Dr. Adrian Moore Department of Pharmacy, Health and Well-being Faculty of Applied Sciences

1. Dr. Adrian MooreDepartment of Pharmacy, Health and Well-beingFaculty of Applied Sciences scientiamdulcehauriens

2. UoS Sciences Complex • >£20 million invested in capital development programmes • new-build projects • refurbishment of existing estate • on-going developments • To provide modern, well-equipped facilities to support high quality teaching and learning, real-life applied research, and support for business and our wider community • One-North East • EU ERDF (innovation, enterprise and business support) • Health and Science Academy “provide the best postgraduate teaching and research facilities to accommodate the professional development needs of the pharmaceutical supply chain and support small companies and business start-up ventures in the region”

3. Computational Methods • Comprehensive investigation of molecules, their reactions and interactions • chemical structures / biological reactions at the molecular level • discovery of new drugs that target particular cells • Pharmacophore or Quantitative Structure Activity Relationship (QSAR) • rationalise existing biological data to make informed choices as to the next series of compounds to make • Rational Drug Design • good quality structural information is available of intended biological target enzyme, receptor protein or strand of DNA/RNA • Molecular mechanics (MM) methods • Quantum mechanical (QM) methods • highly computationally resource intensive • UoScluster computer application framework

4. Synthesis • Traditional; research -> medium scale • Range of current projects • Flow chemistry • wide range of reactions possible • production of libraries or on multi-gram scale • safely use highly reactive/hazardous reagents • easily uses solid-phase reagents • good reproducibility -> reduced scale-up issues • inherent reaction control and selectivity • wide temperature range-> superheating gives faster reactions • reduced scale limitations -> quick reaction evaluation

5. 20 compound library, synthesis and aqueous work-up, high yield and high purity (LCMS), no purification 40 mg of each compound synthesised, mass recovery 87.5 % ± 1.5

6. SPR – Linked to Flow Synthesis • Analyse library molecular substrate/target interactions in real time • proteins • nucleic acids • lipids and membrane-associated molecules • carbohydrates • whole cells • viruses/bacteria • Obtain a wide range of critical, binding-related data • specificity • binding partners • affinity • kinetics • concentration • thermodynamics

7. Analyte Ligand • Chip: • hydrophilic • flexible • low non-specific binding • high binding capacity • easy to activate and use for covalent attachment of ligand • withstands extensive regeneration • Attachment of ligand • amine coupling • ligand thiol coupling • surface thiol coupling • maleimide coupling • aldehyde coupling buffer sample buffer association dissociation

8. Separation Science:Areas of Expertise • All areas of pharmaceutical and biomedical analysis • medicinal plant extracts • biomarker analysis • cleaning validation • drug bioanalysis • preparative isolation of API • preparative isolation of related substances • rapid API screens / related substances • chiral screening • stability screening • confirmation of structure • unknown identification

9. Analytical Science: Capability • HPLC / UHPLC (better efficiency, high throughput) • ELSD (non volatile, nonchromaphoriccompounds) • RI (more volatile, nonchromaphoriccompounds) • fluorescence • diode array • single quad MS cooled auto samplers on MS (to 4C) • UHD-QTOF (high resolution and mass accuracy) • prep-LC • GC-FID, GC-Q, GC-QQQ • SPME, head space, and cold-on-column • MALDI-TOF • CE, CE-MS • proteins and complex/biological analysis especially • atomic absorption, flame photometry, UV, IR, fluorometry, luminometry, Karl Fischer, ion-chromatography, logD, pKa, TGA-DSC, microscopy (SEM, TEM, confocal)

10. Separation Science:Rapid LC Related Substances Method UPLC of paroxetine and all related substances in 1.2 minutes BEH C18 (1.7 mm) (50 mm x 2.1 mm ID); 1 mg ml-1 paroxetine and related substances at ~ 0.002 mg ml-1. UV detection - 295 nm. Mobile phase component A (water – THF – TFA (90:10:0.5, v/v/v)), B (acetonitrile – THF – TFA (90:10:0.5, v/v/v)). Resolution was maintained when using a steep gradient profile throughout and also when the temperature was raised to 80 oC.

11. Complex Samples Unknown identification

12. Complex Samples – Principle Analysis • Metabolic profiling of low MW components in biological fluid samples • Pathological conditions can create metabolic disruptions detectable in the metabolite content of biofluids • variation in concentration and relative proportion • Identify affected pathway -> novel approach to treatment

13. NMR Services • Structure confirmation / determination • spectra with or without interpretation • cooled auto-sampler for biological/unstable samples • fast turnaround routine service • Impurity profiling / identification by NMR / MS • Characterisation of peaks in liquid chromatograms, LC-NMR • characterisation of unstable components in complex mixtures • structure elucidation of minor components in complex mixtures • detailed structural information where LC-MS is inappropriate

14. Identification of Natural Products :Comparison Extracted Product/Standard High resolution MS : M = 610 g mol-1, C27H30O16 1-D NMR 500 MHz LC-1H NMR spectrum of approx. 2 mg of rutin, isolated from extracts of Sophora japonica , in D2O-CH3CN [solvent suppression at d1.90 (CH3CN) and d4.46 (residual water)] Extract Standard

15. S 24795 S 24795completed Phase I ClinicalStudies Synthesisdeveloped to semi-production scale (500 kg) negativeallostericmodulatoratnicotinicreceptors wide-spectrumpro-cognitive, psycho-behaviouralactivity Electrophysiology a7 : 42 ± 2 mMa4b2 : 230 ± 16mM (rat – expressed on Xenopus Laevisoocytes) Bindingstudies a7 > 10000 nM, a4b2 > 10000 nM (a1)2bdg ~ 10000 nM, a3b4 > 10000 nM securitybinding (80 sites) – no negative interactions CaCo2 100% hCMEC/D3 90% Microsomes rat 31%, human 75% in vitroclastogenotoxicity! (minormetaboliterelated, < 0.2% profile)

16. NH Aromatic CH Parent compound Metabolite ; 352 scans (20min) 2 µg Identification of Metabolite:Extraction From Rat Bile MS analysis : +16 → oxidation MS/MS → oxidation on one of the 3 substituted aromatic rings exact position of hydroxylation from LC-NMR

17. WEEK 1 Pharmaceutics • Pre-formulation studies • phase transitions / microscopic changes • polymorphic form studies • solubility screening • pKa, log P/D determination • solution and solid-state stability, degradation studies, degradation product identity • excipient and active compatibility studies • particle size measurements • powder electrostatics • physical characterisation of drug delivery systems and vehicle optimisation • problem solving of existing processes and formulations • Formulation • tablet, capsule, solution, suspension, emulsion, injectable, patch etc. • protein stabilisation and delivery • Aseptic capability – category 2 clean room

18. Dr. Adrian MooreDepartment of Pharmacy, Health and Well-beingFaculty of Applied SciencesUniversity of SunderlandDale Building, Room 1.03Sciences ComplexWharncliffe StreetSunderlandSR1 3SDT : +44 (0)191 515 2554F : +44 (0)191 515 3405E : adrian.moore@sunderland.ac.uk