XXXVII CICLO 

A critical piece in drug discovery and development is conducting DMPK (Drug Metabolism and Pharmacokinetics) studies, often referred to as ADME-T (Absorption, Distribution, Metabolism, Elimination, Toxicity). ADME studies are designed to investigate how a chemical (e.g. a drug candidate) is processed by a living organism and how ADME properties could affect its activity. For a drug candidate, "optimizing" ADME properties before clinical investigation is of utmost importance to ensure it is as effective and safe as possible; it is estimated that close to 50% of drug candidates fail because of unacceptable efficacy due to an unfavourable ADME profile. This course aims to provide PhD students engaged in the early drug discovery process awareness about the importance of preclinical ADME investigation, the strategies and the methodologies that assist to achieve efficiency in the hit-to-lead process.

Biomaterials used in orthopedic applications: advantages and disadvantages. - Tissue engineering may offer new treatment alternatives: scaffolds and cells. - Regenerative medicine: want to replace and repair lost or damaged tissues by stimulating the natural regeneration process. The gap between the damaged fragments has to be filled with functional materials that act as a substrate and as a physical three-dimensional microenvironment for inducing the migration and organization of cells from the native tissue. - Differentiation and maturation signals that positively promote osteogenesis or chondrogenesi.

A food colorant is "any dye, pigment or substance that can impart color to a food, drug, cosmetic or the human body". Starting from the FDA definition, the course will be directed to understand the chemistry of food colorants, their properties, including functional properties, their toxicology. Finally, some innovative perspectives will be analyzed following the question "Eating with the eyes is possible?" 

The course aims at presenting and discussing the major techniques used for the study of proteins structures and functions, and the contributions of structural biology in the study of diseases and in the research of new drugs. 

The aim of the course is to provide technical-practical insights on the distillation at room pressure and distillation under reduced pressure.

Liquid biopsy consists of the detection and isolation of circulating tumor cells, circulating tumor DNA and exosomes, as a source of genomic and proteomic information in patients with cancer.It is a revolutionary technique that is opening previously unexpected perspectives. Newly developed techniques and next-generation sequencing analyses allow a broad application of liquid biopsy in a wide range of settings. Initially correlated to prognosis, liquid biopsy data are now being studied for cancer diagnosis, including screenings, and most importantly for the prediction of response or resistance to given treatments. Still application is far from daily use in clinical application but ongoing research is leading the way to a new era in oncology. 

The course will tackle the issue of drug evaluation from different perspectives, from the need to choose for single individuals with no therapeutic alternatives to the need to choose whether to reimburse a drug for an entire nation or to list a drug on the WHO essential medicines list. Along the way, the drug regulatory framework will be introduced. 

The Università del Piemonte Orientale is a pilot center for the School of INN of the World Health Organization. In this privileged position, the course will introduce international non-proprietary names (INN: i.e. the name that is given to any marketed drug and that does not belong to the company) and the schemes and rules that govern their creation. The course will then have a practical element of reviewing names, which may result in a research publication. 

The course will be an introduction to systematic reviews and meta-analysis, an indispensable tool for research and for Ph.D. students. Alongside theory, the students will be coordinated in a scientific project of meta-analysis that may result in a research publication. 

The chemistry of life is commonly associated to a handful of elements, usually identified with the mnemonic acronym CHNOPS, summarizing the key elements of organic chemistry. However, surprisingly, this small list is much shorter than is believed. The human body contains at least 60 detectable chemical elements and about 25 of them play a role in its healthy functioning. Moreover, more than 45 elements (including radionuclides) are used as metallodrugs in diagnostics or in therapy. A journey through the periodic table will provide an overview of the elements, their role in the living organism and the application of their compounds in medicine.

This short course is targeted at students who wish to learn the basics and challenges of drug screening on information rich cell based models. Aspects of assay design ranging from cell type choice, readout selection, standardization, miniaturization, data analysis, will be covered. Moreover, advantages of targeted versus phenotypic assays in drug screening will be discussed. At the end, participants will be challenged in a competition to propose a cell based-assay that could be potentially relevant for their own research projects/interests. 

Animal models can provide invaluable information to our knowledge of biology and medicine, including the discovery and development of new drugs. At present the overall success rate of drugs during clinical development remains rather low and one commonly raised explanation is flawed preclinical research in animal models. This is especially true in therapeutic areas in which animal models of human diseases are particularly challenging. Based on these assumptions, better design and conduct as well as further development of animal models is certainly essential. The short course highlights, with discussion of successful and unsuccessful case studies, the current challenges and limitations of selected animal models and points at aspects which may be relevant for improving the translational value of such models in drug discovery. Regulatory aspects of animal research studies will also be discussed. At the end, PhD students will be asked to present (10 minute oral presentation) an animal model that could be potentially relevant for their own research projects/interests. 

Human body contains about 1.2 kg of elementary calcium (Ca). The majority of it is immobilized in bones and teeth in a form of phosphate salts, but less than 1% of ionized Ca2+ escapes the trapping and serves as a signaling molecule to conduct and codify signaling information between and inside the cells. Signals conducted by Ca2+ ions as well as proper handling of Ca2+ fluxes inside the cell (called also calcium homeostasis) are fundamental for normal functioning of any cellular type of an organism as many (if not all) cellular processes directly or indirectly depend on Ca2+. When adequate Ca2+ signals are lacking, or when Ca2+ fluxes become too intense, cell or tissue may turn from health to disease. The list of diseases linked to deregulation of Ca2+ homeostasis is impressive and includes genetic and idiopathic diseases from all ategories. Understanding of principles of Ca2+ homeostasis and Ca2+ signaling is important for every person those activity is linked to biology of the cell. In this 8 hours course of "Cellular calcium signaling in Health and Disease" PhD students will learn the mechanisms of maintenance of cellular Ca2+ homeostasis, principles of Ca2+ signaling in healthy and diseased cell and methods used to date to monitor Ca2+ signals inside the cell. The possibility of short practical training in single cell Ca2+ imaging using synthetic Ca2+ probe Fura-2 may also be considered. 

Protein crosstalks as well as protein-ligand interactions play essential roles in many biological processes including signaling pathways, transcriptional regulation and numerous other metabolic reactions. In order to understand the role of such protein interactions in biological processes it is important to investigate the interaction dynamics describing the stoichiometry of the complexes, the binding free energy and their binding cooperativity as inter-molecular communication. These biochemical parameters are complementary to structural biology studies. In particular, the enthalpic and entropic components of the binding free energy directly refer to the mechanistic aspects of the binding and have been widely exploited in drug discovery research pipeline. The aim of this course is the description of practical and theoretical aspects of biophysical methods used for measuring the stoichiometry and affinity of many protein interactions. In particular, the course will focus on the application potential of the following techniques in the field of biochemistry structural biology: i) small-angle X-ray scattering; ii) isothermal titration calorimetry (ITC); iii) differential scanning fluorescence (DSF); iv) surface plasmon resonance (SPR) and v) microscale thermophoresis (MST). 

The natural compounds are an endless source of ideas for the discovery of new drugs to treat diseases, and the total synthesis of complex natural products still represents a big challenge for the organic chemist. While many strategies applied to the synthesis of complex natural products are based on the construction of individual rings or fragments of the natural products followed by a unification step, or by the iterative annulation of one ring onto a preexisting ring, an efficient alternative strategy could be represented by the transannular cyclization reactions. Transannular reactions are defined as those reactions which lead to the formation of covalent bond between atoms on opposite sides of the ring compound. 1 They usually occur in macrocyclic compounds that, to minimize transannular strain (Prelog strain), are constricted in rigid conformations that force some functional groups to be close to each other. 2 This intimacy between functional groups confers entropic advantages to enable transformations that are otherwise difficult in intermolecular and intramolecular settings, 3 making transannular reactions a highly efficient tool for the construction of complex polycyclic architectures. Transannular reactions are classified according to the reaction type involved in the cyclization process [Diels-Alder (TADA), ene reaction, [2+2] and other cycloadditions, Michael addition, aldol condensation, Mannich and miscellaneous reaction]. 4 The transannular cyclisation process is the result of a series of cascade reactions that allow us to obtain a complex polycyclic architecture from an easily accessible macrocyclic compound, representing a shortcut in the synthesis of complex natural products. The entire process can be highly influenced by several factors such as the conformation of the macrocycle and the activation strategy. This course will outline the main features and the applications of transannular ring closure reactions and examples will be given to validate this approach as a versatile, efficient and flexible strategy to access new polycyclic structures on the way to the synthesis of important natural products.

Preclinical models that faithfully recapitulate the genomic and histopathological features of cancer are critical for the development of new treatments. The most commonly used models are two-dimensional cell lines established from primary tumors or fluids. While these have provided some important insights into cancer biology, these cell models have significant limitations. In order to address some of these limitations, spheroids, tumor- derived organoids and microfluidic chips have more recently been used to investigate the role of the three-dimensional environment. Efforts have also been made to develop animal models, including genetically modified mice and patient-derived xenografts. We will highlight strengths and weaknesses of the available in vitro and in vivo models.

For PhD students, the prospect of writing their very own scientific research paper may be both exciting and hard. The goal of this course is to provide effective tools to improve writing skills and manuscript writing process.

The course is devoted to providing PhD students engaged in the early drug discovery process with the concepts, the strategies and the methodologies that assist the identification of high-quality drug candidates. This is of utmost importance, as of the thousands of novel compounds that a drug discovery project leads to, only a fraction of these have sufficient ADMET properties to become a clinical candidate. 

Organic chemistry and pharmacognosy have long been an almost unique discipline, with natural products acting as a major driver of advancement for both areas. With the advent of molecular assays, the development of spectroscopy, and the growing sophistication of current synthetic organic chemistry, a comprehensive technical expertise in both fields has become impossible and natural products research has become sectarian and articulated in several sub-disciplines. In this scenario, the role of organic chemistry has undergone a change, moving from a tool of structure elucidation to a mean to manipulate natural products structures and explore their associated biological space. In this context, organic chemistry is metamorphosing into medicinal chemistry, but the transition is essentially molecular in nature, and has been well managed, with organic chemistry retaining a critical role in the development of a scalable synthesis for natural products difficult and/or expensive to obtain by isolation. 1 On the other hand, there is growing evidence that, rather than magic bullets, natural products are magic shotguns, targeting a host of molecular end-points that are often part of homeostatic feed-back loops difficult to perturb with focused monomolecular agents. 2 In this context, mixtures of products like extracts might play an important role, but working with mixtures rather than monomolecular agents is a challenging task that is only now coming of age. 

The course will give an overview of the molecular mechanisms underlying immune recognition in cancer, the up-to date approaches to harness anti-tumor activities of immune cells and the major challenges that remain to be overcome.

The interdisciplinary course will provide an overview of the immunomodulatory activity of gut microbiota and its impact on cancer onset, progression and response to therapy. Approaches to modulate gut microbiota composition to enhance response to cancer therapy will be also discussed. Particular attention will be given to the study of the dominant members of the gut microbial ecosystem, their spatial distribution among physical niches and their crucial functions in human health.

The course aims to enrich the theoretical knowledge of statistics and probability with suitable data analysis skills; the course focuses also on data visualization and is based on the free and open source software R.

Cancers promote immunological stresses that induce alterations of the myelopoietic output, defined as emergency myelopoiesis, which lead to the generation of different myeloid populations endowed with tumor-promoting activities. New evidence indicates that acquisition of this tumor-promoting phenotype by myeloid cells is the result of a multistep process, encompassing initial events originating into the bone marrow and later steps operating in the tumor microenvironment. The careful characterization of these sequential mechanisms is likely to offer new potential therapeutic opportunities. I will discuss relevant mechanisms of myeloid cells reprogramming that instate immune dysfunctions and limit effective responses to anticancer therapy. 

The aim of this course is to present and show the potentiality of the archetypical click reactions (CuAAC, Bertozzi copper free click chemistry, Staudinger ligation, SuFex, Thiolo-ene reaction) in different areas of research such as drug discovery, bioconjugation, in vivo imaging. In the first part, the molecular mechanisms of these reactions will be presented and discussed, while in the second part of the course will be presented real applications of these techniques.

Innovative bioanalytical approaches to support the development of New Biological Entities (Protein, Antibodies, ADC) Federico Riccardi Sirtori, Ilse De Salve, Elisa Bertotti, Piercesare Balestra The scope of this presentation is to give an overview of the different bioanalytical approaches used to support New iological Entities (NBE) and Antibody Drug Conjugates (ADC) projects. Ligand Binding Assays are considered the gold standard for the determination of the concentration of proteins and antibodies in pharmacokinetic (PK) and toxicokinetic (TK) studies (serum, plasma); different approaches will be presented including ELISA, Gyrolab, Mesoscale Discovery (MSD), and Simoa Quanterix technologies. Immunogenicity risk associated with the development of anti-drug antibodies (ADAs) following administration of biotherapeutics is considered one of the main issues in the development of New Biological Entities. For this reason, different approaches to detect ADA in in-vivo studies will be presented. LC-MS technique is also emerging as an alternative tool for the determination of the concentration of total Antibody in biological samples. In addition, LC-MS/MS techniques can be employed to support in vivo ADC pharmacokinetics studies: they can be dedicated to the quantitation of the conjugated toxin, detecting the active molecule, after the capture of the entire ADC and enzymatic cleavage of the linker. Moreover, LC-MS/MS methods are used to quantify the unconjugated toxin potentially released from the ADC. Finally, cell-based assays are playing a crucial role in the development of NBEs. Different approaches based on flow cytometry will be presented for the understanding of the mechanism of action of novel drugs in in-vitro (target internalization) and in-vivo (receptor occupancy) studies.