Me al
Based
D ugs
Vol.
7,
No.
4,
2000
ISOIATION,
CHARATION
AND
ANIII’UMOUR
PROPIi IES
OFTHE
1,2-PROPYLENEDIAMINETETRAACETATE
ans-DIAQUA-COPPER
(II)
S.
Kamah
,
R.
Vilaplana
,
J.
Mo enC,
K.
Akdi
,
G.
Ga cia-He dugo’
and
F.
Gonzlez-Vilchez
*
Uni e sidad
de
Se illa,
41012
Se i la,
Spain
Dp o.
de
Quimica
Ino ginica,
F.
de
Quimica
Dp o.
de
Biologia
Celula ,
F.
de
Biologia
Abs ac
A
ans-diaquacomplex
o med
by
coppe (ll)
sulpha e
and
he
seques e ing
polyammino-
polyca boxylic
ligand
1,2-p opylenediamine e aace ic
acid
(PDTA)
has
been
isola ed
and
cha ac e ized
by
chemical
analysis,
i ime y,
FT-IR
and
elec onic
spec oscopy.
Po en iome ic
and
elec onic
measu emen s
iden i ied
he
ligand
as
e aden a e,
wo
ni ogen
and
wo
oxygen
a oms
being
bonded
o
he
Cu(II)
in
plana
posi ions.
This
oc ahed al
monome ic
soluble
compound,
is
an
unusual
example
o
a
coppe
(I|)
subs ance
showing
signi ican
in
i o
an i umou
ac i i y
agains
he
human
o a ian
umou
cells
T
G
(IDs0
2.29
k M
a
48
h)
and
impo an
in
i o
an i umou
ac i i y
agains
solid
Sa coma
180
wi h
comple e
eg ession
o
he
umou
a
a
dose
o
12.5
mg/Kg
body
weigh .
In oduc ion
Se e al.compounds
o
ansi ion
pla inum
g oup
me als
wi h
1,2-p opylenediamine-N,N,N’,N’-
e aace ic
acid
(PDTA),
a
me hyl
de i a i e
o
EDTA,
ha e
been
isola ed
and
es ed
o
biological
ac i i y
agains
human
umou s
cells
du ing
ecen
yea s
by
ou
esea ch
g oup[l’3].
Consequen ly,
ou
in e es
has
also
been
add essed
o
he
complexes
o med
by
me al
ions
o he
han
pla inum
as
po en ial
me allopha maceu icals
showing
an i umou
ac ion
ha
di e ed
signi ican ly
om
ha
o
cis-pla in.
In
his
con ex ,
ou
wo k
on
he
chemis y
and
biological
ac ion
o
new
PDTA-Ru(III)
complexes
and
he
in e es ing
esul s
ecen ly
ob ained
[4-8]
conce ning
hei
in e ac ion
agains
DNA
and
human
cells,
encou aged
us
o
unde ake
new
s udies
ocused
on
he
biological
beha iou
shown
by
PDTA
complexes
o med
wi h
coppe
(II),
an
essen ial
ion
p esen ing
no el
in
i o
an ibac e ial,
an i i al
and
an i ungal
ac i i ies
on
a
b oad
ange
o
mic oo ganisms[
9-11],
besides
i
o ms
a
pa
o
he
Cu-Zn
SOD
enzymes.
Ou
in e es
on
hese
ype
o
complexes
is
also
suppo ed
by
he
impo an
ole
played
by
Cu(ll)
complexes
wi h
chela ing
ligands
as
models
o
he
associa i e
complexes
in ol ed
in
he
subs i u ion
eac ions
a
he
coppe
si e
o
hese
enzymes:
indeed
s ongly
seques e ing
ligands
should
a ou
he
o ma ion
o
six-coo dina e
coppe
complexes
p esen ing
he
same
coo dina ion
numbe
as
he
associa i e
complex[
12].
In e es ingly,
complexes
o
he
hexaden a e
PDTA
ha e
been
ound
wo
o de s
o
magni ude
mo e
s able
han
hose
o med
by
analogous
pa en
ligand
(EDTA).
Mo eo e ,
Cu(II)
complexes
wi h
poly unc ional
ligands
such
as
PDTA
has
also
been
impo an
because
PDTA
can
eadily
o m
di e en
shapes
wi h
di e se
coo dina ion
numbe s
and
hus
adap
o
he
subs a e[13].
This
communica ion
deals
wi h
he
syn hesis
o
he
solid
oc ahed al
compound
[Cu(PDTA-
Hz)(H20)2]H20
and
i s
cha ac e iza ion
using
chemical
analyses,
po en iome y,
ib a ional
spec oscopic
and
he mal
s udies.
Finally,
he
ac i i ies
o
his
complex
agains
he
o a ian
umou
cell
line
TG
(in
i o)
and
he
solid
umou
Sa coma
180
(in
i o
ha e
also
been
es ed.
Ma e ials
and
Me hods
Syn hesis
o
he
compound
[CuL(H20)2]H20
(1)
(L
PDTA-H2)
An
aqueous
solu ion
o
CuSO4.SH20
(3.80
g
in
100
ml
o
dis illed
wa e )
was
passed
epea edly
h ough
a
il e ing
lask
su ace
con aining
solid
PDTA
(6.12
g)
un il
he
ini ial
o ma ion
o
a
blue
solu ion.
This
solu ion
was
slowly
e apo a ed
a
oom
empe a u e
un il
p ecipi a ion
o
a
blue
compound
which
was
il e ed
and
washed
wi h
ace one
ollowed
by
die hyl
e he .
The
ob ained
mic oc ys alline
powde
was
Anal.
Calcd.
o
C
iH Ol
IN-Cu:
C,
dissol ed
in
wa e ,
ec ys allized
and
s o ed
in
dessica o
on
CaC
"1
31.2;
H,
5.2;
N,
6.6;
Cu,
15.6;
M
+
421.8.
Found:
C,
31.4;
H,
N,
6.6;
Cu
15.5;
M
/----
422"
Chemicals
Coppe
(ll)sulpha e
pen ahyd a e
was
used
as
p o ided
by
Sigma.
All
chemicals
ob ained
om
comme cial
supplie s
we e
used
wi hou
u he
pu i ica ion.
The
ligand
PDTA
was
syn hesized
in
ou
labo a o y
ollowing
a
published
me hod[14].
Elemen al
analyses
Elemen al
mic oanalyses
we e
pe o med
a
he
Mic oanaly ical
Labo a o y
o
he
Ba celona
Uni e si y.
The
wa e
con en
was
de e mined
by
Ka l-Fische
i ime y
whe eas
ne al
con en
was
de e mined
by
a omic
abso p ion
spec oscopy
on
a
Pe kin
Elme
2380,
a
l0
mA
and
)
o
349.9
nm.
219
F.
Gonzalez-Vilchez
e
al
Isola ion,
cha ac e iza ion
and
An i umou
P ope ies
o
1,2-P opylenediamine e aace a e- ans-diaqua-coppe (II)
Po en iome ic
and
conduc ime ic
s udies
Po en iome ic
and
conduc ime ic
s udies
we e
pe o med
using
a
C ison
Mic oTT
2022
i ime e ,
p o ided
wi h
au obu e e
Mic obu
2030.
An
aqueous
solu ion
o
he
complex
(50
mg/100
ml)
was
i a ed
agains
NaOH
21.5
mM
solu ion.
Elec ical
conduc ime y
o
he
same
solu ion
was
pe o med
on
a
C ison
525
conduc ime e .
Spec oscopic,
magne ic
and
he mal
s udies
In a ed
spec a
we e
eco ded
on
a
Jasco
FT-IR
spec ome e
in
he
4000-400
cm
-1
ange
as
KB
discs
while
a -in a ed
spec a
we e
eco ded
as
Nujol
mulls
suppo ed
be ween
polye hylene
shee s.
Elec onic
u - isible
spec a
we e
measu ed
wi h
a
Jasco
V-550
spec ome e
while
he
magne ic
measu emen
was
ca ied
ou
on
a
Gouy
ype
equipmen
a
298K.
Calib a ing
balance
wi h
he
s anda d
compound
Hg[Co(SCN)a
and
Pascal
cons an s
we e
used
o
diamagne ic
co ec ions.
The
he mal
beha iou
o
he
compl6x
was
s udied
wi h
a
Me le
DSC
ins umen
(un il
300C)
and
a
S an on
he mobalance
(300-500C);
measu emen s
we e
ca ied
ou
wi h
25
mg
o
sample
and
a
hea ing
a e
o
10C/min.
Cy o oxic
and
an i umo
essays
The
cy o oxic
e ec
o
he
coppe
complex
has
been
e alua ed
agains
human
cance
cells
o
he
o a ian
umou
cell
line
TG.
The
cells
we e
ou inely
main ained
as
monolaye s
in
cell
cul u e
lasks
(T-25
cm
2,
Nunc,
Sweden)con aining
Dulbeco’s
minimal
essen ial
media
supplemen ed
wi h
10%
e al
bo ine
se um
(FBS)
2
mM
glu amin,
100
U/ml
penicillin
s ep omycin
and
1%
py u a e.
The
sample
es ed
was
dissol ed
in
wa e
a
pH
7.0
and
hen
added
o
he
app op ia e
cell
lasks
o
ob ain
inal
concen a ions
o
l,
l0
and
100
k g/ml.
Cells
we e
incuba ed
o
120
h
a
37C
unde
g ow h
condi ions
(5%
CO2;
95%
02;
He aeus
incuba o )
and
washed
wice
wi h
p ewa med
phospha e
bu e ed
saline
(PBS,
37C).
The
co esponding
cul u es
we e
exposed
o
ipsin/EDTA
solu ion
(0.1%
ipsin,
0.4%
EDTA
in
PBS)
a
37C
o
2
ain.
Cul u e
medium
( wice
olume
o
ipsin
solu ion)
was
added
o
he
cells
be o e
cen i uga ion
(1500
pm,
5
min)
and
cell
pelle s
we e
esuspended
in
he
cul u e
medium
and
coun ed
in
a
Thomas
chambe
unde
in e ed
ligh
mic oscope
e e y
24
h.
The
da a
we e
s a is ically
analysed
using
he
s uden
-
es .
Di e ences
we e
conside ed
signi ican
when
p<
0.001.
The
an i umou
ac i i y
o
1
was
es ed
in
i o
agains
Sa coma
180
(S180)
by
he
NIO
(Spain),
acco ding
o
he
p o ocol
es ablished
by
he
D ug
Syn hesis
and
Chemical
B anch
o
NCI,
Be hesda,
USA[
15].
Animals
we e
BDF1
emale
mice
wi h
weigh s
wi hin
a
3g
alue
ange
and
a
minimum
weigh
o
17
g.
The
numbe
o
animals
was
6
pe
es
g oup.
The
he apeu ic
ac i i y
o
he
complex
was
ob ained
om
he
T/C
pe cen age
which
is
desc ibed
as
T/C
%
(100)
x
Mean
li e
span
o
ea ed
mice/Mean
li e
span
o
un ea ed
mice;
umou
ee
su i o s
we e
excluded.
The
minimum
alue
o
T/C
o
ac i i y
is
115.
I
T/C
>
125
he
complex
is
conside ed
as
a
candida e
o
u he
es ing
agains
di e en
selec ed
u nou
sys ems.
Resul s
and
Discussion
Po en iome ic
S udy
Figu e
a
co esponds
o
he
po en iome ic
i a ion
o
an
aqueous
solu ion
o
he
coppe
complex.
A
sha p
inc ease
o
pH
o
he
consump ion
o
2
g-equi ,
o
alkali
wi h
an
in lec ion
poin
appea ing
a
pH
6.6
can
be
obse ed.
Fu he mo e,
a
sudden
change
is
also
obse ed
in
he
elec ical
conduc i i y
o
he
solu ion
(Figu e
lb).
All
hese
ac s
demons a ed
he
simul aneous
neu aliza ion
o
wo
ee
-COOH
g oups
o
he
same
s engh .
These
esul s
suppo
he
e aden a e
cha ac e
o
he
PDTA,
ha
con ains
wo
coo dina ed
ca boxyla e
g oups
and
wo
ee
ca boxylic
g oups.
Molecula
weigh
deduced
om
he
i a ion
co esponds
closely
wi h
he
one
(MW:
420)
expec ed
o
he
complex.
In a ed
spec oscopy
The
FT-in a ed
spec um
o
compound
I
show
in ense
bands
a
2920
cm
"l
and
2926
cm
"1,
assigned
o
symme ic
s e ching
ib a ions
(Vs)
o
he
C-H
bond
o
me hylene
g oups
a ached
o
coo dina ed
and
ee
ca boxylic
g oups.
The
obse ed
spli ing
is
demons a i e
o
he
equal
numbe
o
ee
and
coo dina ed
ca boxylic
g oups
16,17]..
The
la e
assignemen
is
u he
suppo ed
in
he
complex
by
he
s ong
cha ac e is ic
bands
a
1730
cm
"l
and
1590
cm
"l
(Vas,
C=O
bond
o
ee
and
coo dina ed
ca boxylic
g oups)
whe eas
he
s
o
C=O
bond
o
coo dina ed
ca boxyla e
g oups
appea ed
a
1400
cm
1
The
di e ence
o
190
cm
"l
be ween
Vas
and
s
o
coo dina ed
ca boxyla e
g oups
indica ed
ha
he
complex
is
o
p edominan ly
ionic
cha ac e .
The
coo dina ed
wa e
in
1
p esen s
di e en
peaks
a
990
cm
-1
( ocking),
760
cm
l
(wagging)
and
a ound
600
cm
l
(Vas)
and
440
cm
-1
(Vs)
[18]
whe eas
none
o
hese
ib a ions
appea
in
he
in i’a ed
spec um
o
compound
1
when
i
was
hea ed
a
220C,
he
empe a u e
a
which
bo h
coo dina ed
and
la ice
wa e
a e
los ,
as
es ablished
by
he mal
s udy
o
he
complex.
Simila
ac s
ha e
been
checked
ecen ly
in
new
u henium
compounds
o med
wi h
iminodiace ic
acid
I19].
220
Me al
Based
D ugs
Vol.
7,
No.
4,
2000
equi ,
alk/mol
o
compound
Figu e
I.
Po en iome ic
(a)
and
conduc ime ic
(b)
s udy
o
[Cu(PDTA-
H)(H20)21H,O.
Mola
conduc ance
(298K)
changes
on
A
190
S
cm
2
(I
g-equi .
alkali)
o
A
142
S
cm
2
(2
g
equi ,
alkali).
:-....,
,o
"
’’"4
b
100
200
300
400
(C)
Figu e
2.
DSC
s udy
(cu e
a),
di e en ial
he mal
analysis
(cu e
b)
and
he mog a ime ic
analysis
(cu e
c)
o
1.
Wa e
no mally
gi es
b oad
abso p ion
a
3500
cm
1.
1
shows
a
single
peak
a
3440
cm
I,
a ibu ed
o
he
la ice
wa e
molecules
mos
likely
bonded
o
anionic
ligands
h ough
hyd ogen
bonds
[20].
This
ac
is
he
cause
o
he
sa elli e
peaks
obse ed
a
a ound
2600
cm
1.
Indeed,
his
is
he
case
in
compound
1,
as
checked
in
he
in a ed
spec a
o
se e al
samples
o
he
complex
aken
a e
hea ing
a
140C.
In
all
cases,
he
spec a
only
show
a
weak
abso p ion
a
abou
3540
cm
-1.
Table
p esen s
hese
and
o he
peaks
obse ed
in
he
in a ed
spec um
o
he
1.
UV- isible
spec al
and
magne ic
s udies
The
elec onic
spec a
o
an
aqueous
solu ion
o
he
complex
(8.0
mM)
shows
a
sligh ly
assymme ic
single
band
a
13700
cm
1
(,
730
nm),
wi h
an
ex inc ion
coe icien
alue
o
67.75.
These
and
o he
pa ame e s
o
his
absop ion
a e
simila
o
hose
obse ed
o
he
analogous
Cu-EDTA
complex[
21]
and
his
band
may
be
consequen ly
a ibu ed
o
he
spin-allowed
ansi ion
2E_
_.._>
2T2g,
indica ing
a
e agonally
dis o ed
oc ahed al
symme y
o
he
complex.
Two
wa e
molecules
a e
loca ed
in
ans
posi ion
on
he
oc ahed al
Z
axis
a
longe
dis ances
om
he
me al
ion
wi h
he
emaining
ou
coo dina ed
dono
a oms
lying
on
he
plane.
Table
1.
IR
da a
(cm
l)
o
[CuL(H20)2]H20
COOH
COO"
CH2
H20
Cu-N
Cu-OH2
(C=O)
(C-H)
h
yd
1730
Vas
1590
Vas
2920
3440
s
400
600
Vas
840
1400
s
2926
1630
b
1120
440
s
s
and
Vas"
symme ic
and
asymme ic
s e ching
ib a ions;
Vb:
bending
ib a ion.
221
F.
Gonzalez-Vilchez
e
al
Isola ion,
cha ac e iza ion
and
An i umou
P ope ies
o
1,2-P opylenediamine e aace a e- ans-diaqua-coppe (II)
The
magne ic
momen
o
he
complex
has
been
calcula ed
a
298K
and
he
ob ained
alue
(1.83
BM)
di e s
om
ha
heo e ical
one
expec ed
o
he
spin-momen
alue
a
his
empe a u e
(1.94
BM),
as
p edic ed
o
an
E
e m
(second
o de
Zeeman
e ec
and
pa ial
cancela ion
o
he
o bi al
angula
momen ).
The
expe imen al
alue
o
he
magne ic
momen
con i ms
he
e agonal
dis o ion
deduced
om
he
elec onic
spec a.
4
C
on
oi
100
0
54
48
72
96
120
Time
(hou s)
Figu e
3.
De e mina ion
o
he
e ec s
exe ed
by
he
coppe (ll)
complex
on
he
g ow h
kine ics
o
he
TG
o a ian
ca cinoma
human
cance
cell
line.
TG
cells
we e
exposed
o
di e en
doses
o
he
coppe (ll)
complex
(see
expe imen al
pa
o
de ails).
The mal
s udy
The
DSC
s udy
pe o med
be ween
20C
and
350C
o
he
complex
indica es
a
medium
and
ex ended
endo he mic
e ec
a
empe a u es
be ween
85C
and
140C
(Fig.
2a).
This
e ec
is
absen
in
he
PDTA
DSC
s udy
and
migh
be
a ibu ed
o
he
dehyd a ion
o
he
uncoo dina ed
wa e
molecules.
Mo eo e ,
a
mo e
p onounced
and
spli ed
endo he mic
e ec
appea ed
a
200-220C,
indica ing
he
elimina ion
o
wo
coo dina ed
wa e
molecules.
The
deca boxila ion
p ocess
clea ly
begins
a
240C
and
con inues
du ing
all
he
empe a u e
ange.
Figu e
2b
shows
he
DTA
cu e
ob ained
be ween
20C
and
500C
while
Fig.
2c
p esen s
he
simul aneous
TGA
cu e
a
same
empe a u es.
In
he
i s
case
(cu e
b),
he
elimina ion
o
he
coo dina ed
wa e
obse ed
in
he
DSC
s udy
is
con i med
by
his
echnique
h ough
an
endo he mic
spli ed
e ec
egis e ed
be ween
200C
and
230C.
The
deca boxyla ion
p ocess
appea s
h ough
an
in ense
exo he mic
double
e ec
egis e ed
be ween
230C
and
330C.
The
i s
acu e
e ec
a
240-
290C
coincides
wi h
he
weigh
loss
de ec ed
in
he
TGA
s udy
(cu e
c)
o
he
deca boxila ion
o
he
wo
ee
ca boxylic
g oups
and
o e laps
wi h
a
second
sha p
and
less
in ense
e ec
a
310C
ha
co esponds
o
he
deca boxila ion
o
he
coo dina ed
ca boxyla e
g oups.
Final
py olysis
akes
place
a
empe a u es
o e
330C.
An i umou
ac i i y
The
inhibi ing
e ec s
o
he
1
agains
he
human
o a ian
umou
cell
line
TG
a e
shown
in
Fig.
3
which
p esen s
he
e olu ion
wi h
ime
o
he
o al
numbe
o
cul u e
cells
in
absence
(con ol)
o
p esence
o
he
complex.
Th ough
his
s udy
i
is
possible
o
de e mine
he
in luence
o
di e en
doses
o
he
coppe
complex
on
he
g ow h
kine ics
o
he
cells
cul u e.
In
his
way,
he
g ow h
kine ic
o
hecul u e
ea ed
wi h
doses
o
b g
ml
"1
and
10
b g
ml
"1
o
complex
is
signi ican ly
lowe
(p<
0.001)
han
ha
shown
in
con ol
cul u es
a e
48
h
o
he
ea men .
Simila
esul s
a e
ob ained
24
h
a e
adminis a ion
o
he
highe
dose
o
compound
(100
l. g
ml’l).
I
can
be
obse ed
ha
he
g ow h
kine ics
o
he
umou
cells
dec eases
wi h
inc easing
doses
o
1
in
such
a
way
ha
he
p oli e a ion
p ocess
is
almos
comple ely
cancelled
by
his
complex
a e
96
h
o
ea men
e en
a
he
lowe
doses
handled.
222
Me al
Based
D ugs
Vol.
7,
No.
4,
2000
80
IG
/,I
/"
!/"
,./
;,, .nd
11E/ml
10
I/ml
Dose
Figu e
4.
Va ia ion
o
he
cellula
cycle
du a ion
o
he
TG
o a ian
ca cinoma
human
cance
cell
line
wi h
a
dose
o
1.
The
e ec
p oduced
by
he
coppe
complex
on
he
i al
cellula
cycle
was
also
s udied.
Fig.
4
shows
he
cellula
cycle
du a ion
co esponding
o
ea ed
and
con ol
TG
cell
lines.
A
doses
o
l g
ml
"1,
an
impo an
induced
inc easing
o
he
TG
cellula
cycle
is
obse ed
(30
h)
i_
compa ed
o
ha
o
he
con ol
cells
(23
h).
The
leng h
o
he
cycle
o
ea ed
cells
a
a
dose
o
10
I. g
ml-1
was
also
signi ican ly
enhanced
(62
h).
Howe e ,
he
dose
o
100
I. g
ml
"1
weas
s ongly
cy o oxic
and
dea h
o
he
ea ed
cells
was
obse ed.
The
ob ained
esul s
demons a e
ha
a
signi ican
enhancemen
o
he
i al
cellula
cycle
is
p oduced
a
all
doses
lowe
han
100
g
ml
"1
On
he
basis
o
hese
s udies
one
can
conside
ha
1
beha es
as
a
po en ial
an i umou
compound
showing
p onounced
in
i o
cy o oxici y.
The
an i umou
ac i i y
agains
S
180
has
been
e alua ed
a
doses
o
3.0,
6.0
and
12.5
mg/Kg
body
weigh .
Table
2
shows
he
esul s
ob ained.
The
oxic
dose
was
ound
equal
o
35
mg/Kg
body
weigh .
F om
Table
2,
a
no ewo hy
ac i i y
agains
S180
o
he
complex
is
deduced
a
all
doses,
wi h
comple e
emission
o
he
umou
a
a
dose
o
12.5
mg/Kg
body
weigh
wi h
100
%
su i o s.
The
complex
is
well
ole a ed
and
shows
a
lack
o
oxici y
a
doses
lowe
han
35
mg/Kg
bodyweigh .
Thus,
1
beha es
as
a
ema kable
an i umou
subs ance
agains
S180
and
should
be
es ed
agains
o he
umou
sys ems
also
because
i s
no able
an ip oli e a i e
e ec
agains
TG
human
umou
cells
line.
The
mechanism
o
he
an i umou
ac i i y
cells
is
unde
p og ess.
Table
2.
An i umou
ac i i y
o
1.
agains
S180
(In
i o
Dosage
MLS(*)
Numbe
o
mice
T/C
%
(mg/Kg
body
weigh )
T/C
su i o s
a e
6
mon hs
3.0
28/40
70.0
6.0
48/40
120.0
12.5
All
ali e
Six
(l
00%)
(*)
T/C
=6/6;
MLS:
Mean
li e
span.
Single
injec ion
doses
we e
adminis e ed
i.p
Acknowledgemen s
We
hanks
inancial
suppo
om
CICYT,
Spain,
p ojec
PB97-0738.
SK
and
KA
a e
indeb ed
o
AECI
(Spain)
o
Ph.D.
schola ships.
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1.
Gonz ilez-Vilchez,
F.,
Vilaplana,
R.,
(1986),
In:
T ends
in
Cance
Resea ch,
Ba be a,
E.
(ed.)
Vizcaya:
UPV,
pp.
194-198.
2.
Vilaplana,
R.,
Gonz ilez-Vilchez,
F.,
Gu ie ez-Puebla,
E.,
Ruiz-Vale o,
C.,
lno g.
Chim.
Ac a,
1994,
224,
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Basallo e,
M.G.,
Vilaplana,
R.,
Gonz ilez-Vilchez,
F.,
Polyhed on,
1994,
13,
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Vilaplana,
R.,
Rome o,
M.A.,
Qui 6s,
M.,
Salas,
J.M.,
Gonz ilez-Vilchez,
F.,
Me al
Based
D ugs,
1995,
2,
211.
223

F.
Gonzalez-
Vilchez
e
al
Isola ion,
cha ac e iza ion
and
An i umou
P ope ies
o
1,2-P opylenediamine e aace a e- ans-diaqua-coppe (II)
11.
12.
13.
14.
15.
16.
17.
Ca ballo,
M.,
Vilaplana,
R.,
M quez,
G.,
Conde,
M.,
Bedoya,
F.,
Gonzilez-Vilchez,
F.,
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F.,
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J.
1997,
328,
559.
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F.,
Vilaplana,
R.,
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G.,
Messo i,
L.,
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M.J.,
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F.,
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D.R.,
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1999,
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E.,
Ve o i,
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Alessio,
E.,
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F.,
Vilaplana,
R.,
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A.,
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L.,
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2000,
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Z.H.,
Rau ,
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Me al
Based
D ugs,
1996,
3,
211.
Roy,
T.G.,
Haza i,
S.K.S.,
Dey,
B.K.,
Chak abo y,
S.,
Tiekink,
E.R.T.,
Me al
Based
D ugs,
1999,
6,
345.
Pan e a,
M.,
Va adino a,
T.,
Tu el,
I.,
Me al
Based
D ugs,
1998,
5,
19.
Lu,
Z.,
Duan,
C.,
Tian,
Y.,
You,
X.,
lno g.
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1996,35,
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M.,
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1993,
17,
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F.P.,
Ga an,
F.L.,
J.
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1959,
81,
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1953,
13,
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D.T.,
Paulsen,
P.J.,
J.
Am.
Chem.
Soc.,
1959,
$1,816.
Sawye ,
D.T.,
Mckinnie,
J.M.,
J.
Am.
Chem.
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1960,
82,
4191.
18.
Sa o i,
G.,
Fu lani,
C.,
Damiani,
A.,
J.
Ino g.
Nucl.
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1958,
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119.
19.
Vilaplana,
R.,
Ma inez,
R.,
F i sky,
I.O.,
Messo i,
L.,
Mish a,
L.,
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F.,
2000,
submi ed.
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J.,
Nakamo o,
K.,
Kobayashi,
M.,
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Hoa d,
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Smi h,
G.S.,
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M.,
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Ad ances
in
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Chemis y
o
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Compounds,
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Ki schne
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MacMillan,
N.Y.,
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Recei ed"
July
6,
2000-
Accep ed"
Augus
14,
2000
Recei ed
in
e ised
came a- eady
o ma "
Oc obe
13,
2000
224