RESUMO
A cross-linking technique involving application of Bacteriochlorophyll Derivative WST-11 mixed with dextran (WST-D) to the epithelium-debrided cornea and illumination with Near Infrared (NIR), has been identified as a promising therapy for stiffening pathologically weakened corneas. To investigate its effect on corneal collagen architecture, x-ray scattering and electron microscopy data were collected from paired WST-D/NIR treated and untreated rabbit corneas. The treated eye received 2.5 mg/mL WST-D and was illuminated by a NIR diode laser (755 nm, 10 mW/cm2). An increase in corneal thickness (caused by corneal oedema) occurred at 1-day post-treatment but resolved in the majority of cases within 4 days. The epithelium was fully healed after 6-8 days. X-ray scattering revealed no difference in average collagen interfibrillar spacing, fibril diameter, D-periodicity or intermolecular spacing between treated and untreated specimens. Similarly, electron microscopy images of the anterior and posterior stroma in healed WST-D/NIR corneas and untreated controls revealed no obvious differences in collagen organisation or fibril diameter. As the size and organisation of stromal collagen is closely associated with the optical properties of the cornea, the absence of any large-scale changes following treatment confirms the potential of WST-D/NIR therapy as a means of safely stiffening the cornea.
Assuntos
Bacterioclorofilas/farmacologia , Substância Própria/efeitos dos fármacos , Substância Própria/efeitos da radiação , Raios Infravermelhos , Animais , Colágeno/metabolismo , Substância Própria/metabolismo , Substância Própria/ultraestrutura , Lasers , Microscopia Eletrônica , CoelhosRESUMO
Photodynamic therapy (PDT) is an approved medical technique to treat certain forms of cancer. It has been used to complement traditional anticancer modalities such as surgery, chemotherapy or radiotherapy, and in certain cases, to replace these treatments. One critical parameter of PDT is the photosensitizer (PS); historically, a purely organic macrocyclic tetrapyrrole-based structure. This short review surveys two recent clinical examples of metal complexes, namely TOOKAD®-Soluble and TLD-1433, which have ideal photophysical properties to act as PDT PSs. We highlight the important role played by the metal ions in the PS for PDT activity.
Assuntos
Antineoplásicos/química , Complexos de Coordenação/química , Neoplasias/terapia , Fármacos Fotossensibilizantes/química , Antineoplásicos/farmacologia , Bacterioclorofilas/química , Bacterioclorofilas/farmacologia , Terapia Combinada , Complexos de Coordenação/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Solubilidade , Relação Estrutura-AtividadeRESUMO
PURPOSE: The aim of this study is to determine the effect of variation of the exposure time of near-infrared irradiation on corneal stiffening after a bacteriochlorophyll derivative (WST11) with dextran (WST-D) application. METHODS: One hundred four paired eyes of 3-month-old New Zealand White rabbits were included in this study. Fifty-two eyes (ex vivo n = 34, in vivo n = 18) were mechanically deepithelialized, treated topically with WST-D, and irradiated at 10 mW/cm using a diode laser at 755 nm for 1, 5, or 30 minutes. Untreated fellow eyes served as controls. Corneoscleral rings were removed immediately after treatment (ex vivo), or 1 month after treatment (in vivo). Corneal strips were cut and underwent biomechanical stress-strain measurements. RESULTS: Ex vivo, the mean tangent elastic modulus was significantly higher in the treatment groups than in the control groups for 1, 5, and 30 minutes of irradiation, respectively, 6.06 MPa, 95% confidence interval (CI, 4.5-7.6) versus 14.02 MPa, 95% CI (10.2-17.8), n = 11, 4.8 MPa, 95% CI (3.9-5.7) versus 15.03 MPa, 95% CI (12-18.1), n = 11, and 7.8 MPa, 95% CI (5.6-10.02) versus 16.2 MPa, 95% CI (13.6-18.9), n = 11; P < 0.001 for all comparisons. In vivo, the mean elastic moduli in the treatment groups were significantly higher for 5 and 30 minutes of irradiation but not for 1 minute of irradiation, respectively, 11.4 MPa, 95% CI (8.5-14.2), versus 17.1 MPa, 95% CI (14.5-19.7), n = 5; P < 0.001, and 9.4 MPa, 95% CI (5.1-13.8) versus 16 MPa, 95% CI (13.1-19), n = 5; P < 0.01, and 11.3 MPa, 95% CI (6-16.6) versus 12.2 MPa, 95% CI (7.5-16.8), n = 5; P = 0.7. CONCLUSIONS: WST-D/near-infrared treatment using shortened irradiation time (1 minute ex vivo and 5 minutes in vivo) results in significant corneal stiffening, and this might provide an alternative to the currently applied riboflavin/ultraviolet A cross-linking.
Assuntos
Bacterioclorofilas/farmacologia , Córnea/efeitos dos fármacos , Reagentes de Ligações Cruzadas , Dextranos/farmacologia , Raios Infravermelhos , Fármacos Fotossensibilizantes/farmacologia , Animais , Fenômenos Biomecânicos , Colágeno/metabolismo , Córnea/metabolismo , Córnea/fisiopatologia , Paquimetria Corneana , Substância Própria/metabolismo , Combinação de Medicamentos , Coelhos , Fatores de TempoRESUMO
Purpose To examine the hypothesis that vascular-targeted photodynamic therapy (VTP) with WST11 and clinically relevant parameters can be used to ablate target tissues in a non-tumor-bearing large-animal model while selectively sparing blood vessels and collagen. Materials and Methods By using an institutional animal care and use committee-approved protocol, 68 ablations were performed in the kidneys (cortex and medulla) and livers of 27 adult pigs. Posttreatment evaluation was conducted with contrast material-enhanced computed tomography in the live animals at 24 hours. Immunohistochemistry was evaluated and histologic examination with hematoxylin-eosin staining was performed at 4 hours, 24 hours, and 7 days. Intravenous infusion of WST11 (4 mg per kilogram of body weight) was followed by using near-infrared illumination (753 nm for 20 minutes) through optical fibers prepositioned in target tissues by using a fixed template. Treated areas were scanned, measured, and statistically analyzed by using the Student t test and two-way analysis of variance. Results Focal WST11 VTP treatment in the liver and kidney by using a single optical fiber resulted in well-demarcated cylindrical zones of nonthermal necrosis concentrically oriented around the light-emitting diffuser, with no intervening viable parenchymal cells. The radius of ablated tissue increased from approximately 5 mm at 150 mW to approximately 7 mm at 415 mW (P < .01). Illumination through fiber triads at 1-cm separation resulted in confluent homogeneous necrosis. Patterns of acute injury within 24 hours were consistent with microcirculatory flow arrest and collagen preservation (demonstrated with trichrome staining). In the peripheral ablation zone, blood vessels at least 40 µm in diameter were selectively preserved and remained functional at 7 days. Ablated tissues exhibited progressive fibrosis and chronic inflammatory cell infiltrates. No histologic changes consistent with thermal injury were observed in blood vessels or collagen. The renal hilum and collecting system did not show treatment effect, despite treatment proximity. Conclusion WST11 VTP induces nonthermal tissue ablation in target tissue while preserving critical organ structures and bystander blood vessels within solid organs. (©) RSNA, 2016 Online supplemental material is available for this article.
Assuntos
Bacterioclorofilas/farmacologia , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fotoquimioterapia/métodos , Espécies Reativas de Oxigênio/metabolismo , Animais , Meios de Contraste , Feminino , Imuno-Histoquímica , Rim/diagnóstico por imagem , Fígado/diagnóstico por imagem , Modelos Animais , Necrose , Fibras Ópticas , Suínos , Tomografia Computadorizada por Raios XRESUMO
Since photodynamic therapy emerged as a promising cancer treatment, the development of photosensitizers has gained great interest. In this context, the photosynthetic pigments, chlorophylls and bacteriochlorophylls, as excellent natural photosensitizers, attracted much attention. In effect, several (bacterio) chlorophyll-based phototherapeutic agents have been developed and (or are about to) enter the clinics. The aim of this review article is to give a survey of the advances in the synthetic chemistry of these pigments which have been made over the last decade, and which are pertinent to the application of their derivatives as photosensitizers for photodynamic therapy (PDT). The review focuses on the synthetic strategies undertaken to obtain novel derivatives of (bacterio)chlorophylls with both enhanced photosensitizing and tumorlocalizing properties, and also improved photo- and chemical stability. These include modifications of the C- 17-ester moiety, the isocyclic ring, the central binding pocket, and the derivatization of peripheral functionalities at the C-3 and C-7 positions with carbohydrate-, peptide-, and nanoparticle moieties or other residues. The effects of these modifications on essential features of the pigments are discussed, such as the efficiency of reactive oxygen species generation, photostability, phototoxicity and interactions with living organisms. The review is divided into several sections. In the first part, the principles of PDT and photosensitizer action are briefly described. Then the relevant photophysical features of (bacterio)chlorophylls and earlier approaches to their modification are summarized. Next, a more detailed overview of the progress in synthetic methods is given, followed by a discussion of the effects of these modifications on the photophysics of the pigments and on their biological activity.
Assuntos
Bacterioclorofilas/química , Bacterioclorofilas/farmacologia , Clorofila/química , Clorofila/farmacologia , Descoberta de Drogas/métodos , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/farmacologia , Animais , Bacterioclorofilas/uso terapêutico , Clorofila/uso terapêutico , Humanos , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêuticoRESUMO
With the knowledge that the dominant photodynamic therapy (PDT) mechanism of 1a (WST09) switched from type 2 to type 1 for 1b (WST11) upon taurine-driven E-ring opening, we hypothesized that taurine-driven E-ring opening of bacteriochlorophyll derivatives and net-charge variations would modulate reactive oxygen species (ROS) photogeneration. Eight bacteriochlorophyll a derivatives were synthesized with varying charges that either contained the E ring (2a-5a) or were synthesized by taurine-driven E-ring opening (2b-5b). Time-dependent density functional theory (TDDFT) modeling showed that all derivatives would be type 2 PDT-active, and ROS-activated fluorescent probes were used to investigate the photogeneration of a combination of type 1 and type 2 PDT ROS in organic- and aqueous-based solutions. These investigations validated our predictive modeling calculations and showed that taurine-driven E-ring opening and increasing negative charge generally enhanced ROS photogeneration in aqueous solutions. We propose that these structure-activity relationships may provide simple strategies for designing bacteriochlorins that efficiently generate ROS upon photoirradiation.
Assuntos
Bacterioclorofilas/síntese química , Espécies Reativas de Oxigênio/metabolismo , Bacterioclorofilas/farmacologia , Modelos Moleculares , Processos Fotoquímicos , Fotoquimioterapia , Relação Estrutura-AtividadeRESUMO
The aim of this study was to assess the red autofluorescence from bacterial species related to dental caries and periodontitis in the presence of different nutrients in the growth medium. Bacteria were grown anaerobically on tryptic soy agar (TSA) supplemented with nutrients, including magnesium-porphyrins from spinach and iron-porphyrins from heme. The autofluorescence was then assessed at 405 nm excitation. On the TSA without additives, no autofluorescence was observed from any of the species tested. On the TSA containing sheep blood, red autofluorescence was observed only from Parvimonas micra. When the TSA was supplemented with blood, hemin, and vitamin K, red autofluorescence was observed from Actinomyces naeslundii, Bifidobacterium dentium, and Streptococcus mutans. Finally, on the TSA supplemented with spinach extract, red autofluorescence was observed from Aggregatibacter actinomycetemcomitans, A. naeslundii, Enterococcus faecalis, Fusobacterium nucleatum, Lactobacillus salivarius, S. mutans, and Veillonella parvula. We conclude that the bacteria related to dental caries and periodontal disease exhibit red autofluorescence. The autofluorescence characteristics of the tested strains depended on the nutrients present, such as metalloporphyrins, suggesting that the metabolic products of the oral biofilm could be responsible for red autofluorescence.
Assuntos
Bactérias Anaeróbias/efeitos dos fármacos , Bactérias Anaeróbias/efeitos da radiação , Bacterioclorofilas/farmacologia , Meios de Cultura/química , Placa Dentária/microbiologia , Fluorescência , Heme/farmacologia , Bactérias Anaeróbias/metabolismo , Bifidobacterium/fisiologia , Biofilmes/efeitos dos fármacos , Biofilmes/efeitos da radiação , Cárie Dentária/diagnóstico , Cárie Dentária/microbiologia , Luz , Periodontite/microbiologia , Streptococcus mutans/fisiologiaRESUMO
We examined the effect of the oxygenation level on efficacy of two photosensitizing agents, both of which target lysosomes for photodamage, but via different photochemical pathways. Upon irradiation, the chlorin termed NPe6 forms singlet oxygen in high yield while the bacteriopheophorbide WST11 forms only oxygen radicals (in an aqueous environment). Photokilling efficacy by WST11 in cell culture was impaired when the atmospheric oxygen concentration was reduced from 20% to 1%, while photokilling by NPe6 was unaffected. Studies in a cell-free system revealed that the rates of photobleaching of these agents, as a function of the oxygenation level, were correlated with results described above. Moreover, the rate of formation of oxygen radicals by either agent was more sensitive to the level of oxygenation than was singlet oxygen formation by NPe6. These data indicate that the photochemical process that leads to oxygen radical formation is more dependent on the oxygenation level than is the pathway leading to formation of singlet oxygen.
Assuntos
Bacterioclorofilas/química , Hepatócitos/efeitos dos fármacos , Oxigênio/química , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Porfirinas/química , Espécies Reativas de Oxigênio/química , Animais , Bacterioclorofilas/farmacologia , Linhagem Celular Tumoral , Hepatócitos/citologia , Hepatócitos/metabolismo , Luz , Camundongos , Fotodegradação , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologiaRESUMO
Photodynamic therapy (PDT) involves photosensitizing agents that, in the presence of oxygen and light, initiate formation of cytotoxic reactive oxygen species (ROS). PDT commonly induces both apoptosis and autophagy. Previous studies with murine hepatoma 1c1c7 cells indicated that loss of autophagy-related protein 7 (ATG7) inhibited autophagy and enhanced the cytotoxicity of photosensitizers that mediate photodamage to mitochondria or the endoplasmic reticulum. In this study, we examined two photosensitizing agents that target lysosomes: the chlorin NPe6 and the palladium bacteriopheophorbide WST11. Irradiation of wild-type 1c1c7 cultures loaded with either photosensitizer induced apoptosis and autophagy, with a blockage of autophagic flux. An ATG7- or ATG5-deficiency suppressed the induction of autophagy in PDT protocols using either photosensitizer. Whereas ATG5-deficient cells were quantitatively similar to wild-type cultures in their response to NPe6 and WST11 PDT, an ATG7-deficiency suppressed the apoptotic response (as monitored by analyses of chromatin condensation and procaspase-3/7 activation) and increased the LD(50) light dose by > 5-fold (as monitored by colony-forming assays). An ATG7-deficiency did not prevent immediate lysosomal photodamage, as indicated by loss of the lysosomal pH gradient. However, unlike wild-type and ATG5-deficient cells, the lysosomes of ATG7-deficient cells recovered this gradient within 4 h of irradiation, and never underwent permeabilization (monitored as release of endocytosed 10-kDa dextran polymers). We propose that the efficacy of lysosomal photosensitizers is in part due to both promotion of autophagic stress and suppression of autophagic prosurvival functions. In addition, an effect of ATG7 unrelated to autophagy appears to modulate lysosomal photodamage.
Assuntos
Apoptose/efeitos da radiação , Luz , Lisossomos/metabolismo , Lisossomos/efeitos da radiação , Proteínas Associadas aos Microtúbulos/deficiência , Aminas/metabolismo , Animais , Apoptose/efeitos dos fármacos , Proteína 7 Relacionada à Autofagia , Bacterioclorofilas/química , Bacterioclorofilas/farmacologia , Linhagem Celular Tumoral , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Núcleo Celular/efeitos da radiação , Núcleo Celular/ultraestrutura , Forma Celular/efeitos dos fármacos , Forma Celular/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Fluorescência , Proteínas de Fluorescência Verde/metabolismo , Lisossomos/efeitos dos fármacos , Camundongos , Proteínas Associadas aos Microtúbulos/metabolismo , Permeabilidade/efeitos dos fármacos , Permeabilidade/efeitos da radiação , Fagossomos/efeitos dos fármacos , Fagossomos/efeitos da radiação , Fagossomos/ultraestrutura , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , Transporte Proteico/efeitos dos fármacos , Transporte Proteico/efeitos da radiação , Proteínas Recombinantes de Fusão/metabolismo , Frações Subcelulares/efeitos dos fármacos , Frações Subcelulares/metabolismo , Frações Subcelulares/efeitos da radiação , Vacúolos/efeitos dos fármacos , Vacúolos/metabolismo , Vacúolos/efeitos da radiaçãoRESUMO
PURPOSE: We evaluated the efficacy and safety of photochemical corneal stiffening by palladium bacteriochlorin 13'-(2-sulfoethyl)amide dipotassium salt (WST11) and near infrared (NIR) illumination, using ex vivo and in vivo rabbit eye models. METHODS: Corneas of post mortem rabbits and living rabbits were pretreated topically with 2.5 mg/mL WST11 in saline or in 20% dextran T-500 (WST-D), washed and illuminated with an NIR diode laser (755 nm, 10 mW/cm(2). Studies with corneas of untreated fellow eyes served as controls. Tensile strength measurements, histopathology, electron spin resonance, and optical spectroscopy and fluorescence microscopy were used to assess treatment effects. Comparative studies were performed with standard riboflavin/ultraviolet-A light (UVA) treatment. RESULTS: WST11/NIR treatment significantly increased corneal stiffness following ex vivo or in vivo treatment, compared to untreated contralateral eyes. The incremental ultimate stress and Young's modulus of treated corneas increased by 45, 113, 115%, and 10, 79, and 174% following 10, 20, and 30 minutes of incubation with WST11, respectively. WST-D/NIR had a similar stiffening effect, but markedly reduced post-treatment edema and shorter time of epithelial healing. WST11/NIR and WST-D/NIR generate hydroxyl and superoxide radicals, but no singlet oxygen in the cornea. Histology demonstrated a reduction in the keratocyte population in the anterior half of the corneal stroma, without damage to the endothelium. CONCLUSIONS: Treatment of rabbit corneas, with either WST11/NIR or WST-D/NIR, increases their biomechanical strength through a mechanism that does not involve singlet oxygen. The WST-D/NIR treatment showed less adverse effects, demonstrating a new potential for clinical use in keratoconus and corneal ectasia after refractive surgery.
Assuntos
Bacterioclorofilas/farmacologia , Córnea , Fototerapia/métodos , Resistência à Tração/efeitos dos fármacos , Resistência à Tração/efeitos da radiação , Animais , Bacterioclorofilas/farmacocinética , Fenômenos Biomecânicos/efeitos dos fármacos , Fenômenos Biomecânicos/fisiologia , Córnea/efeitos dos fármacos , Córnea/fisiologia , Córnea/efeitos da radiação , Ceratócitos da Córnea/efeitos dos fármacos , Ceratócitos da Córnea/fisiologia , Ceratócitos da Córnea/efeitos da radiação , Substância Própria/efeitos dos fármacos , Substância Própria/fisiologia , Substância Própria/efeitos da radiação , Espectroscopia de Ressonância de Spin Eletrônica , Endotélio Corneano/efeitos dos fármacos , Endotélio Corneano/fisiologia , Endotélio Corneano/efeitos da radiação , Raios Infravermelhos/uso terapêutico , Lasers Semicondutores , Modelos Animais , Fotodegradação/efeitos dos fármacos , Fármacos Fotossensibilizantes/farmacologia , Coelhos , Espectrometria de Fluorescência , Estresse Mecânico , Resistência à Tração/fisiologiaRESUMO
Vascular-targeted photodynamic therapy (VTP) takes advantage of intravascular excitation of a photosensitizer (PS) to produce cytotoxic reactive oxygen species (ROS). These ROS are potent mediators of vascular damage inducing rapid local thrombus formation, vascular occlusion, and tissue hypoxia. This light-controlled process is used for the eradication of solid tumors with Pd-bacteriochlorophyll derivatives (Bchl) as PS. Unlike classical photodynamic therapy (PDT), cancer cells are not the primary target for VTP but instead are destroyed by treatment-induced oxygen deprivation. VTP initiates acute local inflammation inside the illuminated area accompanied by massive tumor tissue death. Consequently, in the present study, we addressed the possibility of immune response induction by the treatment that may be considered as an integral part of the mechanism of VTP-mediated tumor eradication. The effect of VTP on the host immune system was investigated using WST11, which is now in phase II clinical trials for age-related macular degeneration and intended to be evaluated for cancer therapy. We found that a functional immune system is essential for successful VTP. Long-lasting systemic antitumor immunity was induced by VTP involving both cellular and humoral components. The antitumor effect was cross-protective against mismatched tumors, suggesting VTP-mediated production of overlapping tumor antigens, possibly from endothelial origin. Based on our findings we suggest that local VTP might be utilized in combination with other anticancer therapies (e.g., immunotherapy) for the enhancement of host antitumor immunity in the treatment of both local and disseminated disease.
Assuntos
Formação de Anticorpos/efeitos dos fármacos , Bacterioclorofilas/farmacologia , Imunidade Celular/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Fotoquimioterapia , Animais , Vasos Sanguíneos/efeitos dos fármacos , Linhagem Celular Tumoral , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Feminino , Citometria de Fluxo , Imuno-Histoquímica , Interferon gama/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias/imunologia , Fármacos Fotossensibilizantes/farmacologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologiaRESUMO
Photodynamic therapy (PDT) mediated with vascular acting photosensitizer Tookad (Pd-bacteriopheophorbide) was investigated as an alternative modality for treating prostate cancer. Photodynamic effects on the prostate gland and its adjacent tissues were evaluated in a canine model. Interstitial prostate PDT was performed by irradiating individual lobes with a cylindrical diffuser fiber at various drug/light doses. The sensitivity of the adjacent tissues to Tookad PDT was determined by directly irradiating the surface of the bladder, colon, abdominal muscle and pelvic plexus with a microlens fiber at various drug/light doses. The prostate and adjacent tissues were harvested one-week after the treatment and subjected to histopathological examination. PDT-induced prostate lesions were characterized by marked hemorrhagic necrosis. The bladder, colon, abdominal muscle and pelvic plexus appeared to be sensitive to PDT although the Tookad PDT-induced responses in these tissues were minimal compared to that of the prostate gland at the same dose levels. Nevertheless, the protection of the adjacent tissues should be taken into consideration during the total prostate ablation process due to their sensitivity to PDT. The sensitivity of the prostatic urethra is worth further investigation. Direct intraurethral irradiation might provide an ideal means to determine the sensitivity of the prostatic urethra and might lead to transurethral PDT protocols for the management of benign prostatic hyperplasia (BHP).
Assuntos
Bacterioclorofilas/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Próstata/efeitos dos fármacos , Próstata/efeitos da radiação , Abdome/efeitos da radiação , Animais , Colo/efeitos dos fármacos , Colo/efeitos da radiação , Cães , Relação Dose-Resposta a Droga , Relação Dose-Resposta à Radiação , Masculino , Neurônios/efeitos dos fármacos , Neurônios/efeitos da radiação , Bexiga Urinária/efeitos dos fármacos , Bexiga Urinária/efeitos da radiaçãoRESUMO
The biodistribution of two near-infrared fluorescent agents was assessed in vivo by time-resolved diffuse optical imaging. Bacteriochlorophyll a (BC) and cypate-glysine-arginine-aspartic acid-serine-proline-lysine-OH (Cyp-GRD) were administered separately or combined to mice with subcutaneous xenografts of human breast adenocarcinoma and slow-release estradiol pellets for improved tumor growth. The same excitation (780 nm) and emission (830 nm) wavelengths were used to image the distinct fluorescence lifetime distribution of the fluorescent molecular probes in the mouse cancer model. Fluorescence intensity and lifetime maps were reconstructed after raster-scanning whole-body regions of interest by time-correlated single-photon counting. Each captured temporal point-spread function (TPSF) was deconvolved using both a single and a multiexponental decay model to best determine the measured fluorescence lifetimes. The relative signal from each fluorophore was estimated for any region of interest included in the scanned area. Deconvolution of the individual TPSFs from whole-body fluorescence intensity scans provided corresponding lifetime images for comparing individual component biodistribution. In vivo fluorescence lifetimes were determined to be 0.8 ns (Cyp-GRD) and 2 ns (BC). This study demonstrates that the relative biodistribution of individual fluorophores with similar spectral characteristics can be compartmentalized by using the time-domain fluorescence lifetime gating method.
Assuntos
Sondas Moleculares/análise , Imagem Corporal Total/métodos , Animais , Bacterioclorofilas/química , Bacterioclorofilas/metabolismo , Bacterioclorofilas/farmacologia , Linhagem Celular Tumoral , Fluorescência , Humanos , Injeções Intravenosas , Masculino , Camundongos , Camundongos Nus , Sondas Moleculares/química , Transplante de Neoplasias , Neoplasias/patologia , Peptídeos/química , Peptídeos/metabolismo , Peptídeos/farmacologia , Espectrofotometria Infravermelho , Fatores de TempoRESUMO
BACKGROUND AND OBJECTIVE: To determine the optimal magnetic resonance imaging (MRI) methodology to assess photodynamic therapy (PDT)-induced histopathological responses in the prostate. STUDY DESIGN/MATERIALS AND METHODS: Laparotomy was performed in five healthy dogs. Cylindrical diffuser was placed in the prostates to deliver light of 50-300 J/cm at 150 mW/cm and 763 nm to activate IV-injected Tookad (1 mg/kg b.w.). Fast spin echo (FSE) T2-weighted, post-contrast-enhanced T1-(CE-T1) and diffusion weighted images (DWI) were obtained pre- and 2 days, 7 days, and 1 month post-PDT. Radiological-histopathological correlation was performed at 7 days (n = 4) and 1 month (n = 1) after PDT. A qualitative assessment of signal changes and apparent diffusion coefficient (ADC) mapping was performed. RESULTS: At 2 or 7 days post-PDT, there was good spatial correlation between PDT-induced hemorrhagic necrosis and unenhanced regions on CE-T1 images. There was a rapidly and persistently enhancing rim corresponding to edema and inflammation. FSE T2 and DWI showed altered signal but did not clearly define necrosis in all cases. At 1 month, it was hard to correlate MR images to histopathologic changes as they represented a mixture of necrosis and developing fibrosis, which led to a mixed signal intensity and less demarcated contrast enhancement. CONCLUSIONS: At 7 days after PDT, gadolinium DTPA contrast-enhanced MRI is superior to DWI and T2 imaging in assessing the boundary of Tookad PDT-induced tissue necrosis in the normal canine prostate.
Assuntos
Bacterioclorofilas/farmacologia , Imageamento por Ressonância Magnética , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Próstata/efeitos dos fármacos , Animais , Tecido Conjuntivo/patologia , Meios de Contraste , Imagem de Difusão por Ressonância Magnética , Cães , Edema/diagnóstico , Edema/patologia , Epitélio/patologia , Fibrose , Gadolínio DTPA , Hemorragia/diagnóstico , Hemorragia/patologia , Aumento da Imagem , Leucócitos Mononucleares/patologia , Masculino , Necrose , Projetos Piloto , Próstata/patologia , Fatores de TempoRESUMO
OBJECTIVE: Our aim is to investigate the effects of photodynamic therapy (PDT) on peripheral nerve conductivity. BACKGROUND DATA: Interstitial PDT has been demonstrated as a promising treatment modality for prostate cancer. However, the sensitivity of nerves, in the immediate vicinity of the prostate gland, to PDT procedures has not been studied. This study attempts to establish an in situ canine model to evaluate direct PDT effect on peripheral nerves. METHODS: PDT was performed by irradiating the cutaneous branches of the saphenous nerve at 763 nm with light doses of 50-200 J/cm2 after i.v. infusion of the photosensitizer Tookad (0-2 mg/kg). Evoked compound-action potentials (CAP) were recorded directly from the surface of the saphenous nerve. The latencies to onset and conduction velocities were determined during PDT and 1-week post-PDT. RESULTS: Nerve and surrounding tissue damage corresponded well with drug/light doses. With Tookad doses of 2 mg/kg, treatment with 50 J/cm2 induced little change in saphenous nerve conduction properties. However, treatment with 100 J/cm2 resulted in localized nerve injury and decreases in nerve conduction velocities, and treatment with 200 J/cm2 severely damaged the nerve. CONCLUSIONS: This canine model adequately demonstrates effects of Tookad PDT on peripheral nerves. Direct irradiation of 100-200 J/cm2 can alter nerve conduction and induce nerve damage. Therefore, possible side effects of interstitial PDT on the pelvic plexus need to be investigated in future studies.
Assuntos
Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/efeitos da radiação , Nervos Periféricos/efeitos dos fármacos , Nervos Periféricos/efeitos da radiação , Fotoquimioterapia , Animais , Bacterioclorofilas/farmacologia , Cães , Relação Dose-Resposta a Droga , Masculino , Modelos Animais , Condução Nervosa/efeitos dos fármacos , Condução Nervosa/efeitos da radiação , Fármacos Fotossensibilizantes/farmacologiaRESUMO
Generation of reactive oxygen species (ROS) is the hallmark of important biological processes and photodynamic therapy (PDT), where ROS production results from in situ illumination of certain dyes. Here we test the hypothesis that the yield, fate, and efficacy of the species evolved highly depend on the dye's environment. We show that Pd-bacteriopheophorbide (Pd-Bpheid), a useful reagent for vascular targeted PDT (VTP) of solid tumors, which has recently entered into phase II clinical trials under the code name WST09 (trade name TOOKAD), forms appreciable amounts of hydroxyl radicals, superoxide radicals, and probably hydrogen peroxide in aqueous medium but not in organic solvents where singlet oxygen almost exclusively forms. Evidence is provided by pico- and nanosecond time-resolved spectroscopies, ESR spectroscopy with spin-traps, time-resolved singlet oxygen phosphorescence, and chemical product analysis. The quantum yield for singlet oxygen formation falls from approximately 1 in organic solvents to approximately 0.5 in membrane-like systems (micelles or liposomes), where superoxide and hydroxyl radicals form at a minimal quantum yield of 0.1%. Analysis of photochemical products suggests that the formation of oxygen radicals involves both electron and proton transfer from (3)Pd-Bpheid at the membrane/water interface to a colliding oxygen molecule, consequently forming superoxide, then hydrogen peroxide, and finally hydroxyl radicals, with no need for metal catalysis. The ability of bacteriochlorophyll (Bchl) derivatives to form such radicals upon excitation at the near infrared (NIR) domain opens new avenues in PDT and research of redox regulation in animals and plants.
Assuntos
Bacterioclorofilas/química , Espécies Reativas de Oxigênio/química , Bacterioclorofilas/farmacologia , Radical Hidroxila/química , Radical Hidroxila/metabolismo , Cinética , Micelas , Fotoquímica , Fotoquimioterapia , Teoria Quântica , Espécies Reativas de Oxigênio/metabolismo , Análise Espectral/métodosRESUMO
New negatively charged water-soluble bacteriochlorophyll (Bchl) derivatives were developed in our laboratory for vascular-targeted photodynamic therapy (VTP). Here we focused on the synthesis, characterization and interaction of the new candidates with serum proteins and particularly on the effect of serum albumin on the photocytotoxicity of WST11, a representative compound of the new derivatives. Using several approaches, we found that aminolysis of the isocyclic ring with negatively charged residues markedly increases the hydrophilicity of the Bchl sensitizers, decreases their self-association constant and selectively increases their affinity to serum albumin, compared with other serum proteins. The photocytotoxicity of the new candidates in endothelial cell culture largely depends on the concentration of the serum albumin. Importantly, after incubation with physiological concentrations of serum albumin (500-600 microM), WST11 was found to be poorly photocytotoxic (>80% endothelial cell survival in cell cultures). However, in a recent publication (Mazor, O. et al. [2005] Photochem. Photobiol. 81, 342-351) we showed that VTP of M2R melanoma xenografts with a similar WST11 concentration resulted in approximately 100% tumor flattening and >70% cure rate. We therefore propose that the two studies collectively suggest that the antitumor activity of WST11 and probably of other similar candidates does not depend on direct photointoxication of individual endothelial cells but on the vascular tissue response to the VTP insult.
Assuntos
Bacterioclorofilas/farmacologia , Proteínas Sanguíneas/fisiologia , Fotoquimioterapia , Bacterioclorofilas/biossíntese , Bacterioclorofilas/efeitos da radiação , Sobrevivência Celular/efeitos da radiação , Endotélio Vascular/citologia , Endotélio Vascular/efeitos da radiação , Soroalbumina Bovina/efeitos da radiaçãoRESUMO
AIM: Understanding the modes and activities of metal bacterial chlorophylls as PHD sensitizers with DNA. METHODS: The modes and activities of the interaction of DNA and metal complexes of bacterial chlorophyll, which have been prepared by extraction and synthesis reaction, have been discussed according to the ultraviolet-visual spectrum and nucleic acid electrophoresis. RESULTS: It indicates that the system of DNA and metal complexes have enchanced the interaction by the ultraviolet-visual spectrum. At the same time, it also indicates that metal complexes of bacterial chlorophyll and DNA have different combining way and have strong cutting effect in illumination by the nucleic acid electrophoresis. CONCLUSION: This paper proved that metal bacterial chlorophylls as PHD sensitizers have great advantage.
Assuntos
Bacterioclorofilas/química , DNA/metabolismo , Compostos Organometálicos/química , Bacterioclorofilas/síntese química , Bacterioclorofilas/farmacologia , Cobre/química , Eletroforese , Células HL-60 , Humanos , Células K562 , Níquel/química , Compostos Organometálicos/síntese química , Compostos Organometálicos/farmacologia , Ligação Proteica , Espectrofotometria Ultravioleta , Zinco/químicaRESUMO
TOOKAD (WST09) is a new, long-wavelength palladium bacteriopheophorbide photosensitizer that targets tissue vasculature. The cutaneous phototoxicity of TOOKAD was assessed in normal rat and pig animal models and in patients in a Phase-I trial of TOOKAD-mediated photodynamic therapy (PDT) for recurrent prostate cancer. Controlled skin exposures were administered using solar-simulated light at various times after drug administration. Two different spectral ranges were used. In the first, the UV portion of the spectrum was removed (UV(-)) because UV irradiation in nondrugged control animals produced an erythema response at incident energy densities (J/cm(2)) lower than those required to induce a PDT response. In the second, the full solar spectrum (UV(+)) was used, and the potentiation by the photosensitizer of the UV-mediated minimum erythema dose was assessed. Results showed that the PDT skin response was negligible at clinical drug doses of 2 mg/kg for any period after administration at light doses of 128 J/cm(2) in the animal models. In patients, there was no observed UV(-) skin response at doses of up to 2 mg/kg, drug-light intervals of 1-3 h or greater and light exposures up to 128 J/cm(2). At higher drug doses in the rat and pig models, the duration of skin phototoxicity was found to be approximately 3 h and less than 1 h, respectively. Using the full spectrum of solar-simulated light, the presence of TOOKAD did not measurably enhance the UV(+)-induced erythema in the rats, pigs or patients.
Assuntos
Bacterioclorofilas/farmacologia , Modelos Animais , Fármacos Fotossensibilizantes/farmacologia , Raios Ultravioleta , Animais , HumanosRESUMO
Various forms of cellular stress induce adaptive responses through poorly understood mechanisms. In maintaining homeostasis, endothelial cells respond and adapt to changes in oxidative stress that prevail in the circulation. Endothelial cells are also the target of many oxidative stress-based vascular therapies. The objectives of this study were to determine whether endothelial cells adapt to oxidative stress induced upon the photosensitization of WST11 (a water-soluble Pd-bacteriochlorophyll derivative being developed as a photodynamic agent) and to study possible cellular mechanisms involved. The hallmark of WST11-based photodynamic therapy is the in situ generation of cytotoxic reactive oxygen species causing vascular shutdown, hypoxia, and tumor eradication. Here we demonstrated that photodynamic therapy also induces adaptive responses and tolerance following a sublethal preconditioning of endothelial cells with the same (homologous) or different (heterologous) stressor. A link among p38 MAPK activity, expression of hsp70 and hsp27, and homologous adaptation to reactive oxygen species induced by photosensitized WST11 was established. In addition to characterization of some key proteins involved, our observations provide a beneficial new working tool for the studies of mechanisms involved in oxidative stress and adaptation using light-controlled photosensitization.