INVESTIGATION OF CHEMICAL DIFFERENCES IN MEDICATIONS OBTAINED FROM DIVERSE SOURCES USING NOVEL SPECTROSCOPIC AND STATISTIC APPROACHES

Generic medications are those medicines manufactured by a pharmaceutical company without a license from the company that has first invented and patented the same drug, when the related patent and other exclusivity rights have expired. Only studies of bioequivalence are requested as requirements to introduce a new generic medication in clinics, making easier and financially attractive for many pharmaceutical companies to participate in this typology of market. Organised criminality is strongly attracted by this market both for the high profitability, and for the high similarity to the production and trafficking of illegal controlled substances associated to the extreme difficulties faced by the law enforcement authorities in effectively investigating the online market, because of its anonymity. A significant paradigm continuously frequented in pharmacology is the confliction between views on generic medications that can be used interchangeably with the original medicines. Several clinical studies conducted in certain medical areas have shown as the generic medications present an overlapping therapeutic equivalence to the original ones. On the contrary, for certain other generics, both pharmacokinetics and pharmacodynamics issue have been reported. Also, in some cases issues on the stability of generics have been raised. Despite the numerous research articles and reviews published on the matter of clinical equivalence among generic and original medications, no study to the scientific community has been presented on an analytical evaluation of the chemical composition of the different generic drugs that could shed some lights on the reason of the different clinical performances reported. The main aim of this research was to develop a non-destructive quick qualitative analytical methodology to be able to discriminate differences in the chemical composition from generic medicines that have been reported not presenting similar therapeutic equivalence in clinical comparison studies, obtained from authorised pharmacies and non-authorised online sellers. From the cardio-vascular area, digoxin (with the related cardiac glycosides digitoxin and digoxigenin) and amlodipine (in its different salts maleate, mesylate and besylate used in therapies). In the gastroenterology area, omeprazole both in its racemic and isomeric forms, have been selected as samples to be analysed. In the antihistamine area, cetirizine, in its racemic and isomeric forms, equally for the same reasons as before, have been considered. As starting analytical approaches, voltammetry, FT-IR, Raman spectroscopies and NMR have been considered and a statistical data analysis approach of the analytical data obtained based on multivariate analysis such as principal component analysis, cross validation, correlation scatter plots and factor loadings has been implemented. This work has matched the aims initially set, generating novel methods of analysis to investigate differences in the chemical composition within different groups of generic medications. This study has led to the creation and interpretation of new knowledge, through a systematic acquisition and understanding of a substantial body of scientific literature and through original research, and adjusting the project design in the light of unforeseen problems, conceptualizing, designing and implementing the research project for the generation of novel knowledge

The test method of composting in vermiculite is unaffected by the priming effect

Glucose, starch, and cellulose can increase the biodegradation of the compost used as a solid matrix in the biodegradation test under composting conditions (priming effect). The enhanced evolution of carbon dioxide determines an overestimation of the biodegradation of the starch- and cellulose-based materials and, in some cases, values higher than 100% can be reached. The work has verified that using activated vermiculite, an inorganic matrix, the priming effect can be reduced, improving the reliability of the test method. Glucose, the most effective primer, causes the attainment of biodegradation values significantly higher than 100% in mature compost while this does not happen in activated vermiculite. Since all the initial carbon present in the activated vermiculite is converted into CO2 within the test period, we conclude that a substantial priming effect cannot occur for the lack of organic carbon. Furthermore, by measuring in parallel both the consumption of glucose and the CO2 evolution, the yield of CO2 production ( ) was determined. In no case a value higher than 1, a clear indication of priming effect, was found

Degradability of plastics. Standard methods developed in Italy

Degradability of plastics. Standard methods developed in Ital

Activated Vermiculite, a Solid Bed for Testing Biodegradability under Composting Conditions

Vermiculite, a clay mineral, can be activated and used as a solid matrix in place of mature compost in the controlled composting test, a respirometric essay widely applied to assess biodegradability of plastics. The results obtained with two materials (cellulose and a starch-based blend) indicate that activated vermiculite affects neither the biodegradation rate nor the final biodegradation level. On the other hand, possible metabolic intermediates and polymeric residues left after biodegradation can be recovered more easily from activated vermiculite than from mature compost, a very complex organic matter. Therefore, at test termination it was possible to determine the carbon balance taking into account both the evolved CO2 and a polymeric residue extracted from vermiculite, totalling 101% of the carbon present originally in the test material. This work was partly financed by the European Commission (DGXII) with the SMT “Labelling biodegradable products” project SMT4-CT97-2187

Detection of toxicity released by a biodegradable plastic after composting in activated vermiculite

The composting test method based on activated vermiculite is a comprehensive system for the assessment of the environmental impact of biodegradable plastics. It allows, in a single test, (i) the measurement of the mineralization of the polymer under study; (ii) the retrieval of the final polymeric residues and (iii) determination of the biomass (to make a final mass balance); (iv) detection of breakdown products of the original polymer. In this study it is shown that the vermiculite test method is also suitable to perform ecotoxicological studies. The Flash test is a method based on kinetic measurement of bioluminescence by Vibrio fischeri, and was applied to evaluate the toxicity of compost samples and vermiculite samples after the biodegradation of a polyurethane (PU) based plastic material. Toxicity was detected in vermiculite samples contaminated by 4,4′ diamino diphenyl methane (MDA), a toxic breakdown product released by the PU moiety, as shown by HPLC. On the other hand, neither toxicity nor the presence of MDA was detected in mature compost. A recovery experiment previously performed had shown a 10% MDA recovery yield from mature compost. The possibility of testing the ecotoxicity of extracts obtained from mineral matrix after biodegradation makes the vermiculite test system particularly interesting for the overall assessment of the environmental impact of biodegradable plastics

Isolation and characterisation of thermophilic micro-organisms able to grow on cellulose acetate

Cellulose acetate (CA) can be successfully used for producing compostable plastics. It is biodegradable under composting conditions, but neither the mechanism of degradation nor the involved microorganisms are known. We isolated five thermophilic strains from compost which were identified by molecular characterisation as belonging to the family Thermomonosporaceae, probably to the genus Actinomadura. The strains are able to grow at 50 °C on minimal plates supplemented with CA (DS=2) or microcrystalline cellulose in powder. The degradation of CA is visible by the clarification of the area surrounding the mycelium. The strains grow at 37 °C on cellulose but not on CA. At 20 and 58 °C no growth is detected on both substrates. The strains are viable after a 2-week incubation period at 65 °C. Neutral and basic pHs are the best for CA degradation, while poor results are obtained under acidic conditions. The best degradation of CA is obtained using nitrate as nitrogen source. Solid-state respiration tests, performed using sterile vermiculite as a matrix, confirmed that the strains are able to mineralise the CA. The strains failed to grow in CA under liquid conditions

Real-world experience with obeticholic acid in patients with primary biliary cholangitis

Background & aims: Obeticholic acid (OCA) is the second-line treatment approved for patients with primary biliary cholangitis (PBC) and an inadequate response or intolerance to ursodeoxycholic acid. We aimed to evaluate the effectiveness and safety of OCA under real-world conditions.Methods: Patients were recruited into the Italian PBC Registry, a multicentre, observational cohort study that monitors patients with PBC at national level. The primary endpoint was the biochemical response according to Poise criteria; the secondary endpoint was the biochemical response according to normal range criteria, defined as normal levels of bilirubin, alkaline phosphatase (ALP), and alanine aminotransferase (ALT) at 12 months. Safety and tolerability were also assessed.Results: We analysed 191 patients until at least 12 months of follow-up. Median age was 57 years, 94% female, 61 (32%) had cirrhosis, 28 (15%) had histologically proven overlap with autoimmune hepatitis (PBC-AIH). At 12 months, significant median reductions of ALP (-32.3%), ALT (-31.4%), and bilirubin (-11.2%) were observed. Response rates were 42.9% according to Poise criteria, and 11% by normal range criteria. Patients with cirrhosis had lower response than patients without cirrhosis (29.5% vs. 49.2%, p = 0.01), owing to a higher rate of OCA discontinuation (30% vs. 12%, p = 0.004), although with similar ALP reduction (29.4% vs. 34%, p = 0.53). Overlap PBC-AIH had a similar response to pure PBC (46.4% vs. 42.3%, p = 0.68), with higher ALT reduction at 6 months (-38% vs. -29%, p = 0.04). Thirty-three patients (17%) prematurely discontinued OCA because of adverse events, of whom 11 experienced serious adverse events. Treatment-induced pruritus was the leading cause of OCA discontinuation (67%).Conclusions: Effectiveness and safety of OCA under real-world conditions mirror those in the Poise trial. Patients with cirrhosis had lower tolerability. Overlap PBC-AIH showed higher ALT reduction at 6 months compared with patients with pure PBC.Lay summary: Obeticholic acid (OCA) was shown to be effective in more than one-third of patients not responding to ursodeoxycholic acid in a real-world context in Italy. Patients with cirrhosis had more side effects with OCA, and this led to suspension of the drug in one-third of patients. OCA was also effective in patients who had overlap between autoimmune hepatitis and primary biliary cholangitis. (C) 2021 The Author(s). Published by Elsevier B.V. on behalf of European Association for the Study of the Liver (EASL)

Incidence of Hepatocellular Carcinoma in Patients With HCV-Associated Cirrhosis Treated With Direct-Acting Antiviral Agents

Background & Aims: Studies have produced conflicting results of the incidence of hepatocellular carcinoma (HCC) in patients with hepatitis C virus–associated cirrhosis treated with direct-acting antivirals (DAAs). Data from clinics are needed to accurately assess the occurrence rate of HCC in patients with cirrhosis in the real world. Methods: We collected data from a large prospective study of 2,249 consecutive patients (mean age = 65.4 years, 56.9% male) with hepatitis C virus–associated cirrhosis (90.5% with Child-Pugh class A and 9.5% with Child-Pugh class B) treated with DAAs from March 2015 through July 2016 at 22 academic and community liver centers in Sicily, Italy. HCC occurrence was evaluated by Kaplan-Meier curves. Cox regression analysis was used to identify variables associated with HCC development. Results: A sustained virologic response (SVR) was achieved by 2,140 patients (total = 95.2%; 95.9% with Child Pugh class A and 88.3% with Child Pugh class B; P <.001). Seventy-eight patients (3.5%) developed HCC during a mean follow-up of 14 months (range = 6–24 months). At 1 year after exposure to DAAs, HCC developed in 2.1% of patients with Child-Pugh class A with an SVR and 6.6% of patients with no SVR and in 7.8% of patients with Child-Pugh class B with an SVR and 12.4% of patients with no SVR (P <.001 by log-rank test). Albumin level below 3.5 g/dL (hazard ratio = 1.77, 95% confidence interval = 1.12–2.82, P =.015), platelet count below 120 × 109/L (hazard ratio = 3.89, 95% confidence interval = 2.11–7.15, P <.001), and absence of an SVR (hazard ratio = 3.40, 95% confidence interval = 1.89–6.12, P <.001) were independently associated increased risk for HCC. The mean interval from exposure to DAAs to an HCC diagnosis was 9.8 months (range = 2–22 months) and did not differ significantly between patients with (n = 64, 9.2 months) and without (n = 14, 12.0 months) an SVR (P =.11). A larger proportion of patients with an SVR had a single HCC lesion (78% vs 50% without an SVR; P =.009) or an HCC lesion smaller than 3 cm (58% vs 28% without an SVR; P =.07). Conclusions: In an analysis of data from a large prospective study of patients with hepatitis C virus–associated compensated or decompensated cirrhosis, we found that the SVR to DAA treatment decreased the incidence of HCC over a mean follow-up of 14 months

Development and Validation of a Scoring System to Predict Response to Obeticholic Acid in Primary Biliary Cholangitis

Background & aims: Obeticholic acid (OCA) is the only licensed second-line therapy for primary biliary cholangitis (PBC). With novel therapeutics in advanced development, clinical tools are needed to tailor the treatment algorithm. We aimed to derive and externally validate the OCA response score (ORS) for predicting the response probability of individuals with PBC to OCA. Methods: We used data from the Italian RECAPITULATE (N = 441) and the IBER-PBC (N = 244) OCA real-world prospective cohorts to derive/validate a score including widely available variables obtained either pre-treatment (ORS) or also after 6 months of treatment (ORS+). Multivariable Cox regressions with backward selection were applied to obtain parsimonious predictive models. The predicted outcomes were biochemical response according to POISE (alkaline phosphatase [ALP]/upper limit of normal [ULN]<1.67 with a reduction of at least 15%, and normal bilirubin), or ALP/ULN<1.67, or normal range criteria (NR: normal ALP, alanine aminotransferase [ALT], and bilirubin) up to 24 months. Results: Depending on the response criteria, ORS included age, pruritus, cirrhosis, ALP/ULN, ALT/ULN, GGT/ULN, and bilirubin. ORS+ also included ALP/ULN and bilirubin after 6 months of OCA therapy. Internally validated c-statistics for ORS were 0.75, 0.78, and 0.72 for POISE, ALP/ULN<1.67, and NR response, which raised to 0.83, 0.88, and 0.81 with ORS+, respectively. The respective performances in validation were 0.70, 0.72, and 0.71 for ORS and 0.80, 0.84, and 0.78 for ORS+. Results were consistent across groups with mild/severe disease. Conclusions: We developed and externally validated a scoring system capable to predict OCA response according to different criteria. This tool will enhance a stratified second-line therapy model to streamline standard care and trial delivery in PBC