Photodetection and photodynamic treatment of ovarian micrometastases : benefits of hypericin-loaded polymeric nanocarriers

Le photodétection par fluorescence et la thérapie photodynamique, exploitant les propriétés des photosensibilisateurs, sont des méthodes alternatives prometteuses qui pourraient améliorer l'issue thérapeutique de cancers à taux de récidive élevé tel que celui de l'ovaire. Ce travail de thèse s'est focalisé sur la valorisation d'une molécule naturelle extraite du millepertuis ("Hypericum perforatum"), l'hypéricine. Certaines limitations des photosensiblisateurs, comme le manque de sélectivité et les difficultés de formulation, persistent et ralentissent leur développement en clinique. Leur incorporation dans des nanoparticules polymériques a été proposée comme stratégie permettant à la fois l'injection des photosensibilisateurs et l'augmentation de leur sélectivité. L'intérêt des NPs pour l'accumulation sélective dans les tumeurs a été étudié et comparé "in vivo" sur deux modèles, un modèle rongeur classique et la membrane chorioallantoïdienne de l'embryon de poulet. La comparaison des résultats obtenus appuie la pertinence du modèle alternatif aux mammifères, simple et économique pour le criblage de photosensibilisateurs

Nanoparticles for photodynamic therapy of cancer

Currently, the limiting factor in cancer chemotherapy is still the lack of selectivity of anticancer drugs towards neoplastic cells. Photosensitizers (PS), the active com pounds used for photodynamic therapy (PDT), have the intrinsic advantage of dis tributing primarily in highly regenerative tissues after intravenous or topical ad ministration. Therefore, they will accumulate preferentially in tumor tissue when present. In addition, these molecules are inactive as such; indeed, the anticancer effect is only attained after irradiation with light at the right wavelength. Compared to other current cancer therapies such as surgery, radiotherapy or chemotherapy, PDT is an effective and selective means of suppressing diseased tissues without altering the surrounding healthy tissue. It also offers a unique opportunity to reach unseen metastasis. As fluorescent molecules, PS may also be used as a tool, namely photodetection (PD), to reveal tum.or tissues that remain unseen by other conventional methods

Screening of nanoparticulate delivery systems for the photodetection of cancer in a simple and cost-effective model

AIMS: In urology, fluorescence-based imaging methods have been proven to significantly improve the detection of small, barely visible tumors and reduce the recurrence rate. Under ethical and economical pressure, new effective screening systems have to be developed to exploit and assess novel strategies for fluorescence photodetection in other areas. For this purpose, the chorioallantoic membrane (CAM) of the developing chick embryo is an attractive alternative model to the mammalian models. MATERIALS & METHODS: Hypericin encapsulated into nanoparticles for the photodetection of ovarian metastases was evaluated in the CAM model with respect to vascular extravazation and tumor targeting and compared with free drug following intravenous administration. RESULTS: To validate the CAM model as a valuable screening system for photodetection of cancer, we drew a comparison with results obtained on a conventional rodent model. CONCLUSION: Rodent and CAM models led to the same conclusion regarding the benefits of nanoencapsulation to improve selective accumulation of drug in ovarian micrometastases

Benefits of nanoencapsulation for the hypercin-mediated photodetection of ovarian micrometastases

The high recurrence and lethality of ovarian cancer at advanced stages is problematic, especially due to the development of numerous micrometastases scattered throughout the abdominal cavity. Fluorescence photodetection (PD) used in combination with surgical resection of malignant tissues has been suggested to improve recovery. Based on promising in vivo results for the detection of bladder cancer, hypericin (Hy), a natural photosensitizer (PS), stands as a good candidate for the photodetection of ovarian cancer. However, due to its hydrophobicity, systemic administration of Hy is problematic. Polymeric nanoparticles (NPs) help to overcome these delivery and stability problems and enable intravenous administration of Hy. In this study, Hy-loaded NPs of polylactic acid were produced with the following properties: (i) mean size of 268 nm, (ii) negative zeta potential, (iii) low residual surfactant and (iv) drug loading of 3.7 % (w/w). The potential of hypericin-loaded nanoparticles for the fluorescence photodetection of ovarian metastases in Fischer 344 rats bearing ovarian tumours was compared to free drug. The selectivity of Hy administered with both formulations was assessed first by fluorescence endoscopy, and then quantified after tissue extraction. The results showed an improved selective accumulation of Hy in ovarian micrometastases when NPs were used

Hypericin-loaded nanoparticles for the photodynamic treatment of ovarian cancer

A photodynamic approach has been suggested to improve diagnosis and therapy of ovarian cancer. As Hypericin (Hy), a natural photosensitizer (PS) extracted from Hypericum perforatum, has been shown to be efficient in vitro and in vivo for the detection or treatment of other cancers, Hy could also be a potent tool for the treatment and detection of ovarian cancer. Due to its hydrophobicity, systemic administration of Hy is problematic. Thus, polymeric nanoparticles (NPs) of polylactic acid (PLA) or polylactic-co-glycolic acid (PLGA) were used as a drug delivery system. Hy-loaded NPs were produced with the following characteristics: (i) size in the 200-300 nm range, (ii) negative zeta potential, (iii) low residual PVAL and (iv) drug loading from 0.03 to 0.15% (w/w). Their in vitro photoactivity was investigated on the NuTu-19 ovarian cancer cell model derived from Fischer 344 rats and compared to free drug. Hy-loaded PLA NPs exhibited a higher photoactivity than free drug. Increasing light dose or incubation time with cells induced an enhanced activity of Hy-loaded PLA NPs. Increased NP drug loading had a negative effect on their photoactivity on NuTu-19 cells: at the same Hy concentration, the higher was the drug loading, the lower was the phototoxic effect. The influence of NP drug loading on the Hy release from NPs was also investigated

Challenges and Opportunities in the Oral Delivery of Recombinant Biologics

International audienceRecombinant biological molecules are at the cutting-edge of biomedical research thanks to the significant progress made in biotechnology and a better understanding of subcellular processes implicated in several diseases. Given their ability to induce a potent response, these molecules are becoming the drugs of choice for multiple pathologies. However, unlike conventional drugs which are mostly ingested, the majority of biologics are currently administered parenterally. Therefore, to improve their limited bioavailability when delivered orally, the scientific community has devoted tremendous efforts to develop accurate cell-and tissue-based models that allow for the determination of their capacity to cross the intestinal mucosa. Furthermore, several promising approaches have been imagined to enhance the intestinal permeability and stability of recombinant biological molecules. This review summarizes the main physiological barriers to the oral delivery of biologics. Several preclinical in vitro and ex vivo models currently used to assess permeability are also presented. Finally, the multiple strategies explored to address the challenges of administering biotherapeutics orally are described

Quenching-induced deactivation of photosensitizer by nanoencapsulation to improve phototherapy of cancer

Photodynamic therapy has emerged as a promising alternative to current cancer treatment. However, conventional photosensitizers have several limitations due to their unsuitable pharmaceutical formulations and lack of selectivity. Our strategy was to exploit the advantages of nanoparticles and the quenching-induced deactivation of the model photosensitizer hypericin to produce "activatable" drug delivery systems. Efficient fluorescence and activity quenching were achieved by increasing the drug-loading rate of nanoparticles. In vitro assays confirmed the reversibility of hypericin deactivation, as the hypericin fluorescence and photodynamic activity were recovered upon cell internalization

The chick embryo and its chorioallantoic membrane (CAM) for the in vivo evaluation of drug delivery systems

Mammalian models are frequently used for preclinical evaluation of new drug delivery systems (DDS). However, valid mammalian models are expensive, time-consuming, and not easy to set up and evaluate. Furthermore, they are often linked to administrative burden with respect to ethical and legal aspects. The present review outlines the possibilities and limitations of using the hen's embryo, and specifically its chorioallantoic membrane (CAM), as an alternative to mammalian models for the evaluation of DDS. Features of the CAM, the anatomy of the embryo, and the blood were investigated to assess properties of the drug carriers such as toxicity and biocompatibility, as well as the activity, toxicity, biodistribution and pharmacokinetics of the drug. The simplicity, rapidity, and low cost of the different assays that can be performed with chick embryos strengthen the interest of routinely using this model in pharmaceutical technology research. It is concluded that there is a big potential for using chick embryos in screening procedures of formulation candidates, thus establishing an intermediate step between in vitro cellular tests and preclinical mammalian models

Abstract B11: The early autophagy inhibitor verteporfin moderately enhances the antitumor activity of gemcitabine in a pancreatic ductal adenocarcinoma model

Abstract Autophagy, a cellular self-eating process that is activated by several cancer drugs and appears to function as a protective mechanism, is a promising therapeutic target. Pancreatic ductal adenocarcinoma (PDAC) is highly resistant to chemotherapy, and has been described as requiring elevated autophagy for growth. To date, all preclinical reports and clinical trials investigating pharmacological inhibition of autophagy have used chloroquine or hydroxychloroquine, which block autophagy at a late stage. Verteporfin is a newly discovered autophagy inhibitor that blocks autophagy at an early stage by inhibiting autophagosome formation. Here, we report that PDAC cell lines show variable sensitivity to verteporfin in vitro and that verteporfin inhibits autophagy stimulated by gemcitabine, the current standard treatment for PDAC. Pharmacokinetic and efficacy studies in a BxPC-3 xenograft mouse model demonstrate that verteporfin accumulated in tumors at autophagy-inhibiting levels but did not reduce tumor volume or increase survival as a single agent. However, in combination with gemcitabine, verteporfin moderately reduced tumor growth and enhanced survival compared to gemcitabine alone. Our results do not agree with the premise that autophagy inhibition is effective against PDAC as a single-modality treatment, but they support autophagy inhibition as an approach to sensitize PDAC to gemcitabine. Citation Format: Elizabeth Donohue, Anitha Thomas, Norbert Maurer, Irina Manisali, Magali Zeisser-Labouebe, Natalia Zisman, Hilary J. Anderson, Murray Webb, Marcel Bally, and Michel Roberge. The early autophagy inhibitor verteporfin moderately enhances the antitumor activity of gemcitabine in a pancreatic ductal adenocarcinoma model. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr B11.</jats:p