CB 3AA3

Lecture 7-8 Jan 28-29, 2021

Saman Sadeghi
Sadegs10@mcmaster.ca
Molecular Imaging

MOLECULAR In vivo
BIOLOGY Imaging
Imaging Modalities
Imaging Modalities
 Abnormal cells with
pathological phenotypes

Molecular expression

Probes / ligands may

be detected and allow

Therapy
Diagnosis with labeled
compounds

Identification Therapy
of targets Therapy response
for drugs planning
 Radiopharmaceuticals
• A radiopharmaceutical is a drug labeled with a
radionuclide targeting biological process
• The overall chemical structure determines biological
properties
• the radionuclide determines imaging and/or
therapeutic properties
PET Tracer Development
PET tracer development
 Production of positron-emitting
radioactive isotope
• Have to hit the target nucleus without enough
force to overcome the binding energy
Radioactive transformations
 Positron emission process
• positrons travel through human tissues, they give up
their kinetic energy mainly by Coulomb interactions
with electrons
• rest mass of the positron is the same as that of the
electron => large deviations in direction with each
Coulomb interaction, and they follow a tortuous
path
 18F - Physical properties
18
9F T1/2 = 109.8 minutes

b+ 96.73% Eb+max = 635 keV

EC 3.27% Eg = 511 keV

8O
18

Methods of preparation:
18O ( p , n ) 18F 16O ( a, 2n ) 18F
20Ne ( d , a ) 18F
16O ( 3He , p ) 18F 20Ne ( 3He , ap ) 18F
16O ( a , pn ) 18F 18F
 History of cyclotrons
• Conceived in 1920s, first operated in 1932
• Stanley Livingston (L) and Ernest O. Lawrence
1934, 27 inch cyclotron
 Major Components of a cyclotron

• Vacuum chamber
• Ion source
• Magnetic field and RF voltage for high energy beam generation
• Beam extraction and proton generation
• Target
 RADIOCHEMISTRY LAB
• Radiosynthesis lab QC setup
 Radiosynthesis

Radiosynthesizer Rotary
evaporator

Hot cell

Semi-prep HPLC
 Automated synthesizers

Steel et al. J Label Compd Radiopharm 50: 308-311 (2007)

FDG, 10 min reaction, 9 sec residence time
Hotcells and Synthesizers
Preclinical Imaging
Molecular Imaging

Chemistry
Immunocompromised Animals
Imaging and Validation
In vitro Studies
 68Ga-DOTATATE

• Neuroendocrine tumors
• Low glucose consumption but high
expression of
somatostatine
receptors
 68Ga-DOTATATE

• Relapse detection
• In conjunction with FDG
Theranostics

• Targeted radionuclide therapy

• (Ga-68-DOTATATE PET/CT) to detect the
presence of a molecular target
• Somatostatin receptor positive selected for
therapeutic intervention (Lu-177-DOTATATE)
 Targeting Of Mitochondrial
Metabolism
• Functionally map mitochondrial
networks in oxidative and non-
oxidative
• Similar metabolic signatures in
HNSCC and Triple Negative Breast
Cancer, going beyond FDG

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 Antibody and Fab radiolabeling

Reference
Fu, R.; et al. Antibody Fragment and Affibody ImmunoPET Imaging Agents: Radiolabelling Strategies and
Applications. Biopolymers. 2019
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 Radiolabelled Antibody Recruiting Molecules

• Form of cancer therapy that Target Binding Antibody Binding

Terminus Terminus
activates the immune
system to “attack” the
tumour
• Using small molecules Cancer Cancer
containing a known antigen cell cell
to recruit endogenous Radiolabelled ARM
antibodies to the tumour
• Antibody - labelled cancer
cells become “visible” to Delivery of
cytotoxic radiation
the immune system
• Concentration of ARM
required for cell death is
similar to that required for Immune-mediated
radiotherapy Clearance

Spiegel et al. J Am Chem Soc. 2009, 131(47), 17090–17092

 Strategy For Radiolabeling

Cysteine Residues Lysine Residues

DFO-Bz-SCN DOTA-NHS

Advantage: Site-selective Advantage: >20 available lysine

Disadvantage: Antibody Instability Disadvantage: Can reduce affinity
Disadvantage: Max ~8 cysteine

40
Pretargeted Imaging

Fast
Selective
Bio-orthogonal
 Experimental Strategy

1 Antibody Conjugation
and Validation

2 Radiolabelled Tetrazine
Probe Development

3 In vivo Testing

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 Strategic Overview
Creating a Novel Radiolabelled anti-CD133 antibody

Conjugation Radiolabelling

Probe
Development

In vitro In vivo

Evaluation