Presented by Juliana Maynard on 13th May 2020

1. In-vivo imaging to
understand the
Biodistribution of a
candidate compound
Juliana Maynard
May 2020

2. © 2019 Medicines Discovery Catapult. All rights reserved.
Introduction: MDC Science Focus
Building the link between compound - target - biological effect in translatable
systems to reduce pre-clinical attrition of drug discovery projects
Translating clinical information back into the pre-clinical phase to ensure
better models, better biomarkers and a patient selection strategy
Pre-clinical imagingComplex Cell Models
Target Engagement & Validation
Bioanalytical Technologies
Biomarkers

3. © 2019 Medicines Discovery Catapult. All rights reserved.
Imaging in drug discovery
Imaging technologies provide visualization of biological functions to accelerate disease understanding
and drug development
Organ accumulation & biodistribution (PK)
Target expression & engagement
(biomarkers, stratification)
Biological activity
(PD, optimal biological dose)
Toxicities/Drug interactions
(safety)
Surrogacy for clinical endpoint
(predictive efficacy)
PK,PK/PDandbiologicalmodelling
Multimodal and multiscale imaging solutions enable quantification and
understanding of:

4. © 2019 Medicines Discovery Catapult. All rights reserved.
WHO WHY HOW WHAT WHERE WORK WITH US
Organ Accumulation & Biodistribution
Tracking where your compounds travel in an experimental animal or human
What is the challenge? Determining the
percentage of an administered dose that is
deposited into specific organs throughout the
body at specific time points.
What is needed? Fully quantitative new and
novel approaches to understand the
pharmacokinetics of small molecules and
complex medicines
What is the solution? In-vivo and ex-vivo pre-
clinical imaging to characterise and validate
compounds towards clinical translation

5. © 2019 Medicines Discovery Catapult. All rights reserved.
What can imaging do?At the Medicines Discovery Catapult:
Imaging expertise
in multi-parametric
imaging modalities:
Positron Emission Tomography (PET)
Gamma counting biodistribution analysis
Computed Tomography (CT)
High Frequency Ultrasound (HFUS)
Bioluminescence/ NIR imaging (IVIS)
Mass Spectrometry Imaging (DESI, MALDI)
In-Vivo/Ex-vivoanatomical
andfunctionalimaging
Home Office PPL and radiological licence
Access to novel radiochemistry &commercial radiotracers
Access to animal models across disease areas
Access to pre-commercial technologies
Multi-modal enhanced interpretation of functional and molecular data
MolecularPathology
Advanced Microscopy (super-
resolution/confocal/multiphoton)
Digital Spatial Profiler (Nanostring DSP)
Photoacoustic imaging (PIA)

6. © 2019 Medicines Discovery Catapult. All rights reserved.
How can we use imaging and why do we need to ?
-Drug distribution reflects the extent to which it is present in extravascular tissues and the rate and
degree is complicated and depends on blood flow, capillary permeability and protein binding
5 required steps required for effective medicines
Circulation
Accumulation
Penetration
Internalisation
Release
Tracking of each process from the systemic to
subcellular level becomes essential in
Characterisation
How can we measure this?
Immunofluorescence
Imaging
Mass Spectrometry
Positron Emission Tomography
Near Infrared Imaging
Mass Spectrometry Imaging

7. © 2019 Medicines Discovery Catapult. All rights reserved.
Longitudinal Cy-7 Biodistribution
NIR imaging holds considerable promise for in-vivo imaging allowing deep biological penetration and the ability
to image whole body distribution with good signal to noise
Labelling of a biomolecule (peptide, protein, or other) with a NIR dye makes it possible to track it, and study
distribution of the molecule
NIR imaging tumour integrin expression NIR imaging antibody tumour distribution
Wu et al, 2006

8. © 2019 Medicines Discovery Catapult. All rights reserved.
89Zr PET distribution to assess off target toxicity
89Zr PET distribution
Off target toxicity and non specific binding can often cause issues with promising therapeutics
Aim: 89Zr imaging full body distribution and assess non-specific binding of an antibody as a promising ADC
Hypothesis: Due to the nature of the
target binding may occur in external organs
Study: Full body 89Zr labelled distribution
study at 4 longitudinal timepoints
Conclusion: No non-specific binding
observed in any tissues demonstrating the
potential for addition of a cytotoxic payload
Longitudinal time points

9. © 2019 Medicines Discovery Catapult. All rights reserved.
89Zr longitudinal imaging to assess tumour penetration
89Zr PET distribution
-Penetration and distribution of biologics depend on a variety of factors, such as tissue density, antibody affinity,
antigen density per cell and physio-chemical properties
-89Zr PET imaging is a tool for assessing penetration kinetics of antibody distribution
High levels of 89Zr accumulation observed in the tumour throughout time
TUMOUR
TUMOUR
TUMOUR
TUMOUR
Longitudinal Time Points

10. © 2019 Medicines Discovery Catapult. All rights reserved.
89Zr PET to explore whether Microbubble Enhances
Therapeutic Delivery to Tumour
Two major obstacles facing cancer nanomedicine are the
tendency of nanoparticles to be taken up by normal tissues and
organs and the nanoparticles' inability to efficiently penetrate
solid tumours
University of Leeds ThMb & HIFU platform
MDC collaboration with the University of Leeds to progress their
delivery platform into clinical development
Can we show superiority of platform and understand
mechanism?
Deliverable: Enhanced tumour delivery of microbubbles as a
drug carrier demonstrated for first time.
Also extensive biodistribution characteristics identified.
Future enablement of clinical development
A full understanding of the mechanism of drug delivery and
distribution properties meet a key demand in regulatory
guidance

11. © 2019 Medicines Discovery Catapult. All rights reserved.
Blood Brain Barrier Penetration
The blood–brain barrier (BBB) prevents the brain uptake
of most pharmaceuticals
Certain small molecule drugs may cross the BBB via lipid-
mediated free diffusion (mw <400kDa)
These chemical properties are lacking in the majority of
small molecule drugs, and all large molecule drugs
Nevertheless, drugs can be reengineered for BBB
transport, based on the knowledge of the endogenous
transport systems within the BBB
There are several ways to test whether a compound
crosses the BBB

12. © 2019 Medicines Discovery Catapult. All rights reserved.
Mass Spectrometryimaging
High spatial resolution approach to define the drug
distribution in very small brain structures
NIRimagingPET imaging
A therapeutic antibody and its ability to cross theBBB
89Zr PET to assess BBB Penetration of an Ab
NIR fluorophores and bioluminescent probes
for
brain imaging
Shihong et al; 2017
Hart & Patel

13. © 2019 Medicines Discovery Catapult. All rights reserved.
BBB Spheroid permeabilized with Triton
Mass Spectrometry Imaging (MSI)
Desorption ElectroSpray Ionisation Matrix Assisted Laser Desorption Ionisation

14. © 2019 Medicines Discovery Catapult. All rights reserved.
Measuring Tissue Drug Concentrations
Compound penetration:
• DESI-MSI for drug distribution
• Overlaid with H&E stain
• MALDI-MSI for improved
resolution and confirmation of
localisation

15. © 2019 Medicines Discovery Catapult. All rights reserved.
Evaluation of BBB penetration of Polymeric
Nanoparticle
Doxorubicin 50 umIdarubicin
50µm
DAPI
Occludin 1
BBB Spheroid Model
BBB Spheroid permeabilized with Triton
BBB Spheroid
Polymeric Nanoparticle-Rhodamine
0
50
100
150
200
250
0 50 100 150 200 250
Fluorescence(A.U)
Distance (microns)
0
50
100
150
200
250
0 50 100 150 200 250
Fluorescence(A.U)
Distance (microns)
BBB Spheroid permeabilized with Triton

16. © 2019 Medicines Discovery Catapult. All rights reserved.
Summary
• Imaging offers huge potential as a platform for non-invasively measuring the
pharmacokinetics and characterisation of small molecules, biologics and non-biological
complex medicines
• Exploitation of current capabilities and development of emerging opportunities and
techniques will facilitate faster translation into clinical development
• Discovery and Research Driven; Challenging and optimising the use of imaging and
testing
• Moving from compounds to cell tracking and more – transformational opportunities
“Reimagining how
drugs can benefit
patients to improve
the efficacy”
“ cutting edge
expertise with a
fresh bold
approach – make a
difference to a
patients life”
“Delivering results
for patients as
efficiently as
possible”
“translating
academic research
into clinical practice
– providing
patients with the
latest thinking”

17. © 2019 Medicines Discovery Catapult. All rights reserved.
Get in touch nowAcknowledgements
Medicines Discovery
Catapult
• Peter Simpson
• Sally Price
• Duygu Yilmaz
• Isabel Peset-Martin
• Philippa Hart
• Benedetta Arno
• Neill Gingles
• Gemma Forrest
• Gayle Marshall
• Emma Jones
Our collaborators
• University of Leeds
• AstraZeneca
• Sutura Therapeutics
• LifeArc

18. © 2019 Medicines Discovery Catapult. All rights reserved.
Get in touch now
Juliana.Maynard@md.catapult.org.uk
@meddisccat
info@md.catapult.org.uk
01625 238734
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Thank you for
listening!