In-lab synthesized turn-off fluorescence sensor for estimation of Gemigliptin and Rosuvastatin polypill appraised by Spider diagram, AGREE and whiteness metrics.

Gemigliptin-Rosuvastatin single-pill combination is a promising therapeutic tool in the effective control of hyperglycemia and hypercholesterolemia. Organic sensors with high quantum yields have profoundly significant applications in the pharmaceutical industry, such as routine quality control of marketed formulations. Herein, the fluorescence sensor, 2-Morpholino-4,6-dimethyl nicotinonitrile 3, (λex; 226 nm, λem; 406 nm), was synthesized with a fluorescence quantum yield of 56.86% and fully characterized in our laboratory. This sensor showed high efficiency for the determination of Gemigliptin (GEM) and Rosuvastatin (RSV) traces through their stoichiometric interactions and simultaneously fractionated by selective solvation. The interaction between the stated analytes and sensor 3 was a quenching effect. Various experimental parameters and the turn-off mechanism were addressed. The adopted approach fulfilled the ICH validation criteria and showed linear satisfactory ranges, 0.2-2 and 0.1-1 µg/mL for GEM and RSV, respectively with nano-limits of detection less than 30 ng/mL for both analytes. The synthesized sensor has been successfully applied for GEM and RSV co-assessment in their synthetic polypill with excellent % recoveries of 98.83 ± 0.86 and 100.19 ± 0.64, respectively. No statistically significant difference between the results of the proposed and reported spectrophotometric methods in terms of the F- and t-tests. Ecological and whiteness appraisals of the proposed study were conducted via three novel approaches: the Greenness Index via Spider Diagram, the Analytical Greenness Metric, and the Red-Green-Blue 12 model. The aforementioned metrics proved the superiority of the adopted approach over the previously published one regarding eco-friendliness and sustainability. Our devised fluorimetric turn-off sensing method showed high sensitivity, selectivity, feasibility, and rapidity with minimal cost and environmental burden over other sophisticated techniques, making it reliable in quality control labs.

Culturable microbiological profile of a non-sterile drugs pharmaceutical production environment

Abstract For places where non-sterile drug production occurs, regulatory bodies recommend monitoring of the environmental bioburden. This procedure provides information regarding possible microbiological risks to which the products may be exposed, so that subsequent action measures may be implemented. The aim of the present work was to quantify and characterize the microorganisms present in Grade D (ISO 8) cleanrooms of a Brazilian pharmaceutical industry, identifying any possible seasonal climatic influences on these environments. Sampling was performed by surface and air monitoring, over 12 months during the year 2019, in rooms that were in operation. For both sampling methods, no statistically significant differences in bacteria and fungi counts were found between months or seasonal periods. Microorganisms that presented higher incidence included Staphylococcus epidermidis (15%) and Micrococcus spp. (13%), common to the human microbiota, and the fungi Cladosporium sp. (23%) and Penicillium sp. (21%), typical of the external environment. The results showed that microbial contamination in the Grade D cleanrooms was within the permissible maximum levels and remained similar throughout the year. Microbiological quality control in the clean areas of the pharmaceutical industry investigated was considered effective, with regular maintenance being necessary to keep bioburden levels controlled.

Automated Good Manufacturing Practice-compliant generation of human monocyte-derived dendritic cells from a complete apheresis product using a hollow-fiber bioreactor system overcomes a major hurdle in the manufacture of dendritic cells for cancer vaccines.

BACKGROUND: Although dendritic cell (DC)-based cancer vaccines represent a promising treatment strategy, its exploration in the clinic is hampered due to the need for Good Manufacturing Practice (GMP) facilities and associated trained staff for the generation of large numbers of DCs. The Quantum bioreactor system offered by Terumo BCT represents a hollow-fiber platform integrating GMP-compliant manufacturing steps in a closed system for automated cultivation of cellular products. In the respective established protocols, the hollow fibers are coated with fibronectin and trypsin is used to harvest the final cell product, which in the case of DCs allows processing of only one tenth of an apheresis product. MATERIALS AND RESULTS: We successfully developed a new protocol that circumvents the need for fibronectin coating and trypsin digestion, and makes the Quantum bioreactor system now suitable for generating large numbers of mature human monocyte-derived DCs (Mo-DCs) by processing a complete apheresis product at once. To achieve that, it needed a step-by-step optimization of DC-differentiation, e.g., the varying of media exchange rates and cytokine concentration until the total yield (% of input CD14+ monocytes), as well as the phenotype and functionality of mature Mo-DCs, became equivalent to those generated by our established standard production of Mo-DCs in cell culture bags. CONCLUSIONS: By using this new protocol for the Food and Drug Administration-approved Quantum system, it is now possible for the first time to process one complete apheresis to automatically generate large numbers of human Mo-DCs, making it much more feasible to exploit the potential of individualized DC-based immunotherapy.

Using Extractables Data of Sterile Filter Components for Scaling Calculations.

Users of single-use (SU) components need extractables data to demonstrate material safety for regulatory bodies before incorporation in the biopharmaceutical process. In this context, the correct use of such extractables data is key to deriving realistic risk assessments for SU devices. In this paper, a standardized extractables approach was used that provides comprehensive extractables information including identity and quantity. The combination of extractables data obtained from different components of a sterile filter capsule, such as the filter cartridge and housing, and the scaling thereof is presented. A sterile filter type including polyethersulfone membrane was extracted with pure water and pure ethanol at 40°C for 24 h. The organic extractables were identified and their concentration quantified using state-of-the-art analytical methods such as gas chromatography-mass spectrometry for semivolatile compounds together with headspace sampling for volatile compounds, and liquid chromatography coupled with high-resolution mass spectrometry. The extractables detected were assigned to the materials of filter construction. The evaluation showed that the extractables quantities per device depended on the surface areas of the contact materials, such as the filter membrane, and also on the plastic parts. This paper confirms the validity of a so-called component approach and a scaling concept to calculate extractables data for SU filters of different sizes with short-term contact.LAY ABSTRACT: In the biopharmaceutical industry, a large number of SU system combinations with a variety of different sizes are used. Suppliers of such diverse SU systems and assemblies cannot perform extraction studies for all of the different configurations and sizes individually. It is acceptable in this industry to use component approaches and scaling concepts to provide extractables data for SU systems and assemblies derived from a dedicated extraction experiment. This paper shows the applicability of a so-called component approach and of a scaling concept to calculate extractables data for sterile polyethersulfone membrane filters and filter capsules of different sizes. Selected extraction conditions allow scaling calculations according to underlying physical principles. The extractions were performed under short-term contact, for example, 24 h, to ensure that the release of extractables was diffusion-controlled. The results demonstrated that extractables quantities depend on the surface area of the contact material. Membrane-related compounds were scalable with the membrane area, whereas polypropylene (PP)-related compounds were scalable with the PP contact area.

Across-the-World Automated Optimization and Continuous-Flow Synthesis of Pharmaceutical Agents Operating Through a Cloud-Based Server.

The power of the Cloud has been harnessed for pharmaceutical compound production with remote servers based in Tokyo, Japan being left to autonomously find optimal synthesis conditions for three active pharmaceutical ingredients (APIs) in laboratories in Cambridge, UK. A researcher located in Los Angeles, USA controlled the entire process via an internet connection. The constituent synthetic steps for Tramadol, Lidocaine, and Bupropion were thus optimized with minimal intervention from operators within hours, yielding conditions satisfying customizable evaluation functions for all examples.

Development of Flow Imaging Analysis for Subvisible Particle Characterization in Glatiramer Acetate.

Proteins, peptides, colloids, and polymers present a rapidly growing field of pharmaceutical industry. Bringing these products into market, however, is a huge regulatory challenge, especially because many of these therapeutics are intended for parenteral administration. Physicochemical properties of such products--size, shape, surface potential, and extent of particle-particle interaction-have to be well understood and monitored throughout manufacturing, release, and stability testing. First and foremost, size distribution of subvisible particles (SVP) in these products should be reliably measured. We present development of a flow imaging method to assess SVP in the polypeptide injectable therapeutic product-glatiramer acetate (Copaxone(®)). Flow imaging comprises optical inspection of a flowing liquid and allows quantitation of particles in the range of 1-500 µm. The challenges of method development are discussed and the method performance characteristics--accuracy, precision, linearity, and specificity--are demonstrated.

Focused beam reflectance method as an innovative (PAT) tool to monitor in-line granulation process in fluidized bed.

Fluidized bed granulation is a commonly used unit operation in the pharmaceutical industry. But still to obtain and control the desired granule size is challenging due to many process variables affecting the final product. Focused beam reflectance measurement (FBRM, Mettler-Toledo, Switzerland) is an increasingly popular particle growth analysis technique. FBRM tool was installed in two different locations inside a fluidized bed granulator (GPCG2, Glatt, Binzen) in order to monitor the granulation growth kinetics. An experimental design was created to study the effect of process variables using FBRM probe and comparing the results with the one's measured by sieve analysis. The probe location is of major importance to get smooth and robust curves. The excess feeding of binder solution might lead to agglomeration and thus to process collapse, however this phenomenon was clearly detected with FBRM method. On the other hand, the process variables at certain levels might affect the FBRM efficiency by blocking the probe window with sticky particles. A good correlation was obtained (R(2) = 0.95) between FBRM and sieve analysis mean particle size. The proposed in-line monitoring tool enables the operator to select appropriate process parameters and control the wet granulation process more efficiently.

Inverse spatially offset Raman spectroscopy for deep noninvasive probing of turbid media.

A new type of highly sensitive spatially offset Raman spectroscopy (SORS) developed for deep noninvasive probing of stratified turbid media is described. The technique, termed inverse SORS, permits much greater depths to be interrogated than those accessible with the conventional SORS approach. This is achieved by enhancing the sensitivity of the technique through the elimination of spectral distortions inherent to the conventional SORS methodology. The method also permits the use of higher laser powers in applications where intensity limits exist, such as when probing human tissue in vivo. In addition, the new approach possesses a much higher degree of flexibility, enabling on-the-spot tailoring of experimental conditions such as the magnitude and number of spatial offsets to individual samples. The scheme uses a reverse SORS geometry whereby Raman light is collected through fibers at the center of the probe and laser radiation is delivered to the sample through a beam in the shape of a ring. The method is demonstrated on a layered powder sample and several practical examples of its uses, presented for the first time, are also given. Potential applications include disease diagnosis, noninvasive probing of pharmaceutical products and chemicals through packaging, probing of polymers, biofilms or paints, and homeland security screening.

Substructure-based support vector machine classifiers for prediction of adverse effects in diverse classes of drugs.

Unforeseen adverse effects exhibited by drugs contribute heavily to late-phase failure and even withdrawal of marketed drugs. Torsade de pointes (TdP) is one such important adverse effect, which causes cardiac arrhythmia and, in some cases, sudden death, making it crucial for potential drugs to be screened for torsadogenicity. The need to tap the power of computational approaches for the prediction of adverse effects such as TdP is increasingly becoming evident. The availability of screening data including those in organized databases greatly facilitates exploration of newer computational approaches. In this paper, we report the development of a prediction method based on a support machine vector algorithm. The method uses a combination of descriptors, encoding both the type of toxicophore as well as the position of the toxicophore in the drug molecule, thus considering both the pharmacophore and the three-dimensional shape information of the molecule. For delineating toxicophores, a novel pattern-recognition method that utilizes substructures within a molecule has been developed. The results obtained using the hybrid approach have been compared with those available in the literature for the same data set. An improvement in prediction accuracy is clearly seen, with the accuracy reaching up to 97% in predicting compounds that can cause TdP and 90% for predicting compounds that do not cause TdP. The generic nature of the method has been demonstrated with four data sets available for carcinogenicity, where prediction accuracies were significantly higher, with a best receiver operating characteristics (ROC) value of 0.81 as against a best ROC value of 0.7 reported in the literature for the same data set. Thus, the method holds promise for wide applicability in toxicity prediction.

Noninvasive Raman spectroscopy of human tissue in vivo.

We report the first transcutaneous Raman spectrum of human bone in vivo obtained at skin-safe laser illumination levels. The spectrum of thumb distal phalanx was obtained using spatially offset Raman spectroscopy (SORS), which provides chemically specific information on deep layers of human tissue, well beyond the reach of existing comparative approaches. The spectroscopy is based on collecting Raman spectra away from the point of laser illumination using concentric rings of optical fibers. As a generic analytical tool this approach paves the way for a range of uses including disease diagnosis, noninvasive probing of pharmaceutical products, biofilms, catalysts, paints, and in dermatological applications.

Paclitaxel production by plant-cell-culture technology.

Plant cells catalyze multiple-step reactions of secondary metabolite biosynthesis, and selectively synthesize chiral compounds with polycyclic structures. Taking advantage of this characteristic, we studied the production of the anticancer drug paclitaxel, which is currently produced in limited supply. Callus culture investigations indicate that woody plant medium supplemented with 10(-5) mol L(-1) 1-naphthylacetic acid and without the NH4+ -type ion is the best condition for growth of the callus. The accumulation of paclitaxel and related taxanes in Taxus plants is thought to be a biological response to specific external stimuli. Several signal transducers were screened; taxane biosynthesis was strongly promoted by methyl jasmonate (MeJA) and silver thiosulfate (STS) as an anti-ethylene compound. Of ten taxane-type diterpenoids isolated from T. baccata suspension-cultured cells treated with MeJA, five have a phenylisoserine side-chain at the C-13 position of the taxane skeleton. Time-course analysis revealed two regulatory steps in taxane biosynthesis: the taxane-ring formation step and the acylation step of the C-13 position. Methyl jasmonate promoted the formation of the taxane-ring. The production of paclitaxel reached a maximum level of 295 mg L(-1) in a large-scale culture of T. x media cells using a two-stage process.

AutoGenomics, Inc.

AutoGenomics has created an automated multiplexing microarray platform to make genomic and proteomic analyses routine and efficient for clinical and research laboratories. While the emergence of microarrays has advanced genomic analyses, a number of underlying issues, such as cross-hybridization, poor spot morphology and intrinsic fluorescence of the solid substrate, have yet to be fully resolved. Current methods use discrete instrumentation, are manual and require highly skilled labor, which leads to inconsistent results. AutoGenomics' automated platform uses a three-dimensional BioFilmChip microarray to circumvent these issues, providing optimal spot morphology and utilizing solution-based hybridization with allele-specific primer extension to improve single-base discrimination. AutoGenomics is developing applications for the early detection and management of complex disease states in oncology, cardiology, and mental disorders. Customers include clinical reference laboratories, hospitals, academic institutions, and pharmaceutical and biotech companies. Founded in 1999, the company is headquartered in Carlsbad, California, USA.

[Gas sterilization with gas hydrogen peroxide: a new technology in the pharmaceutical industry]. / Sterilizzazione con perossido di idrogeno in fase gassosa: una nuova tecnologia a disposizione dell'industria farmaceutica.

This article shows the bactericidal capacity of hydrogen peroxide vapor, particularly if used through VHP vaporizer (Vaporizer Hydrogen Peroxide) which was recently developed and used in the pharmaceutical sector. After a short description technologies and of the characteristics of the chemical agents used in these technics, will illustrate the machine (VHP), its functioning and its possible application fields. The review ends with the illustration of a number of experiments, tests and validation trials which stand out from technical point of view.

Metallurgical, manufacturing and surface finish requirements for high purity stainless steel system components.

The stainless steel piping systems that distribute water for injection (WFI) and other critical fluids in pharmaceutical and biotechnology plants will be the subject of this paper. We will confine our focus to the two major components of such systems--tubing and fittings. It should be noted that the discussions here can certainly be applied in the selection of other components such as valves, filter housings, pumps, and tanks used in these industries. The purpose of this paper will be to guide user choices with regard to raw material selection, surface preparation, and surface analysis within the context of system requirements. A suggested material specification for stainless steel tubing and fittings will also be presented.