Presented at the Global Medicinal Chemistry and GPCR Summit. To find out more, visit: www.global-engage.com Alexander Alanine, CEO of Bacteva, introduces the Totally Integrated Medicines Engine (TIME), designed to speed up drug discovery with integrated microfluidics.

1. alexander.alanine@bactevo.com
Next Generation Drug Discovery:
T.I.M.E. an Integrated Micro-Droplet Based Platform
bactevo
Global Medicinal Chemistry Leaders Summit
www.bactevo.com

2. The Need
* Scannell JW, Bosley J (2016) When Quality Beats Quantity: Decision Theory, Drug Discovery and the R
** Swinney DC, Anthony J (2011) How were new medicines discovered? Nature Reviews Drug Discovery 10
*** Kelvin Stott (Novartis), Linked-In(18 November , 2017) Pharma’s broken business model: industry on the
eproducibility Crisis. PLoS ONE 11(2): e0147215. doi:10.1371
, 507-519 | doi:10.1038/nr
brink of terminal decline
What’s next?
K. Stott (18.11.17 Pharma’s broken business model: industry on the brink of terminal decli
www.bactevo.com2

3. The Need
* Scannell JW, Bosley J (2016) When Quality Beats Quantity: Decision Theory, Drug Discovery and the Reproducibility Crisis. PLoS ONE 11(2): e0147215. doi:10.1371
** Swinney DC, Anthony J (2011) How were new medicines discovered? Nature Reviews Drug Discovery 10, 507-519 | doi:10.1038/nr
*** Kelvin Stott (Novartis), Linked-In(18 November , 2017) Pharma’s broken business model: industry on the brink of terminal decline
What’s next?
The Bactevo Solution
‘Total information’ patient-relevant screening
– Millions diverse small molecules, made, screened and analysed in aweek
– Primary screening can be performed on patientcells
– All ADMET and activity screens performedsimultaneously
K. Stott (18.11.17 Pharma’s broken business model: industry on the brink of terminal decline
www.bactevo.com3

4. Bactevo: Systems Approach: Parallel & Rapid
8 years
Gaining years of additional marketexclusivity
- Exploratory medicine
- Rolling NDA-based clinical studies
- Indication expansion
- Higher commercial returns
Scalable
Target
ID
Chemistry
HTS
Lead Selection Lead Optimisation
0-2 years 1-1.5 years
Traditional Process: Linear & Slow
2 years 3-4 years
months
multiplepatient
derived cells
www.bactevo.com4
Creating a Paradigm Shift in New Medicines Discovery

5. Bactevo Therapeutics Discovery Platform
Totally Integrated Medicines Engine (TIME)
uHTS
Microfluidics
‘Total SAR’
Synthetic
biospace
libraries
Lead profiling,
ADME, safety
In vivo ready
Therapeutic
Drugs
Cheminformatics
Bioinformatics
Machinelearning
Game-Changing Platform
www.bactevo.com5

6. Bactevo Therapeutics Discovery Platform
Totally Integrated Medicines Engine (TIME)
uHTS
Microfluidics
In vivo ready
Therapeutic
Drugs
Game-Changing Platform
www.bactevo.com6

7. The Screening Platform
Can Screen in HT Close to the Disease
Versatile micro-droplets used as reaction vessels
• Conditions inside micro-droplet open to huge variation,
including gas permeability, hydrophobicity, pH etc.
Anything with an assaya
• Micro-droplet stabil
Cells
ble read-out can be placed in micro-droplets
ity > 1 month, survive PCR
Enzymes or Proteins Organisms or organoids/organisms
Throughput > 500 Million Micro-droplets/day: Platform Totally Scalable
www.bactevo.com7

8. Micro-droplet Encapsulation:
Homogenous & Ultra High Throughput
Human cells in droplets
Parallel droplet generation setup
High-Throughput, Monodisperse Microfluidics
www.bactevo.com8

9. Examples of Cell Growth in Micro-droplets
E. coli + GFP Streptomyces
Hoescht
Mitotracker Red CMXRos
Bacteria and
Jurkat
co-culture
Viable co-culture ofmammalian
and bacterialcells
Bacterial cells in micro-droplets Staining HEK cells in micro-droplets
HepG2 cells in micro-droplets
Jurkat
only
Bacterial
cells only
Mammalian cells viable for > 7-14 days, Bacteria Viable >1 Month
www.bactevo.com

10. Versatile : Phenotype, Morphological, Biochemical via Growth Changes orFluorescence
Library
(synthetic, Nat. product)
Indicator cell
(mammalian or bacterial)
Cells Expressing
Reporters
Engineered reporter cell lines
• Cell markers
• Gene expression or reporters
linked to FP or enzyme
• FRET for PPI
Co-encapsulate the
Target with reagent
Dye following cell encapsulation
• Functional dyes (Mito tracker)
• Fluorescent products
• Antibodies
• Proteins
Label the Target
after Incubation
Uniquely label at end of assay
• Antibody staining
• Functional dyes
• Counter stains
The Screening Platform
All High Content Readouts ArePossible
www.bactevo.com10

11. Vast Efficiencies in Screening Ability
Huge Chemical Diversity Can Be Rapidly Explored
www.bactevo.com11
• Pictured 3280 x 384 well plates (1.26 million wells)
• Droplets represent >52,000-fold reduction in volumes
• 1.26 million droplets < 5 minutes screening!
Image: 20 M Droplets vs. 1.26 M wells (384 wellplate)

12. Bactevo Therapeutics Discovery Platform
Totally Integrated Medicines Engine (TIME)
uHTS
Microfluidics
‘Total SAR’
Synthetic
biospace
libraries
In vivo ready
Therapeutic
Drugs
Game-Changing Platform
www.bactevo.com12

13. www.bactevo.comCONFIDENTIAL
http://mappingignorance.org/2014/07/04
Brown, D.G. at al. J.Med Chem. 2016, 59, 4443−4458
Synthetic Small Molecule Library Design
Rule of 3 core design, for Lead-Like Improved Drug Leads
13

14. Generate Total SAR (TSAR) on EVERY Compound
Encoded Synthetic Libraries: Lead-Like design
Scalable libraries & SAR data in phenotypic screening
• Drug-designed synthetic libraries
− Lead-like (<375 Da) compounds created indays
• Smart Design
– Topology-rich sp3-rich cores, tuneable properties: Mw, PSA, logP,fsp3.
– Optimal drug-like space for bioactivity(Rule-of-3)
– Diverse Chemotypes: harness knowledge from internal natural productlibraries
• Organic solvent and low pH chemistry enabled: unleashes vast synthetic library scope
• Single compound tag-less, NOT mixtures
– Screening at high concentrations up to high µm & dose-response screening(EC50)
− True active and inactive dose-response
− N-fold repeats (high coverage)
ALL Compounds dose-response screened: Active, Inactive, Normalisers
www.bactevo.com14

15. Chemical Diversity with Combinatorial Libraries
DNA Encoding Library (DEL)Technology
A
B
C
Pool 1
X
Y
Z
A
B
C
AX
AY
AZ
BX
BY
BZ
CX
CY
CZ
AX
Screen by affinity binding:
Usually depicted as… To scale…
• Very large libraries possible (> 109)
• fMole amounts of each compound inmix.
• Throughput high, protein demand low
• Activity trackable via DNA-tag
• DNA-tag limits assay choices
Huge libraries, Complex Compound Mixtures
Starting core/ linker=
www.bactevo.com15

16. Synthetic Library Screening & Decoding
Retain Link Between Synthesis Code and Phenotype Effect
+
Encapsulate in droplets
Compound carrier & readout in proximity
Payload released from carrier in droplet
Observe phenotypic change
in target cells/ assay
Cells
Detect effect
By fluorescence
Cellular Assays Conducted in Sealed Microdroplets
µ Fluidic
chip
www.bactevo.com16
FACS
(FADS)

17. Dose Response During Primary Screen
Load Defined Amounts of Payload on Beads forConcentrations
in Droplets
High Loading
Detect Positive
Effect
No Effect at Low
concentration
Quantity of scaffold loaded
determines concentration in
droplets (vol. 0.5 nl)
Low Loading
Encapsulate
Dose Response Enables Ranking of Hits Based on Potency
www.bactevo.com17

18. Dose Response During Primary Screen
Load Defined Amounts of Payload on Beads forConcentrations
in Droplets
High Load
Detect Positive
Effect
No Effect at Low
concentration
Quantity of scaffold loaded
determines concentration in
droplets (vol. 0.5 nl)
Low Loading
E
ing
ncapsulate
Dose Response Enables Ranking of Hits Based on Potency
www.bactevo.com18

19. Broad Synthetic Methods for Library Construction
✓ No major barriers to further synthetic reactions
in organic or aqueous conditions.
o Library generation and screening in a week
Growing Synthetic Repertoire
✓ >10 different chemistry methods and counting…
(C-N, O-C, C-C, C-O, cycloaddition, SnAr)
Day 1 Day 3 Day 4Day 2 Day 5
Reaction #1
Reaction #2
Reaction #3
Encapsulate
and screen
Screening
Synthesis
Rapid Methods Development: Aqueous or Organic Phase Chemistry
Sparse library sampling with Multiple cores
X
Y Z
www.bactevo.com19

20. Harnessing The Best Elements
Lead-like ‘medicinal’ Chemistry LibraryDesign
www.bactevo.com20
Attributes Bactevo DELT HTS Collections
No. of compounds accessible in a singlelibrary 1000 million 1000 million 2 million
Rapid library synthesis (<1 week) ✓ ✓
Chemistry in non-aqueous/organicsolvents ✓ ✓
Tolerance to DNA damaging conditions (lowpH) ✓ ✓
Low Library storage demands & synthesiscosts ✓ ✓
Compounds at bio-relevantconcentrations ✓
(μM - mM) (pM)
✓
(μM - mM)
PhenotypicScreening ✓ ✓
Target Screening ✓ ✓ ✓
No steric hindrance of ligand bytag ✓ ✓
Low cell demand/ reagentusage ✓
Simultaneous assay screening incl. safety ✓ ✓

21. Bactevo Therapeutics Discovery Platform
Totally Integrated Medicines Engine (TIME)
uHTS
Microfluidics Lead profiling,
ADME, safety
In vivo ready
Therapeutic
Drugs
Game-Changing Platform
www.bactevo.com21

22. HepG2 cells in dropletsExtracts tissue and screen
1g of tissue = 10 million droplet assays
10x repeats!
Lead Profiling & Toxicity
Integrated ADMET Screening: Drug SafetyPredictions
Safety: Diverse mammalian cell lines
• Isolated organ cells from model species (safety &PK)
• IPSC’s
• Patient derived tissue (biopsy)
• Cross-species analysis (rat, dog, mini-pig, NHP,human)
in vitro ADMET: VariousAssays
• Cytoxicity (membrane, apoptosis, ion flux, necrosis)
• Mitochondrial function
• Cell cycle
• Gene induction
• Cell proliferation
Mammalian cells viable up to 14 days, Bacteria 30 days
Dead cells
by scatter
pattern or
with dyes
www.bactevo.com22

23. Activity
Safety
Parallel Primary Screens: Acceleration in Discovery Process
Clinical leads
ADMET profiled
Multi-Parameter SAR Drives Discovery Process
Navigation of Multi-Parametric Space Toward Lead Candidates
ADME
High Quality Data Sets & Machine Learning = Safe, Efficacious NME’s Faster
Typical ADMET properties:
Typically slow, iterative - few compounds
Parallel & integrated in T.I.M.E. platform
www.bactevo.com23

24. Mitochondrial Disorders
Internal Programs - Mitochondrial Diseases Plus…
Inducing Mitochondrial Biogenesis
• Severe mitochondrial diseases are classed as rare (1 in 4000), but are alsoa
key gateway to many other diseases
• Increasing Mitochondrial Biogenesis increases mitochondrial load andfunction
• UK has co-ordinated response, best and largest patient cohorts in the worldand
key leaders in the basic research and clinical analysis (collaboration with
WTCMR, Newcastle – Prof. Sir Doug Turnbull)
• Discovery of novel compounds which increase mitochondrialbiogenesis
MELAS,
LHON
Parkinson's
Disease
Alzheimer’s
disease
Hearing
Loss
Diabetes
Immuno-
oncology
HMD
Regen.
Medicine
Mitochondrial
Biogenesis
www.bactevo.com24

25. Mitochondrial Biogenesis Increase Detected
IncreasingMitochondrialstain(TMRM)
• Assay compares DNA content to Mitochondria content of a cell
• Hits show increased TMRM staining compared to DNA staining
• Hits confirmed by different secondary screening methods
With Library
in droplets
Mammalian cells
alone in droplets
Internal Programs – Mitochondrial Diseases
Phenotypic Actives Identified
Enhanced
Mitochondrial
Function
25
Hoescht
TMRM
Increasing Hoescht (DNA) staining

26. www.bactevo.com
Game-Changing Platform
Bactevo Therapeutics Discovery Platform
Totally Integrated Medicines Engine (TIME)
26
uHTS
Microfluidics
‘Total SAR’
Synthetic
biospace
libraries
Lead profiling,
ADME, safety
In vivo ready
Therapeutic
Drugs
Cheminformatics
Bioinformatics
Machinelearning

27. Harnessing the Chemical Universe to Treat Disease
Smarter, Faster Drug Discovery
27 www.bactevo.com
Gene Space
(personalised medicine)
Disease Space
(patient cell screening)
Chemical Space
(Smart libraries)
Machine-learning
& Informatics
Bactevo Accelerates Drug Discovery
by the Integration of Chemical Space, Disease Relevant Biology & MachineLearning
Drug Space
(ADMET)

28. www.bactevo.com
Presented at the Global Medicinal
Chemistry and GPCR Summit.
To find out more, visit:
www.global-engage.com