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School of Advanced StudiesDoctorate course in Chemical Sciences
Cycle XXScientific-Sector CHIM/03
Dr.ssa Grazia Papini
New metal complexes supported by scorpionate and macrocyclic ligands:
chemistry and biological studies
TutorProf. Giancarlo Gioia Lobbia
Well-known Scorpionate Ligands
Bis(pyrazolyl)borates Tris(pyrazolyl)borates Tetrakis(pyrazolyl)borates
“Nitrogen heterocycles other than pyrazole can be used, such as imidazole, triazole, benzotriazole, thioimidazole, ecc.”
Bis(imidazolyl)borates
Tris(imidazolyl)borates
Tetrakis(imidazolyl)borates
NN N
B
NNN
H
H N
N N
NN N
B
NNN
H
N
N NB
N
NN
H
H NN
N
N
N NB
N
NN
H
Bis(3-R-2-thioxo-imidazolyl)borates
Poly(triazolyl)borates Poly(benzotriazolyl)borates
N
N
N
N
B
H
H
R5
R4 R4
R5
μ2-N,N’ μ2-N,N’
N
N
N
N
BR5
R4 R4
R5
N
NR4
R5
N
N
R5
R4
μ4-N4 μ4-N4
N
N
N
N
B
H
R5
R4 R4
R5
N
NR4
R5
μ3-N,N’,N” μ3-N,N’,N”
NN N N
B
H
H
S S
R R
3-S,S’,B-H 3-S,S’,B-H 2-S,S’ 2-S,S’
Scorpionate ligands Scorpionate ligands with EWG with EWG
substituentssubstituents
[H2B(pzCOOEt,Me)2]-
N
NB
N
N
OO
OO
H H
[H2B(tzNO2)2]-
NN
N
N
NN
B
HH
O2N NO2K+
[H2B(pzNO2)2]-
N
N
N
N
B
HH
O2N NO2K+
[H2B(pzCF3)2]-
N
N
N
N
B
HH
F3C CF3K+
[H3B(pzCF3)]-
B
H
H
H
NN
CF3
Na
H. V. R. Dias, S. Alidori, G. Gioia Lobbia, G. Papini, M. Pellei, C. Santini Inorg. Chem. (2007)
M. Pellei, S. Alidori, G. Papini, G. Gioia Lobbia, J. D. Gorden, H. V. Rasika Dias, C. Santini, Dalton Trans. (2007)
S. Alidori, M. Pellei, C. Pettinari, C. Santini, B. W. Skelton, A. H. White, Inorg. Chem. Commun., (2004). G. Bandoli, A. Dolmella, G. Gioia Lobbia, G. Papini, M. Pellei, C. Santini Inorg. Chim. Acta (2006)
M. Pellei, F. Benetollo, G.Gioia Lobbia, S. Alidori, and C. Santini, Inorg. Chem., (2005)
“Other modifications include changing the substituents on the heterocyclic ring.”
“In addition, tripodal ligands can have central atoms other than boron, such as carbon, phosphorus, or silicon….”
“…..and bearing a coordinating moiety (R') such as acetate, dithioacetate, sulfonate, ethoxide, ”
Si
NN
NN
N N
Me
R R'
R
R'
R'
RP
N
N
N
N
O
RR
N
N
R
R' R'R'C
N
N
N
N
R
R3R3
N
N
R3
R5 R5R5
R4 R4
RC(pzx)3 RSi(pzx)3(pzx)3PO
NN
NN
CH
CO
O
bdmpza
NN
NN
CH
CS
S
bdmpzta bdmpzs
NN
NN
C H
SO
OLiO
Rhenium complexes
Versatile chemistry: several oxidation states accessible (from -I to VII); different coordination numbers (from 4 to 8); various donor set available
The similarity between technetium and rhenium chemistry, determined a widespread use of the latter as a technetium surrogate to perform macroscopic chemistry of potential radiopharmaceuticals. In this way, a ‘‘cold’’material (the natural isotopic mixture of 185Re and 187Re) can be advantageously manipulated instead of the radioactive nuclide 99gTc (t1/2 = 2.12 · 105 y, Eβ = 292 keV).
Rhenium has two β- emitters isotopes 186Re (β-max = 1.07 MeV; t1/2 = 90 h) and 188Re
(β-max = 2.10 MeV; t1/2 = 17 h) which are of great interest to nuclear medicine due to
their physical and nuclear properties finalized to a potential application in the radiopharmaceutical
Bioactive molecule
LinkerM
The “metal - fragment” strategy
Stable building -block
Labile groups
Re(V) complexes
NN
NN
CH
CO
OLi
NN
NN
C H
SO
OLiO
N N
NN
CO2Na
Cl
ReCl Cl
Cl
O-
Metal fragment
X Y
N
ReN Cl
Cl
O
O
biomolecule
N
ReN X
Y
O
O
S
NHNH
O
OH
O
NN
NN
Re O
Cl
Cl
MeOO
MeO[ReOCl4][NBu4]
CH3CN (Et3N)
N
N
N
N
Re
OClCl
COO
MeOH (Et3N)
[ReOCl4][NBu4]
N
N
N
N
Re
OORCl
SO2ON
N
N
N
Re
OORCl
COO
N N
NN
O
OH
NN
NN
CH
CO
OLi
NN
NN
C H
SO
OLiO
[ReOCl4][NBu4]
ROH(Et3N)
[ReOCl4][NBu4]
ROH (Et3N)
Metal Fragments
M. Porchia, G. Papini, C. Santini, G. Gioia Lobbia, M. Pellei, F. Tisato, G. Bandoli, A. Dolmella, Inorg. Chem. 44 (2005) 4045
Mixed coordination sphere complexes
CH2Cl2 (Et3N)
[ReOCl4][NBu4]
NN
NNE
LiO
HO(CH2)nOH
E= CO,SO2
n= 2, 3
N
N
N
N
Re
O OO
EO
(CH2)n
N
N
N
N
Re
OOMeCl
EO
N
N
N
N
Re
OClCl
EO
N
N
N
N
Re
OOEtCl
EO
Et3N
OH(CH2)nOH
Structure of the complex[Re(O)(bdmpza)(OCH2CH2O)]
Structure of the complex [Re(O)(bdmpza)(OCH2CH2CH2O)]
M. Porchia, G. Papini, C. Santini, G. Gioia Lobbia, M. Pellei, F. Tisato, G. Bandoli, A. Dolmella, Inorg. Chem. 44 (2005) 4045
N
N
N
N
Re
OOMeCl
COO
N
N
N
N
Re
OClCl
COO
N
N
N
N
Re
OOEtCl
COO
Et3N
HOOC(CH2)COOH
N
N
N
N
Re
OOO
COO
O O
N N
N NCO2LiCH2Cl2 (Et3N)
[ReOCl4][NBu4]
HOOC(CH2)COOH
Structure of the complex[Re(O)(bdmpza)(mal)]
Marina Porchia,Grazia Papini, Carlo Santini, Giancarlo Gioia Lobbia, Maura Pellei, Francesco Tisato, Giuliano Bandoli, Alessandro Dolmella Inorganica Chimica Acta 359 (2006) 2501–2508.
Potential Nitridorhenium complexes
N
N
N
N
N
N
B
H
RR
R
N
N
N
N
NN
N
NN
B
H
RR
R
:
:
:
N
N
N
NB
H H
R R
N
N
N
N
NN
: :
B
H H
R R
Cl
ReCl Cl
Cl
N
Nitridorhenium precursors
ReH2O2
H2ORe2O7 ReCl3PPh3
PPh3, [PhNHNH2]HClHCl PPh3
EtOH
ReH2O2
H2ORe2O7 [NBu4][ReO4] HCl g
NaN3
[NBu4]OH
Cl
ReCl Cl
Cl
N
Cl
RePh3P Cl
PPh3
N
Pre-carbene ligands
2 BzBr
X
N
N
N
XN
B
H H
Bz Bz
Br-
KBH4 +
XN
NH
2135°C X
N
N
N
XN
B
H H
K+
KBH4 +
XN
NH
3170°C
X
N
N
N
XN
N
XN
B
H
K+X
N
N
N
XN
N
XN
B
BzBz
Bz
H 2
2Br-3 BzBr
G. Papini, C. Santini, G. Gioia Lobbia, M. Pellei, G. Bandoli, A. Dolmella J. Organomet. Chem. (2008) submitted
R2
OO
R1
+ H
O
H
+ R3 NH2
NH4Ac
N
NR1
R2
R3
MeOH/H2O
Liu J., Chen J., Zhao J., Zhao Y., Li L., Zhang H., Synthesis 17 (2003) 2661–2666.
H
OO
H
+ H
O
H 2) Mes-NH2, NH4AcN
N
Mes
1) CH3COOH, 75°C
H
OO
H
+ tBu-NH2
2) HCOH, NH3AcN
N
t-Bu
1) MeOH/H2O, 70°C
N
N
24h
N
N
N
N
N
N
B
H
Mes
Mes Mes
+ (CH3)2S.HBBr2
CH2Cl2
2Br -
N
N24h
N
N
N
N
N
N
B
H
t-Bu
t-Bu t-Bu
+ (CH3)2S.HBBr2
CH2Cl2
2Br -
X
N
N
N
XN
B
H H
Bz Bz
Li +
THF, n-BuLi
- 78°C
X
N
N
N
XN
B
H H
Bz Bz
Br-
N
N
N
N
N
N
B
RR
R
:
:
:
H
Li +THF, n-BuLi
- 78°CN
N
N
N
N
N
B
RR
R
H 2
2Br-
[NBu4][ReNCl4] MIXTURE OF UNCHARACTERIZABLE
PRODUCTS
R = Bz, t-Bu, Mes
Silver(I) carbene complexes
CH2Cl2, 24h
N
N
N
N
N
N
B
H
R
R R
Ag2O
NN
NN
B
H
R R
NN
NN
B
H
RR
AgAg
NN
R
NN
R
Ag Br
R = Bz, t-Bu, Mes
2Br-
N
N
N
N
NN
B
H H
Br
Bz Bz
Ag2O
CH2Cl2
N
N
N
N
NN
BH
H
Bz
Bz
N
N
N
N
NN
BH
H
Bz
Bz
Ag
Ag
G. Papini, C. Santini, G. Gioia Lobbia, M. Pellei, G. Bandoli, A. Dolmella J. Organomet. Chem. (2008) submitted
Carbene transfer reactions
CH2Cl2, 2h
Au(SMe2)Cl
NN
NN
B
H
R R
NN
NN
B
H
RR
AgAg
NN
R
NN
R
Ag Br
R = Bz, t-Bu, Mes
NN
NN
B
H
R R
NN
NN
B
H
RR
AuAu
NN
R
NN
R
Au Br
N
N
N
N
NN
BH
H
Bz
Bz
N
N
N
N
NN
BH
H
Bz
Bz
Ag
Ag
CH2Cl2, 2h
Au(SMe2)Cl
N
N
N
N
NN
BH
H
Bz
Bz
N
N
N
N
NN
BH
H
Bz
Bz
Au
Au
G. Papini, C. Santini, G. Gioia Lobbia, M. Pellei, G. Bandoli, A. Dolmella J. Organomet. Chem. (2008) submitted
Rhenium derivatives
NBu4ReNCl4
N N
N
NN
N
B
H
H
R
RNN
N
N N
N
B
H
H
R
R
Re
N
N N
N
NN
N
B
H
H
R
R
Re
NCl
ClN
N
N
N
NN
BH
H
Bz
Bz
N
N
N
N
NN
BH
H
Bz
Bz
Ag
Ag
CH2Cl2
B
N
N
N
N
N
H
N
R
R
R
Re
Cl
N
NBu4ReNCl4
NN
NN
B
H
R R
NN
NN
B
H
RR
AgAg
NN
R
NN
R
Ag BrCH2Cl2
Copper and Ruthenium derivatives
NN
NN
B
H
R R
NN
NN
B
H
RR
AgAg
NN
R
NN
R
Ag Br
N
N
N
N
NN
BH
H
Bz
Bz
N
N
N
N
NN
BH
H
Bz
Bz
Ag
Ag
N N
N
NN
N
B
H
H
Bz
Bz
Ru
Cl [Ru(p-cymene)Cl2 ]
2CH
2Cl2
N
N
N
N
BH
Bz
Bz
N
N
Bz
RuCl
[Ru(p-cymene)Cl 2] 2
CH 2Cl 2
N
N
N
N
NN
BH
H
Bz
Bz
N
N
N
N
NN
BH
H
Bz
Bz
Cu
Cu
Cu(SMe 2)Br
CH 3CN
NN
NN
B
H
R R
NN
NN
B
H
RR
CuCu
NN
R
NN
R
Cu Br
Cu(SMe2 )BrCH
3CN
Copper derivatives
It is an essential trace metal for living organisms
Copper complexes’ activity is extremely wide
Copper has a well-documented coordination chemistry
Several radioactive copper isotopes are available nowadays for biomedical purposes both for radioimaging and targeted radiotherapy
isotope half-life imaging(emission, energy, abundance)
therapy(emission, energy, range in tissue)
application
Cu-60 20 min PET(b+, 873 keV, 93%)
Radiolabelling small molecules
Cu-61 3.3 h PET(b+, 527 keV, 62%)
Radiolabelling small molecules
Cu-62 9.7 min PET(b+, 1315 keV, 98%)
Radiolabelling small molecules
Cu-64 12.7 h PET(b+, 278 keV, 19%)
Radiolabelling small molecules, peptides and antibodies
Cu-66 5.4 min (b-, 190 keV; 0.95 mm) Radiolabelling small moleculesfor therapy
Cu-67 62.0 h SPECT(g, 185 keV, 48%)
(b-, 190 keV; 0.95 mm) Radiolabelling small molecules, peptides and antibodies
Fichna et al, Bioconjugate Chem., 14 (2003) 3-17
Copper(I) derivatives
Cu
P P
HO OHHO
HO OH
OH
NCCH3CH3CN
PF6
HO P
OH
OH
P(CH2OH)3
Cells line of ovarian carcinoma (2008) and cis-platino resistent carcinoma cells (C13)
NN N
N
Cu
P PHO
HO
HO
OHOH
OH
C=OO
O2N NO2
H
NNN N
NN
Cu
P PHO
HO
HO
OHOH
OH
C=OO
O2N NO2
H
NNN N
NN
Cu
P PHO
HO
HO
OHOH
OH
C=OO
H
NN N
N
Cu
P PHO
HO
HO
OHOH
OH
C=OO
H
NN N
N
Cu
P PHO
HO
HO
OHOH
OH
C=SS
H
C. Marzano, M. Pellei, D. Colavito, S. Alidori, G. Gioia Lobbia, V. Gandin, F. Tisato, and C. Santini, J. Med. Chem., 4949 (2006) 7317
“CuP4” tipe species
[Cu(CH3CN)4][PF6] + 4 thp [Cu(thp)4][PF6]
[Cu(CH3CN)4][PF6] + 2 bhpe [Cu(bhpe)2][PF6]
P POH
OH
HO
HO
P POH
OH
HO
HOCu
HO OH
OHHO
[Cu(thp)4][PF6]
P POH
OH
HO
HO
P POH
OH
HO
HOCu
[Cu(bhpe)2][PF6]
31P-NMR = - 5.35 (q), -145.14 (septet)[Cu(thp)4]+ m/z = 560 (6)[Cu(thp)3]+ m/z = 436 (65)[Cu(thp)2]+ m/z =312 (100)
31P-NMR = + 9.67 (dbr), - 144.05 (septet) [Cu(bhpe)2]+ m/z = 492 (100)
C. Marzano, V. Gandin, M. Pellei, D. Colavito, G. Papini, G. Gioia Lobbia, M. Porchia, F. Tisato and C. Santini, J. Med. Chem. 51 (2008) 798-808.
bhpe
P
O
O O
Br
Br+
P
P
OO O
OOO
+ P
Br
OO O
P OO O
+
H2P PH2 PPHO
OH
HO
OH
HO
OH
P PHO
HO
OH
OH
180°C
LiAlH4
HCOH, HCl
EtOH
NaHCO3
C. Marzano, V. Gandin, M. Pellei, D. Colavito, G. Papini, G. Gioia Lobbia, M. Porchia, F. Tisato and C. Santini , J. Med. Chem. 1 (2008) 798-808.
Citotoxic activities
Compound
IC50 (µM) ± S.D.
HL60 A549 MCF-7 Daudi HepG2 A375 CaCo2 HCT-15 HeLa
[Cu(thp)4][PF6] 0.60±0.02 9.11±2.71 11.08±0.52 6.94±0.18 1.26±0.10 4.58±2.41 1.08±0.12 2.00±0.03 8.21±1.50
[Cu(bhpe)2][PF6] 47.40±2.92 57.60±2.19 49.71±2.03 65.5±1.22 78.23±1.11 68.21±1.23 52.50±0.81 57.36±1.31 62.41±1.33
thp 68.63±2.44 72.91±2.44 64.23±4.29 >100 98.71±3.63 88.70±3.88 >100 >100 >100
bhpe 83.72±3.23 >100 >100 >100 71.71±1.64 73.21±1.22 84.11±2.22 91.71±4.01 >100
KPF6 >100 >100 >100 >100 >100 >100 >100 >100 >100
Cisplatin 15.91±1.51 29.21±1.92 19.04±1.51 23.97±2.51 21.50±1.41 20.33±1.33 35.42±1.40 25.34±1.31 10.50±1.51
A549 = lung cancer
CaCo2, HCT-15 = colon cancer
Hela = cervix cancer
MCF-7 = breast cancer
HL60 = leukemia
Daudi = lymphoma
HepG2 = epatoma
A375 = melanoma
IC50 values represent the drug concentrations that reduced the mean absorbance at 570 nm to 50% of those in the untreated control wells.
Compound2008
IC50 [µM]C13
IC50 [µM] R.F.
[Cu(thp)4][PF6] 1.48±0.21 2.88±1.07 1.9
Cisplatin 12.69±1.72 89.18±4.50 7.02
CompoundA431
IC50 [µM]A431/Pt
IC50 [µM] R.F.
[Cu(thp)4][PF6] 14.37±1.41 13.26±0.80 0.92
Cisplatin 22.06±1.62 57.76±1.81 2.61
CompoundLoVo
IC50 [µM]LoVo-MDRIC50 [µM] R.F.
[Cu(thp)4][PF6] 1.54±0.03 2.9±0.1 1.88
Doxorubicin 1.46±2.30 44.89±0.90 30.74
Human ovarian adenocarcinoma cells
Human cervix squamous carcinoma cells
Human colon adenocarcinoma cells
Cytotoxic activity of [Cu(thp)4][PF6] onto three additional cell line pairs, two of which (2008/C13* ovarian cancer cells and A431/A431-Pt cervix carcinoma cells) selected for their resistance to cisplatin and one (LoVo/LoVoMDR) for its resistance to doxorubicin.
Cross-resistance profiles were evaluated by means of the resistance factor (RF), which is defined as the ratio between IC50
values calculated for the resistant cells and those arising from the sensitive ones.
Comparison of IC50 values detected by MTT, NR and TB test after incubation of 2008 cells with [Cu(thp)4][PF6] for different exposure times
TB test reveals damage to cell membrane
MTT test mainly reflects damage to mitochondria
The NR assay indicates damage to lysosomes and Golgi apparatus
Lysosomes/Golgi apparatus are more sensitive to complex treatment. On the contrary, the scarce permeability to vital dye indicates that plasma membrane function is still maintained until the late phase of cell death. Lysosomal damage represents the early cellular event associated with copper(I) complex cytototoxicity.
Cell cycle phases
G1 = GAP 1S = Synthesis (DNA replication)G2 = GAP 2M = mitosis (nuclear and cytoplasmic division)I = Interphase
Percentage of cells in different cell cycle phases as function of time exposure of
[Cu(thp)4][PF6], vs control untreated cells
3 h
12 h
24 h
48 h
3 h
12 h
24 h
48 h
----------2008 untreated cells
----------2008 cells treated with IC50 of [Cu(thp)4][PF6] 24h 48h
Ctr Complex 3 p-Value Ctr Complex 3 p-Value
Apoptosis % 4.24±0.71 2.43±0.66 <0.001 1.21±0.73 15.39±0.96 <0.001
G1 % 70.92±1.82 60.9±1.35 <0.001 65.57±1.21 41.03±1.39 <0.001
G2/M% 20.29±1.11 33.5±1.28 <0.001 32.62±1.46 37.03±1.12 <0.001
----------2008 untreated cells----------2008 cells treated with IC50 of [Cu(thp)4][PF6]
Forward scattering (index of cell size) vs side scattering (index of cell granularity) as a function
of time in 2008 cells
Flow cytometric profiles of 2008 cells untreated (panel A) and treated with 3.125 (panel B) or 6.25 (panel C) µM of copper(I) complex for 24 h and stained with TMRM (10 nM).
Copper(I) complex induced a massive increase of the TMRM fluorescence reflecting a dramatic alteration of mitochondrial membrane potential that might be correlated with the induction of a G2/M phase cell cycle arrest.
Mitochondrial energization of treated tumor cells as the retention of a mitochondrial
selective cationic fluorescent probe,
tetramethyl rhodamine methyl ester (TMRM).
The coordination of mono-phosphine ligands to copper(I) gives rise to a metallodrug able to inhibit the growth of tumor cells via cell G2/M cell cycle arrest and paraptosis accompanied with the loss of mitochondrial transmembrane potential.
Potential Cu(I) radiopharmaceuticals
0.0
2.0
4.0
6.0
8.0
10.0
0 20 40 60 80 100 120
% C
ell
Ass
oci
ate
d R
ad
ioa
ctiv
ity
Time (min)
1
3
4
2
Sodium acetatebuffer
TPA64Cu(II)Cl2
64Cu
P
HO
HO
OH
P
OH
OH
HO
P PHO
OHOH HOHO
OH
NNP
NNN
PN
NN
PN NN
P
N
64Cu
N
N
N
N
64Cu
P
CO2
HO
HO
OH
P
OH
OH
HO
NN
N
N
NN
64Cu
P
CO2
HO
HO
OH
P
OH
OH
HO
THPSodium acetatebuffer
Sodium acetatebuffer
Ligand
In vitro cell experiments
Cell uptake behavior of complexes 1-4 into EMT-6 mammary carcinoma cells. Error bars not seen
are within symbols.
(2)
(1)
(3) (4)
Biodistribution Studies
The uptake and retention of activity was high in many non-target tissues lung and liver
Poor blood clearance suggestes breakdown of the complex and binding of 64Cu to serum proteins in vivo.
The heart uptake was high at all time points and there was no clearance from the myocardium over 24 h post-injection potentially due to themonocationic nature of the complex
Tumor uptake of complex 1 was highest at 1 h and decreased slowly over 24 h. In the same EMT-6 tumor model, uptake of 64Cu-ATSM and 64Cu-PTSM (both of which are clinically tested agents) into the tumor at 40 min post-injection showed lower uptake than that of 1
Tumor uptake of complex 1 is significantly higher than that for [64Cu((EtOCH2CH2)2PCH2CH2P(CH2CH2EtO)2)]+
0.0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
40.0
45.0
blood lung liver(all) spleen kidney muscle heart brain bone tumor
%ID
/g
Organ
1 hour
4 hour
24 hour
64Cu
P
HO
HO
OH
P
OH
OH
HO
P PHO
OHOH HOHO
OH
Biodistribution was carried out on 16-18 g female BALB/c mice implanted with EMT-6 cells subcutaneously into the left flank. Tumors were allowed to grow for 14 days (approx 0.3 – 0.7 cm3), at which time the animals received 0.20 MBq (~5 μCi) of complex 1 in 100 μL of saline via lateral tail vein injection. Mice were examined at 3 time points (n = 4 per group at 1, 4 and 24 hours).
S. Alidori, G. Gioia Lobbia, G. Papini, M. Pellei, M. Porchia, F. Refosco, F. Tisato, J.S. Lewis, C. Santini Journal of Biological Inorganic Chemistry, 13 (2008) 307-315.
axial
coronal
1 h 2 h 24 h
ScaleOff-ScaleHigh
Low
ScaleOff-ScaleHigh
Low
0
1
2
3
4
5
0 5 10 15 20 25
SU
V
Time (h)
Tumor
Muscle
Liver
Kidney
Heart
Small animal
PET Imaging
Selected axial and coronal images obtained using co-registration techniques demonstrating the uptake of 1 at 1, 2 and 24 h post injection in a mouse with an EMT-6 tumor (arrow) implanted on the flank.The EMT-6 tumors can be easily visualized at all
time points
Standard uptake values (SUVs) of 1 in selected organs in EMT-6 tumor bearing mice over 24 h (n = 4).
The uptake in the EMT-6 tumor at 1 h which remained static over 24 h
New N-, P- donor ligands
N N
P
P
P
P
O
O
O
O
O
O
O
O
O
O
O
O
NH
NH
P P
O O
O
O
O
O
NN
N N
P
OO
O
CH3
CH3 P
O O
O
CH3
CH3
P
O
O
O
CH3
CH3
P
O
O
OCH3
CH3
NN
P
OO
O
CH3
CH3 P
O O
O
CH3
CH3
N N
P
O
O
O
CH3
CH3
P
O
O
OCH3
CH3
H2N
NH2
P
O
O
O
P
O
O
O Br
P
OO
O
P
OO
O Br
Cyclam
Cyclen
NN
P
OO
O
CH3
CH3 P
O O
O
CH3
CH3
N N
P
O
O
O
CH3
CH3
P
O
O
OCH3
CH3
NN
P
P
N N
P
P
OHOH
OH
OH
OH
OH
OH
OH
1. HCHO, HCl, EtOH
2. NaHCO3
NN
PH2
PH2
N N
PH2
PH2
LiAlH4
1. n-BuLi
2. RX
N
NN
N
P
PRP
P
R
R
R
R
RR
R
New macrociclic ligands
N H 2HO O C
SH
H CH O
H 2O
HNH O O C
S
1) Na/NH3
2) NH4Cl, pH = 2
NHHOOC
SH
HN COOH
HS
N H 2HO O C
SH
H CH O
H 2O
HNH O O C
S
1) Na/NH3
2) NH4Cl, pH = 2
NHHOOC
SH
HN COOH
HS
P. Blondeau, C. Berse, D. Gravel, Can. J. Chem. 45 (1967) 49.
G. Papini, S. Alidori, J. S. Lewis, D. E. Reichert M. Pellei,, G. Gioia Lobbia, G. B. Biddlecombe, C. J. Anderson, C. Santini J. Med. Chem. (2008) submitted
NH
HOOC
SH
HN
COOH
HS
1) NaHCO3
N+HOOC
S
N+COOH
S
+2) HCl, pH = 2
(CH2)3Br
Br
13-membered NEC-SP
N+HOOC
S
N+COOH
S
12-membered NEC-SE
NH
HOOC
SH
HN
COOH
HS
1) NaHCO3+
2) HCl, pH = 2(CH2)2
Br
Br
NH
HOOC
SH
HN
COOH
HS
1) MeOH/H2O/NaOH+
2) HCl, pH = 2(CH2)4
Br
Br
14-membered NEC-SB
N+HOOC
S
N+COOH
S
H
H
H
H
H
HH
H
H
H
H
H
Copper(II) complexes
G. Papini, S. Alidori, J. S. Lewis, D. E. Reichert, M. Pellei, G. Gioia Lobbia, G. B. Biddlecombe, C. J. Anderson, C. Santini J. Med. Chem. (2008) submitted
Cu(CH3COO)2
H2O
Cu(CH3COO)2
H2O
Cu(CH3COO)2
H2O
N+HOOC
S
N+COOH
S
13-membered NEC-SP
N+HOOC
S
N+COOH
S
12-membered NEC-SE
14-membered NEC-SB
H
H
H
H
H
HH
H
Cu(NEC-SP)
Cu(NEC-SE)
Cu(NEC-SB)
S S
NHNH
CuOO O O
S S
NHNH
CuOO O O
S S
NHNH
CuOO O O
N+HOOC
S
N+COOH
S
H
H
H
H
64Cu complexes
Biodistribution data The retention of activity in tissues is
similar to that observed with 64Cu-cyclam
and 64Cu-monooxo-tetrazamacrocyclic
complexes, but, on comparison with 64Cu-TETA and 64Cu-DOTA, the uptake
and retention of and are orders-of-
magnitude higher.
The poor clearance suggests that the
complexes are rapidly degraded in
blood and tissues and the 64Cu is
sequestered by proteins, and
remaining trapped in these tissues
hindering clearance.
64Cu(II)Cl2
Sodium acetate buffer
Macrociclic ligands
N+HOOC
S
N+COOH
S
H
HH
H
64Cu-Complexes
S S
NHNH
64CuOO O O
0
2
4
6
8
10
12
14
16
18
7 8 9 Cu-64-Cyclam Cu-64-TETA Cu-64-DOTA Cu-CB-TE2A
%ID
/org
an
Complex
Blood
Liver
Kidney
Perspectives
1) NaOH, pH = 5.5
2) sulfo-NHS, EDC
NHHOOC
S
HN COOH
S
NH
S
HN
S
O
ON
O
O
SO3-
O
ON
O
O
SO3-
1) pH = 7.5
2) BM-NH2 . HCl
NH
S
HN
S
OO
OH
HO
H
HO
H
OHNH
H
H
HOO H
OH
H
OH
H
HOHN H
H
OH
CuAc
H2O
N
S
N
S
OO
OH
HO
H
HO
H
OHNH
H
H
HOO H
OH
H
OH
H
HOHN
H
H
OH
Cu
M
BFCA
NHHOOC
SH
HN COOH
HS
NHHOOC
S
HN COOH
S
X
XCl Cl
NHHOOC
S
HN COOH
S
NHROOC
S
HN COOR
S
ROH, HCl
Conclusions
The monooxo Re(V) core is conveniently stabilized by tripodal scorpionate ligands comprising carboxylate or sulfonate tails, giving a series of intermediate Re(O)(NNO)Cl(X) (X = Cl, OR). To these entities various bidentate ligands (BID) can be attached to produce "3 + 2" mixed ligand compounds.
Hydrophilic ‘cold’ Cu(I)-complexes have shown significant antiproliferative activity in vitro on a series of tumor cell lines, also resistance to cisplatin, showing a different pathway of action from that of cisplatin.
Hydrophylic ‘hot’ 64Cu(I) monophosphine complexes were evaluated as a basis for a new class of copper radiopharmaceuticals. [64Cu(thp)4]+ = building-block for new radiopharmaceuticals, perhaps the first time such a method has been used in the
production of Cu-radiopharmaceuticals.
Novel macrocyclic ligands, based on the L,L-ethylenedicysteine skeleton, have been prepared in view of the attractive opportunity to use them as bifunctional chelators for copper nuclides. This is the first report of 64Cu labeled to this form (N2S2)
macrocyclics. Although the in vivo biodistribution of complexes suggests dissociation of the 64Cu from the chelates, these new ligands platform offers the potential as a basis
for further development to improve the in vivo stability.
Partners and AcknowledgementsPartners and Acknowledgements
Prof. Giuliano BandoliProf. Alessandro Dolmella
Dr.ssa Cristina MarzanoDip. di Scienze Farmaceutiche
Università di Padova
Dr. Franco BenetolloICIS-CNR, Padova
Prof. Rasika Dias Department of Chemistry and BiochemistryThe University of Texas at Arlington (USA)
Dr. Francesco Tisato Dr.ssa Marina PorchiaDr. Fiorenzo Refosco, Dr.ssa Cristina BolzatiICIS-CNR, Padova
Prof. Giancarlo Gioia LobbiaProf. Carlo Santini
Dr.ssa Maura PelleiDr. Simone Alidori
Prof. Jason S. LewisCarolyn J. Anderson
