ADME SOLUTIONS 1 confidential CORE TECHNOLOGIES PRODUCTS ADME

ADME SOLUTIONS 1 confidential

CORE TECHNOLOGIES/PRODUCTS ADME • Line of automated workstations for drug screening in discovery and development. • Installed base at big pharma, strong current interest, clear product expansion route. Structural Signature Technology • Breakthrough technology for rapid assessment of structural changes to proteins. Proteomics • First true adjustable third dimension. • Very significant potential application base in both biotech and pharma. Initial commercial feasibility underway. • Studies in clinical marker discovery underway. 2 confidential

Technologies 3 confidential

Spectrum of Solutions Automated Discovery Workstation Automated Solubility Workstation Automated Log. D Workstation Solubility Log. D Permeability Blood-Brain Barrier Permeability Metabolic Stability Plasma Protein Binding 4 Available R&D Completed confidential Included Option

ADME in Discovery and Development (1) • Discovery: – – Objective: Samples: Throughput: Accuracy: Fail hits early Dry and DMSO-dissolved Up to 200 samples/day (typically) (Supposedly) not high – rank order is sometime sufficient • Development: – Objective: – Samples: – Throughput: – Accuracy: 5 confidential Optimize lead (better ADME parameters), salt & excipient selection, QSAR, etc. Dry Typically up to 100/day, but increasing High, must use gold-standard validated assays

ADME in Discovery and Development (2) • State-of-the-art today: – – Discrete instruments, each performing a single assay Data consistency between different assays non-existent Significant interaction required from trained personnel No process engineering methodologies: • When assay is not required, the system is idle • When a system is down, there is no replacement • To double the capacity, each system must be duplicated • Automated Discovery Workstation: The modern approach to the entire lab 6 confidential

Automated Discovery Workstation - ADW • • Not an instrument: It is an integrated solution (lab-on-a-bench) Machine work – people think Turnkey, automated, multi-assay workstation Performs (presently) discovery/development assays: – – True Log. D/Log. P vs. p. H True aqueous solubility vs. p. H Identity and purity of compound Compound concentration w/o standard • Scalable, Flexible, Extendable technology • Standardized, validated data 7 confidential same sample

ADW - Automated Discovery Workstation ANALIZA’s In-house Development System (2002) 8 confidential

Two Key ADME Assays 1. Log. D/Log. P • P is the partition coefficient between octanol and a buffer, that is obtained under equilibrium • Important for multiple ADME processes in discovery and development, and for QSAR lead optimization • Very extensive database exists well validated method • Direct Log. P assay (shake flask = gold standard): Add solute Phases Mix Settle Sample and analyze Octanol sat. w/buffer Ctop Buffer sat. w/octanol Cbot 10 confidential

ADW: Log. D/Log. P Assay 11 confidential

Two Key ADME Assays 2. Solubility • Solubility is the concentration of the compound in solution in equilibrium with its solid • Important for bioavailability and other ADME processes • Very sensitive to measurement methodology • Current profiling solubility indicators are frequently different from the true values • Direct solubility assay (shake flask = gold standard) • Unique features: – No sample weighting – No assay calibration required to calculate solubility as mass/volume 12 confidential

ADW: Solubility Assay 13 confidential

Validation: Log. D Accuracy Log. DADW = 0. 002(± 0. 008) + 1. 011(± 0. 005)*Log. Dmanual N = 179; r 2 = 0. 9960; standard error of estimate = 0. 1022 14 confidential

Validation: Solubility Accuracy 15 confidential

Validation: Precision Allopurinol in 0. 15 M Na. Cl in 0. 01 M Universal buffer, p. H 6. 6 16 confidential

ADW Control and Analysis Software 17 confidential

Log. D Assay: Main screen with optional information windows 18 confidential

Solubility Assay: Setup, imported data 19 confidential

Data Viewer: Full featured archival data re-display and re-analyze GUI, which runs from any computer on the network 20 confidential

Data Viewer: Display of archived raw PDA data – sample spectra You Never Lose Your Data! 21 confidential

ADW Hardware • Robotic liquid handler (Hamilton 4200, four computercontrolled syringes, custom valves). • Custom computer-controlled valve system (Rheodyne). • UV/VIS photo-diode array spectrophotometer (Groton Technologies) • Chemiluminescence total nitrogen detector (Antek Instruments) • Custom electronics, bar-code reader, and high-end control computer • Vacuum and HPLC pumps • Deckware: plate and filter holders, buffer racks, etc. 22 confidential

ADW Hardware: Assay Specific Log. D • Customized plate vortexer • Customized plate sealer • Refrigerated plate centrifuge Solubility • Custom filter plate holders • Customized filter plate presser • Customized shaker • Buffer containers holder • Temperature option (heated shaker, on-deck water bath ) 23 confidential

ADW Supplies Consumables: • Log. D systems: – Loaded in 96 deep well plates, bar-coded, QC’ed – Ready-to-use in 6 standard p. H levels • Solubility: – Filter vials Optional/Maintenance: • Nitrogen-detector calibration plates • Extra filter plates • Repair kits, parts Always Available: A direct hot-line to our scientists and system developers – you always get a timely answer, advice, and dedicated service 24 confidential

ADW Key Specifications ATTRIBUTE ASSAY VALUE Sample Requirement Log. D Solubility Dry DMSO-dissolved 150 µl Range Log. D Solubility -3

The ADW Solution Vs. Competition Attribute Competition Turnkey solution Yes Typically No Number of Assays 5 - small incremental cost per additional assay retains capital investment 1 Assay Methodology Direct gold standard assays True data only Typically indirect – limited applicability range and potential incompatibility with development data Degree of Automation High – Typical user interface is about 1 hr. per 24 hr. operation Varied – Typically requires extensive interaction Operator Level Technician Varied – Typically a scientist Archival Database Unique, enterprise-based, web-shared, graphical raw data and analysis tool set Typically final data reporting Flexibility Yes – Reconfigurable easily between assays = full workstation utilization No – When the specific assay is not required, the workstation is idle Process Engineering Methodology 26 ADW Yes – Modular, extendable, production-line approach No – Every instrument performs one assay confidential

ADW: The Integrated ADME Environment Assay Library Integrity (Discovery) Solubility Log. D Metabolic Stability Permeability Plasma Protein Binding Blood-Brain Barrier 27 confidential Description Estimated Time to Market • Identity (LC/MS) • Purity (PDA, LC/MS) • Concentration (N-det) • 6 months • Available, 6 months • Available • True (dry) • Kinetic (DMSO) • Available True (DMSO) Available Standard LC/MS assay 6 months Novel high throughput assay • Log. P & PSA based 3 months • Surface tension based 1 year • Microdialysis • Novel direct free/bound assay • 12 months • 18 months • Log. D/Log. K • Novel high throughput assay • Available (software 3 months) • 12 months

AUTOMATED SOLUBILITY WORKSTATION ASol. W in Pre-Formulation/Development 28 All Slides Confidential confidential

Solubility • Solubility is the concentration of the compound in solution in equilibrium with its solid • Solubility takes several forms and is a function of multiple parameters (crystalline form, temperature, p. H, co-solvent, etc. ) • Important for bioavailability • Very sensitive to measurement methodology • Current profiling solubility indicators in early discovery are frequently different from the true values obtained during development • A direct solubility assay is defined by the shake flask method and is recognized as the gold standard 29 confidential

Who Are The Users? • Discovery: – Obtain a measure of solubility of relevance to HTS and early ADMET • Can we determine if the compound will crash out of solution? – Obtain ADMET information of relevance for selecting amongst hits • Development: – Early lead characterization – Lead optimization – Formulation excipient selection 30 confidential

Who Are The Users? (2) • Will the compound crash out? solubility • Everyone else needs good data: – For promoting a compound: binning may be sufficient – For lead optimization: – For formulation development: – For anything regulatory: 31 confidential accurate data

Alternative Techniques for Measuring Solubility A Comparison (1/2) Feature True Solubility – dry Yes/No Kinetic Solubility – DMSO Yes/No Shake Flask Method Yes/No Automation Yes/No Measure Solubility: Direct/Indirect Standard Curve: Required/Not required 32 confidential Absorbance Light Scattering ASol. W

Alternative Techniques for Measuring Solubility A Comparison (2/2) Feature Multiple p. H Levels Yes/No Multiple Temperatures Yes/No Multiple Co-Solvents Yes/No Timed Sampling Yes/No Additional Steps: Pre-concentration, Dilution curves Rely on: Chromophores Rely on: Scattering definition 33 confidential Absorbance Light Scattering ASol. W

The ASol. W Assay 34 confidential

ASol. W with Whatman’s Mini. Prep Filter 35 confidential

Options for Measuring Solubility ASol. W: Dedicated Instrument 36 ADW: Multi-Assay Instrument confidential

ASol. W – Automated Solubility Workstation ANALIZA’s ASol. W In-house development system 37 confidential

ASol. W: Assay setup 38 confidential

ASol. W: Data Check 39 confidential

ASol. W: Runtime Panel 40 confidential

ASol. W: Accuracy 41 confidential

ASol. W: Precision Allopurinol in 0. 15 M Na. Cl in 0. 01 M Universal buffer, p. H 6. 6 42 confidential

Total Nitrogen Detection • Exceptions: – – 43 N 2 – nothing is detected! Nitrosamine (for GC) Azides (1/3 response) Double bonds (partial response) confidential Separate calibration

ASol. W: Assay Using Nitrogen Content 44 confidential

Basic Issue: What is the Effect of DMSO? 45 confidential

Basic Issues: What is the Effect of DMSO? 46 confidential

Basic Issue: What is the Effect of DMSO? Typically: • DMSO is a fact of life during screening • DMSO solubility is > or >> than true solubility • But just because we relaxed one experimental parameter does not mean that we can dispense with everything else • Make sure you know what you are using the data for – especially in lead promotion decisions 47 confidential

ASol. W Hardware • Robotic liquid handler (Hamilton 4000, two computercontrolled syringes, custom valves). • Custom computer-controlled valve system (Rheodyne). • Chemiluminescence total nitrogen detector (Antek Instruments CLND 8600) • Custom electronics, bar-code reader, and high-end control computer. • Custom filter plate compressor • 4 -filter plate shaker/heater incubator • Miscellaneous accessories: Vacuum pumps, HPLC pump • Deck accessories: Filter plates, buffer racks 48 confidential

ASol. W Options, Kits, and Consumables Options: • Temperature option (to 37 deg. C) Kits: • Starter filter pack (included) • Maintenance kit • Additional filter plates, buffer/co-solvent bottles, etc. Consumables: • Filter packs • Nitrogen-free buffers 49 confidential

Why ADME with ANALIZA? • • We are experienced We provide innovations with gold-standard assays We don’t just automate We report archival-quality data while minimizing material consumption and at high throughput • We guard your investment and offer flexibility and extendibility to our workstations • We pride ourselves with a personal, responsive, firstclass organization before and after the installation 50 confidential

Thank You 51 confidential