Physico-Chemical Laboratory All New Chemistry – Yours to Explore Physico-Chemical Laboratory

Mission and vision The laboratory has been established in 2016 as a part of Life Chemicals Inc. which facilities are located in representative office in Kyiv (Ukraine). Our activities are carried out to attain international regulatory compliance, such as ASTM (American Society for Testing and Materials), The European Chemicals Agency (ECHA), OECD (Organization for Economic Cooperation and Development). Obtained results can also be used further for the purposes of product(s) development, research and development, and technical support. Physico-chemical testing forms a substantial part of the data package as the integral requirement of product registration for drugs, agrochemicals, biocides, chemicals and animal health products particularly in non-EC countries and can be substantial in designing the appropriate toxicological and ecotoxicological test packages.

Mission and vision Activities of the laboratory are not confined only to the physico-chemistry and sample preparation; it also includes: Absorption Plasma Proteins Cytochrome P450 Selected tests of ADME/Tox (Adsorption, Distribution, Metabolism, Excretion, Toxicity) - helping our clients to make and stick to their goals with early stage preclinical drug development Chiral chromatography and resolution of racemates – helping to catch up the exact enantiomer of what you need to come up with a highest biological activity in screening experiments

Platform Microplate readers HPLC systems: Agilent 1260 Infinity II with 600 bar pump, well-plate autosampler (G1367D) and diode-array detector(G4212B). Agilent 1100 with variable wavelength detector, PDR-Chiral Advanced Laser Polarimeter (ALP), and Agilent 1200 Fraction Collector (G1364C) Agilent 1100/1200 with well-plate autosampler, diode-array, fluorescent detector and Softa 1400 evaporative light scattering detector НР 1090 with diode-array detector Microplate readers Perkin-Elmer, Victor 3, 1420 Multilabel Counter with Wallak Software Biotek, Epoch, microplate spectrophotometer with Gen5 Software. Two Eppendorf 5810R refrigerated centrifuges Genevac HT-4 Series II Evaporation System, plate shakers etc. Thermo Scientific® -80◦C Benchtop Freezer( ULT 185-5-V); Large collection of chiral, analytical and semi-preparative HPLC columns High throughput sample preparation platform which includes automatic Viaflo (Voyager) Integra/Thermo electronic pipettes, Merck Millipore filtering plates, microtiter plates etc.

Workflow Physico-chemical Laboratory The workflows of the Laboratory are subdivided on following modules: Physico-chemical Laboratory Physico-chemical test measurements Bio-mimetic chromatography and Rapid Equilibrium Dialysis (ADME) Enzyme research (ADME) Chiral chromatography, resolution of racemates and semi preparative HPLC

Physico-chemical tests Workflow Physico-chemical tests • Thermodynamic Solubility in water (HPLC, PBS) • Kinetic Solubility in water (HPLC, in the presence of DMSO) • Thermodynamic Solubility (UV plate reader, Turbidimetric) • Partition Coefficient (LogP/LogD) (HPLC, EU A8/OECD 117; Shake-Flask, EU A8/OECD 107; Shake-Flask/HPLC, in-house) • Ionization Constants (pKa) (UV microplate spectrophotometer) • Solubility in biorelevant media (FED STATE SIMULATED INTESTINAL FLUID (FeSSIF) and FASTED STATE SIMULATED INTESTINAL FLUID (FaSSIF)) • Solubility in organic solvents • Stability in organic solvents • Storage stability • Thermal stability

Bio-mimetic chromatography and Rapid Equilibrium Dialysis (ADME) Workflow Bio-mimetic chromatography and Rapid Equilibrium Dialysis (ADME) • Absorption (permeability) • Volume of distribution (log Vss) • Brain tissue binding • Overall tissue binding • Lung tissue binding • Mucus binding • Plasma protein binding on HPLC • Unbound volume of distribution (log Vdu) • Plasma protein binding in human, mouse and rat plasma (Rapid Equilibrium Dialysis)

Workflow Enzyme research CYP 450 inhibition assays (ADME) (IC50, inhibition type determination, Ki) Test on microsomes with Glo™ Assays Test on essential CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 with Glo™ Assays Test on microsomes with HPLC assay and fluorogenic substrates Test on essential CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 with HPLC assay and fluorogenic substrates Blood coagulation assays Inhibition assays on thrombin, Xa factor, plasmin with chromogenic substrates (IC50, inhibition type determination, Ki) Blood Clotting assay (fibrinogen-thrombin reaction, monitoring by clot formation)

Workflow Chiral chromatography, resolution of racemates and semi preparative HPLC Enzymatic resolution of racemates (Contract research) Chiral chromatography Enantiomeric excess determination (ee) Separation of enantiomers using chiral stationary phase Semi-prepartive separation of complex samples require special separation conditions