RESUMO
Squamous cell carcinoma (SCC) is a common nonmelanoma skin cancer. Radiotherapy plays an integral role in treating SCC due to its characteristics, such as diminished intercellular adhesion, heightened cell migration and invasion capabilities, and immune evasion. These problems lead to inaccurate tumor boundary positioning and radiotherapy tolerance in SCC treatment. Thus, accurate localization and enhanced radiotherapy sensitivity are imperative for effective SCC treatment. To address the existing limitations in SCC therapy, we developed monoglyceride solid lipid nanoparticles (MG SLNs) and enveloped them with the A431 cell membrane (A431 CM) to create A431@MG. The characterization results showed that A431@MG was spherical. Furthermore, A431@MG had specific targeting for A431 cells. In A431 tumor-bearing mice, A431@MG demonstrated prolonged accumulation within tumors, ensuring precise boundary localization of SCC. We further advanced the approach by preparing MG SLNs encapsulating 5-aminolevulinic acid methyl ester (MLA) and desferrioxamine (DFO) with an A431 CM coating to yield A431@MG-MLA/DFO. Several studies have revealed that DFO effectively reduced iron content, impeding protoporphyrin IX (PpIX) biotransformation and promoting PpIX accumulation. Simultaneously, MLA was metabolized into PpIX upon cellular entry. During radiotherapy, the heightened PpIX levels enhanced reactive oxygen species (ROS) generation, inducing DNA and mitochondrial damage and leading to cell apoptosis. In A431 tumor-bearing mice, the A431@MG-MLA/DFO group exhibited notable radiotherapy sensitization, displaying superior tumor growth inhibition. Combining A431@MG-MLA/DFO with radiotherapy significantly improved anticancer efficacy, highlighting its potential to serve as an integrated diagnostic and therapeutic strategy for SCC.
Assuntos
Carcinoma de Células Escamosas , Membrana Celular , Nanopartículas , Radiossensibilizantes , Neoplasias Cutâneas , Animais , Camundongos , Nanopartículas/química , Humanos , Linhagem Celular Tumoral , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/radioterapia , Radiossensibilizantes/química , Radiossensibilizantes/farmacologia , Radiossensibilizantes/administração & dosagem , Membrana Celular/metabolismo , Ácido Aminolevulínico/química , Ácido Aminolevulínico/farmacologia , Ácido Aminolevulínico/administração & dosagem , Lipídeos/química , Ensaios Antitumorais Modelo de Xenoenxerto , Desferroxamina/química , Desferroxamina/farmacologia , Camundongos Nus , Feminino , Camundongos Endogâmicos BALB C , Espécies Reativas de Oxigênio/metabolismo , Apoptose/efeitos dos fármacos , LipossomosRESUMO
In 2017, the FDA authorized 5-aminolevulinic acid (5-ALA) for intraoperative optical imaging of suspected high-grade gliomas. This was the first authorized optical imaging agent for brain tumor surgery to enhance the visualization of malignant tissue. Herein we report the synthesis of a racemic and enantiopure fluorinated analog of 5-ALA, i.e., 3-fluoro-5-aminolevulinic acid (3F-5-ALA). We anticipate that these studies will provide the foundation for the future construction of a fluorine-18-labeled 5-ALA PET tracer to be used for functional and metabolic imaging of gliomas.
Assuntos
Ácido Aminolevulínico , Halogenação , Ácido Aminolevulínico/química , Ácido Aminolevulínico/síntese química , Ácido Aminolevulínico/análogos & derivados , Estereoisomerismo , Estrutura Molecular , Radioisótopos de Flúor/química , Tomografia por Emissão de PósitronsRESUMO
5-Aminolevulinic acid (ALA) and its derivatives, serving as the endogenous precursor of the photosensitizer (PS) protoporphyrin IX (PpIX), successfully applied in tumor imaging and photodynamic therapy (PDT). ALA and its derivatives have been used to treat actinic keratosis (AK), basal cell carcinoma (BCC), and improve the detection of superficial bladder cancer. However, the high hydrophilicity of ALA and the conversion of PpIX to heme have limited the accumulation of PpIX, hindering the efficiency and potential application of ALA-PDT. This study aims to evaluate the PDT activity of three rationally designed series of ALA-HPO prodrugs, which were based on enhancing the lipophilicity of the prodrugs and reducing the labile iron pool (LIP) through HPO iron chelators to promote PpIX accumulation. Twenty-four ALA-HPO conjugates, incorporating amide, amino acid, and ester linkages, were synthesized. Most of the conjugates, exhibited no dark-toxicity to cells, according to bioactivity evaluation. Ester conjugates 19a-g showed promoted phototoxicity when tested on tumor cell lines, and this increased phototoxicity was strongly correlated with elevated PpIX levels. Among them, conjugate 19c emerged as the most promising (HeLa, IC50 = 24.25 ± 1.43 µM; MCF-7, IC50 = 43.30 ± 1.76 µM; A375, IC50 = 28.03 ± 1.00 µM), displaying superior photodynamic anticancer activity to ALA (IC50 > 100 µM). At a concentration of 80 µM, the fluorescence intensity of PpIX induced by compound 19c in HeLa, MCF-7, and A375 cells was 18.9, 5.3, and 2.8 times higher, respectively, than that induced by ALA. In conclusion, cellular phototoxicity showed a strong correlation with intracellular PpIX fluorescence levels, indicating the potential application of ALA-HPO conjugates in ALA-PDT.
Assuntos
Ácido Aminolevulínico , Antineoplásicos , Ensaios de Seleção de Medicamentos Antitumorais , Fotoquimioterapia , Fármacos Fotossensibilizantes , Humanos , Ácido Aminolevulínico/farmacologia , Ácido Aminolevulínico/química , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/síntese química , Antineoplásicos/farmacologia , Antineoplásicos/química , Antineoplásicos/síntese química , Relação Estrutura-Atividade , Estrutura Molecular , Piridonas/farmacologia , Piridonas/química , Piridonas/síntese química , Linhagem Celular Tumoral , Protoporfirinas/química , Protoporfirinas/farmacologia , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Sobrevivência Celular/efeitos dos fármacos , Pró-Fármacos/farmacologia , Pró-Fármacos/química , Pró-Fármacos/síntese químicaRESUMO
Heme is a critical biomolecule that is synthesized in vivo by several organisms such as plants, animals, and bacteria. Reflecting the importance of this molecule, defects in heme biosynthesis underlie several blood disorders in humans. Aminolevulinic acid synthase (ALAS) initiates heme biosynthesis in α-proteobacteria and nonplant eukaryotes. Debilitating and painful diseases such as X-linked sideroblastic anemia and X-linked protoporphyria can result from one of more than 91 genetic mutations in the human erythroid-specific enzyme ALAS2. This review will focus on recent structure-based insights into human ALAS2 function in health and how it dysfunctions in disease. We will also discuss how certain genetic mutations potentially result in disease-causing structural perturbations. Furthermore, we use thermodynamic and structural information to hypothesize how the mutations affect the human ALAS2 structure and categorize some of the unique human ALAS2 mutations that do not respond to typical treatments, that have paradoxical in vitro activity, or that are highly intolerable to changes. Finally, we will examine where future structure-based insights into the family of ALA synthases are needed to develop additional enzyme therapeutics.
Assuntos
5-Aminolevulinato Sintetase , Anemia Sideroblástica , Doenças Genéticas Ligadas ao Cromossomo X , 5-Aminolevulinato Sintetase/química , 5-Aminolevulinato Sintetase/genética , 5-Aminolevulinato Sintetase/metabolismo , Ácido Aminolevulínico/química , Ácido Aminolevulínico/metabolismo , Anemia Sideroblástica/enzimologia , Anemia Sideroblástica/genética , Animais , Heme , Humanos , Relação Estrutura-AtividadeRESUMO
Silver-indium-sulfide quantum dots (AIS QDs) have potential applications in many areas, including biomedicine. Their lack of regulated heavy metals, unlike many commercialized QDs, stands out as an advantage, but the necessity for alloyed or core-shell structures and related costly and sophisticated processes for the production of stable and high quantum yield aqueous AIS QDs are the current challenges. The present study demonstrates the one-step aqueous synthesis of simple AgInS2 QD compositions utilizing for the first time either a polyethyleneimine/2-mercaptopropionic acid (AIS-PEI/2MPA) mixture or only 2-mercaptopropionic acid (AIS-2MPA) as the stabilizing molecules, providing a AgInS2 portfolio consisting of cationic and anionic AIS QDs, respectively, and tuneable emission. Small AIS QDs with long-term stability and high quantum yields (19-23%) were achieved at a molar ratio of Ag/In/S 1/10/10 in water without any dopant or a semiconductor shell. The theranostic potential of these cationic and anionic AIS QDs was also evaluated in vitro. Non-toxic doses were determined, and fluorescence imaging potential was demonstrated. More importantly, these QDs were electrostatically loaded with zwitterionic 5-aminolevulinic acid (ALA) as a prodrug to enhance the tumor availability of ALA and to improve ALA-induced porphyrin photodynamic therapy (PDT). This is the first study investigating the influence of nanoparticle charge on ALA binding, release, and therapeutic efficacy. Surface charge was found to be more critical in cellular internalization and dark toxicity rather than drug loading and release. Both QDs provided enhanced ALA release at acidic pH but protected the prodrug at physiological pH, which is critical for tumor delivery of ALA, which suffers from low bioavailability. The PDT efficacy of the ALA-loaded AIS QDs was tested in 2D monolayers and 3D constructs of HT29 and SW480 human colon adenocarcinoma cancer cell lines. The incorporation of ALA delivery by the AIS QDs, which on their own do not cause phototoxicity, elicited significant cell death due to enhanced light-induced ROS generation and apoptotic/necrotic cell death, reducing the IC50 for ALA dramatically to about 0.1 and 0.01 mM in anionic and cationic AIS QDs, respectively. Combined with simple synthetic methods, the strong intracellular photoluminescence of AIS QDs, good biocompatibility of especially the anionic AIS QDs, and the ability to act as drug carriers for effective PDT signify that the AIS QDs, in particular AIS-2MPA, are highly promising theranostic QDs.
Assuntos
Ácido Aminolevulínico/farmacologia , Antineoplásicos/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Pontos Quânticos/química , Ácido Aminolevulínico/síntese química , Ácido Aminolevulínico/química , Ânions/síntese química , Ânions/química , Antineoplásicos/síntese química , Antineoplásicos/química , Cátions/síntese química , Cátions/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Índio/química , Imagem Óptica , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/química , Prata/química , Sulfetos/química , Células Tumorais Cultivadas , Água/químicaRESUMO
Photodynamic therapy (PDT) is an effective procedure for the treatment of lesions diseases based on the selectivity of a photosensitising compound with the ability to accumulate in the target cell. Atherosclerotic plaque is a suitable target for PDT because of the preferential accumulation of photosensitisers in atherosclerotic plaques. Dendrimers are hyperbranched polymers conjugated to drugs. The dendrimers of ALA hold ester bonds that inside the cells are cleaved and release ALA, yielding PpIX production. The dendrimer 6m-ALA was chosen to perform this study since in previous studies it induced the highest porphyrin macrophage: endothelial cell ratio (Rodriguez et al. in Photochem Photobiol Sci 14:1617-1627, 2015). We transformed Raw 264.7 macrophages to foam cells by exposure to oxidised LDLs, and we employed a co-culture model of HMEC-1 endothelial cells and foam cells to study the affinity of ALA dendrimers for the foam cells. In this work it was proposed an in vitro model of atheromatous plaque, the aim was to study the selectivity of an ALA dendrimer for the foam cells as compared to the endothelial cells in a co-culture system and the type of cell death triggered by the photodynamic treatment. The ALA dendrimer 6m-ALA showed selectivity PDT response for foam cells against endothelial cells. A light dose of 1 J/cm2 eliminate foam cells, whereas less than 50% of HMEC-1 is killed, and apoptosis cell death is involved in this process, and no necrosis is present. We propose the use of ALA dendrimers as pro-photosensitisers to be employed in photoangioplasty to aid in the treatment of obstructive cardiovascular diseases, and these molecules can also be employed as a theranostic agent.
Assuntos
Ácido Aminolevulínico/farmacologia , Apoptose/efeitos dos fármacos , Células Espumosas/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Fármacos Fotossensibilizantes/farmacologia , Ácido Aminolevulínico/química , Animais , Linhagem Celular , Técnicas de Cocultura , Células Espumosas/fisiologia , Humanos , Macrófagos/fisiologia , Camundongos , Fármacos Fotossensibilizantes/químicaRESUMO
Photodynamic therapy (PDT) is a promising approach for the targeted treatment of cancer and various other human disorders. An effective, clinically approved approach in PDT involves the administration of 5-aminolevulinic acid (ALA) to generate elevated levels of the natural photosensitiser protoporphyrin IX (PpIX). The development of prodrugs of ALA is of considerable interest as a means to enhance the efficiency and cell selectivity of PpIX accumulation for PDT applications. In this work a novel peptide-targeted dendrimeric prodrug of 5-aminolevulinic acid (ALA) 13 was synthesised which displays nine copies of ALA on a core structure that is linked to a homing peptide for targeted delivery to a specific cancer cell type. The synthesis was accomplished effectively via a flexible, modular solid phase and solution phase route, using a combination of solid phase peptide synthesis and copper-catalysed azide-alkyne cycloaddition chemistry. The prodrug system shows a sustained and enhanced production of protoporphyrin IX (PpIX) in the MDA-MB-231 cell line that over-expresses the epidernal growth factor receptor (EGFR+) in comparison to equimolar ALA and the corresponding non-targeted ALA dendrimer (nine copies of ALA). This study provides a proof of concept for the development of a new generation of prodrugs for ALA-based photodynamic therapy that can deliver an enhanced ALA payload to specific tissue types.
Assuntos
Ácido Aminolevulínico/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Pró-Fármacos , Protoporfirinas/metabolismo , Ácido Aminolevulínico/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Desenho de Fármacos , Humanos , Estrutura Molecular , Fármacos Fotossensibilizantes/química , Relação Estrutura-AtividadeRESUMO
Polyamidoamine PAMAM dendrimer generation 3 (G3) was modified by attachment of biotin via amide bond and glucoheptoamidated by addition of α-D-glucoheptono-1,4-lacton to obtain a series of conjugates with a variable number of biotin residues. The composition of conjugates was determined by detailed 1-D and 2-D NMR spectroscopy to reveal the number of biotin residues, which were 1, 2, 4, 6, or 8, while the number of glucoheptoamide residues substituted most of the remaining primary amine groups of PAMAM G3. The conjugates were then used as host molecules to encapsulate the 5-aminolevulinic acid. The solubility of 5-aminolevulinic acid increased twice in the presence of the 5-mM guest in water. The interaction between host and guest was accompanied by deprotonation of the carboxylic group of 5-aminolevulinic acid and proton transfer into internal ternary nitrogen atoms of the guest as evidenced by a characteristic chemical shift of resonances in the 1H NMR spectrum of associates. The guest molecules were most likely encapsulated inside inner shell voids of the host. The number of guest molecules depended on the number of biotin residues of the host, which was 15 for non-biotin-containing glucoheptoamidated G3 down to 6 for glucoheptoamidated G3 with 8 biotin residues on the host surface. The encapsulates were not cytotoxic against Caco-2 cells up to 200-µM concentration in the dark. All encapsulates were able to deliver 5-aminolevulinic acid to cells but aqueous encapsulates were more active in this regard. Simultaneously, the reactive oxygen species were detected by staining with H2DCFDA in Caco-2 cells incubated with encapsulates. The amount of PpIX was sufficient for induction of reactive oxygen species upon 30-s illumination with a 655-nm laser beam.
Assuntos
Amidas/química , Ácido Aminolevulínico/farmacologia , Biotina/química , Dendrímeros/química , Sistemas de Liberação de Medicamentos , Poliaminas/química , Ácido Aminolevulínico/química , Células CACO-2 , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Dendrímeros/síntese química , Fluorescência , Humanos , Espaço Intracelular/metabolismo , Poliaminas/síntese química , Espectroscopia de Prótons por Ressonância Magnética , Protoporfirinas/farmacologia , Espécies Reativas de Oxigênio/metabolismoRESUMO
Oral lichen planus (OLP) is a chronic inflammatory and immune-mediated disease affecting the oral mucosa. OLP presents with asymptomatic, lacelike white stripes and/or symptomatic red, ulcerated mucous membranes. Eating, drinking and oral hygiene procedures may be painful resulting in reduced quality of life (QOL). The histopathological picture is a zone of cellular infiltrate, mainly CD8+ cells, in the superficial layer of the connective tissue and signs of liquefaction degeneration of the basal membrane. Conventional treatment is corticosteroids. Local and systemic side effects are common, and patients may develop drug resistance. The intention with this article is to demonstrate the heterogeneity in photodynamic therapy (PDT) of OLP. A search in PubMed, Embase (Ovid) and Medline (Ovid) identified seventeen clinical studies investigating PDT of OLP. Only five were randomised controlled studies and the study groups varied from 5 to 50 patients. Five different photosensitisers or precursors were tested. Both broadband spectrum lamps, lasers and light-emitting-diodes (LEDs), with wavelengths from 420 nm to 682 nm, were used. The number of treatment sessions varied from one to ten. The patients were followed up for 0 to 48 months, but in thirteen studies the post treatment observation time was ≤6 months. Single arm studies demonstrated improvement of OLP except in one study. In all controlled studies except for one, PDT was superior or equal to conventional treatment. The majority of patients experienced a slight burning sensation during light activation, but no serious adverse events were reported. Only few studies examined the effect of PDT on a cellular level.
Assuntos
Ácido Aminolevulínico/uso terapêutico , Líquen Plano Bucal/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Ácido Aminolevulínico/química , Humanos , Líquen Plano Bucal/patologia , Fármacos Fotossensibilizantes/químicaRESUMO
BACKGROUND: The BLUE 400 filter system (Carl Zeiss Meditec, Oberkochen, Germany) has provided visualization of 5-ALA-induced fluorescence-guided surgery for more than 20 years. Nevertheless, constraints, e.g., limited background discrimination during hemostasis, obstruct fluency of surgery. A novel filter with improved background visualization was developed, requiring validation regarding fluorescence discrimination. The aim of this article is to determine diagnostic accuracy and perception of protoporphyrin IX (PpIX) discrimination of a novel filter system with higher background illumination (BLUE 400 AR) compared with the gold standard, BLUE 400. METHODS: A surgical microscope equipped with both BLUE 400 and BLUE 400 AR was used. Comparisons were performed on a biological basis and on the visual perception of margins. High-resolution images were compared during and after surgery by senior neurosurgeons. In a predefined biopsy algorithm, four biopsies per patient at tumor margins of PpIX fluorescence and adjacent brain were acquired using BLUE 400 AR only from regions intended for resection and assessed for cell count and density. RESULTS: Thirty-two patients with malignant gliomas were included in this study. BLUE 400 AR markedly enhanced the brightness of the surgical field, allowing superior discrimination of brain anatomy. A total of 128 biopsies from fluorescence margins were collected. Positive predictive value (PPV) was 98.44% (95% CI, 90.06-99.77%) for malignant glioma. Residual median cell density in non-fluorescent tissue was 13% (IQR 13 to 31). Perception of the location of fluorescent margins on HD images was equivalent for both filter combinations. CONCLUSIONS: BLUE 400 AR demonstrated superior background compared with conventional BLUE 400 in malignant glioma surgery but comparable fluorescence margins and PPV. Therefore, BLUE 400 AR can be considered safe and effective in supporting malignant glioma surgery.
Assuntos
Ácido Aminolevulínico/química , Neoplasias Encefálicas/cirurgia , Glioma/cirurgia , Procedimentos Neurocirúrgicos/instrumentação , Procedimentos Neurocirúrgicos/métodos , Fármacos Fotossensibilizantes/química , Protoporfirinas/química , Cirurgia Assistida por Computador/instrumentação , Cirurgia Assistida por Computador/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Contagem de Células , Feminino , Fluorescência , Humanos , Masculino , Margens de Excisão , Microscopia de Fluorescência , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos ProspectivosRESUMO
Photodynamic therapy is a non-invasive method where light activates a photosensitizer bound to cancer cells, generating reactive oxygen species and resulting in cell death. This study assessed the oncolytic potential of photodynamic therapy, comparing European Medicines Agency and United States Food and Drug Administration-approved 5-aminolevulinic acid (5-ALA) to a metalloporphyrin, Pd(T4), against a highly invasive uveal melanoma cell line (C918) in two- and three-dimensional models in vitro. Epithelial monolayer studies displayed strong oncolytic effects (>70%) when utilizing Pd(T4) at a fraction of the concentration, and reduced pre-illumination time compared to 5-ALA post-405 nm irradiance. When analyzed at sub-optimal concentrations, application of Pd(T4) and 5-ALA with 405 nm displayed cumulative effects. Lethality from Pd(T4)-photodynamic therapy was maintained within a three-dimensional model, including the more resilient vasculogenic mimicry-forming cells, though at lower rates. At high concentrations, modality of cell death exhibited necrosis partially dependent on reactive oxygen species. However, sub-optimal concentrations of photosensitizer exhibited an apoptotic protein expression profile characterized by increased Bax/Bcl-2 ratio and endoplasmic stress-related proteins, along with downregulation of apoptotic inhibitors CIAP-1 and -2. Together, our results indicate Pd(T4) as a strong photosensitizer alone and in combination with 5-ALA against C918 cells.
Assuntos
Ácido Aminolevulínico/farmacologia , Antineoplásicos/farmacologia , Melanoma/tratamento farmacológico , Metaloporfirinas/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Neoplasias Uveais/tratamento farmacológico , Ácido Aminolevulínico/química , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Luz , Metaloporfirinas/química , Necrose/tratamento farmacológico , Fármacos Fotossensibilizantes/química , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína X Associada a bcl-2/metabolismoRESUMO
Photodynamic therapy (PDT) of bacterial strains presents an attractive potential alternative to antibiotic therapies in search of the solution for the chemoresistance problem. The efficacy of the treatment is dependent on the interaction of photochemically active substances called photosensitizers (PSs) with the bacterial cell wall or their intracellular accumulation. In addition to exogenous PSs, other molecules such as 5-aminolevulinic acid (5-ALA), a natural precursor of heme, are gaining interest. When provided exogenously to cells, 5-ALA uptake results in the overproduction of various photoactive porphyrins. The pattern of their intracellular accumulation and release to the surroundings depends on incubation conditions such as the applied 5-ALA concentration, cell density and incubation duration. The detection of endogenously synthesized porphyrins in samples of Salmonella enterica cells and supernatants was accomplished after 4 h and 20 h incubation periods by means of fluorescence spectroscopy. The relative proportions of different types of porphyrins were assessed by modeling the registered spectra with the fluorescence spectra of standard porphyrins. After the shorter incubation period, the dominant porphyrins in the supernatant medium were coproporphyrins. The longer incubation period shifted the relative proportion of intracellular porphyrins from protoporphyrin IX towards water-soluble porphyrins such as uroporphyrin I, which interfered with additional by-products. The time-dependent changes in compositions of both intracellular and extracellular porphyrins imply that 5-ALA-induced sensitization might have triggered a complex protective mechanism of bacterial cells. Thus, identification and evaluation of the relative amounts of porphyrins, which accumulate in bacterial cells and are extruded outside after different time periods, could provide access to valuable information, working towards more efficient applications of 5-ALA-based antibacterial PDT.
Assuntos
Ácido Aminolevulínico/química , Fármacos Fotossensibilizantes/química , Porfirinas/química , Salmonella enterica/química , Espectrometria de Fluorescência , Ácido Aminolevulínico/farmacologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/metabolismo , Salmonella enterica/efeitos dos fármacosRESUMO
Photodynamic diagnosis (PDD) can improve diagnostic accuracy by using PDD agents such as 5-aminolevulinic acid (ALA). However, the weakness and photobleaching of fluorescence of PDD agents may lead to insufficient fluorescence visibility for the detection of cancer during resection operations. We focused on the "fluorescence enhancement effect" resulting from the addition of polyethylene glycol-modified titanium dioxide nanoparticles (TiO2-PEG NPs) to address these problems. The results showed that the combined administration of TiO2-PEG NPs and ALA could enhance and prolong fluorescence in bladder cancer cells, similar to in the mixture alone. It was suggested that the fluorescence enhancement was related to the accumulation of TiO2-PEG NPs in cells via endocytosis, causing the light scattering and enhancement of fluorescence. This fluorescence enhancement effect could be applicable for PDD.
Assuntos
Ácido Aminolevulínico/farmacologia , Neoplasias/diagnóstico , Titânio/farmacologia , Neoplasias da Bexiga Urinária/diagnóstico , Ácido Aminolevulínico/química , Linhagem Celular Tumoral , Fluorescência , Humanos , Nanopartículas/química , Neoplasias/patologia , Polietilenoglicóis/química , Titânio/química , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/patologiaRESUMO
Metastatic melanoma (MM) has a poor prognosis and is attributed to late diagnoses only when metastases has already occurred. Thus, early diagnosis is crucial to improve its overall treatment efficacy. The standard diagnostic tools for MM are incisional biopsies and/or fine needle aspiration biopsies, while standard treatments involve surgery, chemotherapy, or irradiation therapy. The combination of photodynamic diagnosis (PDD) and therapy (PDT) utilizes a photosensitizer (PS) that, when excited by light of a low wavelength, can be used for fluorescent non-destructive diagnosis. However, when the same PS is activated at a higher wavelength of light, it can be cytotoxic and induce tumor destruction. This paper focuses on PS drugs that have been used for PDD as well as PDT treatment of MM. Furthermore, it emphasizes the need for continued investigation into enhanced PS delivery via active biomarkers and passive nanoparticle systems. This should improve PS drug absorption in MM cells and increase effectiveness of combinative photodynamic methods for the enhanced diagnosis and treatment of MM can become a reality.
Assuntos
Melanoma/diagnóstico por imagem , Melanoma/tratamento farmacológico , Fármacos Fotossensibilizantes/uso terapêutico , Neoplasias Cutâneas/diagnóstico por imagem , Neoplasias Cutâneas/tratamento farmacológico , Ácido Aminolevulínico/química , Ácido Aminolevulínico/farmacocinética , Ácido Aminolevulínico/uso terapêutico , Antracenos , Biópsia por Agulha Fina , Portadores de Fármacos/síntese química , Diagnóstico Precoce , Humanos , Indóis/química , Indóis/farmacocinética , Indóis/uso terapêutico , Isoindóis , Luz , Metástase Linfática , Melanoma/patologia , Imagem Molecular/métodos , Nanopartículas/administração & dosagem , Nanopartículas/química , Perileno/análogos & derivados , Perileno/química , Perileno/farmacocinética , Perileno/uso terapêutico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacocinética , Neoplasias Cutâneas/patologiaRESUMO
Fluorescence-guided surgery (FGS) has been established as a powerful technique for glioblastoma resection. After oral application of the prodrug 5-aminolevulinic acid (5-ALA), protoporphyrin IX (PpIX) is formed as an intermediate of the heme-biosynthesis cascade and accumulates within the tumor. By intraoperative fluorescence microscopy, the specific PpIX fluorescence can be used to differentiate the tumor from healthy brain tissue. To investigate possible limitations of fluorescence diagnosis, the complementary use of molecular and elemental mass-spectrometry imaging (MSI) is presented. Matrix-assisted laser-desorption-ionization mass spectrometry (MALDI-MS) is used to examine the distribution of PpIX and heme b in human brain tumors. MALDI-MS/MS imaging is performed to validate MS data and improve the signal-to-noise ratio (S/N). Comparing the imaging results with histological evaluation, increased PpIX accumulation in areas of high tumor-cell density is observed. Heme b accumulation are only found in areas of blood vessels and hemorrhage, confirming the hampered transformation from PpIX to heme b in glioblastoma tissue. Investigation of non-neoplastic brain tissue and glioblastoma resected without external 5-ALA administration as control samples with true-negative fluorescence verified the absence of PpIX accumulation. Analysis of necrotic tumor tissue and gliosarcoma, one rare type of glioma appearing nonfluorescent during FGS, as case examples with false-negative-fluorescence diagnosis, revealed the absence of significant amounts of PpIX, indicating an impairment of PpIX formation. Molecular analysis is complemented by quantitative laser ablation-inductively coupled plasma (LA-ICP) MSI correlating heme b and Fe distribution. Mathematical pixel-by-pixel correlation of molecular and elemental data revealed a positive correlation with heteroscedasticity for the spatially resolved heme b signal intensities and Fe concentrations.
Assuntos
Neoplasias Encefálicas/diagnóstico por imagem , Fluorescência , Glioblastoma/diagnóstico por imagem , Imagem Óptica , Ácido Aminolevulínico/química , Ácido Aminolevulínico/metabolismo , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/cirurgia , Glioblastoma/metabolismo , Glioblastoma/cirurgia , Humanos , Terapia a Laser , Espectrometria de Massas , Microscopia de Fluorescência , Conformação Molecular , Pró-Fármacos/química , Pró-Fármacos/metabolismo , Protoporfirinas/química , Protoporfirinas/metabolismoRESUMO
The administration of 5-aminolevulinic acid (ALA) to generate enhanced intracellular levels of endogenous porphyrins is currently one of the most important approaches for photodynamic therapy (PDT) and photodiagnosis (PDD). Despite the great promise of ALA-based PDT, the physicochemical behaviour and chemical reactivity of ALA are problematic, and a variety of chemical approaches have been brought to bear to improve cellular delivery, enhance porphyrin production, and generate ALA prodrugs that have appropriate stability for convenient clinical use, as well as selectivity for cancerous tissues. While there has been considerable success, there are still a number of challenges to be addressed and opportunities to be exploited through application of chemical insight in this area.
Assuntos
Ácido Aminolevulínico/farmacologia , Antineoplásicos/farmacologia , Neoplasias/diagnóstico , Neoplasias/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Ácido Aminolevulínico/administração & dosagem , Ácido Aminolevulínico/química , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Humanos , Neoplasias/patologia , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/químicaRESUMO
Gold nanoparticles (AuNPs) have been extensively studied within biomedicine due to their biocompatibility and low toxicity. In particular, AuNPs have been widely used to deliver photosensitiser agents for photodynamic therapy (PDT) of cancer. Here we review the state-of-the-art for the functionalisation of the gold nanoparticle surface with both photosensitisers and targeting ligands for the active targeting of cancer cell surface receptors. From the initial use of the AuNPs as a simple carrier of the photosensitiser for PDT, the field has significantly advanced to include: the use of PEGylated modification to provide aqueous compatibility and stealth properties for in vivo use; gold metal-surface enhanced singlet oxygen generation; functionalisation of the AuNP surface with biological ligands to specifically target over-expressed receptors on the surface of cancer cells and; the creation of nanorods and nanostars to enable combined PDT and photothermal therapies. These versatile AuNPs have significantly enhanced the efficacy of traditional photosensitisers for both in vitro and in vivo cancer therapy. From this review it is apparent that AuNPs have an important future in the treatment of cancer.
Assuntos
Ácido Aminolevulínico/farmacologia , Antineoplásicos/farmacologia , Ouro/farmacologia , Nanopartículas Metálicas/química , Neoplasias/tratamento farmacológico , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Ácido Aminolevulínico/administração & dosagem , Ácido Aminolevulínico/química , Antineoplásicos/administração & dosagem , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Ouro/química , Humanos , Neoplasias/patologia , Fármacos Fotossensibilizantes/administração & dosagem , Fármacos Fotossensibilizantes/químicaRESUMO
Treatment of cancer cells by clinically approved hexyl ester of 5-aminolevulinic acid (ALA-Hex) induces accumulation of fluorescent porphyrins in tumors. This allows fluorescence photodiagnosis (PD) of bladder cancer by blue light illumination. However, PD of other cancers is hampered by acute toxicity of the compound limiting its use to local applications. We have designed and synthesized a new prodrug of ALA-Hex that tackles the stability-activity paradox of amino-modified 5-ALA prodrugs. The glucuronide prodrug Glu-ALA-Hex demonstrates excellent stability under physiological conditions and activation in the presence of the target enzyme. ß-glucuronidase-triggered release of 5-ALA is programmed to yield fluorescence in tumor environment with elevated ß-glucuronidase activity, a characteristic of many solid tumors. Glu-ALA-Hex produces similar levels of fluorescence as ALA-Hex in breast cancer MCF7 cells in vitro but with much lower non-specific cell toxicity.
Assuntos
Ácido Aminolevulínico/farmacologia , Antineoplásicos/farmacologia , Inibidores Enzimáticos/farmacologia , Glucuronidase/antagonistas & inibidores , Fármacos Fotossensibilizantes/farmacologia , Pró-Fármacos/farmacologia , Ácido Aminolevulínico/síntese química , Ácido Aminolevulínico/química , Antineoplásicos/síntese química , Antineoplásicos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Fluorescência , Glucuronidase/metabolismo , Humanos , Células MCF-7 , Estrutura Molecular , Fármacos Fotossensibilizantes/síntese química , Fármacos Fotossensibilizantes/química , Pró-Fármacos/síntese química , Pró-Fármacos/química , Relação Estrutura-Atividade , Células Tumorais CultivadasRESUMO
Osmotic stress induced with 1 M sorbitol inhibited δ-aminolevulinic acid dehydratase (ALAD) and aminolevulinic acid (ALA) synthesizing activities in etiolated maize leaf segments during greening; the ALAD activity was inhibited to a greater extent than the ALA synthesis. When the leaves were exposed to light, the ALAD activity increased for the first 8 h, followed by a decrease observed at 16 and 24 h in both sorbitol-treated and untreated leaf tissues. The maximum inhibition of the enzyme activity was observed in the leaf segments incubated with sorbitol for 4 to 8 h. Glutamate increased the ALAD activity in the in vitro enzymatic preparations obtained from the sorbitol-treated leaf segments; sorbitol inhibited the ALAD activity in the preparations from both sorbitol-treated and untreated leaves. It was suggested that sorbitol-induced osmotic stress inhibits the enzyme activity by affecting the ALAD induction during greening and regulating the ALAD steady-state level of ALAD in leaf cells. The protective effect of glutamate on ALAD in the preparations from the sorbitol-treated leaves might be due to its stimulatory effect on the enzyme.
Assuntos
Pressão Osmótica/efeitos dos fármacos , Folhas de Planta/efeitos dos fármacos , Sintase do Porfobilinogênio/antagonistas & inibidores , Sorbitol/farmacologia , Zea mays/efeitos dos fármacos , Ácido Aminolevulínico/química , Ácido Aminolevulínico/metabolismo , Relação Dose-Resposta a Droga , Folhas de Planta/metabolismo , Sintase do Porfobilinogênio/metabolismo , Relação Estrutura-Atividade , Zea mays/metabolismoRESUMO
Actinic keratosis (AK) is a premalignant epithelial disease that develops on skin chronically exposed to ultraviolet-light. For outdoor workers, AK is an occupational hazard with a potential for malignant transformation into squamous cell carcinoma. Various topical treatments are used to treat AK's. Photodynamic therapy (PDT) promises the highest cure rates. We performed a prospective single center trial evaluating effects of PDT treatment for field cancerization due to occupational exposure to ultraviolet light irradiation in patients unresponsive to the treatment regimen with topical diclofenac gel. For PDT, a nanoemulsified 5-aminolevulinic acid (5-ALA) formulation was combined with narrow band red light irradiation with a peak wavelength of 635 nm and a total light intensity of 37 J/cm2. Clearance rate and adverse events were recorded. Thirty patients with a mean age of 77±10.5 years (26 males and 4 females) were included in the trial. They received between one and nine PDT sessions. Mean pain during PDT was scored at 6.8±2.8 using the visual analogue scale. Patients exhibited a mild immediate erythema after PDT. Among the patients, who tolerated PDT, a complete clearance was achieved in 93.3%, and >90% clearance - in 6.7%. Pain management was performed by cold-air analgesia. In conclusion, PDT with nanoemulsified 5-ALA and narrow band red light irradiation is an effective and safe treatment for occupational field cancerization without systemic toxicity. Adherence to this treatment is much better than to topical treatment.