92296 Chât Chenay-Malabry,âtenay-Malabry, France; France; e- [email protected]: [email protected] The indolo [2, 3-b] quinoline system is present in many marine alkaloids that have demonstrated interesting biological activity (for a review on biological activities, see: Lavrado, J., et al. Curr. Med. Chem. 2010, 17, 2348–2370). Smiles rearrangement, an organic intramolecular nucleophilic aromatic substitution reaction, provides a powerful access to the synthesis of a variety of heterocyclic compounds (Figure 24). Taking advantage of the combination of Smiles rearrangement and radical chemistry, preparation of new indolo [2, 3-b] quinoline derivatives and their biological evaluation

Pharmaceuticals 2019, 12, 179 43 of 47

The indolo [2, 3-b] quinoline system is present in many marine alkaloids that have demonstrated interesting biological activity (for a review on biological activities, see: Lavrado, J., et al. Curr. Med. Chem. 2010, 17, 2348–2370). Smiles rearrangement, an organic intramolecular nucleophilic aromatic substitutionPharmaceuticals reaction,2019, 12, x providesFOR PEER aREVIEW powerful access to the synthesis of a variety of heterocyclic compounds 43 of 47 (Figure 24). Taking advantage of the combination of Smiles rearrangement and radical chemistry, preparationwill be presented of new (Allart-Simon, indolo [2, 3-b] quinoline I., et al. Molecules derivatives 2016 and, 21 their, 878–889; biological Pudlo, evaluation M., et willal. Chem. be presented Comm.

(Allart-Simon,2012, 48, 2442–2444). I., et al. Molecules 2016, 21, 878–889; Pudlo, M., et al. Chem. Comm. 2012, 48, 2442–2444).

Figure 24. Strategy for the synthesis of a variety of heterocyclic compounds. Figure 24. Strategy for the synthesis of a variety of heterocyclic compounds 7.35. Isobile Acids, Their Toxicity and Eﬀect on Activation of farnesoid X receptor (FXR) (P35) Yin7.35. Lu Isobile1, Sandra Acids, Van Their de Toxicity Wiel 2 and and JohnEffect Gilmer on Activation1 of farnesoid X receptor (FXR) (P35) 1YinSchool Lu 1, of Sandra Pharmacy Van and de Wiel Pharmaceutical 2 and John Sciences, Gilmer 1 Trinity College Dublin, D02 PN40 Dublin 2, Ireland 2 a Tytgat School Institute of Pharmacy for and Liver Pharmaceut and Intestinalical Sciences, Research, Trinity College Department Dublin, D02 of PN40 Gastroenterology Dublin 2, Ireland. and Hepatology,b Tytgat Institute Amsterdam for Liver Gastroenterology and Intestinal andResear Metabolism,ch, Department Academic of Gastroenterology Medical Center, and Amsterdam, Hepatology, The Netherlands;Amsterdam Gastroenterology [email protected] and Metabolism, Academic Medical Center, Amsterdam, The Netherlands; email: [email protected] Bile acids have conventionally been thought of as mere digestive surfactants that assist in the absorptionBile acids of fats have and conventionally fat soluble vitamins. been thought In the past of as two mere decades, digestive it has surfactants emerged that that bile assist acids in play the importantabsorption functionsof fats and as fat signalling soluble moleculesvitamins. In in the mammalian past two physiologydecades, it has and emerged that, for example,that bile acids they canplay regulate important bile functions acid metabolism. as signalling FXR, molecules a member in ofmammalian nuclear receptor physiology superfamily, and that, plays for example, a crucial rolethey incan regulating regulate bilebileacid acid homeostasis metabolism. (Kullak-Ublick, FXR, a member G.A., of nuclear et al. Physiology receptor. superfamily,2008, 23(5), 286–295) plays a andcrucial an role abundance in regulating of evidence bile acid suggests homeostasis that FXR (Kullak-Ublick, may be a useful G.A., target et al. Physiology in the prevention. 2008, 23(5), of colon 286– carcinogenesis295) and an abundance (Modica, S.,of evidence et al. Cancer suggests Res. 2008 that, 23FXR, 9589–9595). may be a Ituseful can be target hypothesised in the prevention that isobile of acidscolon (thecarcinogenesis 3β-OH isomers (Modica, of the S., normally et al. Cancer occurring Res. 2008 bile acids), 23, 9589–9595). can activate It thecan FXR be hypothesised and regulate FXRthat functionisobile acids through (the their 3β-OH presence isomers in theof bowelthe normally and isobile occurring acids can bile modify acids) thecan toxicity activate of the bileFXR acids and (Figuresregulate 25FXR and function 26). through their presence in the bowel and isobile acids can modify the toxicity of theWe bile explored acids (Figures new synthetic 25 and approaches26). for producing isobile acids from natural bile acids, and a panelWe of isobileexplored acids new and synthetic normal approaches bile acids were for producing prepared inisobile order acids to help from in thenatural characterization bile acids, and of theira panel eﬀ ectof isobile on cellular acids toxicity and normal and on bile FXR acids activation. were prepared in order to help in the characterization of theirWe effect reached on cellular the conclusion toxicity thatand amongon FXR the activation. natural bile acids, CDCA was found to be the most toxic, followed by LCA, CA and DCA. UDCA was least toxic, and this was as expected from many studies on the relative toxicity of the bile acids, corresponding to the order of hydrophobicity of the

Pharmaceuticals 2019, 12, 179 44 of 47 bile acids. In general, the beta compounds (isobile acids) were less toxic than the alpha compounds. Only in the case of LCA was there no difference in the toxicity of the epimeric pairs. Only isoCDCA was found as a true FXR agonist on the basis of FXR FRET assay, whereas CDCA showed no significant Pharmaceuticals 2019, 12, x FOR PEER REVIEW 44 of 47 effPharmaceuticalsect in FRET, 2019 which, 12, x was FOR surprising.PEER REVIEW 44 of 47

Figure 25. A measurement of toxicity of different bile acids. Caco-2 cells were treated with 100 µM FigureFigure 25. 25.A A measurement measurement of of toxicity toxicity of diofﬀ differenterent bile bile acids. acids. Caco-2 Caco-2 cells cells were were treated treated with with 100 µ 100M bile µM bile acid solution for 24 h, and cells were then treated with 10 μl of 2.5 mg/ml MTT reagent and acidbile solution acid solution for 24 h,for and 24 cellsh, and were cells then were treated then with treated 10 µ withl of 2.5 10 mg μl /mlof 2.5 MTT mg/ml reagent MTT and reagent incubated and incubated for 2 h. forincubated 2 h. for 2 h.

FigureFigure 26. 26. SchematicSchematic representation representation the the mechan mechanismism of of BAS BAS upon upon FXR FXR activation. activation. Figure 26. Schematic representation the mechanism of BAS upon FXR activation. We are most grateful to Dr. John O Brien for NMR spectroscopy, Gary Hessman and Brian Talbot We reached the conclusion that among0 the natural bile acids, CDCA was found to be the most for MSWe spectroscopy reached the and conclusion Tarek Moustafa that among for his the collaboration natural bile inacids, the FXRCDCA FRET was assay. found to be the most toxic, followed by LCA, CA and DCA. UDCA was least toxic, and this was as expected from many toxic, followed by LCA, CA and DCA. UDCA was least toxic, and this was as expected from many studies on the relative toxicity of the bile acids, corresponding to the order of hydrophobicity of the 7.36.studies Synthesis on the and relative Biological toxicity Activities of the of bile New acids, 2-Mercapto-(2-Oxoindolin-3-ylidene)acetonitrile corresponding to the order of hydrophobicity of the bileDerivatives acids. In (P36) general, the beta compounds (isobile acids) were less toxic than the alpha compounds. bile acids. In general, the beta compounds (isobile acids) were less toxic than the alpha compounds. Only in the case of LCA was there no difference in the toxicity of the epimeric pairs. Only isoCDCA BorisOnly Letribot, in the case Régis of LCA Delatouche, was there Jean-Ren no differenceé Chérouvrier, in the toxicity Lisianne of Domon the epimeric and Val pairs.érie ThiOnlyéry isoCDCA was found as a true FXR agonist on the basis of FXR FRET assay, whereas CDCA showed no was found as a true FXR agonist on the basis of FXR FRET assay, whereas CDCA showed no significantLa Rochelle effect UniversityLIENSs in FRET, which UMR was surprising. 7266, 17000-F La Rochelle, France; [email protected] significant effect in FRET, which was surprising. We are most grateful to Dr. John O′Brien for NMR spectroscopy, Gary Hessman and Brian Talbot TheWe occurrence are most grateful of the 3-alkenyl-oxindoleto Dr. John O′Brien skeletonfor NMR inspectroscopy, various natural Gary and Hessman synthetic and products Brian Talbot has for MS spectroscopy and Tarek Moustafa for his collaboration in the FXR FRET assay. generatedfor MS spectroscopy interest of many and Tarek groups Moustafa on account for ofhis its collaboration useful biological in the properties FXR FRET and assay. its versatility as a useful intermediate in the synthesis of many compounds (Bort, A., et al. Sci. Rep. 2018, 8, 4370). As part 7.36. Synthesis and Biological Activities of New 2-Mercapto-(2-Oxoindolin-3-ylidene)acetonitrile of7.36. our Synthesis ongoing researchand Biological on oxindole Activities derivatives, of New 2-Mercapto-(2-Oxoindolin-3-ylidene)acetonitrile we previously designed, synthesized and evaluated Derivatives (P36) newDerivatives series of (P36) bis-oxindoles as potent kinase inhibitors (Beauchard, A., et al. Bioorg. Med. Chem. 2006, 1, Boris6434–6443). Letribot, In Régis an eﬀort Delatouche, to identify Jean-René new pharmacological Chérouvrier, inhibitors Lisianne of Domon disease-relevant and Valérie protein Thiéry kinases Boris Letribot, Régis Delatouche, Jean-René Chérouvrier, Lisianne Domon and Valérie Thiéry Lawith Rochelle increased UniversityLIENSs potency and UMR selectivity, 7266, 17000-F we launched La Rochelle, a research France; e-mail: program [email protected] dealing with the synthesis ofLa rare Rochelle substituted UniversityLIENSs 2-(2-oxoindolin-3-ylidene)acetonitrile UMR 7266, 17000-F La Rochelle, derivatives France; e-mail: bearing [email protected] a sulphur atom directly The occurrence of the 3-alkenyl-oxindole skeleton in various natural and synthetic products has attachedThe on occurrence the external of doublethe 3-alkenyl-oxindole bond. Few 3-(thiazol-5-ylidene)indoline-2-one skeleton in various natural and derivatives synthetic productsI have been has generated interest of many groups on account of its useful biological properties and its versatility as reportedgenerated in interest the literature of many (Havrylyuk, groups on D.,account et al. ofJ. its Med. useful Chem biological. 2012, 55,properties 8630–8641; and Erben,its versatility F., et al. as a useful intermediate in the synthesis of many compounds (Bort, A., et al. Sci. Rep. 2018, 8, 4370). As RSCa useful Adv. intermediate2014, 4 (21), 10879–10893).in the synthesis To of access many thecompounds required (Bort, 2-oxoindolylacetonitrile A., et al. Sci. Rep. 2018 derivatives, 8, 4370).III As part of our ongoing research on oxindole derivatives, we previously designed, synthesized and (Figurepart of 27 our), we ongoing decided research to explore on aoxindole new route deriva basedtives, on thewe applicationpreviously indesigned, synthesis synthesized of the reagent and evaluated new series of bis-oxindoles as potent kinase inhibitors (Beauchard, A., et al. Bioorg. Med. 4,5-dichloro-1,2,3-dithiazoliumevaluated new series of bis-oxindoles chloride as (Appel potent salt) kinase1. The inhibitors work described (Beauchard, in this A., posteret al. Bioorg. represents Med. Chem. 2006, 1, 6434–6443). In an effort to identify new pharmacological inhibitors of disease-relevant furtherChem. 2006 application, 1, 6434–6443). of Appel In saltan effort in the to conception identify new of novelpharmacological heterocyclic inhibitors rings to accessof disease-relevant to original protein kinases with increased potency and selectivity, we launched a research program dealing with bioactiveprotein kinases compounds. with increased potency and selectivity, we launched a research program dealing with the synthesis of rare substituted 2-(2-oxoindolin-3-ylidene)acetonitrile derivatives bearing a sulphur the synthesis of rare substituted 2-(2-oxoindolin-3-ylidene)acetonitrile derivatives bearing a sulphur atom directly attached on the external double bond. Few 3-(thiazol-5-ylidene)indoline-2-one atom directly attached on the external double bond. Few 3-(thiazol-5-ylidene)indoline-2-one derivatives I have been reported in the literature (Havrylyuk, D., et al. J. Med. Chem. 2012, 55, 8630– derivatives I have been reported in the literature (Havrylyuk, D., et al. J. Med. Chem. 2012, 55, 8630– 8641; Erben, F., et al. RSC Adv. 2014, 4 (21), 10879–10893). To access the required 2- 8641; Erben, F., et al. RSC Adv. 2014, 4 (21), 10879–10893). To access the required 2- oxoindolylacetonitrile derivatives III (Figure 27), we decided to explore a new route based on the oxoindolylacetonitrile derivatives III (Figure 27), we decided to explore a new route based on the application in synthesis of the reagent 4,5-dichloro-1,2,3-dithiazolium chloride (Appel salt) 1. The application in synthesis of the reagent 4,5-dichloro-1,2,3-dithiazolium chloride (Appel salt) 1. The

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 45 of 47

work described in this poster represents further application of Appel salt in the conception of novel Pharmaceuticals 2019, 12, x FOR PEER REVIEW 45 of 47 heterocyclic rings to access to original bioactive compounds. work described in this poster represents further application of Appel salt in the conception of novel Pharmaceuticals 2019, 12, 179 R 45 of 47 N 3 heterocyclic rings to access to original bioactive compounds.N S O Cl S NC S SH R N 3 R N S R R O O Cl 2 S O NC O N S N N SH R H This work R1 R H R I O II III 2 O O N N R = H; 5-Br; 5-Cl; 5-NO (Z)-3-(4-oxo-4,5-dihydro-thiazol-5-ylidene)indolin-2-one H This work N2 R1 H I II III Figure 27. The (Z)-3-(4-oxo-4,5-dihydro-thiazol-5-ylidene)indolin-2- one pattern I and targets R = H; 5-Br; 5-Cl; 5-NO synthesis(Z)-3-(4-oxo-4,5-dihydro-thiazol-5-ylidene)indolin-2-one 3-(4-chloro-5H-1,2,3-dithiazol-5-yliden)indolin-2-one II and (2-oxoindolin-3-ylidene)-2-2 mercaptoacetonitrile III. Figure 27. 27. The (Z)-3-(4-oxo-4,5-dihydro-thiazol-5-The (Z)-3-(4-oxo-4,5-dihydro-thiazol-5-ylidene)indolin-2-ylidene)indolin-2- one pattern oneI and patterntargets Isynthesisand 3-(4-chloro-5 targets synthesisH-1,2,3-dithiazol-5-yliden)indolin-2-one 3-(4-chloro-5H-1,2,3-dithiazol-5-yliden)indolin-2-one II and (2-oxoindolin-3-ylidene)-2-II and Thanks to the “Ligue Nationale Contre le IIICancer, comité 17” for financial support and to the (2-oxoindolin-3-ylidene)-2-mercaptoacetonitrilemercaptoacetonitrile III. . “Conseil Départemental de Charente-Maritime” for PhD grant (BL). Thanks to the “Ligue Nationale Contre le Cancer, comité 17” for financial support and to the Thanks to the “Ligue Nationale Contre le Cancer, comité 17” for financial support and to the “Conseil7.39. Building Départemental a Diverse and de Experimentally-Curated Charente-Maritime” for Fragment PhD grant Library (BL). (P39) “Conseil Départemental de Charente-Maritime” for PhD grant (BL). 1 1 2 3 7.37.Steve Building Brough a Diverse, Andrew and Lowerson Experimentally-Curated , Steven LaPlante Fragment , Patrick Library McCarren (P39) and Michael Serrano- 7.39.Wu 3 Building a Diverse and Experimentally-Curated Fragment Library (P39) Steve Brough 1, Andrew Lowerson 1, Steven LaPlante 2, Patrick McCarren 3 and Michael Serrano-Wu 3 Steve1 Key Brough Organics 1, Andrew Limited, LowersonCornwall, United 1, Steven Kingdom. LaPlante 2, Patrick McCarren 3 and Michael Serrano- 1 Wu2 Key 3NMX Organics Research Limited, and Solutions, Cornwall, Montreal, UK CA. 2 NMX Research and Solutions, Montreal, CA 13 Broad Institute, Cambridge, MA, USA; e-mail: [email protected] 3 Key Organics Limited, Cornwall, United Kingdom. 2 Broad Institute, Cambridge, MA, USA; [email protected] NMXFragment Research libraries and Solutions, are commonly Montreal, assembled CA. by rule of three filtering followed by manual Fragment libraries are commonly assembled by rule of three filtering followed by manual curation. 3curation. Broad However, Institute, Cambridge, the robust MA,experimental USA; e-mail: data [email protected] that ensures the proper physicochemical attributes However,needed for the high-concentration robust experimental screening data thatis often ensures lacking the and proper replaced physicochemical instead by in attributessilico calculations needed Fragment libraries are commonly assembled by rule of three filtering followed by manual forof uncertain high-concentration predictive screening value. A is fragment often lacking collection and replaced with experimentally-determined instead by in silico calculations aqueous of curation. However, the robust experimental data that ensures the proper physicochemical attributes uncertainsolubility predictivewill address value. a major A fragment source of collection false positives with experimentally-determined and attrition in fragment screening aqueous solubilitylibraries: needed for high-concentration screening is often lacking and replaced instead by in silico calculations willaggregation, address a stability major source and ofsolubility. false positives 1H NMR and spectral attrition data in fragment in aqueous screening buffer libraries: will further aggregation, enable of uncertain predictive1 value. A fragment collection with experimentally-determined aqueous stabilitypractitioners and solubility.to rapidly buildH NMR fragment spectral pools data and in aqueousinitiate screening. buffer will further enable practitioners to solubility will address a major source of false positives and attrition in fragment screening libraries: rapidlyDiversity build fragment selection pools methods and initiate in shape, screening. scaffold, fingerprint and predicted property space aggregation, stability and solubility. 1H NMR spectral data in aqueous buffer will further enable combinedDiversity with selection industry-standard methods in shape,substructure scaffold, filtering fingerprint were and used predicted to select property over 2500 space Key combined Organics practitioners to rapidly build fragment pools and initiate screening. withcompounds industry-standard for experimental substructure profiling. filtering NMR were and usedLC–MS to select analysis over allowed 2500 Key the Organics careful compoundsselection of Diversity selection methods in shape, scaffold, fingerprint and predicted property space forhighly-soluble experimental fragments profiling. with NMR desirable and LC–MS physicochemical analysis allowed and thestability careful characteristics. selection of highly-soluble Importantly, combined with industry-standard substructure filtering were used to select over 2500 Key Organics fragmentsthe curated with molecules desirable were physicochemical enriched in cyclic and scaffolds stability commonly characteristics. found Importantly, in drug candidates the curated and compounds for experimental profiling. NMR and LC–MS analysis allowed the careful selection of moleculesspanned chemical were enriched space that in cyclic minimally scaffolds overlapped commonly with found existing in drug commercial candidates collections. and spanned This chemical poster highly-soluble fragments with desirable physicochemical and stability characteristics. Importantly, spacesummarizes that minimally the experimental overlapped and with cheminformatic existing commercial features collections. of this next This generation poster summarizes Key Organics the the curated molecules were enriched in cyclic scaffolds commonly found in drug candidates and experimental‘BIONET Premium and cheminformatic Fragment Library features′. (Bradley, of this C., next et al. generation J. Chem. Inf.Model. Key Organics 2006, 46 ‘BIONET, 1060–1068; Premium Baell, spanned chemical space that minimally overlapped with existing commercial collections. This poster FragmentJ. B., et al. J. Library Med. Chem.0. (Bradley, 2010, 53, C., 2719–2740; et al. J. Chem. Lagorce, Inf.Model. D., et2006 al. BMC, 46, 1060–1068; Bioinformatics Baell, 2008 J., B., 9, et396; al. Kazius,J. Med. summarizes the experimental and cheminformatic features of this next generation Key Organics Chem.J., et al.2010 J. Med., 53, Chem. 2719–2740; 2005, Lagorce,48, 312–320; D., etBruns, al. BMC R. F., Bioinformatics et al. J. Med. 2008Chem., 9 2012, 396;, 55, Kazius, 9763–9772; J., et al. LiJ.surek, Med. ‘BIONET Premium Fragment Library′. (Bradley, C., et al. J. Chem. Inf.Model. 2006, 46, 1060–1068; Baell, Chem.M, et al.2005 Mol., 48 Divers., 312–320; 2010, Bruns, 14(2), R.401–408; F., et al. Taylor,J. Med. R. Chem. D., et 2012al. J., Med.55, 9763–9772; Chem. 2014 Lisurek,, 57 (14), M, 5845–5859). et al. Mol. Divers.J. B., et al.2010 J. Med., 14(2) Chem., 401–408; 2010, Taylor, 53, 2719–2740; R. D., et Lagorce, al. J. Med. D., Chem. et al. 2014BMC, 57Bioinformatics (14), 5845–5859). 2008, 9, 396; Kazius, J., et al. J. Med. Chem. 2005, 48, 312–320; Bruns, R. F., et al. J. Med. Chem. 2012, 55, 9763–9772; Lisurek, M, et al. Mol. Divers. 2010, 14(2), 401–408; Taylor, R. D., et al. J. Med. Chem. 2014, 57 (14), 5845–5859).

Build Strategy

Pharmaceuticals 2019, 12, x FOR PEER REVIEW 46 of 47 Pharmaceuticals 2019, 12, 179 Build Strategy 46 of 47

7.38.7.40. Overcoming tumor necrosis factor (TNF)-Related ApoptosisApoptosis InducingInducing LigandLigand (TRAIL)(TRAIL) ResistanceResistance inin Cancer Stem Cells: Computer-Aided Discovery of c-FLIP Inhibitors Cancer Stem Cells: Computer-Aided Discovery of c-FLIP Inhibitors Kok Yung Lee 1, Andrea Brancale 1, Richard Clarkson 2 and Andrew Westwell 1 Kok Yung Lee 1, Andrea Brancale 1, Richard Clarkson 2 and Andrew Westwell 1 1 School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Cardiff CF10 3NB, UK 1 School of Pharmacy and Pharmaceutical Sciences, Cardiff University, Cardiff CF10 3NB, United Kingdom. 2 2 EuropeanEuropean Cancer Cancer Stem Stem Cell Cell Research Research Institute, Institute, Cardiff CardiUniversity,ff University, Cardiff CF24 Cardi 4HQ,ff UnitedCF24 Kingdom; 4HQ, UK; e- leek10@cardimail: [email protected]ff.ac.uk The ability of cancer stem cells to self-renew indefinitelyindefinitely and didifferentiatefferentiate into multiple tumour cell typestypes has made their elimination critical to completely eradicate tumours (Battle, E.; Clevers, H. Nat MedMed.. 2017, 23 (10), 1124–1134). TRAIL (TNF-related apoptosis inducing ligand) is a death ligandligand thatthat selectivelyselectively inducesinduces apoptosisapoptosis in cancercancer cells,cells, but clinicalclinical trialstrials utilisingutilising recombinant TRAIL or TRAILTRAIL agonistsagonists havehave eventually eventually failed failed due due to to the the development development of TRAILof TRAIL resistance resistance post-treatment post-treatment (de Miguel,(de Miguel, D., etD., al. etCell al. Cell Death Death Differ Differ. 2016. 2016, 23 (5), 23, 733–747;(5), 733–747; Herbst, Herbst, R. S.; R. et S.; al. etJ Clinal. J OncolClin Oncol. 2010. ,201028 (17), 28, 2839–46)(Figure(17), 2839–46)(Figure 28). c-FLIP 28). c-FLIP (cellular (cellular FLICE-like FLICE-like inhibitory inhibito protein)ry inhibitsprotein) TRAIL-mediated inhibits TRAIL-mediated apoptosis andapoptosis its overexpression and its overexpression has since been has identified since been as one identified of the mechanisms as one of the of TRAIL mechanisms resistance of presentTRAIL inresistance cancer stem present cells in (French, cancer stem R.; et cells al. Mol (French, Cancer R.;. 2015et al., 14Mol, 209). Cancer As. 2015 a strategy, 14, 209). to overcome As a strategy TRAIL to resistance,overcome TRAIL this project resistance, aims to this develop project new aims small to devel moleculeop new inhibitors small molecule of c-FLIP inhibitors that re-sensitises of c-FLIP cancer that stemre-sensitises cells to TRAIL-mediatedcancer stem cells to apoptosis. TRAIL-mediated apoptosis.

Figure 28. The role of c-FLIP in the extrinsic apoptotic pathway.

Twenty didifferentfferent compoundscompounds werewere identifiedidentified throughthrough inin silicosilico screeningscreening asas potentialpotential leadlead compounds targeting DEAD-box protein 1 (DED1) of c-FLIP. HeLaHeLa cellscells werewere treatedtreated withwith thesethese compounds inin combination combination with with TRAIL TRAIL to test to fortest their for abilitytheir ability to sensitise to sensitise HeLa cells HeLa to TRAIL-mediated cells to TRAIL- apoptosis.mediated apoptosis. Caspase-mediated Caspase-mediated apoptosis apoptosis was confirmed was confirmed through rescue through with rescue a pan-caspase with a pan-caspase inhibitor. Threeinhibitor. of twenty Three compoundsof twenty compounds in combination in combinat with TRAILion with successfully TRAIL successfully induced apoptosis induced in apoptosis HeLa cells in rescuableHeLa cells by rescuable caspase inhibitor. by caspase Further inhibitor. work Further is in progress work tois confirmin progress c-FLIP to inhibitorconfirm c-FLIP activity inhibitor of these compoundsactivity of these through compounds co-immunoprecipitation through co-immunoprecip while analoguesitation while of the analogues best performing of the best lead performing compound arelead synthesised. compound are synthesised.

7.39.7.41. University of Nottingham Managed ChemicalChemical Compound Collection (MCCC)(MCCC) (P41)(P41) Lubna Hashmi,Hashmi, PeterPeter Fischer Fischer and and Michael Michael Stocks Stocks

Centre for for Biomolecular Biomolecular Sciences, Sciences, University University of Nott of Nottingham,ingham, Nottingham, Nottingham, Nottingha Nottinghamshiremshire NG7 2RD, NG7 United 2RD, UK;Kingdom; [email protected] e-mail: [email protected]

An AAffordableﬀordable andand AutomatedAutomated CompoundCompound ManagementManagement FacilityFacility Pharmaceuticals 2019, 12, 179 47 of 47

Compound library for high and medium throughput screening; • >85K Structurally diverse drug-like molecules; • Molecules obey Lipinski “rule of 5”; # No reactive functional groups. # Purity Analysis and Structural Conﬁrmation

Samples stored at optimum condition as 10mM DMSO stock solutions; • Dedicated LC–MS system to ensure purity of compounds. • Medicinal and Computation Chemistry Support

Collaborative projects welcome. • For further information please contact: Dr Michael Stocks; Head of Division of Medicinal Chemistry and Structural Biology; [email protected]

8. Conclusions The meeting attracted 114 delegates with successful growth of both the network membership and the geographical range of attendee home countries, including France, Germany, Russia, Spain, Portugal, Ireland, Italy and the United Kingdom. The programme comprised a discipline-leading line-up of presenters from both academic and industrial sectors (including talks from 2 plenaries, 12 keynotes and 6 young researchers, and 41 posters), spanning a broad range of topics related to medicinal chemistry. For the ﬁrst time, the inclusion of a pre-conference workshop expanded attendee knowledge and skills in the area of quantitative pharmacology. In addition, attendees of the conference dinner enjoyed some words from Emeritus Prof. Malcolm Stevens FRS (UoN), who appropriately wore a bowtie dyed with the original mauveine made by William Henry Perkin. A number of prizes were awarded, with the best poster prize (sponsored by the Royal Society of Chemistry) going to Mirjana Antonijevic (UNICAEN, France), the best young researcher oral presentation to Scott Grossman (School of Pharmacy, University of Nottingham) and the MDPI Pharmaceuticals travel award to Jorge Grilo (University of Lisbon). The 28th Annual GP2A Medicinal Chemistry Conference will take place between 26 and 28 August 2020 at the University of La Rochelle, France. The GP2A committee and the local organizing committee thank the Division of Biomolecular Science and Medicinal Chemistry, School of Pharmacy (UoN), and all the sponsors and exhibitors for supporting this annual event.

Funding: This research received no external funding. Conﬂicts of Interest: The authors declare no conﬂict of interest.

© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).