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Les bactériophages sont des virus naturels très répandus dans l’environnement qui ciblent spécifiquement les bactéries. Leur utilisation en médecine, connue sous le terme phagothérapie, consiste à les isoler, les caractériser, les cultiver, puis les purifier pour traiter des infections bactériennes. Il existe actuellement un renouveau pour la thérapie phagique, et sa mise en œuvre présuppose de disposer de phages actifs de qualité pharmaceutique. D’un point de vue réglementaire, le statut des phages n’est pas encore clairement défini par les autorités, mais la mise à disposition de phages produits par l’industrie pharmaceutique et les programmes de développement académiques, comme le programme PHAGEinLYON, constituent un tournant dans le déploiement de la phagothérapie.

Bacterial resistance against antibiotics is an emergent medical issue. The development of novel therapeutic approaches is urgently needed and, in this context, bacteriophages represent a promising strategy to fight multi resistant bacteria. However, for some applications, bacteriophages cannot be used without an appropriate drug delivery system which increases their stability or provides an adequate targeting to the site of infection. This review summarizes the main application routes for bacteriophages and presents the new delivery approaches designed to increase phage's activity. Clinical successes of these formulations are also highlighted. Globally, this work paves the way for the design and optimization of nano and micro delivery systems for phage therapy.

Background Acute myeloid leukemia mainly affects adult patients. Complete remission for patients younger than 60 years, who are candidates for standard induction therapy, is achieved in 60%-80% of cases. However, the prognosis is still poor for older patients, who are unfit for intensive chemotherapy, and only a few therapies are available. Hypomethylating agents, such as decitabine, are approved for such patients. The current dosing regimen consists of one administration per day, for 5 days, each 4 weeks. Methods Here, we present the synthesis of a decitabine prodrug, combined with its encapsulation into a lipid-based nanocapsule formulation. Decitabine (C12)2 was synthetized, then loaded into nanocapsules. Its stability in phosphate buffer ans human plasma was checked. Its activity was evaluated by Cell proliferation assays and cell-cycle analysis on human erythroleukemia cells. Then its pharmacokinetics was determined on a rat model. Results Decitabine (C12)2 was obtained with a yield of 50%. Drug loading into nanocarriers of 27.45±0.05 nm was 5.8±0.5 mg/mL. The stability of decitabine was improved and its activity on leukemia cells was not altered. Finally, pharmacokinetics studies showed a prolonged mean residence time of the drug. Conclusion Decitabine (C12)2 as a prodrug showed high encapsulation efficiency, a good stability in plasma with no impact on its activity on leukemia cells and improved pharmacokinetics.

A prospective cohort study was conducted to evaluate long-term safety of tedizolid as suppressive antimicrobial treatment in patients with implant-associated bone and joint infection caused by multidrug-resistant gram-positive pathogens. Seventeen patients received tedizolid with a median duration of treatment of 6 months. No patients developed a serious adverse event.

Background Microbial keratitis are severe infectionsgenerally linked to risk factors. High-doses antibiotic eye-drops could be required to avoid severe complications. In such cases, hospital pharmacists are in charge of their production given the lack of such formulations on the market. The stability of these antibiotic eye-drops is generally limited to a couple of days and publications generally do not describe addition of microbial preservatives even though it is a European Pharmacopeia requirement. The aim of this study was to describe a new ceftazidime eye-drops formulation at 50 mg/mL with a antimicrobial additive, benzalkonium chloride at 0.04 mg/mL. Methods Physico-chemical studies of this new formulation were performed by a stability indicating HPLC-UV method validated according to ICH standards, osmolality measurements, pH monitoring and visual examinations. Antimicrobial preservative efficacy was evaluated according to the method from the European Pharmacopeia. Results After 75 days at −20 °C followed by 7 days at 4 °C, or after 7 days at 4 °C, the eye-drops were stable. A degradation trend was finally observed at day 14 at 4 °C. Conclusions A new ceftazidime eye-drops formulation is proposed with a stability of 7 days. Outpatients do not need to return to the hospital pharmacy for repeat dispensing, thus possibly improving treatment compliance.

Purpose: To improve the tumor localization during laparoscopic surgery, we describe an innovative technique involving superselective intra-arterial injection of blue dye in tumoral vessels to color the tumor before surgical enucleation. Materials and Methods: The dye injection was performed at the same time as superselective embolization, immediately before laparoscopic surgery in a hybrid operating room. We used this new treatment sequence on 50 consecutive patients. Results: The selective intra-arterial injection of an emulsion of blue dye and lipiodol was feasible in 46 (92%) cases and well tolerated, followed by superselective embolization of the tumor vessels with glue or coils. The tumor was easily localized during surgery due to the blue coloration. Tumor coloration was not associated with postoperative complication, especially allergic reaction or renal failure. Pathologic analysis of the tumor was not modified by the coloration and all tumors had negative surgical margins. Conclusions: The preoperative dye localization is a feasible, safe, and accurate procedure. This combined approach reduces the difficulty of surgery and increases patient safety.

Background Acute myeloid leukemia mainly affects adult patients. Complete remission for patients younger than 60 years, who are candidates for standard induction therapy, is achieved in 60%–80% of cases. However, the prognosis is still poor for older patients, who are unfit for intensive chemotherapy, and only a few therapies are available. Hypomethylating agents, such as decitabine, are approved for such patients. The current dosing regimen consists of one administration per day, for 5 days, each 4 weeks. Methods Here, we present the synthesis of a decitabine prodrug, combined with its encapsulation into a lipid-based nanocapsule formulation. Decitabine (C12)2 was synthetized, then loaded into nanocapsules. Its stability in phosphate buffer ans human plasma was checked. Its activity was evaluated by Cell proliferation assays and cell-cycle analysis on human erythroleukemia cells. Then its pharmacokinetics was determined on a rat model. Results Decitabine (C12)2 was obtained with a yield of 50%. Drug loading into nanocarriers of 27.45±0.05 nm was 5.8±0.5 mg/mL. The stability of decitabine was improved and its activity on leukemia cells was not altered. Finally, pharmacokinetics studies showed a prolonged mean residence time of the drug. Conclusion Decitabine (C12)2 as a prodrug showed high encapsulation efficiency, a good stability in plasma with no impact on its activity on leukemia cells and improved pharmacokinetics.

International audience; PURPOSE:To describe, in patients treated for infectious keratitis, the microorganisms identified and their antibiotic susceptibility over a period of 18 months.METHOD:Retrospective, descriptive, non-comparative study. Medical and biological data were extracted from the patients' file treated with strengthened antibiotic eye drops at Angers University Hospital between January 2015 and June 2016. The main elements noted were the bacteria involved and their susceptibility to antibiotics. Patients' visual acuity at the start and end of treatment was compared.RESULTS:Forty-eight patients were included. Almost one bacterium was identified in 31 patients, totalling 43 pathogens of 24 different species. The most frequently found microorganisms were Gram-positive cocci (55.8%), including Staphylococcus Aureus (14.0%) and Epidermidis (14.0%). All Gram-negative bacilli amounted to 30.2% of the identified bacteria, including 9.3% of Pseudomonas aeruginosa. None of the Gram-positive cocci were resistant to vancomycin and all Gram-negative bacilli were susceptible to ceftazidime and amikacin. Following treatment with at least one of the three antibiotic eye drops produced by our pharmacy (amikacin at 50mg/mL, ceftazidime at 50mg/mL and vancomycin at 25mg/mL), patients' visual acuity was significantly improved (P=0.043).CONCLUSION:The study helped identify the bacterial ecology of patients admitted for infectious keratitis. Among the bacteria identified, none were found to be resistant to any of the three strengthened antibiotic eye drops produced by the hospital pharmacy. These eye drops allowed for a rapid and effective treatment of patients and the improvement of their visual acuity before even identifying the bacteria.; ObjectifDécrire, les microorganismes responsables des kératites bactériennes.MéthodeLes données médicales et biologiques ont été extraites rétrospectivement des dossiers médicaux des patients traités pour kératites bactériennes par collyres antibiotiques renforcés. L’identification des germes ainsi que leur sensibilité aux antibiotiques ont été notées. Les acuités visuelles des patients avant l’instauration du traitement et à la fin de celui-ci ont été comparées.RésultatsAu total, 48patients ont été inclus. Au moins un germe bactérien a été identifié chez 31patients, soit 43germes parmi 24espèces différentes. Les cocci à Gram positif ont été les germes les plus fréquemment rencontrés (55,8%), dont Staphyloccocus aureus (14,0%), et Epidermidis (14,0%). Les bacilles à Gram négatif ont représenté 30,2% des germes identifiés, incluant Pseudomonas aeruginosa (9,3%). Aucun cocci à Gram positif n’a été résistant à la vancomycine, tous les bacilles à Gram négatif ont été sensibles à la ceftazidime et l’amikacine. Après traitement avec au moins l’un des trois collyres antibiotiques renforcés produits par la pharmacie du CHU d’Angers (amikacine 50mg/mL, ceftazidime 50mg/mL ou vancomycine 25mg/mL), l’acuité visuelle des patients a été significativement améliorée (p=0,043).ConclusionL’écologie bactérienne des patients hospitalisés pour kératite bactérienne et nécessitant un traitement par collyre antibiotique renforcé est désormais connue. Parmi les bactéries identifiées, aucune n’a été résistante à l’un des trois collyres antibiotiques renforcés produits par la pharmacie du CHU. Ces antibiotiques ont permis une amélioration rapide et significative de l’acuité visuelle des patients.

International audience; OBJECTIVES: In this study, a novel analytical method to quantify prion inactivating detergent in rinsing waters coming from the washer-disinfector of a hospital sterilization unit has been developed. The final aim was to obtain an easy and functional method in a routine hospital process which does not need the cleaning product manufacturer services.METHODS: An ICP-MS method based on the potassium dosage of the washer-disinfector's rinsing waters was developed. Potassium hydroxide is present on the composition of the three prion inactivating detergent currently on the French market. The detergent used in this study was the Actanios LDI(®) (Anios laboratories). A Passing and Bablok regression compares concentrations measured with this developed method and with the HPLC-UV manufacturer method.RESULTS: According to results obtained, the developed method is easy to use in a routine hospital process. The Passing and Bablok regression showed that there is no statistical difference between the two analytical methods during the second rinsing step. Besides, both methods were linear on the third rinsing step, with a 1.5ppm difference between the concentrations measured for each method.CONCLUSIONS: This study shows that the ICP-MS method developed is nonspecific for the detergent, but specific for the potassium element which is present in all prion inactivating detergent currently on the French market. This method should be functional for all the prion inactivating detergent containing potassium, if the sensibility of the method is sufficient when the potassium concentration is very low in the prion inactivating detergent formulation.; ObjectifsL’objectif principal de cette étude est de développer une méthode analytique de dosage des détergents désinfectants prionicides dans les eaux de rinçage des laveurs désinfecteurs d’un service de stérilisation. Cette méthode doit être simple à mettre en œuvre et utilisable en routine à l’hôpital pour permettre de s’affranchir des dosages réalisés par les fabricants de détergents désinfectants prionicides.MéthodeUne méthode ICP-MS basée sur le dosage du potassium dans les eaux de rinçage d’un laveur désinfecteur a été développée. En effet, les trois détergents désinfectants prionicides actuellement sur le marché contiennent de l’hydroxyde de potassium. Le détergent utilisé dans l’étude a été de l’Actanios® LDI (Laboratoire Anios). Une régression de Passing et Bablok compare des résultats obtenus par cette nouvelle méthode à d’autres obtenus grâce à une méthode (HPLC-UV) développée par le laboratoire Anios.RésultatsLa méthode développée est simple à mettre en œuvre en routine et la régression de Passing et Bablok réalisée montre que les deux méthodes ont été équivalentes sur les 15dosages réalisés dans la deuxième eau de rinçage. Les deux méthodes ont été linéaires mais avec une différence de 1,5ppm sur la troisième eau de rinçage.ConclusionLa méthode ICP-MS développée est non spécifique du détergent étudié mais spécifique de l’élément potassium présent dans tous les détergents désinfectants prionicides actuellement sur le marché. Elle pourrait donc probablement être appliquée à l’ensemble des détergents désinfectants prionicides après avoir vérifié qu’elle est suffisamment sensible lorsque les concentrations en potassium dans le détergent sont faibles.

Mitomycin C (MMC) is widely used in treatment of non-muscle invasive bladder cancer at a 1 mg/mL concentration, by intravesical instillation. MMC is also used as an ophthalmic procedure in glaucoma care mostly with 0.2 mg/mL concentration. To accelerate syringes provision, it could be interesting to demonstrate the stability of the drug, in order to be able to prepare the chemotherapeutic drug several hours before the chemotherapy administration.: A stability indicating HPLC-UV method was developed and validated according to the ICH guidelines. Concentrations of the MMC stored at 25 °C and 60 % of relative humidity and protected from light in polypropylene syringes (1 mg/mL and 0.2 mg/mL) or glass vials (1 mg/mL) were evaluated for 96 h and compared to the initial observed concentrations.: MMC stability was demonstrated in syringes and glass vials at 1 mg/mL only for 8 h in water for injections and for 10 h at 0.2 mg/mL in 0.9 % sodium chloride solutions, because relative concentrations (95 % confidence interval of the mean of 3 samples) were systematically over 90 % of the initial concentrations. After 96 h the relative concentrations were found below 80 % as compared to initial concentrations, thus indicating instability of these solutions. Degradation products were observed and remained below 3 %.: This study confirms that MMC solutions for ophthalmic application at 0.2 mg/mL or vesical instillation at 1 mg/mL have to be formulated extemporaneously to maintain the desired concentration.

International audience; Micafungin is a costly treatment and packaging of 50mg or 100mg bottles only are available, while doses lower than 5mg and 20mg are often necessary in neonates and paediatrics patients, respectively. The stability of micafungin sodium in polypropylene syringes and glass bottles was studied at different concentrations. Solutions of micafungin diluted with NaCl 0.9% were prepared in glass bottles (20 and 10mg/mL) or syringes (1 and 0.5mg/mL) and stored at 25°C, 60% humidity (RH), in the dark (ICH conditions). Solutions were also exposed to heat (70°C) or alkaline solution (NaOH) in order to force degradation. Samples were analysed at days 1, 5, 8 (for bottles) and also 15 (for syringes) after the preparation and assayed in triplicate. Stability was studied using a stability-indicating high-performance liquid chromatographic method. Syringes stored at 25°C retained over 90% of their initial concentration over the study period. Temperature and alkaline conditions had significant effect on the stability of micafungin, leading to apparition of degradation products. Moreover, sub visible particles were in the specification of the European Pharmacopeia along 15 days. To conclude, micafungin diluted in NaCl 0.9% and stored in polypropylene syringes was chemically stable for at least 15 days at 25°C in the dark.

International audience; Oral paediatric forms development by pharmaceutical industry is still insufficient. The present study was performed to propose an adapted and pleasant formulation of liquid oral formulation of MTX. The solution is composed of injectable methotrexate, water, Ora Sweet(®) and sodium bicarbonate. After 120 days storage, pH remained stable at about 8 in all formulations, insuring no risk of MTX precipitation. MTX content in solution formulation, determined by high performance liquid chromatography measurements, remained in the specifications of >90% of the initial concentration when stored at 4 and 25°C. Forced degradation of MTX by heat and acidic conditions allowed formation and detection of degradation products by the analytical method. Microbial study of the preparation shows that the solution remains in the specifications during all the storage, or after one sample each week during one month, eventually indicating the microbial properties are not affected by patient use. To conclude, we here propose a new MTX liquid formulation stable for at least 120 days.

Background Docetaxel is an antineoplastic agent widely used in combination with others cytotoxic agents in many cancers (breast cancer, non-small cell lung cancer, prostate cancer, etc.). Today, this costly cytotoxic agent is marketed by several pharmaceutical companies who suggest discarding any remainder immediately after use, making it a very costly drug. Purpose The aim of this study was to determine the physical-chemical stability of docetaxel stock solution after the first sampling in the vial. Materials and methods The study was conducted in accordance with European consensus guidelines for the practical stability of anticancer drugs (1) and by two societies GERPAC and SFPC (2) . The physical-chemical stability was assessed on 3 different vials of docetaxel (Taxotere 20 mg/mL). On day 0, 2, 4 and 30 triplicate samples of each vial of docetaxel were assayed by a high performance liquid chromatography (HPLC) method with UV detection at 230 nm (method validated following ICH guidelines). Docetaxel concentration at day 0 was considered to be 100% and if the docetaxel concentrations in samples were greater than 90% in the following days they were considered stable. The reference concentration was degraded by 20% by addition of a quantity of 0.01N NaOH in order to produce and observe primary degradation products. On each vial and on different days, docetaxel UV absorption spectra between 200 and 600 nm, pH and colour change were compared by a visual inspection with reference at T = 0, and finally a turbidimetry method at 350, 410 and 530 nm was used to evaluate the formation of visible and sub-visible particles. Results After 30 days, for each sample, no colour or pH change were observed, all UV spectra and turbidimetry measures were strictly similar. From day 2 to day 30, docetaxel concentrations were not significantly different to the day 0 solution and no degradation products were observed in any samples. According to these results, no significant drug loss was shown during the study period. Conclusions At a storage temperature between 20 to 25°C for 30 days, docetaxel solution at 20 mg/mL was seen to be stable. The sterility of the solution was not tested because the handling environment (Iso 5) was strictly controlled and operator validations are regularly checked. The authors report. References Bardin C, et al . Guidelines for the practical stability studies of anticancer drugs: A European consensus conference. Ann Pharm Fr 2011;69:221-31 Methodological guidelines for stability studies of hospital pharmaceutical preparations, V Sautou et al ., First edition, October 2013, 74p No conflict of interest.