ABSTRACT
Using the principle of "Magic Bullet", a cisplatin-derived platinum(IV) prodrug heterobimetallic Pt(IV)-Ru(II) complex, cis,cis,trans-[Pt(NH3)2Cl2{Ru(tpy-BODIPY)(tpy-COO)}(biotin)]Cl2 (Pt-Ru-B, 2), having two axial ligands, namely, biotin as water-soluble B-vitamin for enhanced cellular uptake and a BODIPY-ruthenium(II) (Ru-B, 1) photosensitizer having N,N,N-donor tpy (4'-phenyl-2,2':6',2Ć¢ĀĀ³-terpyridine) bonded to boron-dipyrromethene (BODIPY), is developed as a "Platin Bullet" for targeted photodynamic therapy (PDT). Pt-Ru-B exhibited intense absorption near 500 nm and emission near 513 nm (λex = 488 nm) in a 10% dimethyl sulfoxide-Dulbecco's phosphate-buffered saline medium (pH 7.2). The BODIPY complex on light activation generates singlet oxygen as the reactive oxygen species (ROS) giving a quantum yield (ΦΔ) of Ć¢ĀĀ¼0.64 from 1,3-diphenylisobenzofuran experiments. Pt-Ru-B exhibited preferential cellular uptake in cancer cells over noncancerous cells. The dichlorodihydrofluorescein diacetate assay confirmed the generation of cellular ROS. Confocal images revealed its mitochondrial internalization. Pt-Ru-B showed submicromolar photocytotoxicity in visible light (400-700 nm) in A549 and multidrug-resistant MDA-MB-231 cancer cells. It remained nontoxic in the dark and less toxic in nontumorigenic cells. Cellular apoptosis and alteration of the mitochondrial membrane potential were evidenced from the respective Annexin V-FITC/propidium iodide assay and JC-1 dye assay. A wound healing assay using A549 cells and Pt-Ru-B revealed inhibition of cancer cell migration, highlighting its potential as an antimetastatic agent.
Subject(s)
Antineoplastic Agents , Biotin , Photochemotherapy , Photosensitizing Agents , Prodrugs , Ruthenium , Humans , Prodrugs/pharmacology , Prodrugs/chemistry , Prodrugs/chemical synthesis , Ruthenium/chemistry , Ruthenium/pharmacology , Photosensitizing Agents/pharmacology , Photosensitizing Agents/chemistry , Photosensitizing Agents/chemical synthesis , Antineoplastic Agents/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/chemical synthesis , Biotin/chemistry , Biotin/pharmacology , Coordination Complexes/pharmacology , Coordination Complexes/chemistry , Coordination Complexes/chemical synthesis , Boron Compounds/chemistry , Boron Compounds/pharmacology , Boron Compounds/chemical synthesis , Porphobilinogen/analogs & derivatives , Porphobilinogen/chemistry , Porphobilinogen/pharmacology , Cell Line, Tumor , Drug Screening Assays, Antitumor , Platinum/chemistry , Platinum/pharmacology , Molecular Structure , Cell Survival/drug effects , Reactive Oxygen Species/metabolism , Apoptosis/drug effects , Cisplatin/pharmacology , Cisplatin/chemistryABSTRACT
Photodynamic Therapy (PDT) is recognized for its exceptional effectiveness as a promising cancer treatment method. However, it is noted that overexposure to the dosage and sunlight in traditional PDT can result in damage to healthy tissues, due to the low tumor selectivity of currently available photosensitizers (PSs). To address this challenge, we introduce herein a new strategy where the small molecule-targeted agent, erlotinib, is integrated into a boron dipyrromethene (BODIPY)-based PS to form conjugate 6 to enhance the precision of PDT. This conjugate demonstrates optical absorption, fluorescence emission, and singlet oxygen generation efficiency comparable to the reference compound 7, which lacks erlotinib. In vitro studies reveal that, after internalization, conjugate 6 predominantly accumulates in the lysosomes of HepG2 cells, exhibiting significant photocytotoxicity with an IC50 value of 3.01 ĀµM. A distinct preference for HepG2 cells over HELF cells is observed with conjugate 6 but not with compound 7. In vivo experiments further confirm that conjugate 6 has a specific affinity for tumor tissues, and the combination treatment of conjugate 6 with laser illumination can effectively eradicate H22 tumors in mice with outstanding biosafety. This study presents a novel and potential PS for achieving precise PDT against cancer.
Subject(s)
Erlotinib Hydrochloride , Liver Neoplasms , Photochemotherapy , Photosensitizing Agents , Porphobilinogen , Humans , Photochemotherapy/methods , Animals , Mice , Porphobilinogen/analogs & derivatives , Porphobilinogen/chemistry , Photosensitizing Agents/pharmacology , Photosensitizing Agents/chemistry , Hep G2 Cells , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Erlotinib Hydrochloride/pharmacology , Erlotinib Hydrochloride/chemistry , Boron Compounds/chemistry , Boron Compounds/pharmacologyABSTRACT
A boron dipyrromethene (BODIPY) derivative bearing a cis-proline residue at the meso-position crystallizes in the form of platelets with strong (i.e., ΦF = 0.34) red fluorescence, but the absorption and emission spectra differ markedly from those for dilute solutions. A key building block for the crystal is a pseudo-dimer where hydrogen bonding aligns the proline groups and separates the terminal chromophores by ca. 25 Ć . Comparison with a covalently linked bichromophore suggests that one-dimensional (1D) excitonic coupling between the terminals is too small to perturb the optical properties. However, accretion of the pseudo-dimer forms narrow channels possessing a high density of chromophores. The resultant absorption spectrum exhibits strong excitonic splitting, which can be explained quantitatively using the extended dipole approach and allowing for coupling between ca. 30 BODIPY units. Fluorescence, which decays with a lifetime of 2.2 ns, is assigned to a delocalized and (slightly) super-radiant BODIPY dimer situated at the interface and populated via electronic energy transfer from the interior.
Subject(s)
Boron , Proline , Boron/chemistry , Boron Compounds , Porphobilinogen/analogs & derivatives , Porphobilinogen/chemistryABSTRACT
Dual emissions at ~700 and 800 nm have been achieved from a single NIR-AZA fluorophore 1 by establishing parameters in which it can exist in either its isolated molecular or aggregated states. Dual near infrared (NIR) fluorescence color lymph node (LN) mapping with 1 was achieved in a large-animal porcine model, with injection site, channels and nodes all detectable at both 700 and 800 nm using a preclinical open camera system. The fluorophore was also compatible with imaging using two clinical instruments for fluorescence guided surgery. Methods: An NIR-AZA fluorophore with hydrophilic and phobic features was synthesised in a straightforward manner and its aggregation properties characterised spectroscopically and by TEM imaging. Toxicity was assessed in a rodent model and dual color fluorescence imaging evaluated by lymph node mapping in a large animal porcine models and in ex-vivo human tissue specimen. Results: Dual color fluorescence imaging has been achieved in the highly complex biomedical scenario of lymph node mapping. Emissions at 700 and 800 nm can be achieved from a single fluorophore by establishing molecular and aggregate forms. Fluorophore was compatible with clinical systems for fluorescence guided surgery and no toxicity was observed in high dosage testing. Conclusion: A new, biomedical compatible form of NIR-dual emission wavelength imaging has been established using a readily accessible fluorophore with significant scope for clinical translation.
Subject(s)
Endoscopy/methods , Fluorescent Dyes/administration & dosage , Lymph Nodes/diagnostic imaging , Optical Imaging/methods , Animals , Endoscopy/instrumentation , Female , Fluorescent Dyes/chemistry , Fluorescent Dyes/toxicity , HeLa Cells , Humans , Intraoperative Care/instrumentation , Intraoperative Care/methods , Intravital Microscopy/methods , Lymphatic Metastasis/diagnosis , Male , Models, Animal , Neoplasms/pathology , Neoplasms/surgery , Optical Imaging/instrumentation , Porphobilinogen/administration & dosage , Porphobilinogen/analogs & derivatives , Porphobilinogen/chemistry , Porphobilinogen/toxicity , Rats , Spectrophotometry, Infrared/instrumentation , Spectrophotometry, Infrared/methods , Sus scrofa , Toxicity Tests, Subacute/methodsABSTRACT
Through a simple 1,3-cycloaddition reaction, three BODIPY-peptide conjugates that target the extracellular domain of the epidermal growth factor receptor (EGFR) were prepared and their ability for binding to EGFR was investigated. The peptide ligands K(N3)LARLLT and its cyclic analog cyclo(K(N3)larllt, previously shown to have high affinity for binding to the extracellular domain of EGFR, were conjugated to alkynyl-functionalized BODIPY dyes 1 and 2 via a copper-catalyzed click reaction. This reaction produced conjugates 3, 4, and 5 in high yields (70-82%). In vitro studies using human carcinoma HEp2 cells that overexpress EGFR demonstrated high cellular uptake, particularly for the cyclic peptide conjugate 5, and low cytotoxicity in light (~1 JĀ·cm-2) and darkness. Surface plasmon resonance (SPR) results show binding affinity of the three BODIPY-peptide conjugates for EGFR, particularly for 5 bearing the cyclic peptide. Competitive binding studies using three cell lines with different expressions of EGFR show that 5 binds specifically to EGFR-overexpressing colon cancer cells. Among the three conjugates, 5 bearing the cyclic peptide exhibited the highest affinity for binding to the EGFR protein.
Subject(s)
Boron Compounds/chemistry , Boron/chemistry , Fluorescent Dyes/chemistry , Peptides, Cyclic/chemistry , Porphobilinogen/analogs & derivatives , Cell Line, Tumor , ErbB Receptors/metabolism , Humans , Ligands , Porphobilinogen/chemistry , Protein Binding , Surface Plasmon Resonance/methodsABSTRACT
Boron-dipyrromethenes (Bodipys), since first reported in 1968, have emerged as a fascinating class of dyes in the past few decades due to their excellent photophysical properties including bright fluorescence, narrow emission bandwidth, resistance to photobleaching, and environment insensitivity. However, typical Bodipys are highly lipophilic, which often results in nonfluorescent aggregates in aqueous solution and also severely limits their bioavailability to cells and tissues. In this work, based on a simple one-atom B Ć¢ĀĀ C replacement in the Bodipy scaffold, we present a new class of carbon-dipyrromethenes (Cardipys for short) fluorescent dyes with tunable emission wavelengths covering the visible and near-infrared regions. These Cardipys not only retain the excellent photophysical properties of conventional Bodipys but also show improved water solubility and photostability due to their cationic character. Moreover, the cationic character also makes them extremely easy to penetrate the cell membrane and specifically accumulate into mitochondria without resorting to any mitochondria-targeted groups. Interestingly, several Cardipys bearing active styryl groups could serve as fluorescent indicators to map cellular trafficking of the glutathione conjugates produced within mitochondria under the catalysis of glutathione S-transferase (GST), thus showing potential in either exploring the detoxification mechanism of the mitochondrial GST/GSH system or evaluating the drug resistance of cancer cells that is closely related with GST activity.
Subject(s)
Carbon/chemistry , Fluorescent Dyes/chemistry , Glutathione/chemistry , Mitochondria/chemistry , Porphobilinogen/analogs & derivatives , A549 Cells , Boron Compounds/chemistry , Cations/chemistry , Cell Membrane/metabolism , Cell Membrane/ultrastructure , Cell Movement , Electrochemical Techniques , Glutathione Transferase/metabolism , Humans , Mitochondria/ultrastructure , Molecular Structure , Optical Imaging , Photochemical Processes , Porphobilinogen/chemistry , Solubility , Solvents/chemistry , Spectrometry, FluorescenceABSTRACT
Senescence-associated diseases have severely diminished the quality of life and health of patients. However, a sensitive assay of these diseases remains limited due to a lack of straightforward methods. Considering that senescence-associated Ć-galactosidase (SA-Ć-Gal) is overexpressed in senescent cells, the detection of SA-Ć-Gal in senescent cells and tissues might be a feasible strategy for the early diagnosis of SA diseases. In this study, a Ć-galactosidase-activatable nanoprobe BOD-L-ĆGal-NPs was developed for the imaging of senescent cells and vasculature in atherosclerotic mice via real-time monitoring of Ć-Gal. BOD-L-ĆGal-NPs was fabricated by encapsulating a newly designed NIR ratiometric probe BOD-L-ĆGal within a poly(lactic-co-glycolic) acid (PLGA) core. Nanoprobe BOD-L-ĆGal-NPs showed good accumulation in arteries, thus successfully visualizing senescent cells and vasculature in atherosclerotic mice by tail vein injection. Our findings indicated that nanoprobe BOD-L-ĆGal-NPs holds great potential for the early diagnosis and therapy of atherosclerosis and other aging-associated diseases.
Subject(s)
Atherosclerosis/diagnosis , Boron/chemistry , Fluorescent Dyes/chemistry , Nanoparticles/chemistry , Porphobilinogen/analogs & derivatives , beta-Galactosidase/analysis , Animals , Atherosclerosis/metabolism , Boron/metabolism , Cellular Senescence , Fluorescent Dyes/metabolism , Male , Mice , Mice, Inbred C57BL , Molecular Structure , Nanoparticles/metabolism , Porphobilinogen/chemistry , Porphobilinogen/metabolism , Rats , Rats, Sprague-Dawley , beta-Galactosidase/genetics , beta-Galactosidase/metabolismABSTRACT
A series of fluorescent boron-dipyrromethene (BODIPY, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) dyes have been designed to participate, as aglycons, in synthetic oligosaccharide protocols. As such, they served a dual purpose: first, by being incorporated at the beginning of the process (at the reducing-end of the growing saccharide moiety), they can function as fluorescent glycosyl tags, facilitating the detection and purification of the desired glycosidic intermediates, and secondly, the presence of these chromophores on the ensuing compounds grants access to fluorescently labeled saccharides. In this context, a sought-after feature of the fluorescent dyes has been their chemical robustness. Accordingly, some BODIPY derivatives described in this work can withstand the reaction conditions commonly employed in the chemical synthesis of saccharides; namely, glycosylation and protecting-group manipulations. Regarding their photophysical properties, the BODIPY-labeled saccharides obtained in this work display remarkable fluorescence efficiency in water, reaching quantum yield values up to 82 %, as well as notable lasing efficiencies and photostabilities.
Subject(s)
Boron Compounds/chemistry , Boron/chemistry , Fluorescent Dyes/chemistry , Porphobilinogen/analogs & derivatives , Fluorescence , Glycosylation , Light , Porphobilinogen/chemistryABSTRACT
In this study, we report for the first time the use of four aza-dipyrromethenes (ADPMs) as photosensitizers for cancer PDT. The synthesis and characterization of the ADPMs and their photodynamic action against B16F10 melanoma cells were assessed. ADPM 2 is the best singlet oxygen generator and the most phototoxic (at 2.5 ĀµM) towards B16F10 cells.
Subject(s)
Antineoplastic Agents/pharmacology , Aza Compounds/pharmacology , Melanoma/drug therapy , Photochemotherapy , Photosensitizing Agents/pharmacology , Porphobilinogen/analogs & derivatives , Animals , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Aza Compounds/chemical synthesis , Aza Compounds/chemistry , Cell Proliferation/drug effects , Cell Survival/drug effects , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Melanoma/pathology , Mice , Molecular Structure , Photosensitizing Agents/chemical synthesis , Photosensitizing Agents/chemistry , Porphobilinogen/chemical synthesis , Porphobilinogen/chemistry , Porphobilinogen/pharmacology , Structure-Activity Relationship , Tumor Cells, CulturedABSTRACT
Pt(II) photosensitizers are emerging as novel Pt anticancer agents for cancer photodynamic therapy (PDT) to avoid uncontrollable toxicity of cisplatin. However, the application of Pt(II) photosensitizers is limited by tumor hypoxia and the poor penetration depth of excitation light. To overcome these drawbacks, exploiting the next generation of Pt anticancer agents is of urgent need. According to theoretical calculations, novel near-infrared (NIR)-absorbing Pt(II)-chelated azadipyrromethene dyes (PtDP-X, where X = N, C, and S) were designed. Importantly, spin-orbit coupling of the Pt atom could promote the intersystem crossing of a singlet-to-triplet transition for converting oxygen to singlet oxygen (1O2), and the azadipyrromethene skeleton could provide a strong photothermal effect. As expected, PtDP-X exhibited intense NIR absorption and synergistic PDT and photothermal effects with low dark cytotoxicity. Furthermore, water-soluble and biocompatible PtDP-N nanoparticles (PtDP-N NPs) were prepared that achieved effective tumor cell elimination with low side effects under 730 nm light irradiation in vitro and in vivo. This pioneering work could push the exploitation of NIR-absorbing metal-chelated azadipyrromethene dyes, so as to promote the positive evolution of phototherapy agents.
Subject(s)
Photosensitizing Agents/chemical synthesis , Platinum Compounds/chemical synthesis , Platinum Compounds/pharmacology , Porphobilinogen/analogs & derivatives , Furans , HeLa Cells , Humans , Infrared Rays , Molecular Structure , Photosensitizing Agents/chemistry , Phototherapy , Platinum Compounds/chemistry , Porphobilinogen/chemistry , Spectrophotometry, InfraredABSTRACT
The ruthenium(II) complexes [RuCl(L1)(L3)]Cl (1), [RuCl(L1)(L4)]Cl (2), [RuCl(L2)(L4)]Cl (3), [RuCl(L1)(L5)]Cl (4), and [RuCl(L2)(L5)]Cl (5) of NNN-donor dipicolylamine (dpa) bases (L4, L5) having BODIPY (boron-dipyrromethene) moieties, NN-donor phenanthroline derivatives (L1, L2), and benzyldipicolylamine (bzdpa, L3) were prepared and characterized by spectroscopic techniques and their cellular localization/uptake and photocytotoxicity studied. Complex 1, as its PF6 salt (1a), has been structurally characterized with help of a single-crystal X-ray diffraction technique. It has a RuN5Cl core with the Cl bonded trans to the amine nitrogen atom of bzdpa. The complexes showed intense absorption spectral bands near 500 nm (ĆĀµ ≈ 58000 M-1 cm-1) in 2 and 3 and 654 nm (ĆĀµ ≈ 80000 M-1 cm-1) in 4 and 5 in 1/1 DMSO/DPBS (v/v). Complex 5 having biotin and PEGylated-disteryl BODIPY gave a singlet oxygen quantum yield (ΦΔ) of Ć¢ĀĀ¼0.65 in DMSO. Complex 5 exhibited remarkable PDT (photodynamic therapy) activity (IC50 ≈ 0.02 ĀµM) with a photocytotoxicity index (PI) value of >5000 in red light of 600-720 nm in A549 cancer cells. The biotin-conjugated complexes showed better photocytotoxicity in comparison to nonbiotinylated analogues in A549 cells. The complexes displayed less toxicity in HPL1D normal cells in comparison to A549 cancer cells. The emissive BODIPY complexes 3 and 5 (ΦF ≈ 0.07 in DMSO) showed significant mitochondrial localization.
Subject(s)
Antineoplastic Agents/pharmacology , Coordination Complexes/pharmacology , Light , Photochemotherapy , Photosensitizing Agents/pharmacology , A549 Cells , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Biotin/chemistry , Biotin/pharmacology , Boron/chemistry , Boron/pharmacology , Cell Line , Cell Proliferation/drug effects , Cell Survival/drug effects , Coordination Complexes/chemical synthesis , Coordination Complexes/chemistry , DNA Cleavage/drug effects , Density Functional Theory , Drug Screening Assays, Antitumor , Humans , Models, Molecular , Optical Imaging , Photosensitizing Agents/chemical synthesis , Photosensitizing Agents/chemistry , Porphobilinogen/analogs & derivatives , Porphobilinogen/chemistry , Porphobilinogen/pharmacology , Ruthenium/chemistry , Ruthenium/pharmacologyABSTRACT
Because BF4- is a labile, non- or weakly coordinating anion, it is generally chosen by chemists who do not want the anion to interfere with the associated cation. Herein, we demonstrate that BF4- actually strongly binds to triazole-appended dipyrromethenes (TADs). In particular, HETCOR NMR experiments and DFT calculations were used to rationalize the results observed with anion titrations. Hence, special care should be taken when considering that BF4- is innocent.
Subject(s)
Boron Compounds/chemistry , Fluorides/chemistry , Porphobilinogen/analogs & derivatives , Triazoles/chemistry , Porphobilinogen/chemistryABSTRACT
Herein we report on a straightforward access method for boron dipyrromethene dyes (BODIPYs)-coumarin hybrids linked through their respective 8- and 6- positions, with wide functionalization of the coumarin fragment, using salicylaldehyde as a versatile building block. The computationally-assisted photophysical study unveils broadband absorption upon proper functionalization of the coumarin, as well as the key role of the conformational freedom of the coumarin appended at the meso position of the BODIPY. Such free motion almost suppresses the fluorescence signal, but enables us to apply these dyads as molecular rotors to monitor the surrounding microviscosity.
Subject(s)
Boron/chemistry , Coumarins/chemistry , Porphobilinogen/analogs & derivatives , Fluorescence , Fluorescent Dyes/chemistry , Molecular Conformation , Porphobilinogen/chemistry , Spectrometry, FluorescenceABSTRACT
This study focuses on the behavior of a new fluorescent marker for labeling individual biomolecules and staining cell organelles developed on a meso-substituted BODIPY platform. Boron(III) complex with meso-4-methoxycarbonylpropylsubstituted 3,3',5,5'-tetramethyl-2,2'-dipyrromethene has been synthesized and identified via visible, UV-, NMR- and MS-spectra X-ray. The behavior of fluorophore in solutions has been studied with various experimental techniques. It has been found that luminophore exhibits a high quantum yield (almost ~100-75%) in the blue-green region (513-520 nm) and has high photostability. In addition, biological analysis indicates that the fluorophore exhibits a tendency to effectively penetrate into cell membranes. On the other hand, the proposed BODIPY can be used to study the significant differences among a large number of pathogens of mycotic infections, as well as to visualize structural changes in the plasma membrane, which is necessary for the clearance of mammalian cells undergoing apoptotic cell death.
Subject(s)
Boron/chemistry , Diagnostic Imaging , Porphobilinogen/analogs & derivatives , Boron Compounds/chemical synthesis , Boron Compounds/chemistry , Candida albicans/cytology , Cell Line, Tumor , Crystallography, X-Ray , Doxorubicin/pharmacology , Electrons , Fusarium/cytology , Humans , Porphobilinogen/chemistry , Solvents/chemistry , Spectrometry, Fluorescence , Subcellular Fractions/metabolism , Ultraviolet RaysABSTRACT
In recent years, new drug discovery approaches based on novel pharmacological concepts have emerged. Allosteric modulators, for example, target receptors at sites other than the orthosteric binding sites and can modulate agonist-mediated activation. Interestingly, allosteric regulation may allow a fine-tuned regulation of unbalanced neurotransmitter' systems, thus providing safe and effective treatments for a number of central nervous system diseases. The metabotropic glutamate type 5 receptor (mGlu5R) has been shown to possess a druggable allosteric binding domain. Accordingly, novel allosteric ligands are being explored in order to finely regulate glutamate neurotransmission, especially in the brain. However, before testing the activity of these new ligands in the clinic or even in animal disease models, it is common to characterize their ability to bind mGlu5Rs in vitro. Here, we have developed a new series of fluorescent ligands that, when used in a new NanoBRET-based binding assay, will facilitate screening for novel mGlu5R allosteric modulators.
Subject(s)
Drug Discovery/methods , Fluorescent Dyes/chemistry , Receptor, Metabotropic Glutamate 5/chemistry , Allosteric Regulation/drug effects , Allosteric Site , Binding Sites , Bioluminescence Resonance Energy Transfer Techniques , Boron Compounds/chemical synthesis , Boron Compounds/chemistry , Calcium/metabolism , Drug Discovery/instrumentation , HEK293 Cells , Humans , Ligands , Porphobilinogen/analogs & derivatives , Porphobilinogen/chemistry , Protein Binding , Receptor, Metabotropic Glutamate 5/genetics , Receptor, Metabotropic Glutamate 5/metabolismABSTRACT
Chlorination procedures are commonly applied in swimming pool water and wastewater treatment, yet also in food, pharmaceutical, and paper production. The amount of chlorine in water needs to be strictly controlled to ensure efficient killing of pathogens but avoid the induction of negative health effects. Miniaturized microfluidic fluorescence sensors are an appealing approach here when aiming at online or at-site measurements. Two meso-enamine-substituted boron dipyrromethene (BODIPY) dyes were found to exhibit favorable indication properties, their reaction with hypochlorite leading to strong fluorescence enhancement. Real-time assays became possible after integration of these fluorescent probes with designed two-dimensional (2D) and three-dimensional (3D) microfluidic chips, incorporating a passive sinusoidal mixer and a microhydrocyclone, respectively. A comparison of the two microfluidic systems, including their abilities to prevent accumulation or circulation of microbubbles produced by the chemical indication reaction, showed excellent fluidic behavior for the microhydrocyclone-based device. After coupling to a miniaturized optical reader for fluorescence detection, the 2D microfluidic system showed a promising detection range of 0.04-0.5 mg L-1 while still being prone to bubble-induced fluctuations and suffering from considerably low signal gain. The microhydrocyclone-based system was distinctly more robust against gas bubbles, showed a higher signal gain, and allowed us to halve the limit of detection to 0.02 mg L-1. The use of the 3D system to quantify the chlorine content of swimming pool water samples for sensitive and quantitative chlorine monitoring was demonstrated.
Subject(s)
Boron Compounds/chemistry , Charcoal/chemistry , Chlorides/analysis , Drinking Water/analysis , Microfluidic Analytical Techniques/instrumentation , Microfluidic Analytical Techniques/methods , Porphobilinogen/analogs & derivatives , Chlorides/isolation & purification , Fluorescence , Fluorescent Dyes/chemistry , Halogenation , Humans , Porphobilinogen/chemistry , Spectrometry, FluorescenceABSTRACT
As traditional phototherapy agents, boron dipyrromethene (BODIPY) photosensitizers have attracted increasing attention due to their high molar extinction coefficients, high phototherapy efficacy, and excellent photostability. After being formed into nanostructures, BODIPY-containing nano-photosensitizers show enhanced water solubility and biocompatibility as well as efficient tumor accumulation compared to BODIPY molecules. Hence, BODIPY nano-photosensitizers demonstrate a promising potential for fighting cancer. This review contains three sections, classifying photodynamic therapy (PDT), photothermal therapy (PTT), and the combination of PDT and PTT based on BODIPY nano-photosensitizers. It summarizes various BODIPY nano-photosensitizers, which are prepared via different approaches including molecular precipitation, supramolecular interactions, and polymer encapsulation. In each section, the design strategies and working principles of these BODIPY nano-photosensitizers are highlighted. In addition, the detailed in vitro and in vivo applications of these recently developed nano-photosensitizers are discussed together with future challenges in this field, highlighting the potential of these promising nanoagents for new tumor phototherapies.
Subject(s)
Antineoplastic Agents/pharmacology , Boron/pharmacology , Neoplasms/therapy , Photosensitizing Agents/pharmacology , Phototherapy , Porphobilinogen/analogs & derivatives , Animals , Antineoplastic Agents/chemistry , Humans , Photosensitizing Agents/chemistry , Porphobilinogen/chemistry , Porphobilinogen/pharmacologyABSTRACT
We report a new boron dipyrromethene (BODIPY)-bridged bisphenoxyl diradicaloid (2), which showed closed-shell diamagnetic character in less polar solvents such as dichloromethane but open-shell diradical character with paramagnetic activity in the very polar solvent N,N-dimethylformamide. X-ray crystallographic analysis of 2 revealed an anti-parallel stacked dimer structure via intermolecular dipole-dipole interaction, and the observed solvent-dependent diradical character can be explained by the different dihedral angles between the phenoxyl units and the BODIPY bridge, and structural flexibility of the molecule in different solvents. Compound 2 also exhibited solvent-dependent optical and electrochemical properties.
Subject(s)
Boron Compounds/chemistry , Models, Molecular , Porphobilinogen/analogs & derivatives , Crystallography, X-Ray , Molecular Structure , Porphobilinogen/chemistryABSTRACT
Direct cell-to-cell transmission of proteopathic α-synuclein (α-syn) aggregates is thought to underlie the progression of neurodegenerative synucleinopathies. However, the specific intracellular processes governing this transmission remain unclear because currently available model systems are limited. For example, in cell culture models of α-syn-seeded aggregation, it is difficult to discern intracellular from extracellular exogenously applied α-syn seed species. Herein, we employed fluorescently labeled α-syn preformed fibrils (pffs) in conjunction with the membrane-impermeable fluorescence quencher trypan blue to selectively image internalized α-syn seeds in cultured primary neurons and to quantitatively characterize the concentration dependence, time course, and inhibition of pff uptake. To study the long-term fates of exogenous α-syn pffs in neurons, we developed a pff species labeled at amino acid residue 114 with the environmentally insensitive fluorophore BODIPY or the pH-sensitive dye pHrodo red. We found that pffs are rapidly trafficked along the endolysosomal pathway, where most of the material remains for days. We also found that brief pharmacological perturbation of lysosomes shortly after the pff treatment causes aberrations in intracellular processing of pff seeds concomitant with an increased rate of inclusion formation via recruitment of endogenous α-syn to a relatively small number of exogenous seeds. Our results validate a quantitative assay for pff uptake in primary neurons, implicate lysosomal processing as the major fate of internalized proteopathic seeds, and suggest lysosomal integrity as a significant rate-determining step in the transmission of α-syn pathology. Further, lysosomal processing of transmitted seeds may represent a new therapeutic target to combat the spread of synucleinopathies.
Subject(s)
Endosomes/metabolism , Hippocampus/metabolism , Lysosomes/metabolism , Neurons/metabolism , Protein Aggregation, Pathological/metabolism , alpha-Synuclein/metabolism , Amino Acid Substitution , Animals , Cells, Cultured , Coloring Agents/analysis , Embryo, Mammalian/cytology , Endocytosis , Endosomes/pathology , Endosomes/ultrastructure , Fluorescent Dyes/analysis , Fluorescent Dyes/chemistry , Green Fluorescent Proteins/chemistry , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Hippocampus/pathology , Hippocampus/ultrastructure , Humans , Hydrogen-Ion Concentration , Lysosomes/pathology , Lysosomes/ultrastructure , Mice , Microscopy, Electron, Transmission , Mutation , Neurons/pathology , Neurons/ultrastructure , Porphobilinogen/analogs & derivatives , Porphobilinogen/analysis , Porphobilinogen/chemistry , Protein Aggregation, Pathological/pathology , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/metabolism , Rhodamines/analysis , Rhodamines/chemistry , Trypan Blue/analysis , alpha-Synuclein/chemistry , alpha-Synuclein/geneticsABSTRACT
Human porphobilinogen deaminase (PBGD), the third enzyme in the heme pathway, catalyzes four times a single reaction to convert porphobilinogen into hydroxymethylbilane. Remarkably, PBGD employs a single active site during the process, with a distinct yet chemically equivalent bond formed each time. The four intermediate complexes of the enzyme have been biochemically validated and they can be isolated but they have never been structurally characterized other than the apo- and holo-enzyme bound to the cofactor. We present crystal structures for two human PBGD intermediates: PBGD loaded with the cofactor and with the reaction intermediate containing two additional substrate pyrrole rings. These results, combined with SAXS and NMR experiments, allow us to propose a mechanism for the reaction progression that requires less structural rearrangements than previously suggested: the enzyme slides a flexible loop over the growing-product active site cavity. The structures and the mechanism proposed for this essential reaction explain how a set of missense mutations result in acute intermittent porphyria.