“Graduate Studies in Radiochemistry at

University of Missouri”

Susan Z. Lever

Biomedical Program Director, MURR

Associate Professor of Chemistry

Things to know about MU #1:Things to know about MU #1:

• Highest concentration of “radiochemistry” research scientists on any campus– Breadth and diversity of research topics

• Vet School, Med School on the same campus– Interdisciplinary research possibilities

• Law School, Business School on campus– Student population has a range of interests

• Journalism School and School of Education– Cross-discipline possibilities

Things to know about MU #2:Things to know about MU #2:

• Highest flux research reactor of any US campus

• High field nmrs, with an 800 MHz on order

• Cyclotron to be installed in 2007

• Small animal imaging capabilities include SPECT, PET, MRI and CT

Radiopharmaceutical Science Program at MU

ChemistryChemistry

BiochemistryBiochemistry

Veterinary Veterinary MedicineMedicine RadiologyRadiology

Internal Internal MedicineMedicine

MURRMURR

Things to know about MU #3:Things to know about MU #3:

• Long history of radiopharmaceutical development

• Established program in archeometry

• World-renowned location for contribution of trace metal analyses to biomedical problems

• Active grant funding and current projects in the above areas

Ceretec - a Neutral Tc-99m Complex

• Ceretec is used for the diagnosis of abnormalities of regional cerebral blood flow

• Epilepsy

• Alzheimer's Disease

Andrew Newberg, University of Pennsylvania

http://www.andrewnewberg.com/ accessed 6/21/05

Blood Flow during Meditation

Design, synthesis and evaluation of novel biological targets for diagnosis and therapy

Metal-LigandConjugate

OrganicLinker

Biomolecule

TumorCellReceptor

N

S

N

SH

O

Thiolactone containing molecule was designed to permit one-step coupling with amine functionalities, and concomitant completion of the tetradentate core required for complexation.

Bifunctional Chelate Approach

Bifunctional Chelate ApproachBifunctional Chelate Approach for Labeling

Peptides, Proteins and Antibodies with Tc

• Presence of the N-Methyl group yields charged complexes upon reaction with technetium.

N

S

N

SH

O

NH2

NN

SH

NH

O

HS

NN

NH

O

S S

Tc

O

Step 1.

Step 2.

O

CH3O

HO

CH3CH3

OH

N

Approaches to radiolabelingdiprenorphine with radioiodine

Diprenorphine

Radioiodination via vinylstannylatedalkylating agents

OTs

OTs

Nuc

Nuc

I* Nuc

I*

Nuc

I* = 127I, 125I, 123I

I*

(n-Bu)3Sn

(n-Bu)3Sn

Nuc:

(n-Bu)3Sn

(n-Bu)3Sn

E

Z

J. L. Musachio and J. R. Lever, Tetrahedron Lett., 30, 3613-3616 (1989)

Lever et al., Synapse 29: 172-182, 1998.

C6-O-iodoallyl-diprenorphine forin vivo studies of opioid receptors

CH3

CH3

HO

O

HO

N

O*I

I* = I127

, I125

, I123

O-IA-DPN

Radiosynthesis of[125/123I]-O-IA-DPN

CH3CH3

HO

O

HO

N

O

n-Bu3Sn

O

HO

N

O

*ICH3

CH3

HO

1. NaI125/123

MeOH / HOAc (95:5)Chloramine-T, 2 min.

2. HPLC / Solid PhaseExtraction

3. Formulation / QC(5% EtOH /saline)

[125I]-O-IA-DPN70 - 80% Radiochemical Yield

1200 - 1700 mCi /mol

[123I]-O-IA-DPN58 - 84% Radiochemical Yield

> 2400 mCi /mol2 Hours Total Time

[125I]-O-IA-DPN70 - 80% Radiochemical Yield

1200 - 1700 mCi /mol

[123I]-O-IA-DPN58 - 84% Radiochemical Yield

> 2400 mCi /mol2 Hours Total Time

[123I]-O-IA-DPN Allows Ex Vivo AutoradiographicLocalization of Opioid Receptors in Mouse Brain (1

hr)

Saline Control Naltrexone Block

SA4503, FESA4503 and Initial Analogs

Elsinga et al., Synapse 43:259-267, 2002.Ishiwata et al., Nucl. Med. Biol. 30:273-284, 2003.

SA4503; R = 11CH3

FESA4503; R = 18FCH2CH2-

R = I

I

E - iodoallyl

Z - iodoallyl

N

N OCH3

OCH3

Synthetic Scheme for Iodoallyl Analogs

R1=H, R2=n-Bu3Sn, 3ZR1=n-Bu3Sn, R2=H, 3E

HO AHO R1

R2B

TsO R1

R2

NN OCH3

O R1

R2

Scheme 1b. Reagents: A, propargyl alcohol/ tri-n-butyltin hydride. B, potassium trimethylsilanolate/ tosylate chloride. C, 4-O-demethyl SA4503. D, iodine.

Scheme 1b

D

NN OCH3

O R1

R2

1 23

4

5

R1=H, R2=n-Bu3Sn, 4ZR1=n-Bu3Sn, R2=H, 4E

R1=H, R2=I, 5ZR1=I, R2=H, 5E

C

+ TsOn-Bu3Sn

In Vitro Binding Affinity (IC50 nM)of Z- and E-Iodoallyl Analogs of SA4503

Sigma-1 Sigma-2

SA4503

FE SA4503

E-IA SA4503

Z-IA SA4503

17.4a

6.48b

24.50 ± 1.61c

15.10 ± 1.03c

1784.1a

2.11b

22.0 ± 2.78c

20.3 ± 1.29c

a Matsuno et al, Eur J Pharm 306:271-279, 1996.b Elsinga et al, Synapse 43:259-267, 2002. c n=4, Mean ± SEM

Radiochemical Synthesis

NN OCH3

OH

OTsR1

R2

NN OCH3

O R1

I2

R2

NN OCH3

O R4

R3

R1=H, R2=n-Bu3Sn, 1ZR1=n-Bu3Sn, R2=H, 1E

R1=H, R2=n-Bu3Sn, 2ZR1=n-Bu3Sn, R2=H, 2E

R1=H, R2=I, 3ZR1=I, R2=H, 3E

Radiochemical Yields with 125I:

(57-58.5%) with highSpecific Activity

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

Whole Brain%ID / g

0 30 60 90 120

Time (min)

Z-IA-SA4503

E-IA-SA4503

(means +/- SD; n = 4-5)

Biodistribution of Biodistribution of 125I-Iodoallyl Analogs I-Iodoallyl Analogs of SA4503 in normal mice:of SA4503 in normal mice:

0.0

0.2

0.4

0.6

0.8

%ID / gWhole Brain

30 min 60 min

p = 0.03

*

Whole brain:Whole brain:Control and BlockedControl and Blocked

Whole BrainWhole Brain

Question: So, is there a chemical way to:1) Decrease Lipophilicity and Retain Affinity?

N

N OCH3

OCH3

I

N

N OCH3

O I

NN OCH3

OCH3

I

NN OCH3

OCH3

I

Z-IA-DM-PE-PIPZ Ki 1 19.9 ± 1.3 nM; 2 455 ± 66

nM (n = 4) 2 / 1 = 23

[125I] -Z-IA-DM-PE-PIPZ Log D7.4 = 2.27 ± 0.006 (n = 5)

E-IA-DM-PE-PIPZ Ki 1 15.1 ± 1.3 nM; 2 1263 ± 166

nM (n = 4) 2 / 1 = 84

[125I] -E-IA-DM-PE-PIPZ Log D7.4 = 2.25 ± 0.008 (n = 5)

New Iodoallyl analogs:

Biodistribution of “New and Improved” Biodistribution of “New and Improved” EE 125I-I-Iodoallyl Analog in normal mice:Iodoallyl Analog in normal mice:

I-125 IA-Sigma CompoundsWhole Brain Comparison at 60 min

(n = 4, means +/- SD )

E-1 E-1 Haldol E-2 E-2 Haldol Z-2 Z-2 Haldol0

1

2

3

%ID / gWhole Brain

Haldol (2.5 / )mol kg

*

**p = 0.03

p < 0.0001

**p < 0.0001

0.5 1.5 2.5 3.5 4.5

Cerebellum

Hypothalamus

Striatum

Frontal Cortex

Pons / Medulla

Colliculi

Saline ControlCold Compound

%ID / g Tissue (60 min)

Haldol (σ1/σ2)1063(BD σ1)- (Br Machσ2)

***

* ,ANOVA Dunnett'sp<0.01

Test drugs i.v. at 2.5 mol / kg. Values are means ± SD; n = 4.

In vivo performance of “New and Improved”Trans IA-Sigma Ligand

Radiopharmaceutical Chemistry

also deals with Radiotherapy

Features of MURR

• Highest neutron flux of any U.S. university reactor

• High reliability -- operates 105% of schedule [91% of clock time]

• Traditionally access samples once/week

• Currently access samples several times each week

The Blue Glow

Selected IsotopesCurrently Supplied by MURR

• Lanthanides [all M3+]– Pm-149, Sm-153, Ho-166, Dy-166, Lu-177

• Transition Metals– Rh-105, Pd-109, Re-186, Ir-192, Au-198

• Main Group Elements– P-32, P-33, Se-75

Nuclear Properties of Various M3+ Radionuclides

Isotope Half-Life Decay Ave Range Approx. Range

(days) - (MeV) (mm water) (cell diameters)

Dy-166 3.40 0.40 0.18 15

Lu-177 6.71 0.50 0.24 20

Sm-153 1.93 0.69 0.35 30

Pm-149 2.21 1.1 0.71 60

Ho-166 1.12 1.9 1.38 117

Y-90 2.67 2.28 1.77 150

153153Sm-EDTMP [Quadramet]Sm-EDTMP [Quadramet] 99m99mTc-MDPTc-MDP 153153Sm-EDTMPSm-EDTMP

NN

PO 3H 2

PO 3H 2

PO 3H 2

PO 3H 2

153153SmSm

+

Story of QuadraMetTM

• Sm-153 identified as a useful nuclide for radiotherapy by MU researchers• Development began in early 1980’s at MU in collaboration with the Dow Chemical

Company [phosphonate ligands]• Successful in palliative treatment of bone cancer in canine patients, with added bonus

of ~ 15% cure rate [MU College of Veterinary Medicine program of Comparative Oncology]

• Clinical trials began in late 1980’s, with doses supplied by MURR for Phase I studies• ~ 80% efficacy, with ~ 25% obtaining full pain remission• Approved in U.S. for pain palliation of metastatic bone cancer in March, 1997

Yttrium-90 Microspheres

Nearly 100% blood supply of a liver tumor comes from hepatic artery

Effective treatment with minimal toxicityOutpatient procedure

Yttrium-90 Microspheres

Average diameter of microspheres - 20 -30 mYttrium is an integral part of the glass matrixDelbert Day (University of Missouri - Rolla) and Gary

Ehrhardt (University of Missouri - Columbia) U.S. Patent 4,789,501 December 6, 1988

Preparation of Active Microspheres

Y-89 microspheres are irradiated in the MU-Research Reactor (MURR), resulting in the conversion of the non-radioactive yttrium to Y-90, a powerful - emitter (64.2 hr half-life, average distance in tissue: 2.5 mm)

Yttrium-90 Microspheres

In a recent study (Dancy et al., J. Nucl. Med. 41: 1673-1681, 2000), patients with hepatocellular carcinoma (HCC) were treated with 100 Gy

Outcome: 79% patients decreased or had stable disease for a median of 10.3 months vs. 1 - 4 months for untreated patients

Yttrium-90 Microspheres become TheraSphere®

March, 2000: MDS Nordion (Kanata, Ont. Canada) was granted a Humanitarian Device Exemption by the U.S. Food and Drug Administration as an alternative to chemotherapy in inoperable cases

Centerfor Single Photon-Emitting

Cancer Imaging AgentsFive year grant award from the National Cancer

Institute of the National Institutes of Health, 8/01/03 - 7/31/08

Complements on-going interdisciplinary research at MU focused on imaging and therapeutic applications of radioisotopes

Includes a career development component, supporting undergraduate research activities.

Departmental Research Interests

AnalyticalAnalytical

InorganicInorganic

OrganicOrganic

PhysicalPhysical

EnvironmentalEnvironmental

MaterialsMaterials

RadiochemistryRadiochemistry

InterdisciplinaryInterdisciplinaryResearchResearch

Radiochemistry ProgramRadiochemistry Program

• Chemistry Department• Radiopharmaceutical Chemistry (Organic/Inorganic)

• Radioanalytical Chemistry

• Actinide Chemistry (Inorganic)

• Radioenvironmental Chemistry

• University of Missouri Research Reactor• Radioisotope Production (medical and biological isotopes)

• Trace Analysis (archaeometry, epidemiology, environmental science)

• Material Science

Jurisson Group ResearchJurisson Group Research

Radiopharmaceutical Chemistry• Inorganic Chemistry

• Tc, Re, Rh, Au, Ln

• Ligand design

• Radiochemistry

• Biochemistry/Biology

Radioenvironmental Chemistry

Research in the Duval Group

UU

U

U

U

U

O

OO

O

O

O

O

O

O

OO

O

OCp‡

Cp‡

Cp‡

Cp‡

N N

N N

Fe

U

O

O

U

O

ON

N

R

L

L

L

LN

NN

N

RuL

LL

U

O

O

L

L

R

N N

N N

M

HN

NH

Mes

Mes

Mes

U

O

O

O

O

O

O

4-

N

N

O

N

R

RR

U

N

OU

O

O

N

N

R

N

R

NR

2-

Probes new coordination environments of actinides that bridge non-aqueous complexes with traditional systems found under environmental conditions.

J. David Robertson

Using the unique properties of the nucleus to study chemical processes/problems which, frequently, cannot be examined by other means.

Investigating the source and impact of fine airborne particulate

Elemental imbalances in the formation of Alzheimer’s disease

“Aging” the plaques and tangles in Alzheimer’s disease

Using red ochre for provenance studies

Evaluating children’s chronic exposure to As and Cr from Cu/Cr/As treated wood structures.

MURR Archaeometry Lab

NAA became the preeminent technique in archaeometric materials analysis following the pioneering work at the national labs in the late 1960s and early 1970s

• unequalled replicability• sensitivity down to sub-ppm level• almost no matrix effect• fully instrumental, multi-element characterization

• Collaborative projects with US archaeologists at academic institutions or museums in 46 states

• Service to cultural resource management (CRM) archaeologists in 15 states

• Collaborations with archaeologists in 15 countries in the Americas, Europe, the Middle East, and Far East

Epidemiological studies have associatedlow selenium intake with increased cancer

risk• Population-Based Studies: Cancer incidence or mortality is inversely proportional to regional indicators of status.

• Case-Control Studies: In some case-control studies, selenium status, measured in a prospectively-collected biologic monitor, is lower in cancer cases compared to matched controls.

• Intervention Trials: Cancer incidence and mortality risks are lower among participants taking a supranutritional selenium supplement compared to control subjects in double-blind, placebo-controlled trials.

http://www.smartdraw.com/

Things to know about MU #4:Things to know about MU #4:

• Active Graduate Student Organization

• Geographically south of the glaciated till plain, so rolling hills heading to the Ozarks

• Numerous caves for spelunking and rivers for floating

• In between Kansas City and St. Louis