Continuous freeze-drying and its relevance to the pharma/biotech industry

The new paradigm of pharmaceutical industry is to move from batch to continuous processes in order to satisfy the stringent requirements of quality, safety and efficiency set by regulatory authorities and reduce production costs. In this perspective, freeze-drying needs to be completely rethought in order to be more integrated in the chain of production of drugs, more flexible to respond to variations in market needs and allowing the monitoring of product quality. The future of freeze-drying, as a downstream process, is therefore to move from batch to continuous. Over the past decades many ideas regarding continuous freeze-drying has been proposed, but none of them has been successfully applied. The objective of this work is to demonstrate the feasibility of an innovative concept to produce lyophilized unit-dose drugs using a continuous process. This novel strategy was demonstrated to improve both yield and vial-to-vial uniformity, giving all those advantages that are typical of continuous technology such as flexibility and elimination of process scale-up from laboratory to industrial scale. The feasibility of continuous freeze-drying has been studied simulating the process using a functional version of the continuous freeze-dryer. Heat transfer during freezing and primary drying was studied reproducing the same conditions occurring in the continuous process. Various process conditions and formulations were investigated in order to better understand the range of applicability of this new process. It has been demonstrated that the cycle duration of the continuous freeze-drying was comparable to that of a conventional batch process, and the aesthetic acceptability of the product was achieved. The continuous freeze-drying technology also impacted positively on inter- and intra-vial heterogeneity. As can be seen Figure 1, the continuous technology gave the most narrow distribution of residual moisture at the end of primary drying. Please click Additional Files below to see the full abstract

Economic analysis of a freeze-drying cycle

Freeze-drying has always been considered an extremely expensive procedure to dehydrate food or pharmaceutical products, and for this reason, it has been employed only if strictly necessary or when the high added value of the final product could justify the costs. However, little effort has been made to analyze the factors that make this technology so unaffordable. In this work, a model was proposed to calculate in detail the operational (OC) and capital costs (CC) of a freeze-drying cycle and an evaluation of the process bottlenecks was made. The main result is that the process itself, contrary to the classic belief, is not the most expensive part of freeze-drying, while the initial investment is the real limiting factor. Under this consideration, the optimization of a freeze-drying cycle should be formulated in order to fit more cycles in the lifespan of the apparatus, instead of merely reducing the power consumption of the machine

Controlled release of ethylene via polymeric films for food packaging

In modern fruit supply chain a common method to trigger ripening is to keep fruits inside special chambers and initiate the ripening process through administration of ethylene. Ethylene is usually administered through cylinders with inadequate control of its final concentration in the chamber. The aim of this study is the development of a new technology to accurately regulate ethylene concentration in the atmosphere where fruits are preserved: a polymeric film, containing an inclusion complex of α-cyclodextrin with ethylene, was developed. The complex was prepared by molecular encapsulation which allows the entrapment of ethylene into the cavity of α-cyclodextrin. After encapsulation, ethylene can be gradually released from the inclusion complex and its release rate can be regulated by temperature and humidity. The inclusion complex was dispersed into a thin polymeric film produced by UV-curing. This method was used because is solvent-free and involves low operating temperature; both conditions are necessary to prevent rapid release of ethylene from the film. The polymeric films were characterized with respect to thermal behaviour, crystalline structure and kinetics of ethylene release, showing that can effectively control the release of ethylene within confined volume

CFD modelling based X-ray microtomography reconstruction of lyophilized products

In this work, 3D non-destructive X-ray micro-CT tomography is used to analyze and reconstruct the internal structure of lyophilized samples, and CFD simulations for calculating their structural properties, i.e., porosity, pore diameter, tortuosity, and permeability

Increased levels of neurotrophins in the cerebrospinal fluid of children with Epstein–Barr virus meningoencephalitis

SummaryObjectiveThe aim of this study was to evaluate the expression of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in the cerebrospinal fluid (CSF) of children with Epstein–Barr virus (EBV)-induced meningoencephalitis (ME) in order to establish a possible correlation with laboratory findings and neurological manifestations.MethodsA prospective observational clinical study was performed on 10 children with viral ME, five of them with EBV-induced ME. As controls, we used CSF samples collected from children admitted with febrile seizures. Neurotrophin levels were measured using an enzyme immunoassay.ResultsSignificantly higher levels of BDNF and NGF were detected in all patients with viral ME compared to controls. Moreover, in patients with EBV-induced ME, the neurotrophin levels were higher than in those with other viral ME. Of note, in children with EBV-induced ME, we found a significant correlation between neurotrophic factor levels and the number of lymphocytes in the CSF (p<0.001). In these patients we also found a significant correlation between BDNF expression and the blood platelet count (p<0.001). Interestingly, two patients with EBV-induced ME showed a correlation between neurotrophin increase and persistent brain abnormalities, such as prolonged alteration of mental status, psychomotor agitation, and athetosis.ConclusionsViral ME induces an early and strong increased biosynthesis of neurotrophic factors. This neurotrophin over-expression is likely to play a key role in the mechanisms of neuronal inflammation and in the severity of brain damage, particularly in EBV-induced ME

Interleukin and neurotrophin up-regulation correlates with severity of H1N1 infection in children: a case–control study

Summary Objective To evaluate the correlation between cytokine and neurotrophin expression and clinical findings, disease severity, and outcome of children with H1N1 influenza infection. Methods A prospective observational clinical study was performed on 15 children with H1N1 infection, 15 controls with lower respiratory tract infections (LRTI), and 15 non-infected children. Plasma levels of interleukin (IL)-1β, IL-6, and neurotrophic factor (nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and glial derived neurotrophic factor (GDNF)) were measured using immunoenzymatic assays. Results Significantly higher levels of IL-1β, IL-6, BDNF, and NGF were detected in patients with H1N1 infection compared to LRTI controls, while there was no significant variation in GDNF in the two groups. IL-1β, IL-6, BDNF, and NGF levels were significantly higher in H1N1 patients with more severe clinical manifestations compared to H1N1 patients with mild clinical manifestations. Of note, IL-6 was significantly correlated with the severity of respiratory compromise and fever, while NGF up-regulation was associated with the duration of cough. No correlation was found between interleukin and neurotrophic factor expression and outcome. Conclusions H1N1 infection induces an early and significant IL-1β, IL-6, BDNF, and NGF up-regulation. The over-expression of these molecular markers is likely to play a neuroimmunomodulatory role in H1N1 infection and may contribute to airway inflammation and bronchial hyper-reactivity in infected children

Image Segmentation and 3D reconstruction for improved prediction of the sublimation rate during freeze drying

[EN] In a freeze drying process, the freezing step determines the pore size distribution within the product, which, in turn, affects the sublimation rate. Traditionally, pore analysis is carried out on SEM images by means of a manual, time-consuming approach. Here, an image segmentation technique was used to automatize this process and improve its reliability. A 3D structure of the cake was then reconstructed from the distribution of the super-pixels. We show that the approach herein proposed can remarkably improve prediction of the sublimation rate with respect to traditional methods.Computational resources were provided by ISCRA-Cineca HPC CLASS-C Grant to L.C.C. (ParticLy - HP10CQRVJV)Capozzi, L.; Arsiccio, A.; Sparavigna, A.; Pisano, R.; Barresi, A. (2018). Image Segmentation and 3D reconstruction for improved prediction of the sublimation rate during freeze drying. En IDS 2018. 21st International Drying Symposium Proceedings. Editorial Universitat Politècnica de València. 411-418. https://doi.org/10.4995/IDS2018.2018.7646OCS41141

DHA-Induced Perturbation of Human Serum Metabolome. Role of the Food Matrix and Co-Administration of Oat β-glucan and Anthocyanins

Docosahexaenoic acid (DHA) has been reported to have a positive impact on many diet-related disease risks, including metabolic syndrome. Although many DHA-enriched foods have been marketed, the impact of different food matrices on the effect of DHA is unknown. As well, the possibility to enhance DHA effectiveness through the co-administration of other bioactives has seldom been considered. We evaluated DHA effects on the serum metabolome administered to volunteers at risk of metabolic syndrome as an ingredient of three different foods. Foods were enriched with DHA alone or in combination with oat beta-glucan or anthocyanins and were administered to volunteers for 4 weeks. Serum samples collected at the beginning and end of the trial were analysed by NMR-based metabolomics. Multivariate and univariate statistical analyses were used to characterize modifications in the serum metabolome and to evaluate bioactive-bioactive and bioactive-food matrix interactions. DHA administration induces metabolome perturbation that is influenced by the food matrix and the co-presence of other bioactives. In particular, when co-administered with oat beta-glucan, DHA induces a strong rearrangement in the lipoprotein profile of the subjects. The observed modifications are consistent with clinical results and indicate that metabolomics represents a possible strategy to choose the most appropriate food matrices for bioactive enrichmen

A potential prognostic marker in primitive lung neuroendocrine tumor: A case report.

Background: The diagnosis and monitoring of primitive lung neuroendocrine tumors (lung pNETs) are usually performed by the measurement of serum chromogranin A (CgA) and urinary 5-hydroxyindolacetic acid (5-HIAA) levels. However, imaging techniques are necessary due to the poor diagnostic efficiency of the laboratory tests. Methods: A total-body computed tomography and bone scintigraphy scans showed multiple hepatic and bone metastases of a 55-year-old man affected by well-differentiated lung pNETs without severe initial symptoms. After diagnosis, he started therapy and was monitored with serum, urinary markers, and imaging techniques. Results: During follow-up, the urinary 5-HIAA levels did not significantly increase, while serum CgA and urinary para-hydroxyphenylacetic acid (pHPAA) levels (urinary organic acid physiologically present in the urines of healthy subjects) showed significant increases related to worsening clinical condition. Conclusions: The early increase in urinary pHPAA levels—usually not dosed in pNET patient monitoring—could be a promising prognostic marker