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Organisation and Research Direction
– The Sydney Cancer Institute is an independent research institute established in February 2003 by:
• Sydney Cancer Centre
• The Central Sydney Area Heath Service (CSAHS)
• The University of Sydney
• The Sydney National Cancer Foundation
– The primary function of the institute is:
“Conducting research to substantially improve cancer outcomes”
Objectives and Research Directions
• The Sydney Cancer Institute’s research programs are based around a clinically driven set of research priorities
• The Sydney Cancer Institute comprises the following divisions
– Molecular Oncology
– Cancer Biology
– Experimental Therapeutics
– Cancer Care and Control
Research Goals• The Sydney Cancer Institute aims to pursue
research excellence, depth and relevance by:
– Integration with clinical cancer medicine by the development of translational research programs
– Providing core research facilities such as:• A clinical cancer register• Statistician and information technology• Shared instruments and operators• A tumour bank available to all SCI researchers
– Providing more structured national and international connections
Scientific Advisory Board• The SCI scientific advisory board includes researchers with a
diverse array of skills and interests
– Prof. W McCarthy (Director Melanoma Foundation)
– Prof. J Bishop (Director Sydney Cancer Centre)
– Prof. B Armstrong (Head School of Public Health, USyd)
– Prof. G Halliday (Department of Dermatology, USyd)
– Prof. P Lay (Centre for Heavy Metals Research, USyd)
– Prof. B Roufogalis (Pro-Dean Pharmaceutical Chemistry, USyd)
– Prof. J Simes (Director NHMRC Clinical Trials Centre, USyd)
– Prof. J Thompson (Director Sydney Melanoma Unit)
– A/Prof. P Buttow (Executive Director, Medical Psychology Unit, USyd)
– A/Prof. M Millward (Head Clinical Research, SCI)
– Dr S Clarke (Senior Research Fellow, SCI)
Affiliations• The SCI has affiliation agreements with:
– The University of Sydney
– Central Sydney Area Health Service
– The Sydney National Cancer Foundation
– The John Wayne Cancer Institute – Los Angeles
– The National Institutes of Health (USA)
– National Cancer Institute (CTEP, USA)
– Cancer Therapeutics Research Group (Singapore)
– National Cancer Centre (Singapore)
– John Hopkins (Singapore)
Research programs• The SCI has a diverse research program including:
– DNA Methylation (Dr Susan Clarke)
– Cancer Genetics and Drug Resistance (Dr Quihan Dong, Dr John Young)
– Viral Oncology (Professor Y Cossart and Dr Carol Thompson)
– Tumour Pathology and Molecular Biology (A/Prof. Soon Lee)
– Skin Cancer Biology (Prof. Gary Halliday)
– Skin Cancer and Photobiology (Dr Vivienne Reeve)
– Cancer Invasion and Metastasis (Dr Guy Lyons)
– Heavy Metals Research Program (Prof. Peter Lay)
– Clinical Pharmacology (A/Prof. Stephen Clarke)
– Pharmaco Oncology (Prof. Basil Roufogalis)
– Gene Therapy (A/Prof John Rasko)
– Molecular Imaging (A/Prof Michael Fulham)
– Clinical Trials (A/Prof. Michael Millwood)
– Early Detection and Diagnosis (Dr Scott Menzies)
Biometals Section, CHMR• Associate Professor Robert Armstrong• Dr. Rachel Codd• Dr Carolyn Dillon• Dr. Ron Fenton• Professor Hans Freeman• Professor Trevor Hambley• Associate Professor Margaret Harding• Dr. Hugh Harris• Associate Professor Brendan Kennedy• Professor Peter Lay• Professor Len Lindoy• Associate Professor Tony Masters• Dr. Lou Rendina
Research Areas/Interests• Anti-Cancer Drugs
- Boron complexes: boron neutron capture therapy (Rendina)
- Co(III) Hypoxic Agents (Hambley)
- Copper Complexes (Dillon, Hambley, Harris, Kennedy, Lay)
- Metallocenes (Dillon, Harding)
- Metal Complexes of Organic Anti-Cancer Drugs (Hambley,
Harding)
- Metalloporphyrin/Fullerenes (Armstrong, Lay)
- Pt(IV) and Pt(II) (Fenton, Hambley)
- Ruthenium (Armstrong, Dillon, Lay)
• Radiopharmaceuticals- Use of copper-64 in macrocyclic systems for the imaging
and therapy of cancer (Lindoy)
- Improved 99Tc Generators (Masters)
Research Areas/Interests• Metal-Induced Cancers and Toxicity
- Cr- and Ni-induced cancers (Codd, Dillon, Harris, Lay)
- As carcinogenesis and toxicity (Dillon, Harris)
• Anti-Inflammatory Drugs- Cu, Zn, Ni, Zn complexes as anti-inflammatories (Dillon,
Hambley, Kennedy, Lay)
• Anti-Diabetics- Cr(III) (Dillon, Harris, Lay)
- Vanadium (Codd, Lay)
Research Areas/Interests• Metalloproteins
- structure of heme proteins and their roles in the immune system, and heart disease (Armstrong, Lay)
• Chelation Therapy and Metabolic Processes- role of transition metal-sialic acid species in metal
homeostasis/disease (Codd)
- cold-adapted 'super-siderophores' for metal chelation
therapies (Codd).
• Diagnostics
- use of vibrational spectroscopy in the diagnosis of cancer
(Armstrong, Lay)
Facilities/Expertise• Structural Biology and Structural Chemistry X-ray, Neutron & Electron Scattering and Diffraction; Synchrotron Techniques; NMR Spectroscopy;
See separate presentation on CSBSC
• Mass SpectrometryElectrospray (including HPLC front end); GC-MS; Maldi
• Vibrational SpectroscopyIR, Raman and Resonance Raman: including tissue mapping
• EPR SpectroscopyL, Q and X band, ENDOR, He cryogenics, Whole cell
• Cell Biology Cytotoxicity; Genotoxicity; Permeability; Imaging, Spectroscopy
• Animal StudiesPharmacology; Pharmacokinetics; Efficacy; Toxicity
Boron Neutron Capture - Boron Neutron Capture - RendinaRendina
10B
1n
11B*
4He
7Li
kinetic
5 5
0 2
3
energy+(~2.28 MeV)
tumour cell
selectivedestruction
0n110B
10B
Clinical BNCT AgentsClinical BNCT Agents
Malignant brain tumours Malignant melanoma
B SH
B
NH3+
O
O–
OHHO
2–
BPA
B–H
BSH
Two new classes:
• Platinum(II)-amine complexes
• Metallo-intercalator complexes
Dinuclear Platinum ComplexesDinuclear Platinum Complexes
Woodhouse, S. L.; Rendina, L. M. Chem. Commun., 2001, 2464.
International Patent PCT/AU02/00943
HCl.H2N
HCl.H2N
NH2
NH2
Pt
Cl
Cl
NH3
1. K2CO3
2. K[PtCl3(NH3)] or cis-[PtCl2I(NH3)]–/Ag+/HCl(aq)
Pt
NH3
Cl
Cl
NH2
NH2
Pt
NH3
Cl
NH3
1. K2CO3
2. trans-[PtCl(dmf)(NH3)2]OTf
Pt
NH3
Cl
NH3
2+
CBH
(OTf)2
Metallo-intercalator ComplexesMetallo-intercalator Complexes
Todd, J. A.; Rendina, L. M. Inorg. Chem., 2002, 41, 3331.
International Patent PCT/AU02/00943.
H
NN
N
PtS
(CH2)n
+
(n = 0-3)
H
NN
N
PtCH2S
+
BHC
Control A375 Melanoma Cells
P S Cl K
Ca Cr Fe Ni
Cu Zn ScatteringJ. Aitken, H. Harris, P. A. Lay, USyd, P. Farmer, UC Irvine
CuDSF-treated A375 Melanoma Cells
P S Cl K
Ca Cr Fe Ni
Cu Zn ScatteringJ. Aitken, H. Harris, P. A. Lay, USyd, P. Farmer, UC Irvine
Cu XANES of Melanoma Cells
8.95 8.96 8.97 8.98 8.99 9 9.01 9.02
Control
CuCl
Melanin 1 (Cu II)
Melanin 2 (Cu I)
J. Aitken, H. Harris, P. A. Lay, USyd, P. Farmer, UC Irvine
Identification of Novel Intercalating Platinum Compounds - Fenton, Aldrich-Wright
CISPLATIN & analogs bind to DNA
New Pt compounds intercalate into DNA - IP protection- PCT filing, Priority date February 22nd 2001
0
2
4
6
8
10
12
L1210 DDP 2008 C13 SKOV3 PC3cell line
IC50
uM
Test compound
Cisplatin
L1210 = mouse leukemia; 2008 = human ovarian tumor PC3= prostate tumorDDP = Cisplatin-resistant; C13 = acquired; SKOV3= intrinsic Cisplatin resistance
EFFICACY of lead compound- Comparison with Cisplatin & in Cisplatin-resistant cell lines
Phase I clinical trials
• toxic effects : hypoglycaemia, metallic after taste
• bone marrow largely unaffected
• liver toxicity does-limiting side-effect
Phase II clinical trials
• breast cancer
• renal cell carcinoma
A/Prof Margaret Harding, School of Chemistry
• poor hydrolytic stability in water pH> 4.0
• Ti accumulates in nucleic rich regions tumour cells
• species formed in vivo and how interaction with DNA occurs not understood
• mechanism is distinct from platinum based drugs
Titanocene Dichloride
Antitumour Metallocenes
Ti V
Nb Mo
Re
Cr Mn
TcZr
Hf Ta WY = Cl, Br, I, NCS, N3
• Each complex has independent mechanism of action
• Current studies focus on cellular distribution, interaction with biomolecules of Mo, Nb complexes
• M = Mo targets thiols
A/Prof Margaret Harding, School of Chemistry
Design water soluble, stable derivs
Streptonigrin
A/Prof Margaret Harding, School of Chemistry
Water, pH 6.5
NN
O
O
H2N
CH3O
CO2H
CH3H2N
OCH3
OH
OCH3
Clinical use human cancers until 1977
•broad spectrum antitumor activity
•lymphoma, melanoma•cancers of the breast, cervix, head, neck
•severe and unpredictable side-effects
Streptonigrin Metal Complexes
A/Prof Margaret Harding, School of Chemistry
N
O
O
H3CO
H2N
N
O OH
affectsDNA binding
topo II recognition site
Redox related to DNA cleavage
ACCELERATED BYMETAL IONS
bipyridylzwitterionlabile metal complexes
N
O
O
H3CO
H2N
N
O OH
NH2
CH3
OCH3HO
OCH3
•Ru(II) complex
•Reductively activated to semiquinone
•Strong DNA binding/cleavage predicted
Improved Tc-99 Generators - Masters Understanding the interactions between alumina and molybdates has led to improvements in the process for generating 99Tc for radio-diagnostics
Schematic outline of 99Mo production process at ANSTO.
Effect of Oxidation State on Cr Genotoxicity
Dillon, C. T.; et al. Chem. Res. Toxicol. 1998, 11, 119-129; Dillon, C. T.; et al. Chem. Res. Toxicol. 2000, 13, 742-748.
0.0 0.1 0.2 0.3 0.4 5 10 15 20 25 300
20
40
60
80 [CrO
4]2-
[CrO(ehba)2]-
[CrO(mampa)]-
[Cr(en)3]3+
[Cr(glygly)2]-
Inci
denc
e of
MN
/100
0 B
N c
ells
[Cr] (mol/dish)
} Cr(VI)
} Cr(V)
} Cr(III)
Exposure of Whole Cells to Cr(VI) and Cr(III)
Cr(III)-Treated Cell
Cr(VI)-Treated Cell
P
K Cr Zn
P K Cr Zn
Min MaxDillon, C. T.; Lay, P. A.; Kennedy, B. J.; Stampfl, A. P. J.; Cai, Z.; Ilinski, P.; Rodrigues,
W.; Legnini, D. G.; Lai, B.; Maser, J. J. Biol. Inorg. Chem. 2002, 7, 640-645.
• Copper indomethacin is a dimeric copper complex containing 4 indomethacin ligands bound to Cu through the carboxylic acid group.
• Cu-algesic (CuIndo) is an effective anti-inflammatory drug commonly used in dogs and horses.
• Importantly, CuIndo is much less TOXIC in dogs than IndoH.
N
O
Cl
CH2CH3O
CH3
OCuO
O
O
CC
O
O
O OCuR
C
C
R
R
R
L
L
R=
L= solvent ligand
Copper IndomethacinLay, Hambley, Kennedy, Dillon
Effect of the Formulation on Gastro-Intestinal Damage
• Equivalent doses of indomethacin (10 mg/kg) were administered to each animal.
• The number of animals tested per treatment ranged from 4-6.
0
25
50 Stomach
CMC micelle powder paste86 paste3 indo (CMC)0
50
100
150 Small Intestine
Ulc
erat
ions
(m
m2 )
Formulation
Efficacy of Cr Dietary Supplements• Chromium supplements are the second biggest market for dietary
supplements (over $1B industry).• Taken to convert fat into muscle in athletes (used instead of steroids for
humans) and food animals.• Used to help prevent diabetes.• There is no compelling evidence (epidemiology, cell work, or
biochemical assays) that Cr supplements convert fat into muscle or help prevent diabetes.
• The FDA has prevented companies from advertising such health benefits.
• There are anti-diabetic effects exhibited in animal studies for certain Cr
complexes and they are believed to have anti-diabetic effects on humans.
Side-Effects of Cr Dietary Supplements• They react with enzyme systems such as glucose oxidase and xanthine
oxidase to form highly genotoxic mixtures of high oxidation state Cr species, which are very damaging to DNA
• The Cr(VI) generated in these enzymatic processes and strongly inhibits phosphatase enzymes, which is likely to be responsible for the anti-diabetic effects of Cr supplements
• High-oxidation-state Cr and V species appear to act in the same way, through similar intermediates in their anti-diabetic effects
0 50 100 150 200
0
25
50
75
100 Cr(VI) Cr(V) V(V)
Activ
ity, %
Concentration, M
Inhibition of PTP (phosphatase enzyme by Cr(VI), Cr(V) and V(V). Mulyani, Levina, Lay, JACS, submitted
Heme Proteins - Armstrong, Harris, Lay
• Characterized NO, CO and O2 adducts of heme proteins, many of which are too unstable to crystallise
• Studied unfolding of cytochrome c• Collaborations with Paul Witting and
Roland Stocker- Studies on indolamine 2,3-dioxygenase (IDO) - important in the immune system
- Studies on the roles of heme proteins in heart disease
Fe-O-O = 146
Np
NpNp
NpFe
O
N
1.832.02
2.05
O
Molecular structure of the active site of horseheart MbO2 at 10 K.
Heme Proteins
Rich, Cheng, Armstrong and Lay
Metal-Sialic Acid Speciation CoddInsight into the nature and role of species formed between sialic acid and transition metal ions.
This polyfunctional carbohydrate is present as theterminal residue in many glycopoteins involved inmetal transport (e.g., transferrin; ceruloplasmin)
Sia Sia2,3 2,3
Gal Gal1,4 1,4
GlcNAc GlcNAc1,2 1,2
Man Man1,3 1,6
Man1,4
GlcNAc1,4
GlcNAc
Asn
Membrane glycoproteins and glyco-lipids: extracellular sugar residues on mammalian plasma membranes
Metal-Sialic Speciation9.76
pH
8.38
7.18
6.21
5.47
4.37
3.14
2.08
1.982 1.980 1.978 1.976
giso
1.982 1.980 1.978 1.976
giso
ba
Cr(V)-sialic acid profile•Development of transition metalEPR Spectroscopy as a diagnostictechnique for glycosylation patterns
•Extension of studies of metal-sialic acid speciation with other biologicallyrelevant transition metal ions(e.g., Cu, Fe, Zn, V).
Cu(II)-sialic acid profile
•Implications for metal transport, homeostasis, and role(s) in sialylglycoprotein-dependent disease
Current Diagnostic TechniquesCurrent Diagnostic TechniquesTriple Assessment & Side EffectsTriple Assessment & Side Effects
• A combination of physical examination, mammogram and fine needle aspiration cytology.
• Standard for breast diagnosis.
• Side effects:– Probable false positive and negative results
from mammogram– Time consuming progress
New Diagnostic For Breast CancerNew Diagnostic For Breast Cancer
• 1H Magnetic Resonance Spectroscopy at IMRR– Simple and quick diagnosis of aspirated breast tissues– The choline-to-creatine peak ratio is being used to compare
various breast disease states
• Vibrational Spectroscopy– Infrared Spectroscopy
• Quick diagnosis by comparing the spectral differences in proteins (amides), lipids and DNA levels of sectioned tissues
– Raman Spectroscopy • By using various laser excitations to compare proteins, lipids
and DNA levels of different breast diseases.• Imaging and mapping provide parallel results often difficult
to determine in pathology.
Cho
Cr
-CH=CH-
-CH2- -CH3
ppm
HODsuppressed
1H Magnetic Resonance Spectroscopy
BenignBenign
MalignantMalignant
Wavenumbers (cm1)
Tra
nsm
issi
on
Am
ide
I
Am
ide
II
Nu
clei
c A
cid
s
Pro
tein
s &
Lip
ids
Collagen Levels
FT-IR Spectra
Benign
Malignant
Benign
t
1455(CH3) + (CH2)proteins
1239 as(PO2
ˉ )Nucleic acids
Wavenumbers (cm1)
Tra
nsm
issi
onFT-IR Spectra
Malignant
Wavenumber (cm1)
Ram
an I
nte
nsi
ty (
a.u
.)3100 2750 1800 80
0
s(CH2)LIPIDS
as(CH2)
as(CH3
)
Am
ide
I
CH
2 & C
H3
def
orm
atio
n
Ph
e
Am
ide
IIIL
IPID
SFT-Raman Spectra
Malignant
Benign
Raman ImagingRaman Imaging• New approach to compare results parallel to the native pathological examinations
• Colour intensity of particular Raman marker bands have potential for identifying the progress of cancer
• Proteins, lipids, DNA and other important components can be detected in tissues that require less preparation than for infrared and magnetic resonance spectroscopy
SummarySummary
• 1H Magnetic Resonance Spectroscopy– Fast and suitable for low-lipid content of breast lesions– Research is in collaboration with The Institute for
Magnetic Resonance Research, University of Sydney
• Vibrational Spectroscopy– Suitable for both high- and low-lipid content of breast
lesions– Inexpensive and non-destructive.– can run in parallel with histopathology
• Multivariate Statistical Analysis– Improve significance for the spectral data– Correlate and classify different breast diseases