Novel Short PEG Chain-Substituted Porphyrins: Synthesis, Photochemistry, and In Vitro Photodynamic Activity against Cancer Cells.

This work presents the synthesis and characterization of metal-free, zinc (II), and cobalt (II) porphyrins substituted with short PEG chains. The synthesized compounds were characterized by UV-Vis, 1H and 13C NMR spectroscopy, and MALDI-TOF mass spectrometry. The origin of the absorption bands for tested compounds in the UV-Vis range was determined using a computational model based on the electron density functional theory (DFT) and its time-dependent variant (TD-DFT). The photosensitizing activity was evaluated by measuring the ability to generate singlet oxygen (ΦΔ), which reached values up to 0.54. The photodynamic activity was tested using bladder (5637), prostate (LNCaP), and melanoma (A375) cancer cell lines. In vitro experiments clearly showed the structure-activity relationship regarding types of substituents, their positions in the phenyl ring, and the variety of central metal ions on the porphyrin core. Notably, the metal-free derivative 3 and its zinc derivative 6 exerted strong cytotoxic activity toward 5637 cells, with IC50 values of 8 and 15 nM, respectively. None of the tested compounds induced a cytotoxic effect without irradiation. In conclusion, these results highlight the potential value of the tested compounds for PDT application.

Photoinduced Skeletal Rearrangement of N-Substituted Colchicine Derivatives.

Colchicine is an active pharmaceutical ingredient widely used for treating gout, pericarditis, and familial Mediterranean fever with high antimitotic activity. The photoisomerization of colchicine deactivates its anti-inflammatory and antimitotic properties. However, despite numerous reports on colchicine derivatives, their photostability has not been investigated in detail. This report reveals the effects of UV-induced rearrangement on the structure and reports the biological activity of new N-substituted colchicine derivatives.

In-Depth Studies of Ground- and Excited-State Properties of Re(I) Carbonyl Complexes Bearing 2,2':6',2″-Terpyridine and 2,6-Bis(pyrazin-2-yl)pyridine Coupled with π-Conjugated Aryl Chromophores.

In the current work, comprehensive photophysical and electrochemical studies were performed for eight rhenium(I) complexes incorporating 2,2':6',2″-terpyridine (terpy) and 2,6-bis(pyrazin-2-yl)pyridine (dppy) with appended 1-naphthyl-, 2-naphthyl-, 9-phenanthrenyl, and 1-pyrenyl groups. Naphthyl and phenanthrenyl substituents marginally affected the energy of the MLCT absorption and emission bands, signaling a weak electronic coupling of the appended aryl group with the Re(I) center. The triplet MLCT state in these complexes is so low lying relative to the triplet 3ILaryl that the thermal population of the triplet excited state delocalized on the organic chromophore is ineffective. The attachment of the electron-rich pyrenyl group resulted in a noticeable red shift and a significant increase in molar absorption coefficients of the lowest energy absorption of the resulting Re(I) complexes due to the contribution of intraligand charge-transfer (ILCT) transitions occurring from the pyrenyl substituent to the terpy/dppy core. At 77 K, the excited states of [ReCl(CO)3(Ln-κ2N)] with 1-pyrenyl-functionalized ligands were found to have predominant 3ILpyrene/3ILCTpyrene→terpy character. The 3IL/3ILCT nature of the lowest energy excited state of [ReCl(CO)3(4'-(1-pyrenyl)-terpy-κ2N)] was also evidenced by nanosecond transient absorption and time-resolved emission spectroscopy. Enhanced room-temperature emission lifetimes of the complexes [ReCl(CO)3(Ln-κ2N)] with 1-pyrenyl-substituted ligands are indicative of the thermal activation between 3MLCT and 3IL/3ILCT excited states. Deactivation pathways occurring upon light excitation in [ReCl(CO)3(4'-(1-naphthyl)-terpy-κ2N)] and [ReCl(CO)3(4'-(1-pyrenyl)-terpy-κ2N)] were determined by femtosecond transient absorption studies.

Radiation- and Photo-Induced Oxidation Pathways of Methionine in Model Peptide Backbone under Anoxic Conditions.

Within the reactive oxygen species (ROS) generated by cellular metabolisms, hydroxyl radicals (HO•) play an important role, being the most aggressive towards biomolecules. The reactions of HO• with methionine residues (Met) in peptides and proteins have been intensively studied, but some fundamental aspects remain unsolved. In the present study we examined the biomimetic model made of Ac-Met-OMe, as the simplest model peptide backbone, and of HO• generated by ionizing radiation in aqueous solutions under anoxic conditions. We performed the identification and quantification of transient species by pulse radiolysis and of final products by LC-MS and high-resolution MS/MS after γ-radiolysis. By parallel photochemical experiments, using 3-carboxybenzophenone (CB) triplet with the model peptide, we compared the outcomes in terms of short-lived intermediates and stable product identification. The result is a detailed mechanistic scheme of Met oxidation by HO•, and by CB triplets allowed for assigning transient species to the pathways of products formation.

Near-Infrared Photoactive Aza-BODIPY: Thermally Robust and Photostable Photosensitizer and Efficient Electron Donor.

We report herein the synthesis of aza-BODIPY substituted with strongly electron-donating p-(diphenylamino)phenyl substituents (p-Ph2 N-) at 3,5-positions. The presence of p-Ph2 N- groups lowers the energy of the singlet excited state (Es ) to 1.48 eV and induces NIR absorption with λabs at 789 nm in THF. The compound studied is weakly emissive with the emission band (λf ) at 837 nm and with the singlet lifetime (τS ) equal to 100 ps. Nanosecond laser photolysis experiments of the aza-BODIPY in question revealed T1 →Tn absorption spanning from ca. 350-550 nm with the triplet lifetime (τT ) equal to 21 µs. By introducing a heavy atom (Br) into the structure of the aza-BODIPY, we managed to turn it into a NIR operating photosensitizer. The photosensitized oxygenation of the model compound-diphenylisobenzofuran (DPBF)-proceedes via Type I and/or Type III mechanism without formation of singlet oxygen (1 O2 ). As estimated by CV/DPV measurements, the p-Ph2 N- substituted aza-BODIPYs studied exhibits oxidation processes at relatively low oxidation potentials (Eox1 ), pointing to the very good electron-donating properties of these molecules. Extremely high photostability and thermal robustness up to approximately 300 °C are observed for the p-Ph2 N- substituted aza-BODIPYs.

Unexpected Reaction Pathway of the Alpha-Aminoalkyl Radical Derived from One-Electron Oxidation of S-Alkylglutathiones.

Laser flash photolysis and high-resolution mass spectrometry were used to investigate the mechanism of one-electron oxidation of two S-alkylglutathiones using 3-carboxybenzophenone (3CB) as a photosensitizer. This report indicates an unexpected reaction pathway of the α-aminoalkyl radical cation (αN+) derived from the oxidation of S-alkylglutathiones. Instead of a common hydrolysis reaction of αN+ reported earlier for methionine and other sulfur-containing aminoacids and peptides, an intramolecular ring-closure reaction was found for S-alkylglutathiones.

Hydrogen Bond-Mediated Conjugates Involving Lanthanide Diphthalocyanines and Trifluoroacetic Acid (Lnpc2@TFA): Structure, Photoactivity, and Stability.

The interaction between lanthanide diphthalocyanine complexes, LnPc2 (Ln = Nd, Sm, Eu, Gd, Yb, Lu; Pc = C32H16N8, phthalocyanine ligand) and trifluoroacetic acid (TFA) was investigated in benzene, and the stability of the resulting molecular system was assessed based on spectral (UV-Vis) and kinetic measurements. Structural Density Functional Theory (DFT) calculations provided interesting data regarding the nature of the bonding and allowed estimating the interaction energy between the LnPc2 and TFA species. Conjugates are created between the LnPc2 and TFA molecules via hydrogen bonds of moderate strength (>N∙∙H··) at the meso- -bridges of the Pc moieties, which renders the sandwich system to flatten. Attachment of TFA is followed by rearrangement of electronic density within the chromophore system of the macrocycles manifested in considerable changes in their UV-Vis spectra and consequently the color of the studied solutions (from green to orange). The LnPc2@TFA conjugates including Nd, Sm, Eu, and Gd appeared evidently less photostable when exposed to UV radiation than the related mother compounds, whereas in the case of Yb and Lu derivatives some TFA-prompted stabilizing effect was noticed. The conjugates displayed the capacity for singlet oxygen generation in contrast to the LnPc2s itself. Photon upconversion through sensitized triplet-triplet annihilation was demonstrated by the TFA conjugates of Nd, Sm, Eu, and Gd.

Synthesis, Photophysics and Redox Properties of Aza-BODIPY Dyes with Electron-Donating Groups.

A series of novel aza-BODIPY dyes substituted with p-(dimethylamino)phenyl groups were synthesized and their spectral and electrochemical properties were compared. In particular, the impact of p-(Me2 N)Ph- groups on these characteristics was of consideration. For two aza-BODIPYs studied, a near-IR absorption band was observed at circa λabs =796 nm. Due to the pronounced intramolecular charge transfer (ICT) exerted by the presence of strongly electron-donating p-(Me2 N)Ph- substituents, the compounds studied were weakly emissive with the singlet lifetimes (τS ) in the picosecond range. Nanosecond laser photolysis experiments of the brominated aza-BODIPYs revealed T1 →Tn absorption spanning from ca. 350 nm to ca. 550 nm with the triplet lifetimes (τT ) ranged between 6.0 µs and 8.5 µs. The optical properties of the aza-BODIPYs studied were pH-sensitive. Upon protonation of the dimethylamino groups with trifluoroacetic acid in toluene, a stepwise disappearance of the NIR absorption band at λabs =790 nm was observed with the concomitant appearance of a blue-shifted absorption band at λabs =652 nm, which was accompanied by a prominent emission band at λfl =680 nm. The transformation from a non-emissive to an emissive compound is associated with the inhibition of the ICT. As estimated by CV/DPV measurements, all aza-BODIPYs studied exhibited two irreversible oxidation and two quasi-reversible reduction processes. All compounds studied exhibit extremely high photostability and thermal stability.

Fluorescent 2-(Pyridin-2-yl)vinyl Pyridine Dyes and Their Thermocontrolled Release.

The generation of unique thermosensitive fluorescent dyes via heteroaromatic Heck cross-coupling and N-pyridin-2-yl nucleophilic substitution was described. To demonstrate thermosensitive properties, the precursor was converted into carbonates or phosphates and heated at various temperatures and for various time periods. Significant changes in the fluorescence intensity and emission wavelengths, between carbonates and the cyclic product, were observed, and it was proved that the dyes may serve as removable fluorescent labels with large Stokes shifts (>80 nm). The application of thermosensitive fluorescent dyes in oligonucleotide labeling has been demonstrated.

Facile Synthesis, Triplet-State Properties, and Electrochemistry of Hexaiodo-Subphthalocyanine.

In view of the ever-growing demand for efficient triplet photosensitizers and photoactive components of various optoelectronic devices, we herein report the synthesis and properties of hexaiodo-subphthalocyanines (I6 SubPcs). The improved five-step route to 4,5-diiodophthalonitrile, which serves as precursor for the synthesis of the I6 SubPcs, is reported. The improved synthesis merely required one chromatographic separation to afford the high-purity target product. Highly desirable photophysical and photochemical properties were induced in the I6 SubPcs due to the presence of six heavy iodine atoms. In particular, high values of the singlet-oxygen quantum yields (ΦΔ ) ranging from 0.83 to 0.9 were measured. The I6 SubPcs investigated proved to be phosphorescent at 77 K in 2-MeTHF with emission band maxima (λP ) located at λ=957 and 970 nm. The excited-triplet-state energies (ET ) were estimated to be approximately 1.30 eV, whereas the triplet lifetimes (τT ) were found to be 27.7 and 30.1 µs. The CV/DPV measurements indicated that both I6 SubPcs exhibited one irreversible oxidation and one quasi-reversible reduction. The spectroelectrochemical measurements pointed to a relative stability and reversibility of the electrochemically formed anion radical, that is, I6 SubPc.- , and an instability of the species formed upon one-electron oxidation, that is, I6 SubPc.+ . Estimation of EHOMO gave a value of approximately -5.8 eV whereas ELUMO was found to be located at around -3.8 eV.

Experimental and theoretical studies on fluvastatin primary photoproduct formation.

Fluvastatin (FLV) belongs to the group of compounds referred to as statins, also known as 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors. Statins act as cholesterol-lowering agents and are among the most frequently prescribed drugs. They upregulate low-density lipoprotein receptors in the liver by binding to the active site of HMG-CoA reductase, which is the key enzyme in cholesterol biosynthesis. Statins have been detected as contaminants in natural waters and are susceptible to degradation upon exposure to light. Fluvastatin is extremely sensitive to light; upon irradiation it forms a range of photoproducts. In this study the fluvastatin molar absorption coefficient and the quantum yield of the drug photodegradation were determined. The FLV photodegradation quantum yield value determined in this work (Φ = 0.13 ± 0.02) was found to be significantly larger than that previously reported in the literature. Our results also showed that the generation of singlet oxygen is not involved in the drug photodecomposition indicating that the excited triplet state of fluvastatin is not populated efficiently. Moreover, experimental methods and DFT calculations were applied to get insight into the possible mechanisms of fluvastatin primary photoproduct formation. Using the transient absorption spectroscopy technique, the transient species formed immediately after the drug excitation were followed, and the scheme for fluvastatin primary photochemistry was suggested. The primary photoproducts were identified on the basis of spectroscopic and spectrometric methods. A new mechanism for photooxygenation leading to the formation of one of the identified photoproducts (FP2) was proposed and a new approach to the formation of the other photoproduct (FP1) was provided. The theoretical mechanistic explanation of the photoproduct formation is in excellent agreement with the experimental data.

A reevaluation of the photolytic properties of 2-hydroxybenzophenone-based UV sunscreens: are chemical sunscreens inoffensive?

The excited states of a set of popular sunscreen agents (2-hydroxybenzophenone, oxybenzone, and sulisobenzone) are studied by using femto- and nanosecond time-resolved spectroscopy. Upon excitation, the compounds undergo an ultrafast excited-state intramolecular proton transfer (ESIPT) reaction as the major energy-wasting process and the rate constant of this reaction is k=2×10(12) s(-1) . The ESIPT yields a keto conformer that undergoes a fast, picosecond internal conversion decay. However, a photodegradative pathway is a monophotonic HO bond breakage that subsequently leads to trace yields of phenoxyl radicals. Because potentially harmful phenoxyl radicals are formed upon irradiation of sunscreen agents, care should be taken about their reactivity towards biologically relevant compounds.

Spectroscopy and Photophysics of Monomethyl-Substituted Derivatives of 5-Deazaalloxazine and 10-Ethyl-5-Deaza-Isoalloxazine.

Steady-state and time-resolved spectra were used to describe the singlet and triplet states of 8-methyl-5-deazaalloxazine (8-Me-5-DAll), 9-methyl-5-deazaalloxazine (9-Me-5-DAll) and 10-ethyl-5-deaza-isoalloxazine (10-Et-5-DIAll). Solvatochromic properties were described using different polarity scales, including Δf and the four-parameter scale proposed by Catalán. The results indicate that the Catalán scale shows a strong influence of solvent acidity (hydrogen-bond donating ability) on the emission properties of 8-Me-5-DAll and 9-Me-5-DAll. These results indicate the importance of intermolecular solute-solvent hydrogen-bonding interactions in the excited state of these compounds. Contrary to deazaalloxazines, solvent acidity affects the absorption spectra of 10-Et-5-DIAll. Fluorescence lifetimes and quantum yields and also transient absorption spectra were determined for all of the compounds studied. Electronic structure and S(0)-S(i), S(0)-T(i ), T(1)-T(i) transitions energies and oscillator strengths were calculated using the TD-DFT methods. Theoretical calculations were compared to experimental data.

High intrinsic barriers against spin-state relaxation in iron(II)-complex solutions.

Slow relaxation: Exergonic high-spin→low-spin relaxation after photoexcitation has been found to be exceedingly slow in a class of iron(II) complexes with hexadentate imine ligands. The thermal activation barriers that arise between the quintet- and singlet-spin manifolds are the highest ever recorded for spin crossover of isolated molecules in free solution (see figure).

Photochemical formation of thiirene and thioketene in 1,2,3-thiadiazoles with phenyl substituents studied by time-resolved spectroscopy.

Photochemistry of 4-phenyl-1,2,3-thiadiazole (PT) and 4,5-diphenyl-1,2,3-thiadiazole (DPT) in solution was studied at room temperature using UV-vis and IR transient absorption spectroscopies (λ(ex) = 266 nm). Ultrafast techniques show a very fast rise (<0.3 ps) of thiirene and thioketene species, formed from 1,2,3-thiadiazoles in the singlet excited state. The remarkable unimolecular stability of thiirenes in solution is observed. On a millisecond time scale thiirenes with phenyl substituents undergo an intermolecular reaction (dimerization of thiirene-thioketene complexes) leading to 1,3-dithiole derivatives.

The structure of model and peptide disulfides markedly affects their reactivity and products formed with singlet oxygen.

Disulfide bonds are critical structural elements in proteins and stabilize folded structures. Modification of these linkages is associated with a loss of structure and function. Previous studies have reported large variations in the rate of disulfide oxidation by hypohalous acids, due to stabilization of reaction intermediates. In this study we hypothesized that considerable variation (and hence selective oxidation) would occur with singlet oxygen (1O2), a key intermediate in photo-oxidation reactions. The kinetics of disulfide-mediated 1O2 removal were monitored using the time-resolved 1270 nm phosphorescence of 1O2. Stern-Volmer plots of these data showed a large variation (∼103) in the quenching rate constants kq (from 2 × 107 for α-lipoic acid to 3.6 × 104 M-1s-1 for cystamine). The time course of disulfide loss and product formation (determined by LC-MS) support a role for 1O2, with mono- and di-oxygenated products detected. Elevated levels of these latter species were generated in D2O- compared to H2O buffers, which is consistent with solvent effects on the 1O2 lifetime. These data are interpreted in terms of the intermediacy of a zwitterion [-S+(OO-)-S-], which either isomerizes to a thiosulfonate [-S(O)2-S-] or reacts with another parent molecule to give two thiosulfinates [-S(O)-S-]. The variation in quenching rates and product formation are ascribed to zwitterion stabilization by neighboring, or remote, lone pairs of electrons. These data suggest that some disulfides, including some present within or attached to proteins (e.g., α-lipoic acid), may be selectively modified, and undergo subsequent cleavage, with adverse effects on protein structure and function.

Tetramethylalloxazines as efficient singlet oxygen photosensitizers and potential redox-sensitive agents.

Tetramethylalloxazines (TMeAll) have been found to have a high quantum yield of singlet oxygen generation when used as photosensitizers. Their electronic structure and transition energies (S0 → Si, S0 → Ti, T1 → Ti) were calculated using DFT and TD-DFT methods and compared to experimental absorption spectra. Generally, TMeAll display an energy diagram similar to other derivatives belonging to the alloxazine class of compounds, namely π,π* transitions are accompanied by closely located n,π* transitions. Photophysical data such as quantum yields of fluorescence, fluorescence lifetimes, and nonradiative rate constants were also studied in methanol (MeOH), acetonitrile (ACN), and 1,2-dichloroethane (DCE). The transient absorption spectra were also analyzed. To assess cytotoxicity of new compounds, a hemolytic assay was performed using human red blood cells (RBC) in vitro. Subsequently, fluorescence lifetime imaging experiments (FLIM) were performed on RBC under physiological and oxidative stress conditions alone or in the presence of TMeAll allowing for pinpointing changes caused by those compounds on the intracellular environment of these cells.

Sensitized photoreduction of selected benzophenones. Mass spectrometry studies of radical cross-coupling reactions.

The hydrogen atom transfer reaction (HAT) between selected benzophenones (benzophenone BP, 3-carboxybenzophenone 3CB, and 4-carboxybenzophenone 4CB) and 2-propanol was reinvestigated focusing on stable product analysis. As expected, the primary species of these HAT's are the respective diphenyl and dimethyl ketyl radicals that eventually undergo several radical coupling reactions leading to stable photoproducts. However, the mechanisms of these free radical reactions remain unclear and open to question. In this report, we focus on the detailed analysis of the stable photoproducts of these reactions using liquid chromatography coupled with high-resolution mass spectrometry (LC-ESI-QTOF-MS/MS). Products of photopinacolization (benzpinacol and two diastereoisomers of 4CB and 3CB dimers) and isomeric radical cross-coupling adducts of respective diphenyl and dimethyl ketyl radicals were separated chromatographically, and their structures were determined by high-resolution MS/MS, and the mechanisms of the reactions are discussed.

Reductive Modification of Carbon Nitride Structure by Metals-The Influence on Structure and Photocatalytic Hydrogen Evolution.

Pt, Ru, and Ir were introduced onto the surface of graphitic carbon nitride (g-C3N4) using the wet impregnation method. A reduction of these photocatalysts with hydrogen causes several changes, such as a significant increase in the specific surface area, a C/N atomic ratio, a number of defects in the crystalline structure of g-C3N4, and the contribution of nitrogen bound to the amino and imino groups. According to the X-ray photoelectron spectroscopy results, a transition layer is formed at the g-C3N4/metal nanoparticle interphase, which contains metal at a positive degree of oxidation bonded to nitrogen. These structural changes significantly enhanced the photocatalytic activity in the production of hydrogen through the water-splitting reaction. The activity of the platinum photocatalyst was 24 times greater than that of pristine g-C3N4. Moreover, the enhanced activity was attributed to significantly better separation of photogenerated electron-hole pairs on metal nanoparticles and structural distortions of g-C3N4.

Understanding structure-properties relationships of porphyrin linked to graphene oxide through π-π-stacking or covalent amide bonds.

Two graphene oxide nanoassemblies using 5-(4-(aminophenyl)-10,15,20-triphenylporphyrin (TPPNH2) were fabricated by two synthetic methods: covalent (GO-CONHTPP) and noncovalent bonding. GO-CONHTPP was achieved through amide formation at the periphery of GO sheets and the hybrid material was fully characterized by FTIR, XPS, Raman spectroscopy, and SEM. Spectroscopic measurements together with theoretical calculations demonstrated that assembling TPPNH2 on the GO surface in DMF-H2O (1:2, v/v) via non-covalent interactions causes changes in the absorption spectra of porphyrin, as well as efficient quenching of its emission. Interestingly, covalent binding to GO does not affect notably neither the porphyrin absorption nor its fluorescence. Theoretical calculations indicates that close proximity and π-π-stacking of the porphyrin molecule with the GO sheet is possible only for the non-covalent functionalization. Femtosecond pump-probe experiments revealed that only the non-covalent assembly of TPPNH2 and GO enhances the efficiency of the photoinduced electron transfer from porphyrin to GO. In contrast to the non-covalent hybrid, the covalent GO-CONHTPP material can generate singlet oxygen with quantum yields efficiency (ΦΔ = 0.20) comparable to that of free TPPNH2 (ΦΔ = 0.26), indicating the possible use of covalent hybrid materials in photodynamic/photothermal therapy. The spectroscopic studies combined with detailed quantum-chemical analysis provide invaluable information that can guide the fabrication of hybrid materials with desired properties for specific applications.