Basics of advanced therapy medicinal product development in academic pharma and the role of a GMP simulation unit.

Following successes of authorized chimeric antigen receptor T-cell products being commercially marketed in the United States and European Union, product development of T-cell-based cancer immunotherapy consisting of cell-based advanced therapy medicinal products (ATMPs) has gained further momentum. Due to their complex characteristics, pharmacological properties of living cell products are, in contrast to classical biological drugs such as small molecules, more difficult to define. Despite the availability of many new advanced technologies that facilitate ATMP manufacturing, translation from research-grade to clinical-grade manufacturing in accordance with Good Manufacturing Practices (cGMP) needs a thorough product development process in order to maintain the same product characteristics and activity of the therapeutic product after full-scale clinical GMP production as originally developed within a research setting. The same holds true for transferring a fully developed GMP-grade production process between different GMP facilities. Such product development from the research to GMP-grade manufacturing and technology transfer processes of established GMP-compliant procedures between facilities are challenging. In this review, we highlight some of the main obstacles related to the product development, manufacturing process, and product analysis, as well as how these hinder rapid access to ATMPs. We elaborate on the role of academia, also referred to as 'academic pharma', and the added value of GMP production and GMP simulation facilities to keep innovation moving by reducing the development time and to keep final production costs reasonable.

[The accidental aspiration and ingestion of petroleum in a "fire eater"]. / Akzidentelle Aspiration und Ingestion von Petroleum bei einem "Feuerschlucker".

A 26-year-old man, practicing for a variety performance as "fire-eater", accidentally inhaled and ingested about 10 ml petroleum. Soon afterwards he developed dyspnoea, an urge to cough, fever up to 39 degrees C and loss of retentiveness. He was treated as an out-patient with doxycycline, 100 mg daily, and aspirin, 500 mg three times daily. While this reduced the dyspnoea, the elevated temperature persisted and he had haemoptysis. Chest x-ray and computed tomography 12 days after the aspiration revealed areas of atelectasis and of liquefaction necroses. Bronchoscopic and cytological examinations showed eosinophilic alveolitis and mucosal necrosis in both main bronchi. The symptoms were improved by two inhalations of beclomethasone four times daily, and systemic treatment with prednisolone, 50 mg daily, together with parenteral antibiotic administration (cefotaxime, 1.0 g twice daily). The focal lung lesions regressed completely within a few weeks. Five months after the aspiration computed tomography merely demonstrated discrete scarring of the previously necrotic lesions. This case illustrates that, even with extensive necrotic lung changes after petroleum aspiration, conservative treatment is justified and likely to be effective.

[Relation between 67-gallium scintigraphy and bronchoalveolar lavage--differential cell count and superoxide anion liberation--in patients with systemic scleroderma and systemic lupus erythematosus]. / Beziehungen zwischen 67-Gallium-Szintigraphie und bronchoalveolärer Lavage--Differentialzellbild und Superoxid-Anion-Freisetzung--bei Patienten mit systemischer Sklerodermie und systemischem Lupus erythematodes.

The aim of the study was to determine the pulmonary 67-gallium uptake, bronchoalveolar lavage (BAL) cell differentiation and the activity of BAL cells, measured as release of superoxide anion (O2-), and to investigate the results whether there are relations. In 11 nonsmoking systemic scleroderma (SS) patients and 11 systemic lupus erythematosus (SLE) patients with lung involvement double-sided BAL were performed, mainly in regions with increased 67-gallium uptake. Release of O2- was measured by INT-assey. BAL cell differentiation was in SS and SLE pathological in 68.2% without side-difference. In contrast to SS, O2(-)-release in SLE depends on BAL cell differentiation and is most increased in normal BAL cell differentiation. There is no correlation between 67-gallium uptake and both BAL cell differentiation and O2(-)-release. The results suggest, that in contrast to SS, BAL cells in SLE with pathological differentiation are less activated than BAL cells with normal differentiation probable due to autoimmunological factors. Pulmonary 67-gallium scan and BAL seem to be independent from each other.

The growth of ordered two-dimensional sheets of ribosomal particles from salt-alcohol mixtures.

A procedure for the in vitro growth of well-ordered two-dimensional sheets from ribosomal particles using salts and salt-alcohol mixtures has been developed. Employing this procedure, ordered two-dimensional sheets of the wild type as well as of mutated 50 S ribosomal subunits from Bacillus stearothermophilus can readily be obtained. These sheets, stained with uranyl acetate or gold-thioglucose, are suitable for three-dimensional image reconstruction. They consist of relatively small unit cells with dimensions of 160 +/- 15 and 365 +/- 20 A. Diffraction patterns of electron micrographs of these sheets contain features to 25 A resolution.

Reconstitution and crystallisation experiments with isolated split proteins from Bacillus stearothermophilus ribosomes.

Six proteins (B-L1, B-L6, B-L10, B-L11, B-L12 and B-L16) were removed from 50S ribosomal subunits of Bacillus stearothermophilus by treatment with ethanol and ammonium chloride. The proteins were isolated in a pure form, and one of them (B-L6) was crystallized. Five of the six proteins (in various combinations) were added back to the core particles, resulting in 50S subunits lacking one protein. The biological activities of these ribosomal particles as determined in the poly(U)-system varied over a wide range, depending on the protein which was omitted. The particles lacking one protein provide useful tools for heavy-atom derivation necessary for our crystallographic studies on the 50S subunits of Bacillus stearothermophilus.