DOI 10.1007/s00706-014-1262-1
ORIGINAL PAPER
Received: 11 March 2014 / Accepted: 10 June 2014 / Published online: 30 July 2014
Springer-Verlag Wien 2014
Abstract The substitution of both coordinated water The kinetic and mechanistic description of the square
molecules of cis-diaqua(1,3-diaminopropane)platinum(II) planar platinum(II) complexes attracted continued attention
complex by a series of amino acids, viz. L-asparagine, due to their intrinsic chemical and bio-medical applications
L-glutamic acid, and glycine was investigated as a function of [3]. In particular, cisplatin and its structural analogs are
ligand concentration, temperature, and pH and found to occur used in cancer chemotherapy. But the major drawback of
in two consecutive reaction steps. The mechanism of the these chloro derivatives of N,N-chelated platinum(II)
substitution reactions appears associative in nature as sup- complexes was their nephrotoxicity. However, the aqua
ported by the large and negative values of DS=. The kinetic variety was found to be superior to others [35].
study has been substantiated by product isolation, UV, IR, Nuclear DNA has been recognized as the principal target
and ESI-MS spectroscopic analysis and rate parameters have for Pt and certain platinated DNA adducts elicit DNA deg-
been evaluated under different reaction conditions. radation and apoptosis (programmed cell death) [6, 7]. Even
though intensive efforts of various research groups in recent
Keywords Platinum(II) 1,3-Diaminopropane past have contributed appreciably towards understanding of
Amino acids Kinetics the anticancer bustle of platinum complexes, most of the
studies paying attention on the structural identification of the
Introduction binding of the DNA constituents to the metal center by means
of NMR, HPLC, ESI-MS, and X-ray technique. Signifi-
In the twenty-first century, the number of patients suffering cantly, very less is known about the mechanistic details with
from cancer is increasing worldwide. Cancer, which is regard to the various steps of adduct formation and the fac-
expected to be the most significant disease in the twenty- tors controlling such adduct formation.
first century [1, 2], is generally known to be the unre- This paper reports a thorough study of the concentration
strained growth of a tumor or cells. In the last half of the of the ligand, temperature, and pH dependencies of the rates
twentieth century, several compelling platinum-containing of substitution of the title complex with amino acids in
anticancer complexes were developed. Unfortunately, these aqueous medium. We preferred the above-mentioned amino
complexes have some toxic side effects, which are intim- acids seeing that platinum complexes can also interact with
idating because of the development of many severe other molecules in a cellular environment, like peptide
secondary complications. Today, widespread effort is being structures or RNAs. Appraisal of the modes of action of
made to look for new anticancer agents with high potency metal-based antitumor drugs therefore requires the study of
and little or no side effects. their interactions with a range of feasible biological targets
including amino acids, hormones, peptides, and proteins.
The study of model species such as essential amino acids
S. Ray D. Nandi A. Chattopadhyay R. Sarkar can give a hand in the elucidation of more amalgamated
P. Karmakar A. K. Ghosh (&)
systems. The results of this study, together with those of the
Department of Chemistry, The University of Burdwan,
Burdwan 713104, West Bengal, India previous investigations, shed light on the kinetic behavior
e-mail: alakghosh2002@yahoo.co.in of the N,N-chelated cationic platinum(II) complexes.
123
1728 S. Ray et al.
Results and discussion characteristic changes when compared to those of the free
ligands. The platinumligand vibrations are expected to
Spectroscopic analysis occur in a very low frequency range (below 600 cm-1) [8].
FT-IR spectra of L-asparagine (L1H), L-glutamic acid
The electronic transitions observed in the spectra produced (L2H), and glycine (L3H)-substituted products, the strong
by UVVis spectroscopy were comparable with what was bands at 437, 443, and 505 cm-1, respectively, are
expected for platinum(II) coordination complexes (Fig. 1). assigned to the PtN stretching vibrations. The PtO
The IR spectra of the compounds were measured in the stretching vibrations are assigned to the bands observed at
region 4,000400 cm-1 (Figs. 2, 3, 4) and showed 497, 526, and 605 cm-1, respectively. The strong bands at
80
437.84
70
497.63
1041.56
1109.07
705.95
1462.04
60
1290.38
1406.11
1238.30
50
%T
40
3261.63
30
3417.86
1681.93
20
10
0
4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500
Wavenumber/cm-1
123
Reactions of essential amino acids 1729
90
443.63
1045.42
596.00
526.57
80
1195.87
70
1404.18
1707.00
%T
1666.50
60
3192.19
3234.62
3435.22
50
40
30
4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500
Wavenumber/cm-1
90
80
777.31
927.76
70
605.65
1195.87
1126.43
2611.62
688.59
891.11
1039.63
60
%T
505.35
3437.15
2966.52
1330.88
50
1500.62
1408.04
3132.40
40
1598.99
30
20
4000 3500 3000 2500 2000 1750 1500 1250 1000 750 500
Wavenumber/cm-1
3,261, 3,234, and 3,132 cm-1 in the experimental FT-IR coordinated COO group has the masym (COO) stretch at
spectrum are assigned to the NH stretching vibrations of lower frequency [9]. The band at 1,682, 1,666, and
the amino groups. The asymmetric COO stretching fre- 1,599 cm-1 is therefore assigned to the masym (COO) of the
quency of the amino acids occurs at 1,5801,660 cm-1 metal-bound carboxyl group. From the individual assign-
when the group is coordinated to metals, whereas a non- ments of different bands it can be presumed that the final
123
1730 S. Ray et al.
123
Reactions of essential amino acids 1731
products are (O,N)-coordinated chelates and amino acids 6.04(COOH), 9.67 a(NH3?) and 2.34 a(COOH), 9.60
behave like a bidentate ligand in the experimental pH. ESI- a(NH3?) [13], respectively, at 25 C. From the pKA
MS of the resulting solutions, obtained from the 1:1 mix- values of the ligands it can be concluded that at pH 4.0,
ture of the substrate complex 1 and three amino acids are ligands L1H, L2H, and L3H remain in the zwitterionic
shown in Figs. 5, 6, and 7. It is clear from these spectra that forms, which take part in the reaction. The pK1 and pK2
the ions at m/z = 400.07, 415.07, and 343.05 (major peak) values for cis-[Pt(1,3-diaminopropane)(H2O)2]2? have
for L1H, L2H, and L3H are the precursor ion species in the been evaluated by Irving-Rossotti method [14] and found
mixture solutions and this is cautiously attributed to [amino to be 6.14 and 7.67, respectively. Hence it can be
acids ? Pt ? (1,3-diaminopropane)] the relative abun- assumed that at pH 4.0, the reactant complex exists as
dance of isotope peaks, which matched with the predictable diaqua ion. The reactions between complex 1 and amino
values. The precursor ions are made known schematically acids involve a two-step consecutive route; we put for-
(Scheme 1). ward that in the first step one aqua ligand is replaced
We proposed that Pt(II) is likely to be coordinated to from cis-[Pt(1,3-diaminopropane)(OH2)2]2? by the LH.
both COO and NH2 considering the strong coordination The second step is slower, where the remaining aqua
ability of these functional groups [1012]. Thus, the ligand is substituted with chelate ring closure. The rate
structure proposed here for product ion species, deduced constant for such a process can be evaluated by assuming
from ESI-MS and IR are generally consistent with those the following scheme:
derived from other experimental methods. k1 k2
A ! B ! C 1
Kinetic studies where A is the diaqua species 1, B is the single substituted
intermediate, and C is the final product 2, [Pt(1,3-diami-
The pKA values of the amino acids L1H, L2H, and L3H nopropane)(L)]?. Formation of C from B is predominant
are 2.02 a(COOH), 8.84 a(NH3?); 2.19 a(COOH), after some time has elapsed.
123
1732 S. Ray et al.
k1obs k1 LH; 4
where LH is the zwitterionic form of amino acids. Based Effects of pH on reaction rate
on the above scheme, a rate expression Eq. (4) can be
derived for the A ? B step: The reaction was studied at four different pH values (2.8,
k1 Pt1;3-diaminopropaneH2 O2 3.4, 4.0, and 4.6). The k(obs) values increased with increase
dB=dt 2 total LH 3
in pH; at fixed concentration of 2 9 10-4 mol dm-3 of
where [Pt(1,3-diaminopropane)(H2O)2? 2 ]total is the complex 1, 6 9 10-3 mol dm-3 glutamic acid, and
concentration of the unreacted complex. Hence, we can 0.1 mol dm-3 NaClO4 ionic strength. The increase in rate
write may be explained based on the acid dissociation equilibria
123
Reactions of essential amino acids 1733
-1
10 k1(obs) /s
C
8
B
3
A
4
0
0 2 4 6 8 10
3 -3
10 [Asparagine]/mol dm
8 C
B
3
A
4
0
0 2 4 6 8 10
3 -3
10 [Glutamic acid]/mol dm
8
D
-1
C
10 k1(obs)/s
B
3
4
A
0
0 2 4 6 8 10
3 -3
10 [Glycine]/mol dm
123
1734 S. Ray et al.
123
Reactions of essential amino acids 1735
0.6 ing to our previous work [17]. Origin software was used for
0.5
computational analysis. Rate data, represented as an aver-
0.4
age of duplicate runs, were reproducible to within 4 %.
0.3
Acknowledgments One of the authors (Sumon Ray) is grateful to
0.0 0.2 0.4 0.6 0.8 1.0 the University Grants Commission, New Delhi, India, for providing
[L1H] / ([L1H] + [M]) financial assistance and authors are also very much gratified to the
Department of Chemistry, University of Burdwan, West Bengal,
Fig. 11 A typical Jobs plot for the reaction between complex 1 and India, for infrastructural amenities.
L1H at pH = 4.0 and ionic strength = 0.1 mol dm-3 NaClO4
References
ion mode. Kinetic measurements were monitored on a
Shimadzu spectrophotometer (UV-2450) equipped with a 1. Kelland L (2007) Nat Rev Cancer 7:573
Shimadzu TCC 240A cell temperature controller (accuracy 2. Ho YP, Au-Yeung CFS, To KWK (2003) Med Res Rev 23:633
0.1 C). 3. Lippert B (ed) (1999) Cisplatin: chemistry and biochemistry of a
cis-Dichloro(1,3-diaminopropane)platinum(II) was syn- leading anticancer drug. Wiley-VCH, Zurich
4. Meijer C, Mulder NH, Timmer-Bosscha H, Sluiter WJ, Meersma
thesized according to an optimized procedure, based on a GJ, de Vries EG (1992) Cancer Res 52:6885
previously reported method for Pt(en)Cl2 [16]. The reactant 5. Fokkema E, Groen HJ, Helder MN, de Vries EG, Meijer C (2002)
complex cis-[Pt(1,3-diaminopropane)(OH2)2](ClO4)2 (1) Biochem Pharmacol 63:1989
was prepared from cis-dichloro(1,3-diaminopropane)plati- 6. Cornelison TL, Reed E (1993) Gynecol Oncol 50:147
7. Kasparkova J, Vojtiskova M, Natile G, Brabec V (2008) Chem-
num(II) by hydrolysis in the presence of two molar istry 14:1330
equivalents of silver perchlorate. The above-stated reactant 8. Apralahti J, Lehikoinen P (1989) Inorg Chim Acta 159:115
complex 1 was then characterized spectrophotometrically. 9. Tyagi P, Gahlot P, Kakkar R (2008) Polyhedron 27:3567
The products of the reaction between complex 1 and amino 10. Goswami AM, De K (2007) Transit Met Chem 32:419
11. Burger K (1996) Biocoordination chemistry: coordination equ-
acids were separately prepared by mixing the reactants at libria in biologically active systems. Ellis Horwood Limited,
pH 4.0 in different molar ratios, namely 1:1, 1:2, 1:3, 1:4, Chichester
and 1:5, and thermostating the mixtures at 60 C for 48 h. 12. Burgenson IE, Kostic NM (1991) Inorg Chem 30:4299
The absorption spectra of the resulting solutions were 13. Fox SW, Foster JF (1957) Introduction to protein chemistry.
Wiley, New York, p 26
recorded and each set was found to exhibit almost identical 14. Irving HM, Rossotti HS (1954) J Chem Soc, p 2904
absorbance at the characteristic wavelengths irrespective of 15. Weyh JA, Hamm RE (1969) Inorg Chem 8:2298
their different molar ratios. The difference in spectra 16. Karmakar P, Ray S, Nandi D, Mandal A, Mondal S, Mallick S,
between the product complexes and the substrate complex Bera BK, Ghosh AK (2013) J Solut Chem 42:441 (and references
therein)
is shown in Fig. 1. The pH of the solutions was adjusted by 17. Karmakar P, Ray S, Nandi D, Mandal A, Mondal S, Mallick S,
adding NaOH/HClO4 at pH 4.0 so that reactant complex 1 Bera BK, Ghosh AK (2013) Synth React Inorg Met Org Nano
exists as diaqua species in the reaction mixture. According Met Chem 43:1563
to Jobs method of continuous variation, metalligand
compositions were found to be 1:1 in each case (Fig. 11).
123