Dalton

Transactions
View Article Online
PAPER View Journal | View Issue

Photophysical, photooxidation, and biomolecule-

Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.

Cite this: Dalton Trans., 2022, 51,

interaction of meso-tetra(thienyl)porphyrins
1646 containing peripheral Pt(II) and Pd(II) complexes.
Insights for photodynamic therapy applications†
Isadora Tisoco,a Maria Carolina Donatoni,b Henrique Fernandes Vieira Victória,c
José Roberto de Toledo,c Klaus Krambrock,c Otávio Augusto Chaves,d
Kleber Thiago de Oliveira b and Bernardo Almeida Iglesias *a

Received 21st October 2021,
Accepted 30th December 2021
DOI: 10.1039/d1dt03565g
rsc.li/dalton
We report the synthesis and characterization of two novel tetra-cationic porphyrins, containing Pt(II) or Pd
(II) polypyridyl complexes attached at the peripheral position of N4-macrocycle. Compounds were charac-
terized through elemental analysis, molar conductivity, cyclic voltammetry, and spectroscopy analysis.
Photophysical and photobiological parameters were also evaluated. Also, the binding capacity of each
porphyrin with human serum albumin (HSA) was determined by UV–Vis, steady-state, and time-resolved
fluorescence spectroscopy, combined with molecular docking calculations. The results suggest that the
interaction of these compounds is spontaneous, weak to moderate, and probably occurs at site III (subdo-
main IB) by non-covalent forces, including van der Waals and H-bonding. Moreover, porphyrins contain-
ing peripheral complexes improve their interactions with biomolecules, show good photostability, gene-
rate reactive oxygen species under white light studied by electron paramagnetic resonance (EPR) analysis,
and promote photo-damage of HSA.

1. Introduction tion mechanisms: type I which produces radical species, e.g.,

Porphyrins are organic macrocycles widely used as sensitizers
in photodynamic processes, including photodynamic therapy
of cancer (PDT) and antimicrobial photodynamic therapy
(aPDT), due to their good photostability, high reactive oxygen
species (ROS) yield, and specific interaction with target bio-
macromolecules.1 Different physicochemical factors determine
both safe-use and applications of different photosensitizers
(PS) in the photobiology, e.g., number of charges, aggregation
state, and hydrophobicity character.2 The ROS formation is a
fundamental role in PDT assays since the combination of
photosensitizer (PS), oxygen (O2), and adequate light will
produce reactive species. In this case, there are two main reac-
a are few reports based on tetra-( pyridyl)porphyrin macrocycle
Department of Chemistry, Federal University of Santa Maria, Av. Roraima, Santa
Maria-RS, Brazil. E-mail: bernardopgq@gmail.com, bernardo.iglesias@ufsm.br
b
Department of Chemistry, Federal University of São Carlos, Rod. Washington Luiz,
São Carlos-SP, Brazil
c
Department of Physics, Federal University of Minas Gerais, Av. Antônio Carlos, Belo
Horizonte-MG, Brazil
d applied for pharmacokinetic studies of potential drugs.19
Department of Chemistry, University of Coimbra, Rua Larga, Coimbra, Portugal
† Electronic supplementary information (ESI) available. See DOI: 10.1039/
d1dt03565g
hydroxyl, superoxide, and hydroperoxide (•OH, O2•−, •OOH,
respectively) by electron/hole transfer, while type II might gen-
erates singlet oxygen (1O2) by energy transfer processes.3 Due
to their high reactivity, ROS can oxidize different biological
substrates and biomacromolecules, including nucleic acids,
lipoproteins, and albumin.4–7
The meso-substituted porphyrins containing thienyl units (S
atoms) can be an excellent alternative for the formation of
novel peripherical cationic metalloporphyrins due to the
effective complex stabilization based on bonds between thiol
group and soft metal ions, e.g., Pt(II), Pd(II), and Ru(II) ions.8
Cationic porphyrins containing polypyridyl peripheral tran-
sition metal complexes such as Pt(II) or Pd(II) are good ROS
generators and their applications in photooxidative processes
have already been reported in the literature.9–12 To date, there
as main core for photodynamic assays.13–18
Some circulatory proteins such as human or bovine serum
albumins (HSA and BSA, respectively) are the most abundant
plasma proteins in the mammal’s bloodstream, being widely
Serum albumin plays several important roles, including regu-
lation of plasma pressure, transport, and biodistribution of

1646 | Dalton Trans., 2022, 51, 1646–1657 This journal is © The Royal Society of Chemistry 2022

Dalton Transactions
different types of both endogenous or exogenous compounds,
including hormones, fatty acids, vitamins, heme type, metab-
olites, and drugs.20 The binding capacity of albumins to metal
ions and coordination compounds reveals considerable inter-
est in its physiological role as a metal-binding protein, being
one of the most intriguing and potentially useful properties
available that impact the solubility and pharmacokinetics of
inorganic compounds.21 Regarding the photooxidation of pro-
teins, different techniques e.g., fluorescence, UV-Vis or elec-
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.
tron paramagnetic resonance (EPR) are often applied. In this
way, it was shown by EPR spin trapping and fluorescence emis-
sion the photooxidation capacity of some tetra-cationic Pd(II)
porphyrin derivatives, indicating that these compounds are
able to produce singlet oxygen (1O2) species under white-light
irradiation resulting in photodamage of BSA, which indicates
a good alternative for photodynamic processes.22
Due to the increase of the importance of porphyrins in PDT
assays, the present work reports the synthesis of two novel
meso-tetra-(2-thienyl)porphyrins coordinated to peripheral Pt
(II) or Pd(II) complexes, namely H2TTP, PtbpyTTP and
PdbpyTTP. Their characterization by elemental analysis, molar
conductivity, cyclic voltammetry, and spectroscopy analysis
were conducted. The photophysical/photobiological properties,
including aggregation, solution stability, photostability, singlet
oxygen quantum yield, and water/octanol coefficient partition
were also determined. Finally, the HSA-binding capacity was
evaluated by spectroscopy methods combined with molecular
docking calculations, and the albumin photooxidation by EPR
analysis.
2. Experimental section
2.1. General
All chemical reagents were of analytical grade and purchased
from Sigma-Aldrich® and Oakwood Chemical® (USA) without
any further purification. The HSA was lyophilized powder and
fatty acid-free (Sigma-Aldrich®; purity ≥ 99%). The concen-
tration of the stock solutions of albumin was confirmed by
UV-Vis analysis through the Beer–Lambert equation with the
molar absorptivity (ε) value of 35 700 M−1 cm−1 at 280 nm in
Tris-HCl buffer ( pH 7.4) solution23 and the water used in all
experiments was milliQ grade.
2.2. Photophysical characterization
UV-Vis absorption spectroscopy analysis was recorded using a
Shimadzu UV-2600 spectrophotometer (data interval, 1.0 nm)
in dimethyl sulfoxide (DMSO) as solvent at 250–800 nm range
(concentration of 2.0 µM). Steady-state fluorescence emission
spectra for the porphyrins in DMSO were measured in a
Horiba Yvon-Jobin Fluoromax Plus (Em/Exc; slit 5.0 mm) at
600–800 nm range (concentration of 1.0 µM). Fluorescence
quantum yield values (Φf ) values of the derivatives H2TTP,
PtbpyTTP and PdbpyTTP were determined by comparing the
corrected fluorescence spectra with those obtained by meso-
tetra( phenyl)porphyrin (TPP) in dichloromethane solution (Φf
This journal is © The Royal Society of Chemistry 2022
View Article Online
Paper
= 0.15, λexc = 522 nm)24 as the Φf standard as the fluorescence
yield and eqn (1) was used to determine the Φf values:
I ð1  10A Þstd η2
ΦF ¼ ΦFstd ð1Þ
Istd ð1  10A Þ η2std
where Φf, I, A, and η are the fluorescence quantum yield, inte-
gral area of fluorescence, absorbance in λexc, and refractive
index of selected solvents (DCM = 1.4241 and DMSO = 1.479).
The subscript “std” refers to the standard molecule.
Fluorescence lifetime decays (τf ) were recorded using time
correlated single photon counting (TCSPC) method with
DeltaHub controller in conjunction with Horiba spectrofluorom-
eter. Data was processed with DAS6 and Origin® 8.5 software
using exponential (mono-exponential) fitting of raw data.
NanoLED (Horiba) source (1.0 MHz, pulse width <1.3 ns at
455 nm excitation wavelength) was used as an excitation source.
In this way, radiative (kr) and non-radiative (knr) constants can be
determined by knowing the fluorescence quantum yield and fluo-
rescence lifetime, as following eqn (2) and (3):25
kr ¼ ϕf =τf ð2Þ
knr ¼ ð1  ϕf Þ=τf ð3Þ
2.3. Aggregation study by UV-Vis analysis
UV-Vis absorption experiments were conducted as a function
of successive increase of porphyrin concentration (0 to 60 μM)
in DMSO solution and changes in the λmax in the 250–700 nm
range were monitored, according to the related literature.15
2.4. Stability in solution by UV-Vis assays
The stability experiments in pure dimethyl sulfoxide (DMSO)
solution and in DMSO(5%)/Tris-HCl mixture buffer ( pH 7.4) of
related porphyrin was also monitored by absorption electronic
UV–Vis measurements at several days (up to one week). All
experiments were performed in duplicate and independently.
2.5. Photostability assays and singlet oxygen quantum yield
(ΦΔ) determination
The photostability experiments in dimethyl sulfoxide (DMSO)
solution of related porphyrin was also monitored by absorp-
tion electronic UV–Vis measurements at different exposure
times (0 to 30 min) under white-light LED array system (400 at
800 nm) at fluence rate of 50 mW cm−2 and a total light
dosage of 90 J cm−2. All experiments were performed in dupli-
cate and independently.
According to typical 1,3-diphenylisobenzofuran (DPBF)
singlet oxygen quencher using in photodegradation experi-
ments, the maximum volume of 1.0 mL which contained
100 μM DPBF in DMSO was mixed with 0.5 mL (50 μM) of por-
phyrins H2TTP, PtbpyTTP and PdbpyTTP. The flask was then
filled with 2.0 mL of DMSO to a final volume of 3.5 mL. In
order to measure singlet oxygen generation, absorption UV-Vis
spectra of each solution were recorded at different exposure
times (0 to 600 s, using red-light diode laser; Thera Laser DMC
– São Paulo; potency of 100 mW) according to the literature.26
Dalton Trans., 2022, 51, 1646–1657 | 1647

Paper
The singlet oxygen production quantum yield (ΦΔ) was calcu-
lated applying eqn (4):
ΦΔ ¼ Φstd
Δ k=k
std
 I std =I ð4Þ
where I /I = (1–10 )/(1–10 ), Φstd
std Astd A
is the singlet oxygen
Δ
quantum yield of standard sample (meso-tetra( phenyl)por-
phyrin TPP in DMF, Φstd 27
Δ = 0.66), k and kstd are the photode-
gradation kinetic constants for porphyrins and TPP (standard),
respectively. Astd and A are the absorbances for TPP and por-
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.
phyrin, respectively.
2.6. Water/n-octanol partition coefficients (log POW)
The partition coefficient of porphyrins was determined using
octanol (3.0 mL) and water (3.0 mL), according to the litera-
ture.28 A mass of 0.5 mg of each porphyrin was added to the
mixture and stirred vigorously for 24 h at room temperature.
Then, the mixture was centrifuged to separate the organic
phase from the aqueous phase. The porphyrin concentrations
as well as their respective absorbances were determined by the
absorption spectrum in the UV-Vis region (250–800 nm), and
the Soret band was chosen for monitoring. The log POW value
was calculated from eqn (5), with Aorg and Aaq being the
maximum absorbances of the Soret band in the organic and
aqueous phases, and Vorg and Vaq being the final volumes of
the organic and aqueous phases, respectively.
log P OW ¼ log½ðAorg =Aaq ÞðV aq =V org Þ ð5Þ
2.7. HSA-binding assays by UV-Vis absorption analysis
UV-Vis absorption spectra for each porphyrin without and in
the presence of successive additions of HSA solution were
obtained at 298.15 K in DMSO(5%)/Tris-HCl buffer ( pH 7.4)
mixture solution, in the 250 to 700 nm range. The porphyrins
concentration was fixed in 1.0 μM and HSA concentration was
in the 0 to 60 μM range. The hyperchromicity (H%), bathochro-
mic shift (Δλ), binding constant (Kb), and Gibb’s free-energy
(ΔG°) values of the studied porphyrins were calculated accord-
ing to the literature, through Benesi–Hildebrand and free-
energy equations (eqn (6) and (7)).25,29
½HSA ½HSA 1 rometer. Data was processed with DAS6 and Origin® 8.5 soft-
¼ þ ð6Þ
jðεa  εf Þj jðεb  εf Þj Kb jðεb  εf Þj
ΔG° ¼ RT ln K b ð7Þ
where [HSA], εf, εb, and εa are the HSA concentration (in µM), the
molar absorptivity of the porphyrins in the absence and presence
of albumin, and apparent molar absorptivity of the porphyrins
(εa = Aobs/[porphyrin]) (in M−1 cm−1), respectively. For ΔG° calcu-
lation, R and T are the gas constant (1.987577 kcal K−1 mol−1)
and temperature (298.15 K), respectively.
2.8. HSA-binding assays by steady-state fluorescence
emission analysis
Additional binding parameters between HSA and each por-
phyrin (H2TTP, PtbpyTTP e PdbpyTTP) were obtained by
1648 | Dalton Trans., 2022, 51, 1646–1657
View Article Online
Dalton Transactions
steady-state fluorescence emission measurements at 298.15,
310.15, and 318.15 K in DMSO(5%)/Tris-HCl buffer ( pH 7.4) in
the 300 to 550 nm range. It is well known that fluorescence
quenching might occurs by static or dynamic processes, which
can be determined by Stern–Volmer analysis. To analyze the
data from the fluorescence quenching experiments, the Stern–
Volmer equation (8) was used:
F0
¼ 1 þ Ksv ½Q ¼ 1 þ kq τ0 ½Q ð8Þ
F
where F0 and F are the fluorescence intensities in the absence
and presence of a quencher, respectively. KSV, kq, τ0 and [Q]
denote Stern–Volmer constant, quenching rate constant, the
original lifetime of HSA (5.67 × 10−9 s)30 and the concentration
of quencher, respectively. According to eqn (8), the KSV values
were calculated from the slope and kq is equal to KSV/τ0.
In order to verify a certain trend in the association of por-
phyrins with the biomolecule, association constant (Ka) values
were obtained by modified Stern–Volmer equation (9):
F0 1 1
¼ þ ð9Þ
F0  F fKa ½Q f
where F0 and F are the steady-state fluorescence intensities of
HSA in the absence and presence of each porphyrin derivative,
respectively. Ka, f and [Q] are the modified Stern–Volmer
binding constants; the fraction of the initial fluorescence that
is accessible to the quencher ( f ≈ 1.00) and porphyrin deriva-
tive concentration, respectively.
In order to obtain quantitative information on the thermo-
dynamic parameters of the binding between HSA and each
porphyrin, van’t Hoff equation (10) was applied.31 Gibbs’ free
energy was calculated using the same eqn (7), but Ka values
instead Kb:
ΔH° ΔS°
ln Ka ¼  þ : ð10Þ
RT R
2.9. Time-resolved fluorescence decay with HSA
Fluorescence lifetime decays (τf ) were recorded using time cor-
related single photon counting (TCSPC) method with
DeltaHub controller in conjunction with Horiba spectrofluo-
ware using exponential (mono-exponential) fitting of raw data.
NanoLED (Horiba) source (1.0 MHz, pulse width <1.2 ns at
284 nm excitation wavelength) was used as an excitation
source. The HSA and porphyrins concentration were fixed in
2.0 µM each in the solution.
2.10. Molecular docking procedure
The crystallographic structure of HSA was obtained from
Protein Data Bank, with access code 1N5U.32 The chemical
structure of the synthetic porphyrins H2TTP, PtbpyTTP, and
PdbpyTTP was built and minimized in terms of energy by
Density Functional Theory (DFT), available in the Spartan’18
software (Wavefunction, Inc., Irvine, CA, USA).33 Molecular
docking calculations were performed with GOLD 2020.2 soft-
This journal is © The Royal Society of Chemistry 2022

Dalton Transactions
ware (Cambridge Crystallographic Data Centre, Cambridge,
CB2 1EZ, UK).34 Hydrogen atoms were added to the albumin
following tautomeric states and ionization data inferred by
GOLD 2020.2 software at pH 7.4. A 10 Å radius around each of
the three main binding pockets (subdomains IIA, IIIA and IB –
known as sites I, II, and III, respectively)32,35 were defined and
explored for in silico calculations. The standard ChemPLP
function was used due to the redocking studies and best
results obtained in previous work for other porphyrins.36 The
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.
figures for the best docking pose were generated with PyMOL
Delano Scientific LLC software.37
2.11. EPR analysis
Electron Paramagnetic Resonance (EPR) technique was used
for the characterization and identification of the reactive
oxygen species (ROS) generated under white-light illumination
of porphyrins H2TTP, PtbpyTTP and PdbpyTTP at concen-
tration of 57 μM, mixed with aqueous solutions of HSA at con-
centration of 114 μM.
In order to obtain which reactive oxygen species the por-
phyrins generate, EPR measurements were performed in
DMSO solutions using the spin trap methodology in a com-
mercial MiniScope MS400 spectrometer (Magnettech,
Germany) operating in X band (microwave frequency approxi-
mately equal to 9.4 GHz). The experimental parameters used
in the measurements were: 10 mW of microwave power, a
modulation field of 100 kHz with an amplitude of 0.2 mT, a
field centered on 337 mT with an application interval of
10 mT, a scan time of 60 s and 4096 spots. All EPR spectra were
measured at room temperature. The spin traps used in this
experiment were TEMP (2,2,6,6-tetramethylpiperidinol, Sigma-
Aldrich®, USA), PBN (N-tert-butyl-α-phenylnitrone, Tokyo
Chemical Industry Company, Japan) and DMPO (5,5-dimethyl-1-
Scheme 1 Synthetic pathways for the tetra-cationic thienyl–porphyrins PtbpyTTP and PdbpyTTP.
This journal is © The Royal Society of Chemistry 2022
View Article Online
Paper
pyrroline n-oxide, Oakwood Chemical®, USA). Aliquots were pre-
pared using 14 of the total volume containing a DMSO solution
with porphyrin at 57 μM, 12 of the total volume containing a
DMSO solution with the spin trap and 14 of the completed volume
with the aqueous protein solution at concentration of 114 µM or
just water. Then, the solutions were placed in glass capillaries
with a volume of 50 μL and later inserted in a quartz tube to
carry out the EPR measurements.
2.12. HSA photooxidation by steady-state fluorescence
emission analysis
The photooxidation assays of HSA were conducted by steady-
state fluorescence emission at room temperature. Stock solu-
tions of HSA (5.0 μM) was prepared in Tris-HCl buffer ( pH 7.4)
containing H2TTP, PtbpyTTP, and PdbpyTTP (at 50 μM in
DMSO) and the solutions were irradiated with white-light
source (fluence rate of 50 mW cm−2 and a total light dosage of
90 J cm−2) in a time span of 30 min. The albumin was excited
at 290 nm and the fluorescence emission intensity was
measured at 334 nm. Also, plots of ln F0/F versus time for HSA
gave a straight line from which the photodegradation rate con-
stant was calculated.
3. Results and discussion
3.1. General characterization
The free-base tetra-(2-thienyl)porphyrin H2TTP was previously
described and fully characterized by Momo and co-workers.38
The tetra-cationic porphyrin synthesis was carried out in dry
DMF with a little excess of cis-[M(bpy)Cl2] complex (M = PtII or
PdII) per thienyl group at 50 °C for 48 h and under an argon
atmosphere, according to the literature (Scheme 1).39 The
Dalton Trans., 2022, 51, 1646–1657 | 1649
 View Article Online

Paper Dalton Transactions

obtained derivatives exhibited dark-brown aspect and yields of
80–90%. These structures were characterized and confirmed
by CHN% analysis, molar conductivity, and cyclic voltammetry
(see the ESI – Fig. S1 and Tables S1 and S2†). Due to a very
high number of peaks in NMR and vibrational spectra, which
can lead to a complicated and erroneous assignment, the
NMR and FT-IR data of the derivatives PtbpyTTP and
PdbpyTTP were not presented.

3.2. Photophysical properties

The UV-Vis spectra of the porphyrins containing peripheral Pt(II)
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.

or Pd(II) complexes in DMSO solution consist of an envelope of

absorption bands in the range of 250–700 nm, arising from the
characteristic absorptions of porphyrin (Soret and Q bands) and
polypyridyl metal derivatives (π → π* and metal-to-ligand charge
transfer transitions, MLCT) entities (Fig. 1a and Table 1).
Porphyrins generally present two fluorescence emission
transitions in the excited state (Fig. 1b). The different values
for the fluorescence quantum yields (Φf ) may be a clue con-
cerning the electronic coupling between the [M(bpy)Cl]+ unit
and the porphyrin ring (Table 1), and the decrease in the fluo-
rescence can be associated with an increase of non-radiative
processes.40 Time-resolved fluorescence was measured by excit-
ing the porphyrins at 455 nm with a NanoLED. The fluo-
rescence lifetimes (τf; Table 1) were obtained through a mono-
exponential fit (I = I0 e−t/τ), and the best fits are presented in
the ESI (Fig. S2–S4†). In this way, it was observed that both
free-base porphyrins without (H2TTP) or with coordination
complexes (PtbpyTTP and PdbpyTTP) presented the lower fluo-
rescence quantum yield (2.0 to 6.0%) and the shorter fluo-
rescence lifetime (2.0 to 2.20 ns range), in agreement to the
literature.40

3.3. Photobiological parameters

Fig. 1 (a) Normalized UV-Vis absorption spectra of porphyrins H2TTP,
PtbpyTTP and PdbpyTTP, in DMSO solution and (b) normalized steady-
state fluorescence emission spectra of porphyrins H2TTP, PtbpyTTP and
PdbpyTTP, in DMSO solution, at λexc = 522 nm.
Table 2 summarizes the 1O2 generation, photostability, photo-
bleaching, and partition coefficients values. In general, both por-
phyrins did not tend to form aggregates in solution (in between
0.5 to 60 µM), being stable for a few days in DMSO solution and
remained photostable at least over a period of 30 min under
white-light LED irradiation (see ESI – Fig. S5–S13†).
For 1O2 quantum yields, the porphyrins H2TTP, PtbpyTTP,
and PdbpyTTP are capable of producing singlet oxygen species
(Table 2). Compared to the standard meso-tetra( phenyl)por-
phyrin (TPP), the presence of thienyl units and peripheral Pt

Table 1 Photophysical parameters of thienyl–porphyrins in DMSO solution

Porphyrin

H2TTP PtbpyTTP PdbpyTTP

λ, nm 426 (5.54); 522 (4.24); 560 (3.99); 280 (5.15); 326 (4.89); 425 (5.52); 522 (4.21); 560 313 (4.88); 426 (5.49); 522 (4.25); 561
(log ε)a 596 (3.82); 660 (3.63) (3.96); 597 (3.78); 660 (3.63) (4.02); 597 (3.88); 659 (3.75)
λEm, nm 667, 730 (6.0) 618, 685 (2.0) 618, 678 (3.0)
(QY, %)b
τf (ns)c 2.02 2.15 2.20
kr (107 s−1)d 2.90 0.95 1.35
knr (107 s−1)d 46.5 45.6 44.0
a
In DMSO solution. b In DMSO solution (excited at 522 nm and using TPP in DCM as standard – Φf = 0.15). c In DMF solution (using NanoLED at
455 nm). d Using eqn (2) and (3).

1650 | Dalton Trans., 2022, 51, 1646–1657 This journal is © The Royal Society of Chemistry 2022
 View Article Online

Dalton Transactions Paper

Table 2 Photobiological data of thienyl–porphyrins (II) or Pd(II) complexes can interfere in the intersystem crossing
by means of spin–orbit coupling, mainly due to the presence
Porphyrin of chemical groups with high electronic density.40 In the
H2TTP PtbpyTTP PdbpyTTP photobleaching study, it is possible to notice that the deriva-
tives PtbpyTTP and PdbpyTTP are less susceptible to photooxi-
ΦΔa 0.33 0.40 0.38
kpo (M−1 s−1)b 0.0014 0.0017 0.0016 dative processes, a fact that is reflected in the ΦPB values (see
kpb (s−1)c 0.87 0.78 0.84 Table 2 and ESI – Fig. S14–S16†). In addition, the log POW was
ΦPB (%)d 17.0 2.45 2.30 measured and the values found for the cationic derivatives
log POWe +2.95 +1.56 +1.51
containing the Pt(II) and Pd(II) polypyridyl complexes are in
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.

a
Singlet oxygen generation, in DMSO solution (using TPP in DMF as accordance with the literature,18,28 with neutral tetra-thienyl
standard – Φf = 0.66). b DPBF photooxidation constant. derivative H2TTP showing a more hydrophobic character when
c
Photobleaching constant determined by photostability assays (white-
LED with fluence rate of 50 mW cm−2 and a total light dosage of 90 J compare to the tetra-cationic derivatives PtbpyTTP and
cm−2 for 30 min). d Photobleaching quantum yield determined by PdbpyTTP (Table 2).
photostability assays (white-LED with fluence rate of 50 mW cm−2 and
a total light dosage of 90 J cm−2 for 30 min). e Partition coefficients in
octanol/water. 3.4. HSA-binding study
The porphyrins under study were also evaluated according to
their interaction with albumin (HSA), due to the importance of
this protein in transporting drugs in the human
bloodstream.30,31 The interactive parameters are presented in
Table 3 The HSA-binding parameters for the interactions with thienyl–
porphyrins in DMSO(5%)/Tris-HCl pH 7.4 buffer mixture solution by Table 3 and the entire set of spectra are listed in the ESI
UV-Vis absorption analysis (Fig. S17 and S18†).
Firstly, by UV-Vis analysis, there is a hyperchromic profile
Porphyrin of the spectra with no significant bathochromic shift. The
H2TTP PtbpyTTP PdbpyTTP
intrinsic binding constant (Kb ≈ 103 M−1) and Gibb’s free-
energy (ΔG° < 0) values indicated that the porphyrins interact
a
H (%) 41.0 35.5 58.5 weakly to moderately and spontaneously with albumin, mainly
Δλ (nm)b 0.0 0.0 0.0
Kb (×103 M−1)c 1.57 ± 0.09 0.63 ± 0.02 1.62 ± 0.07 by non-covalent forces, probably by van der Waals and
ΔG° (kJ mol−1)d −18.20 −15.95 −18.30 H-bonding (Table 3). As an example, the UV-Vis HSA titration
of compound PtbpyTTP is presented in the Fig. 2.
a
Hyperchromicity (H%) = Afinal − Ainitial/Ainitial × 100. b Red shift.
c
Intrinsic binding constant determined by Benesi–Hidelbrand The additional binding parameters obtained by steady-state
equation (6). d Gibb’s free-energy determined by eqn (7). fluorescence emission corroborated with the trend described

Fig. 2 UV–Vis absorption spectra of porphyrin PtbpyTTP with increase HSA concentrations (0 to 60 µM), in a DMSO(5%)/Tris-HCl buffer ( pH 7.4)
solution. The insert shows the plot of [HSA]/(εa − εf ) versus [HSA].

This journal is © The Royal Society of Chemistry 2022 Dalton Trans., 2022, 51, 1646–1657 | 1651
 View Article Online

Paper Dalton Transactions

Table 4 The HSA-binding parameters for the interactions with porphyrins H2TTP, PtbpyTTP, and PdbpyTTP, in DMSO(5%)/Tris-HCl pH 7.4 buffer
mixture solution by fluorescence emission analysis

Porphyrin T (K) Qa (%) KSV b (×104 M−1) kq c (×1012 M−1 s−1) Ka d (×103 M−1) ΔH° e (kJ mol−1) ΔS° e (kJ mol−1 K−1) ΔG° f (kJ mol−1)

H2TTP 298.15 51.30 1.69 ± 0.02 2.98 ± 0.35 4.46 ± 0.13 −20.80 ± 0.39 +0.070 ± 0.01 −20.85
310.15 1.40 ± 0.02 2.47 ± 0.35 4.38 ± 0.17 −21.60 ± 0.39 +0.069 ± 0.01 −21.65
318.15 1.36 ± 0.01 2.40 ± 0.17 4.27 ± 0.18 −22.10 ± 0.39 +0.069 ± 0.01 −22.15
PtbpyTPP 298.15 80.30 4.62 ± 0.04 8.15 ± 0.70 6.66 ± 0.25 −22.70 ± 0.70 +0.073 ± 0.02 −21.85
310.15 5.73 ± 0.06 10.10 ± 0.10 5.88 ± 0.14 −22.40 ± 0.70 +0.072 ± 0.02 −22.40
318.15 6.50 ± 0.07 11.5 ± 0.12 5.58 ± 0.13 −22.25 ± 0.70 +0.072 ± 0.02 −22.85
−21.70 ± 0.12 −20.40
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.

PdbpyTTP 298.15 63.50 1.66 ± 0.02 2.93 ± 0.35 3.67 ± 0.18 +0.068 ± 0.04
310.15 2.32 ± 0.03 4.09 ± 0.53 6.35 ± 0.21 −23.15 ± 0.12 +0.073 ± 0.04 −22.60
318.15 3.25 ± 0.05 5.73 ± 0.88 5.78 ± 0.20 −22.90 ± 0.12 +0.072 ± 0.04 −22.95
a
Quenching (Q%) = F − F0/F0 × 100. b Stern–Volmer quenching constant calculated by eqn (8). c Bimolecular quenching rate constant calculated
by eqn (8). d Modified Stern–Volmer binding constant calculated by eqn (9). e Enthalpy and entropy changes by Van’t Hoff plot calculated by
eqn (10). f Gibbs free-energy values calculated by eqn (7).

above, as well as indicated that the interaction is enthalpically
and entropically driven (Table 4). A ground-state association
(static fluorescence quenching mechanism) was also detected
due to the bimolecular quenching rate kq values are about
three orders of magnitude higher than the diffusion rate con-
stant in water (kdiff ≈ 7.40 × 109 M−1 s−1, at 305 K according to
Smoluchowski–Stokes–Einstein).41 For the metalloporphyrins
the increase in the KSV values with the increasing of tempera-
ture, suggest the possibility to also occur a dynamic process,
probably due to the high steric volume of these compounds.
All results are listed in the Table 4, as an example, the steady-
state fluorescence emission spectra and modified Stern–
Volmer plots for PtbpyTTP porphyrin at three different temp-
eratures are represented in the Fig. 3. All selected spectra and
graphs are listed in the ESI file (Fig. S19–S31†).
In order to identify the main fluorescence quenching
mechanism (static or dynamic), time-resolved fluorescence
decays were obtained for HSA without and in the presence
of H2TTP, PtbpyTTP, and PdbpyTTP in DMSO(5%)/Tris-HCl
pH 7.4 buffer mixture solution and lifetime values are pre-
sented in the Table 5. There is a significant decrease in the
τf value of HSA upon porphyrin addition, indicating that at
high porphyrin concentration a combined static and
dynamic fluorescence quenching mechanism is feasible for
the interaction HSA:H2TTP/PtbpyTTP/PdbpyTTP. All fluo-
rescence decays plots are listed in the ESI (Fig. S32–S35†).
Finally, the number of binding sites (n), calculated by
double-logarithmic equation,25 are between 0.85 and 1.45,
indicating that the proportion HSA : porphyrin is mainly
1 : 1, thus one porphyrin interact with only one single
albumin compound.

Table 5 Time-resolved fluorescence parameters and molecular

docking score (dimensionless) values for the interaction
HSA : porphyrins in DMSO(5%)/Tris-HCl buffer ( pH 7.4) solution

Time-resolved
fluorescence Molecular docking
Fig. 3 (a) Steady-state fluorescence emission spectra for HSA without
and upon successive additions of PtbpyTTP, in a DMSO(5%)/Tris-HCl Compound τf (ns) χ2 Site I Site II Site III
buffer ( pH 7.4) solution at 298.15 K. The concentration of compounds HSA 5.06 ± 0.001 1.04780 — — —
ranged from 0 to 60 μM. Insert graph shows the plot of F0/F versus [ por- HSA:H2TTP 3.89 ± 0.002 1.04934 53.2 4.09 80.9
phyrin] and (b) modified Stern–Volmer plots for HSA:PtbpyTTP adduct HSA:PtbpyTTP 3.74 ± 0.009 1.01757 36.2 1.27 66.8
at three different temperatures. HSA:PdbpyTTP 3.71 ± 0.001 1.04731 42.1 1.35 67.1

1652 | Dalton Trans., 2022, 51, 1646–1657 This journal is © The Royal Society of Chemistry 2022
 View Article Online

Dalton Transactions Paper

To offer an atomic-level explanation on the binding capacity
of each porphyrin into HSA pocket, in silico studies via mole-
cular docking calculations were performed (Fig. 4). In this
way, Table 6 shows the docking score value (dimensionless),
suggesting that the main binding site is subdomain IB (an
external binding region), probably due to the high molecular
surface area of the porphyrins, which impacts the fit capacity
of the compound. Additionally, hydrogen bonding and van der
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.
Waals interactions were detected as the main intermolecular
forces responsible for the association HSA : porphyrins, being
in agreement with the experimental data.
3.5. EPR analysis
ROS generation was identified and quantified via EPR tech-
nique by spin trap methodology for a system upon white-light
illumination (see Experimental section). To evaluate the gene-

Fig. 4 Best docking poses for the interaction (a) HSA:H2TTP, (b) HSA:PtbpyTTP and (c) HSA:PdbpyTTP, into site III. Selected amino acid residue,
H2TTP, PdbpyTTP and PdbpyTTP are in stick representation in cyan, purple, brown and beige, respectively. Elements’ color: hydrogen: white;
oxygen: red; nitrogen: dark blue, sulfur: yellow, chloro: green, Pt(II): gray and Pd(II): dark green.

This journal is © The Royal Society of Chemistry 2022 Dalton Trans., 2022, 51, 1646–1657 | 1653
 View Article Online

Paper Dalton Transactions

Table 6 Molecular docking results for the interaction between HSA: the kinetics of spin adduct generation, it can be noted that all
H2TTP, HSA:PtbpyTTP, and HSA:PdbpyTTP in the site III porphyrins show similar behavior, with an accentuated pro-
duction of singlet oxygen in the first minutes of irradiation
Code Amino acid residues Interaction Distance (Å)
(<5 min). After this initial irradiation time, the radical concen-
H2TTP Arg-117 Hydrogen bonding 3.20 tration slowly decreased, due to the degradation of the spin
Phe-134 van der Waals 1.70
adducts caused by the high concentration of 1O2 generated
Tyr-138 van der Waals 2.90
Ile-142 van der Waals 3.20 during the photo-induced process.22,42 In the presence of the
Phe-149 van der Waals 2.40 HSA protein, it is possible to observe that the production of
Leu-154 van der Waals 3.50
the spin adducts is reduced in relation to the HSA-free por-
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.

Phe-157 van der Waals 3.50

Ala-158 van der Waals 2.00 phyrins (Fig. 5). According to the observed results, it is poss-
Tyr-161 Hydrogen bonding 2.40 ible to claim that the derivatives generate 1O2 and can promote
Phe-165 van der Waals 2.10
oxidation in the HSA protein (Table 7).
Arg-186 van der Waals 2.70
PtbpyTTP Arg-114 van der Waals 3.40 To identify the ROS formed by type I mechanism, the PBN
Arg-117 van der Waals 2.30 spin trap (0.1 M) was used and the analysis revealed the for-
Tyr-138 van der Waals 1.70
mation of a methyl radical (•CH3, aN = 1.49 mT and aH(β) =
Ile-142 van der Waals 1.40
His-146 van der Waals 2.90 0.25 mT) (see ESI – Fig. S36†). This radical is characteristic of
Phe-157 van der Waals 3.60 the interaction of the hydroxyl radical (•OH) with the DMSO
Tyr-161 Hydrogen bonding 3.00
molecule.43–45 However, the concentration of •OH is much less
Arg-186 van der Waals 1.50
Lys-190 Hydrogen bonding 2.00 in relation to the formation of 1O2, expected behavior for this
PdbpyTTP Arg-114 van der Waals 3.60 type of photosensitizer, where the dominant ROS formation
Leu-115 van der Waals 3.20
mechanism is due to type II process (Fig. 6). All porphyrins
Arg-117 van der Waals 3.50
Tyr-138 van der Waals 3.00 showed the formation of the same adducts, with the difference
Ile-142 van der Waals 3.00 that the presence of the HSA protein contributes to a slight
His-146 van der Waals 3.50
increase in the concentration of PBN/•CH3 formed during
Tyr-161 Hydrogen bonding 2.20
Arg-186 Hydrogen bonding 3.50 white-light illumination. Even so, it is reckless to conclude
Lys-190 Hydrogen bonding 2.00 that protein binding improves the generation of ROS by the
type I mechanism.
Complementarily, the DMPO spin trap (0.3 M) was used
ration of 1O2 species, the hydroxy-TEMP spin trap in water (1.0 and added the formation of the superoxide radical (O2•−, aN =
M) was used. The concentration of spin adducts generated 1.33 mT, aH(β) = 1.04 mT and aH(γ) = 0.14 mT) indicating elec-
upon white-light illumination was determined by double inte- tron transfer between the porphyrins and molecular oxygen
gration of the EPR signal (Fig. 5) and compared with the signal present in the medium.46–49 All porphyrins, with or without
intensity of aqueous TEMPOL solution (1.0 mM). Considering HSA protein, exhibited a similar line shape in the EPR spectra
(see ESI – Fig. S37 and S38†).
Finally, Fig. 7 depicts the steady-state fluorescence emission
quenching of HSA in the presence of the studied tetra-cationic
porphyrins upon white-light irradiation conditions for 30 min
(HSA photooxidation assays) used as example compound
PtbpyTTP. The data show that under irradiation, tetra-cationic
porphyrins PtbpyTTP and PdbpyTTP present a similar behav-
ior in the photodegradation constants (kpd) (Table 8). All fluo-
rescence emission spectra are presented in the ESI – Fig. S39
and S40.†

Table 7 Spin adducts generation reaction constant formed by singlet

oxygen (1O2) capture by TEMP generated in the white-light irradiation
conditions using H2TTP, PtbpyTTP, and PdbpyTTP, without and in the
presence of HSA

Porphyrin

H2TTP PtbpyTTP PdbpyTTP

Fig. 5 Concentration of spin adduct formed by singlet oxygen (1O2) Without HSA −1
ks (min ) 0 to 5 min 11.0 10.0 13.0
capture by TEMP as a function of irradiation time for H2TTP, PtbpyTTP and ks (min−1) 0 to 2 min 13.0 12.0 16.0
PdbpyTTP porphyrins, in the absence (solid black, blue and red lines) and With HSA ks (min−1) 0 to 5 min 6.0 7.0 7.0
in the presence of HSA (dashed orange, pink and light-blue lines). ks (min−1) 0 to 2 min 10.0 13.0 11.0

1654 | Dalton Trans., 2022, 51, 1646–1657 This journal is © The Royal Society of Chemistry 2022
 View Article Online

Dalton Transactions Paper

Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.

Fig. 6 Comparison of concentrations of PBN/•CH, PBN/•CH3 and PBN* spin adducts, formed by irradiating porphyrins H2TTP, PtbpyTTP and
PdbpyTTP dissolved in DMSO. In (a) a comparison of the concentration of spin adducts formed for porphyrins without HSA protein is presented. In
the other panels, comparisons for porphyrins (b) H2TTP, (c) PtbpyTTP and (d) PdbpyTTP as a function of the presence or absence of HSA albumin
are presented.

Table 8 Photooxidation rate constants in the white-light irradiation

conditions using H2TTP, PtbpyTTP, and PdbpyTTP, in the presence of
HSA, by steady-state fluorescence emission analysis

Porphyrin

H2TTP PtbpyTTP PdbpyTTP

a
Q (%) 28.6 35.6 34.2
kpd (min−1)b 9.30 16.0 15.0
a
Quenching = F − F0/F0 × 100%. b First-order kinetics profile.

Fig. 7 HSA photooxidation assays (DMSO(5%)/Tris-HCl buffer pH = 7.4
at 298.15 K) in the presence of porphyrin PtbpyTTP using an excitation
wavelength of 290 nm, under white-light irradiation conditions. Inset
plot: first-order kinetic graphical profile.
4. Conclusions
In summary, the results indicated that meso-tetra-cationic(2-
thienyl)porphyrins PtbpyTTP and PdbpyTTP are interesting
scaffolds for the design of novel photosensitizer, presenting
relevant properties for future application as PDT and aPDT
agents. The insertion of the peripheral-coordinated Pt(II) or
Pd(II) complexes resulted in increased interaction with HSA
when compared to the non-cationic porphyrin H2TTP. These
novel meso-tetra-cationic(2-thyenil)porphyrin derivatives inter-
act weakly to moderately and spontaneously with human

This journal is © The Royal Society of Chemistry 2022 Dalton Trans., 2022, 51, 1646–1657 | 1655

Paper
serum albumin, mainly by non-covalent forces, probably by
van der Waals and H-bonding forces, showing good binding
parameters to be feasible their biodistribution in the human
bloodstream. Additionally, there is a great capacity of these
porphyrins to generate ROS upon white-light irradiation con-
ditions, including protein photodamage. Overall, porphyrins
PtbpyTTP and PdbpyTTP can be considered as potential and
promising candidates as bio-supramolecular compounds in
the photobiological applications.
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.
Author contributions
K. T. Oliveira and B. A. Iglesias idealized the work. B. A.
Iglesias conducted the characterization, photophysical and
photobiological assays. H. F. V. Victória, J. R. de Toledo and
K. Krambrock conducted the EPR experiments. O. A. Chaves
conducted the molecular docking calculations. K. T. Oliveira,
B. A. Iglesias, O. A. Chaves and K. Krambrock wrote and
correct the manuscript.
Conflicts of interest
The authors declare that they do not have conflict of interest.
Acknowledgements
This research was supported by the Conselho Nacional de
Desenvolvimento Científico e Tecnológico (CNPq/grants
409150/2018–5 and 304711/2018–7), Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior (CAPES –
Finance Code 001), Fundação de Amparo à Pesquisa do Estado
de São Paulo (FAPESP/grant 2013/07276-1, 2019/27176-8 and
2020/06874-6) and Fundação de Amparo à Pesquisa do Estado de
Minas Gerais (FAPEMIG). O. A. C. thanks Fundação para a
Ciência e a Tecnologia (FCT – Portuguese Foundation for Science
and Technology) for the PhD fellowship 2020.07504.BD.
References
1 K. Lang, J. Mosinger and D. M. Wagnerová, Coord. Chem.
Rev., 2004, 248, 321–350.
2 F. Ricchelli, J. Photochem. Photobiol., B, 1995, 29, 109–118.
3 J. M. Dąbrowski, Adv. Inorg. Chem., 2017, 70, 343–394.
4 B. Halliwell and J. M. C. Gutteridge, Free Radicals in Biology
and Medicine, Oxford Science, Oxford, 3rd edn, 1998, 1–35.
5 W. E. Savive, Aust. J. Chem., 1971, 24, 1285–1293.
6 I. E. Borissevitch, T. T. Tominaga and C. C. Schmitt,
J. Photochem. Photobiol., A, 1998, 114, 201–207.
7 N. S. Lebedeva, E. A. Malkova, T. E. Popova, A. E. Kutyrev,
S. A. Syrbu, E. V. Parfenyuk and A. I. Vyugin, Spectrochim.
Acta, Part A, 2014, 118, 395–398.
8 T. H. O. Leite, G. Grawe, J. Honorato, B. N. Cunha,
O. R. Nascimento, P. S. de Vargas, C. Donatoni,
1656 | Dalton Trans., 2022, 51, 1646–1657
View Article Online
Dalton Transactions
K. T. Oliveira, J. M. S. Lopes, N. M. B. Neto, W. C. Moreira,
L. R. Dinelli and A. A. Batista, Inorg. Chem., 2019, 58, 1030–
1039.
9 V. A. Oliveira, B. A. Iglesias, B. L. Auras, A. Neves and
H. Terenzi, Dalton Trans., 2017, 46, 1660–1669.
10 G. Basso, J. F. Cargnelutti, A. L. Oliveira, T. V. Acunha,
R. Weiblen, E. F. Flores and B. A. Iglesias, J. Porphyrins
Phthalocyanines, 2019, 23, 1041–1046.
11 G. K. Couto, B. S. Pacheco, V. M. Borba, J. C. R. Junior,
T. L. Oliveira, N. V. Segatto, F. K. Seixas, T. V. Acunha,
B. A. Iglesias and T. Collares, J. Photochem. Photobiol., B,
2020, 202, 111725–111737.
12 T. T. Tasso, T. M. Tsubone, M. S. Baptista, L. M. Mattiazzi,
T. V. Acunha and B. A. Iglesias, Dalton Trans., 2017, 46,
11037–11045.
13 A. Naik, R. Rubbiani, G. Gasser and B. Spingler, Angew.
Chem., Int. Ed., 2014, 53, 6938–6941.
14 L. Schneider, M. Kalt, M. Larocca, V. Babu and B. Spingler,
Inorg. Chem., 2021, 60, 9416–9426.
15 C. H. da Silveira, V. Vieceli, D. J. Clerici, R. C. V. Santos and
B. A. Iglesias, Photodiagn. Photodyn. Ther., 2020, 31,
101920.
16 G. G. Rossi, K. B. Guterres, C. H. da Silveira, K. S. Moreira,
T. A. L. Burgo, B. A. Iglesias and M. M. A. de Campos,
Microb. Pathog., 2020, 148, 104455.
17 D. C. Jornada, R. Q. Garcia, C. H. da Silveira, L. Misoguti,
C. R. Mendonça, R. C. V. Santos, L. De Boni and
B. A. Iglesias, Photodiagn. Photodyn. Ther., 2021, 35,
102459.
18 S. C. Pinto, T. V. Acunha, J. M. Santurio, L. B. Denardi and
B. A. Iglesias, Photodiagn. Photodyn. Ther., 2021, 36,
102550.
19 T. Peters, All About Albumin: Biochemistry, Genetics, and
Medical Applications, Academic Press, San Diego, CA, 1996,
pp. 79–131.
20 P. Ascenzi and M. Fasano, IUBMB Life, 2009, 61, 1118–1122.
21 O. A. Chaves, L. B. Menezes and B. A. Iglesias, J. Mol. Liq.,
2019, 294, 111581.
22 F. S. Santos, C. H. da Silveira, F. S. Nunes, D. C. Ferreira,
H. F. V. Victória, K. Krambrock, O. A. Chaves,
F. S. Rodembusch and B. A. Iglesias, Dalton Trans., 2020,
49, 16278–16295.
23 T. Chatterjee, A. Pal, S. Dey, B. K. Chatterjee and
P. Chakrabarti, PLoS One, 2012, 7, e37468.
24 (a) P. Foletto, F. Correa, L. Dornelles, B. A. Iglesias, C. H. da
Silveira, P. A. Nogara, J. B. T. da Rocha, M. A. F. Faustino
and O. E. D. Rodrigues, Molecules, 2018, 23, 2588;
(b) L. H. Z. Cocca, T. M. A. Oliveira, F. Gotardo, A. V. Teles,
R. Menegatti, J. P. Siqueira, C. R. Mendonça,
L. A. M. Bataus, A. O. Ribeiro, T. F. M. Souza,
G. R. L. Souza, P. J. Gonçalves and L. De Boni, J. Photochem.
Photobiol., B, 2017, 175, 1–8; (c) R. N. Sampaio,
W. R. Gomes, D. M. S. Araujo, A. E. H. Machado,
R. A. Silva, A. Marletta, I. E. Borissevitch, A. S. Ito,
L. R. Dinelli, A. A. Batista, S. C. Zílio, P. J. Gonçalves and
N. M. Barbosa Neto, J. Phys. Chem. A, 2012, 116, 18–26.
This journal is © The Royal Society of Chemistry 2022

Dalton Transactions
25 T. V. Acunha, B. M. Rodrigues, J. A. da Silva,
D. D. M. Galindo, O. A. Chaves, V. N. da Rocha,
P. C. Piquini, M. H. Köhler, L. De Boni and B. A. Iglesias,
J. Mol. Liq., 2021, 340, 117223.
26 R. C. Pivetta, B. L. Auras, B. de Souza, A. Neves, F. S. Nunes,
L. H. Z. Cocca, L. D. Boni and B. A. Iglesias, J. Photochem.
Photobiol., A, 2017, 332, 306–315.
27 M. Pineiro, A. L. Carvalho, M. M. Pereira, A. M. D. A. Rocha
Gonsalves and L. Arnaut, Chem. – Eur. J., 1998, 4, 2299–
Published on 03 January 2022. Downloaded by Universidad de Granada on 8/22/2023 9:44:23 PM.
2307.
28 F. M. Engelmann, S. V. O. Rocha, H. E. Toma, K. Araki and
M. S. Baptista, Int. J. Pharm., 2017, 329, 12–18.
29 H. A. Benesi and J. H. Hildebrand, J. Am. Chem. Soc., 1949,
71, 2703–2707.
30 O. A. Chaves, T. V. Acunha, B. A. Iglesias, C. S. H. Jesus and
C. Serpa, J. Mol. Liq., 2020, 301, 112466.
31 O. A. Chaves, L. B. Menezes and B. A. Iglesias, J. Mol. Liq.,
2019, 294, 111581.
32 M. Wardell, Z. Wang, J. X. Ho, J. Robert, F. Rucker, J. Ruble
and D. C. Carter, Biochem. Biophys. Res. Commun., 2002,
291, 813–819.
33 W. J. Hehre, A guide to molecular mechanics and quantum
chemical calculations, Wavefunction, INC., Irvine, USA, 2003.
34 http://www.ccdc.cam.ac.uk/solutions/csd-discovery/com-
ponents/gold/, accessed in October 2021.
35 O. A. Chaves, M. R. L. Santos, M. C. C. de Oliveira,
C. M. R. Sant’Anna, R. C. Ferreira, A. Echevarria and
J. C. Netto-Ferreira, J. Mol. Liq., 2018, 254, 280–290.
36 V. Viecelli, O. A. Chaves, K. Araki, P. R. Martins and
B. A. Iglesias, J. Braz. Chem. Soc., 2020, 31, 2282–2298.
This journal is © The Royal Society of Chemistry 2022
View Article Online
Paper
37 W. L. DeLano, PyMOL User’s Guide, DeLano Scientific LLC,
San Carlos, CA, USA, 2002.
38 P. B. Momo, C. Pavani, M. S. Baptista, T. J. Brocksom and
K. T. de Oliveira, Eur. J. Org. Chem., 2014, 4536–4547.
39 J. A. Naue, S. H. Toma, J. A. Bonacin, K. Araki and
H. E. Toma, J. Inorg. Biochem., 2009, 103, 182–189.
40 L. H. Z. Cocca, F. Gotardo, L. F. Sciuti, T. V. Acunha,
B. A. Iglesias and L. De Boni, Chem. Phys. Lett., 2018, 708,
1–10.
41 M. Montalti, A. Credi, L. Prodi and M. T. Gandolfi,
Handbook of Photochemistry, 3rd edn, CRC Press, Taylor &
Francis, 2006.
42 G. A. Sáfar, R. N. Gontijo, C. Fantini, D. C. Martins,
Y. M. Idemori, M. V. Pinheiro and K. Krambrock, J. Phys.
Chem. C, 2015, 119, 4344–4350.
43 M. G. Steiner and C. F. Babbs, Arch. Biochem. Biophys.,
1990, 278, 478–481.
44 K. Hensley, M. Aksenova, J. M. Carney, M. Harris and
D. A. Butterfield, Amyloid, 8-peptide spin trapping II: evi-
dence for NeuroReport, 1995, vol. 6, pp. 493–496.
45 A. N. Saprin and L. H. Piette, Arch. Biochem. Biophys., 1977,
180, 480–492.
46 A. E. Alegria and P. Riesz, Photochem. Photobiol., 1988, 48,
147–152.
47 M. M. Mossoba and P. L. Gutierrez, Biochem. Biophys. Res.
Commun., 1985, 132, 445–452.
48 P. Bilski, K. Reszka, M. Bilska and C. F. Chignell, J. Am.
Chem. Soc., 1996, 118, 1330–1338.
49 J. A. Silvester, G. S. Timmins and M. J. Davies, Free Radicals
Biol. Med., 1998, 24, 754–766.
Dalton Trans., 2022, 51, 1646–1657 | 1657