The initial pharmaceutical development of an artesunate/amodiaquine oral formulation for the treatment of malaria: a public-private partnership

<p>Abstract</p> <p>Background</p> <p>Artemisinin-based combination therapy is currently recommended worldwide for the treatment of uncomplicated malaria. Fixed-dose combinations are preferred as they favour compliance. This paper reports on the initial phases of the pharmaceutical development of an artesunate-amodiaquine (ASAQ) bilayer co-formulation tablet, undertaken following pre-formulation studies by a network of scientists and industrials from institutions of both industrialized and low income countries.</p> <p>Methods</p> <p>Pharmaceutical development was performed by a research laboratory at the University Bordeaux Segalen, School of Pharmacy, for feasibility and early stability studies of various drug formulations, further transferred to a company specialized in pharmaceutical development, and then provided to another company for clinical batch manufacturing. The work was conducted by a regional public-private not-for-profit network (TropiVal) within a larger Public Private partnership (the FACT project), set up by WHO/TDR, Médecins Sans Frontières and the Drugs for Neglected Disease initiative (DND<it>i</it>).</p> <p>Results</p> <p>The main pharmaceutical goal was to combine in a solid oral form two incompatible active principles while preventing artesunate degradation under tropical conditions. Several options were attempted and failed to provide satisfactory stability results: incorporating artesunate in the external phase of the tablets, adding a pH regulator, alcoholic wet granulation, dry granulation, addition of an hydrophobic agent, tablet manufacturing in controlled conditions. However, long-term stability could be achieved, in experimental batches under GMP conditions, by physical separation of artesunate and amodiaquine in a bilayer co-formulation tablet in alu-alu blisters. Conduction of the workplan was monitored by DND<it>i</it>.</p> <p>Conclusions</p> <p>Collaborations between research and industrial groups greatly accelerated the process of development of the bi-layered ASAQ tablet. Lack of public funding was the main obstacle hampering the development process, and no intellectual property right was claimed. This approach resulted in a rapid technology transfer to the drug company Sanofi-Aventis, finalizing the process of development, registration and WHO pre-qualification of the fixed-dose co-formulation together with DND<it>i</it>. The bi-layered tablet is made available under the names of Coarsucam^®and Artesunate amodiaquine Winthrop^®, Sanofi-Aventis. The issue related to the difficulty of public institutions to valorise their participation in such initiative by lack of priority and funding of applied research is discussed.</p

Molecular Blocking of CD23 Supports Its Role in the Pathogenesis of Arthritis

BACKGROUND: CD23 is a differentiation/activation antigen expressed by a variety of hematopoietic and epithelial cells. It can also be detected in soluble forms in biological fluids. Initially known as the low-affinity receptor for immunoglobulin E (Fc epsilonRII), CD23 displays various other physiologic ligands such as CD21, CD11b/c, CD47-vitronectin, and mannose-containing proteins. CD23 mediates numerous immune responses by enhancing IgE-specific antigen presentation, regulating IgE synthesis, influencing cell differentiation and growth of both B- and T-cells. CD23-crosslinking promotes the secretion of pro-inflammatory mediators from human monocytes/macrophages, eosinophils and epithelial cells. Increased CD23 expression is found in patients during allergic reactions and rheumatoid arthritis while its physiopathologic role in these diseases remains to be clarified. METHODOLOGY/PRINCIPAL FINDINGS: We previously generated heptapeptidic countrestructures of human CD23. Based on in vitro studies on healthy and arthritic patients' cells, we showed that CD23-specific peptide addition to human macrophages greatly diminished the transcription of genes encoding inflammatory cytokines. This was also confirmed by significant reduction of mediator levels in cell supernatants. We also show that CD23 peptide decreased IgE-mediated activation of both human and rat CD23(+) macrophages. In vivo studies in rat model of arthritis showed that CD23-blocking peptide ameliorates clinical scores and prevent bone destruction in a dose dependent manner. Ex-vivo analysis of rat macrophages further confirmed the inhibitory effect of peptides on their activation. Taken together our results support the role of CD23 activation and subsequent inflammatory response in arthritis. CONCLUSION: CD23-blocking peptide (p30A) prevents the activation of monocytes/macrophages without cell toxicity. Thus, targeting CD23 by antagonistic peptide decreases inflammatory markers and may have clinical value in the treatment of human arthritis and allergic reactions involving CD23

Intérêt thérapeutique et formulation galénique des polyphénols dans le traitement des infections et inflammations

La réponse immune pro-inflammatoire peut parfois entraîner des processus pathologiques, comme la destruction de la matrice articulaire dans le cas de la polyarthrite rhumatoide ou une cachexie mortelle dans le cas de la trypanosomose africaine. Dans ces deux pathologies, des nouveaux traitements sont nécessaires pour une meilleure prise en charge des malades. Les flavonols, des molécules naturelles végétales peu toxiques, pourraient apporter cette alternative thérapeutique, mais leurs effets anti-inflammatoires et anti-parasitaires doivent être démontrés et/ou caractérisés. Notre travail consistait à évaluer le potentiel thérapeutique de deux flavonols : la quercétine et la rutine. Nous avons étudié leurs effets à des doses non toxiques pour les cellules humaines sur les médiateurs d'inflammation macrophagiques au niveau génomique et protéique. L'inhibition duTNFa, de l'IL1b et du NO a pu être mise en évidence in vitro. L'effet anti-inflammatoire a été confirmé in vivo sur un modèle d'arthrite chez le rat (un modèle proche de la polyarthrite rhumatoide humaine) où l'amélioration des scores cliniques et de la cachexie était corrélée à la diminution des marqueurs inflammatoires sériques. Par ailleurs, l'effet trypanocide de la rutine et de la quercétine a été démontré et étudié in vitro pour déterminer sa cinétque et les relations dose-réponse. De plus, l'effet trypanocide de la vitamine C a pu être démontré in vitro. Les études in vivo devront vonfirmer l'efficacité de ces molécules dans le cas de la trypanosomose africaine. Une première approche de développement galénique a été réalisée pour améliorer la biodisponibilité et donc les effets thérapeutiques des flavonols.Immune response can become a pathological process, leading to a joint matrix destruction in rheumatoid arthritis or to a chronic cachexia in African trypanosomosis. In both pathologies, new therapeutics are needed for a better patients care. Flavonols, non toxic vegetal compounds, could bring a therapeutic alternative in these diseases, but their anti-inflammatory and anti-parasitic effects should be proved and/or characterized. We evaluated therefore the therapeutic potential of two flavonols, quercetin and rutin, in view of their use in human medicine. The effects of non toxic doses of flavonols on macrophage inflammatory mediators' gene transcription and protein expression were studied. In vitro inhibition of TNFa? il1b and NO was also confirmed on rat adjuvant-induced arthritis model, showing a correlation between clinical signs of inflammation (clinical scores and cachexia) and serum inflammatory mediators. In addition, quercetin and rutin trypanocidal effects were demonstrated and the kinetics and dose-response relationship studied. Furthermore, the in vitro trypanocydal effect of vitamin C was highlighted. In vivo studies should confirm the effectiveness of these molecules in African trupanosomosis. A first galenic approach was also conducted in order to improve the bioavailability and consequent therapeutic effects of flavonols.BORDEAUX2-BU Santé (330632101) / SudocSudocFranceF

Lipid oligonucleotides as a new strategy for tackling the antibiotic resistance

International audienceAntibiotic resistance has become a major issue in public health especially for one of the most used antibiotics; the third-generation cephalosporins. One of the main resistance mechanisms in Enterobacteriaceae, is the production of Extended-Spectrum β-lactamases. Here, we demonstrated that the oligonucleotide therapy is an efficient approach to reduce the resistance of bacteria to antibiotic treatment. Lipid oligonucleotides (LONs) were proved to be efficient strategies in both delivering the oligonucleotide sequences in the prokaryotic cells and decreasing the Minimum Inhibitory Concentration of resistant bacteria to a third generation cephalosporin, the ceftriaxone. Accordingly, we demonstrated the strong antimicrobial potential of this LON strategy targeting the ß-lactamase activity on both clinical and laboratory strains. Our results support the concept that the self-delivery of oligonucleotide sequences via lipid conjugation may be extended to other antimicrobial drugs, which opens novel ways to struggle against the antibiotic resistance

Rutoside decreases human macrophage-derived inflammatory mediators and improves clinical signs in adjuvant-induced arthritis

BACKGROUND: Dietary flavonols may play an important role in the adjunct therapy of chronic inflammation. The availability of therapeutic formulations of pentahydroxyflavone glycoside, rutoside (RU), led us to investigate the ability of this molecule to modulate the release of various proinflammatory mediators from human activated macrophages in vitro and to ameliorate arthritic markers in a rat model. METHODS: RU was added simultaneously to human macrophages during their activation. Cells were then analyzed for inflammation-related gene expression using a specific array, and cell supernatants were collected to measure inflammatory mediators. RU was also injected into adjuvant-induced arthritic rats, and disease progression and body weight were evaluated until 50 days after injection. Sera and peritoneal macrophages were also collected to quantify the RU effect on various inflammatory markers. RESULTS: RU inhibited inflammation-related gene expression in activated human macrophages and the release of nitric oxide, tumor necrosis factor-alpha, interleukin (IL)-1, and IL-6 from these cells. In a rat model, RU inhibited clinical signs of chronic arthritis, correlating with decreased levels of inflammatory cytokines detected in rat sera and macrophage supernatants. CONCLUSION: Thus, RU may have clinical value in reducing inflammatory manifestations in human arthritis and other inflammatory diseases

Correction: Nguyen et al. Oligonucleotide Solid Nucleolipid Nanoparticles against Antibiotic Resistance of ESBL-Producing Bacteria. Pharmaceutics 2022, 14, 299

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Preformulation studies of ceftriaxone for pediatric non-parenteral administration as an alternative to existing injectable formulations.

Ceftriaxone, a third generation cephalosporin, has a wide antibacterial spectrum that has good CNS penetration, which makes it potentially suitable for initial treatment of severe neonatal pediatric infections providing suitable formulation. We evaluated its physicochemical and technical characteristics to assess its potential for development as a non-parenteral dosage form. As ceftriaxone is marked only for injectable use, these data are not available. Using HPLC and Karl Fischer titration, sensitivity of ceftriaxone to water, feasibility and impact of pharmaceutical processes and compatibility with common pharmaceutical excipients were assessed. X-ray diffraction studies gave deeper insight into the mechanisms involved in degradation. Chemometrical analysis of near infrared spectra enabled classification of ceftriaxone powder according to exposure conditions or processes applied. The results showed that ceftriaxone was not highly hygroscopic, could be processed in all climatic zones, but should be packaged protected against humidity. Controlling water presence in formulation was shown critical, as ceftriaxone degraded in the presence of water content above 2.4% w/w. To improve flowability, a critical parameter for dry dosage form development, granulation (wet and dry techniques, providing complete drying and moderate force compaction respectively) was shown feasible. Compression with moderate forces was possible, but grinding and high compression forces significantly affected long term ceftriaxone stability and should be avoided. Based on these results, development of ceftriaxone non-parenteral solid or liquid non-aqueous forms appears feasible