Applied Pharmaceutical Sciences

Preliminary Definitions: Definitions of drug, toxicant, toxin, poison, experimental pharmacology, preclinical and clinical studies. Definition of officinal and magistral preparation, pharmaceutical specialty, generic and equivalent drug. Compassionate use and off-label use. Methods of preclinical and clinical experimental pharmacology. Clinical experimental pharmacology: The research, development and commercialization phases of a new drug. Preclinical development phases, clinical development phases. Drug attrition rate. Marketing authorization. GMP, GLP, GCP, Guidelines and legislation for drugs for human use. Ethics in clinical trials, Nuremberg Code, Declaration of Helsinki and Taipei. Phase I, II, III, and IV studies, types of patients, methods and main objectives. Randomized controlled clinical trials. What is meant by control, placebo, historical control. Sampling and sampling methods. Sampling error and bias. Inclusion and exclusion criteria. Randomization and the methods of randomization. Blindness in the clinical trial, objectives and methods. Activity, efficacy and safety study. Risk-benefit ratio. Superiority and non-inferiority studies. Primary and secondary endpoints. Informed Consent. Role of the ethics committee. Measures of association between risk factor and clinical event. Prevalence (point, period and lifetime). Cumulative incidence and incidence rate. Risk. Relationship between prevalence and incidence. Definition of risk factor and confounding factor. Measures of strength of association between risk factor and clinical event used in clinical trials: absolute risk, relative risk, attributable risk, odds ratio. The number of patients to treat (NNT), relative risk reduction. Causality criteria between risk factor and clinical event. Causality criteria between drug intake and adverse reaction. Karch-Lasagna and Naranjo's algorithm. Types of clinical, etiologic, observational and descriptive studies. Cross-sectional and longitudinal, prospective and retrospective studies. Cohort and case-control studies. Types of literature review, simple and systematic. Meta-analysis. Stages and principles of meta-analysis. Interpreting a forest plot. Prism guideline and its diagram. Distortions in metanalysis, publication, selection, economic support and conflict of interest. Heterogenicity of source studies and its influence in metanalysis. Graphical and statistical method of investigation of heterogeneity. Evidence-based medicine and the pyramid of strength of evidence. Evidence-based practice. Difference between efficacy and effectiveness. Bibliographic research: primary, secondary and tertiary sources. Main databases for bibliographic research in the field of experimental pharmacology. Guide to the use of the most common scientific databases and websites of public and private organizations in the field. Search strategies, by keywords, thesaurus, Boolean operators, filters. PICO format. Use of these strategies to perform a literature search with examples in Pubmed, Cochrane Library and Trip databases. Evaluation of scientific evidence and publications. Preclinical experimental pharmacology: general principles and objectives. In vitro, ex vivo and in vivo studies. In vivo: Animal experimentation, importance, objectives. Ethics in animal experimentation. Regulatory references, 2010 European directive and 2014 Italian legislative decree. The principle of the 3 Rs. Alternative methods to animal experimentation and body in charge of their validation. Concept of animal model and animal species used. In vivo experimental toxicity studies (acute, repeated, chronic). Determination of dose-response relationship, DL50, DE50, DT50, the therapeutic index and margin of safety. Mutagenesis studies. Advantages of using rodent as animal species in experimentation. Concept of outbred and inbred. Animal models in the field of behavioral experimental pharmacology. Criteria of validity of an animal model (face validity, construct validity, predictive validity) and applications. Monitoring of voluntary caged consumption of food, water and aqueous solutions. Monitoring of well-being/illness status. Associative and non-associative learning theory. Habituation. Classical and operant conditioning. The studies of Pavlov, Thorndike and Skinner. The conditioned response. Positive and negative reinforcement. Gratification and punishment. Concept of acquisition, extinction and reinstatement of a behavior. Cages for experiments with classical conditioning. Cages for experiments with operant conditioning. Examples of behavioral tests for the study of anxiety, depression, sociality, memory and learning, and abuse behavior. Genetically modified animals. Applications in agriculture, pastoralism and biomedical research. The transgene and the technique of delivery by microinjection into the pronucleus. Mice with a conditional modification (knock-in and knock-out). The cre-loxP system for generating genetically modified mice. Genomic editing with Cripr-Cas9. Ex vivo and In vitro: Cell cultures. Types of cultures: primary, secondary and cell lines. Adhesion and suspension culture. Organization and operation of a cell culture laboratory. Basic methods, advantages and disadvantages. Applications of cell cultures with special reference to the study of pharmacological/toxic effects of pharmaceutical compounds or potential environmental pollutants. Plating and counting the number of cells in culture with a hematocytometer (Burker chamber). Confluence. Main media for cell culture. Maintenance of environmental conditions (temperature, carbon dioxide, humidity). Cell culture media, the importance of nutrients, growth factors and buffer. Stem cells and the technique of reprogramming specialized cells into iPSCs. Current and potential applications. Organoids and their current and potential applications. Methods applied to preclinical and clinical research for the study of tissue, gene and protein expression in animal and cellular samples. Characteristics of analytical methods (analytical range, sensitivity, limit of detection, specificity or selectivity, interference, robustness). The calibration curve, calibration line. Histochemistry. Examples of specific reactions for each class of macromolecules. Orthochromatic and metachromatic methods. Fluorochromes and fluorescence. Hints at immunohistochemistry/immunofluorescence protocols and microscopic analysis. Methods of studying proteins: purification (separation from other cellular components, determination of degree of purity); study (determination of structure, function, regulation). Spectroscopic determination of protein concentration of a protein lysate. Bradford's assay. Lambert-Beer's law. The chromophores. Horizontal and vertical electrophoresis. General principles, media types and examples of application to pharmacological studies. Methods using antibodies (ELISA, RIA, FRET, flow cytometry, fluorescence immunohistochemistry and electron microscopy). Hints on polyclonal and monoclonal antibodies (production, use, advantages and disadvantages). The western blot: principles and applications for the study of proteins. Hints on post-translational modifications and detection techniques. Methods of studying gene expression. Extraction, isolation and purification of nucleic acids. Semiquantitative and quantitative PCR (polymerase chain reaction). Principles of statistics applied to experimental pharmacology: The scientific method and experimentation. Descriptive statistics and inferential statistics. Definition of population and sample. Reasons for foregoing population analysis in favor of the sample. The representative sample. Qualitative and quantitative variables, dependent and independent variables. Precision and accuracy. Indices of central tendency. Indices of dispersion. The variability of data. Forms of distribution, the gauss curve and its properties. Healthy population normality values. Absolute and relative frequency. Probability and risk. The error and bias. Confidence levels. The standard error of the mean and its difference with the standard deviation. Types of association between variables (spurious, causal and noncausal). Confounding factors. Cause-effect relationship and causality criteria. Definition of hypothesis. Null hypothesis and alternative hypothesis. Verification of the hypothesis. The analysis of variance. The value of F and its significance. Comparison of two groups with the t-test. The t-value and its significance. Statistical significance. P value. Type I and type II errors. The power of a statistical test. Degrees of freedom. Difference between technical and biological replicates. Repeated measures in the same subjects. Replication and pseudoreplication. Main graphical methods of representing data: histogram, bar chart, aerogram, box-plot, line plot, the forest plot.

In order to understand the content of the lectures in this course and to achieve the learning objectives, it is important that the student has a basic knowledge of mathematics, physics, biochemistry, molecular biology and pharmacology at the beginning of the lectures. These prerequisites are not essential, but are strongly recommended in order to pass the exam at the end of the course.

free but strongly recommended because there is no textbook that encapsulates in a unified and organized manner all the topics covered in class. Class attendance is recommended to enable the student not only to acquire specific knowledge pertaining to the course subject matter, but also to develop communication skills and technical language through classroom interaction with the lecturer. In addition, the student will be stimulated to make judgments and critical sense regarding the issues addressed in class.

Test of knowledge through a written multiple-choice test followed by an oral examination to be held on the same day. Written test: about 45 minutes to answer 31 questions. Some questions will ascertain theoretical knowledge, while others will test practical skills through completion of exercises done in class. A grade of not less than 18/30 is required to be admitted to the oral examination. This will be followed by an oral examination in which the following aspects will be evaluated and will contribute to the final exam score: the student's specific preparation on the various course topics, reasoning and judgment skills, and ability to communicate in appropriate technical language. The duration of the oral examination may vary depending on the student's ability; an average duration of about 15 minutes can be assumed. The final grade will ascertain knowledge as follows: 18 = sufficient knowledge; 19-23 = fair knowledge; 24-27 = good knowledge; 28-30 = excellent knowledge; 30 cum laude = outstanding knowledge.

lecture notes Materials made available by the lecturer

Lectures supported by a power point presentation In-class video viewing Exercises carried out in class with the lecturer Exercises to be done alone in class in the presence of the lecturer and verification of the solution and how it was done in class. The teaching method adopted includes face-to-face lectures involving the use of power points, of databases (e.g., PubMed) and the viewing of short scientific films that can stimulate the critical sense promoted by continuous interactive discussions in the classroom. The face-to-face lectures will address the topics in the syllabus, dwelling in particular on the topics that are more difficult for the student to understand and suggesting connections between the various topics in the syllabus that can help the student to approach the study also autonomously.