Drugs from Nature Targeting Inflammation (DNTI) – Ein erfolgreiches interdisziplinäres Österreichisches Netzwerk-Projekt

Entzündungen sind Reaktionen des Organismus auf die Einwirkung von unterschiedlichen Reizen, die physikalisch chemischer Art sein können, oder auf Infektionserreger und maligne Neoplasmen zurückzuführen sind. Entzündungen spielen bei einer Vielzahl von Krankeitsbildern wie z. B. Arthritis, Atherosklerose, dem metabolischen Syndrom, Sepsis, Allergien und Autoimmunerkrankungen oder Krebs eine essentielle Rolle. Im Rahmen eines Nationalen Forschungsnetzwerkes arbeiten Wissenschaftler der Universität Innsbruck, der Universität Wien, der Medizinischen Universität Wien, der Veterinärmedizinischen Universität Wien, der Technischen Universität Wien und der Universität Graz zusammen. Finanziert wird das Projekt durch den Fonds zur Förderung der wissenschaftlichen Forschung (Förderperiode 2008-2014). Zusätzliche Unterstützung erfolgt durch die Standardagentur Tirol. Ziel dieses nationalen Netzwerkes ist die Identifizierung und Charakterisierung von bioaktiven Naturstoffen für die Behandlung entzündlicher Erkrankungen speziell im Bereich des kardiovaskulären Systems. Die Identifizierung und Isolierung von entzündungshemmenden Naturstoffen erfolgt auf Basis von Computertechniken, wie Pharmakophor-Modelling und virtuellem Screening von Naturstoffdatenbanken (molekularer Ansatz) sowie der volksmedizinischen Anwendung von Heilpflanzen (ethnopharmakologischer Ansatz). Wirkstoffkandidaten beider Ansätze werden in In-vivo-Modellen sowie mechanistischen Studien in vitro untersucht. Struktur-Wirkungsbeziehungen isolierter oder synthetisierter Strukturanaloga aktiver Naturstoffe werden analysiert. Stichwörter: DNTI, Entzündung, NaturstoffeDrugs from Nature Targeting Inflammation (DNTI) – A sucessful interdisziplinary Austrian network projectInflammation is the body's response to tissue injury, either physical (mechanical, irradiation), by infectious agents, or by malignant or altered normal cells. Thus, various and at first glance diverse pathological conditions involve inflammation. These include arthritis, atherosclerosis, the metabolic syndrome, sepsis, allergies and auto-immune diseases or even cancer. In the course of an ongoing Austrian research network project involving scientists from the University of Innsbruck, the University of Vienna, the Medical University of Vienna, University of Veterinary Medicine Vienna, Vienna University of Technology and the University of Graz. The project is financed (2008-2014 programming period) by the Austrian Science Fund. Additional support is provided by the standard agency Tyrol. The aim of this project is to identify and characterize natural products (NPs) capable to combat inflammatory processes and disorders associated with inflammation. This goal is approached by a unique combination of strategies e.g. i) virtual screening of NP databases using structure- or ligand-based pharmacophore models to identify bioactive NPs against selected anti-inflammatory targets, ii) parallel pharmacophoric profiling to predict desired but also unwanted bioactivities of single NPs and to provide suggestions for their pharmacological mechanisms of action and iii) exploitation of indigenous knowledge of medicinal plants to select promising candidates for phytochemical and subsequent pharmacological investigation. Keywords: DNTI, inflammation, natural product

Quantitative analysis of anti-inflammatory lignan derivatives in Ratanhiae radix and its tincture by HPLC–PDA and HPLC–MS

AbstractRoot preparations of Krameria lappacea (Dombey) Burdet et Simpson are traditionally used against oropharyngeal inflammation. Besides antimicrobial and astringent procyanidines, lignan derivatives, including ratanhiaphenol I, II, III and (+)-conocarpan, contribute to the activity of Ratanhiae radix, exerting a significant topical anti-inflammatory activity in vivo, and in vitro by inhibiting NF-κB and the formation of inflammatory prostaglandins and leukotrienes. Besides gravimetrical analysis of the ratanhiaphenols I, II and III, the content of these compounds in the herbal drug has never been determined. The developed HPLC method enables the quantification of twelve active lignan derivatives in the roots, and is also suitable for the determination of the constituents in Tinctura Ratanhiae. Separation was achieved on a phenyl-hexyl column material using a solvent gradient consisting of 0.02% aqueous TFA and a mixture of acetonitrile/methanol (75:25, v/v). Sensitivity, accuracy (recovery rates were between 95% and 105.6%), repeatability (RSD≤4.6%), and precision (intra-day precision≤4.8%; inter-day precision≤3.4%) of the method were determined. HPLC–MS experiments in positive and negative electrospray ionization mode confirmed identity and peak purity of analytes. The analysis of several root and tincture samples revealed that (+)-conocarpan and ratanhiaphenol II dominated with contents of 0.49–0.71% and 0.51–0.53% in the roots and 0.66–0.68mg/ml and 0.70–0.71mg/ml in the commercial tinctures, respectively

Apple scab control with grapefruit seed extract: no alternative to chemical fungicides

The growth inhibiting effect of four commercially available grapefruit seed extracts on the causal organism of apple scab Venturia inaequalis was tested. Germination of the conidia of Venturia inaequalis was pronouncedly inhibited by all tested extracts. The commercial products were analyzed by high pressure liquid chromatography and thin layer chromatography. All samples contained at least one preserving agent. These substances were identified as either benzethonium chloride, benzalkonium chloride, methyl parabene or propyl parabene. Freshly prepared extracts from seeds of grapefruits (Citrus paradisi) did not inhibit the germination of Venturia inaequalis. It was therefore concluded that the antifungal effect of grapefruit seed extracts is caused by the added preservatives

Structure-Guided Identification of Black Cohosh (Actaea racemosa) Triterpenoids with In Vitro Activity against Multiple Myeloma

Black cohosh is a well-established medicinal plant and preparations of its rootstock are used for the treatment of mild climacteric complaints. The compounds considered responsible for the therapeutic effect are triterpene glycosides, characterized by a cycloartane scaffold and a pentose moiety. Because some of these triterpenoids were found to exhibit relevant cytotoxic effects against human breast cancer cells, we decided to investigate their activity on multiple myeloma cell lines NCI-H929, OPM-2, and U266. In a systematic approach, we initially tested three known cytotoxic compounds of three different triterpenoid types, revealing the cimigenol-type triterpenoid as the most active constituent. In a second round, seven naturally occurring cimigenol derivatives were compared with respect to their sugar moiety and their substitution pattern at position C-25, leading to 25-O-methylcimigenol-3-O-α-L-arabinopyranoside as the most potent candidate. Interestingly, not only the methyl group at position C-25 increased the cytotoxic effect but also the arabinose moiety at position C-3 had an impact on the activity. The variety of cimigenol derivatives, moreover, allowed a detailed discussion of their structure-activity relationships, not only for their effect on multiple myeloma cells but also with regard to previous studies on the cytotoxicity of black cohosh triterpenoids

Purification of thonningianins A and B and four further derivatives from Thonningia sanguinea by one‐ and two‐dimensional centrifugal partition chromatography

International audienc

Development, validation, and application of a fast, simple, and robust SPE-based LC-MS/MS method for quantification of angiotensin I-converting enzyme inhibiting tripeptides Val-Pro-Pro, Ile-Pro-Pro, and Leu-Pro-Pro in yoghurt and other fermented dairy products

Dairy products are an important part of a nutritionally balanced diet as their constituents can affect the human state of health. By inhibiting the angiotensin I-converting enzyme, the tripeptides Val-Pro-Pro, Ile-Pro-Pro, and Leu-Pro-Pro can lower blood pressure. As these peptides are produced during fermentation, they are found in various dairy products like cheese, yoghurt, etc., but except for cheese only little is known about their content. To investigate how other dairy products contribute to a supply of these antihypertensive peptides, we developed and validated a fast and sensitive assay for quantification of the three tripeptides with LC-MS/MS combined with a simple protocol for extraction and SPE-purification from yoghurt, curd, or other products. Finally, the entire method was successfully applied to survey peptide concentrations in samples from local dairies and thus expands our awareness on the content of antihypertensive peptides in our food. (C) 2019 Elsevier Ltd. All rights reserved.Peer reviewe

Ryania Speciosa: an insecticide for organic orchards?

Ryania speciosa is a grove belonging to the family Flacourtiacea, located in tropical climates in Central- and South- America. The Indigenous population used the plant, which contains toxic alkaloids, against cockroaches. In the forties the ground wood was used as insecticide against different pests. In the first version of annex 2b from the EC regulation 2092/91, Ryania was still listed, but it was subsequently cancelled due to its lack of registration as a pesticide in any European country. In organic farming, only a few insecticides are allowed, but in orchards there is a permanent need to control larvae such as coddling moth, etc. The Italian Ministry of Agriculture initiated a survey to investigate the possibility of developing an 'organic' pesticide based on Ryania. Basic research was done, working on extraction methods, synergists, spectrum of action, degradation, and efficacy in comparison with other modern pesticides including ecotoxicological investigations. Organic fruit growers hope that a company takes on Ryania to pass it through the registration procedure and puts some formulations on the market

A new eudesmane derivative from Leontodon tuberosus

Besides the known compounds 2,4,6-trihydroxyacetophenone 4-O-β -D-glucopyranoside and syringaresinol 4'-O-β -D-glucopyranoside, the novel sesquiterpenoid 1,2-dehydro-3-oxocostic acid β -D-glucopyranoside ester was isolated from Leontodon tuberosus L. and its structure established by mass spectrometry and 1D-and 2D-NMR spectroscopy. Additionally, a number of fatty and phenolic acids was identified in the crude methanolic extract by HPLC-DAD and HPLC-MS. The chemosystematic impact of the new sesquiterpenoid is discussed briefly

Bioassay-Guided Isolation of Anti-Inflammatory Constituents of the Subaerial Parts of Cyperus articulatus (Cyperaceae)

Based on data from a previous ethnobotanical study in northern Angola, phytochemical investigations into the methanolic rhizomes and roots extract of Cyperus articulatus, monitored by in vitro assays, resulted in the recovery of 12 sesquiterpenes, 3 stilbenes, 2 phenolic acids, 1 monoterpene, and 1 flavonoid. Among them, 14 compounds were isolated for the first time from this species. Their inhibitory potential against nitric oxide (NO) production, as well as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression, was evaluated in LPS-treated J774A.1 murine macrophages. Especially, both stilbene dimer trans-scirpusin B and trimer cyperusphenol B showed promising inhibitory activity against the production of the inflammatory mediator, NO, in a concentration-dependent manner (10-1 mu M). The obtained data are the first results confirming the anti-inflammatory potential of C. articulatus and support its indigenous use as a traditional remedy against inflammation-related disorders

Screening of Vietnamese medicinal plants for NF-κB signaling inhibitors: Assessing the activity of flavonoids from the stem bark of Oroxylum indicum

AbstractEthnopharmacological relevanceSeventeen plants used in Vietnamese traditional medicine for the treatment of inflammatory disorders were screened for NF-κB inhibitory activity. Oroxylum indicum, which exhibited activity, was investigated in detail.Materials and methodsForty plant extracts from 17 species were prepared by maceration using dichloromethane and methanol and were tested (10µg/mL) to evaluate their ability to inhibit NF-κB activation using TNF-α-stimulated HEK-293 cells stably transfected with a NF-κB-driven luciferase reporter. The active extract of Oroxylum indicum was subsequently fractionated by different chromatographic techniques. After isolation, all single compounds were identified by spectroscopic methods and assessed for NF-κB inhibitory effects.ResultsThe dichloromethane extracts obtained from Chromolaena odorata leaves and the stem bark of Oroxylum indicum showed distinct inhibitory effects on NF-κB activation at a concentration of 10µg/mL. The active extract of Oroxylum indicum was subjected to further phytochemical studies resulting in identification of four flavonoid aglyca and six flavonoid glycosides. Pharmacological evaluation of the obtained compounds identified oroxylin A as the most active substance (IC50=3.9µM, 95% CI: 3.5–4.4µM), while chrysin and hispidulin showed lower activity with IC50=7.2µM (95% CI: 6.0–8.8µM) and 9.0µM (95% CI: 7.9–10.2µM), respectively. Interestingly, in this study the activity of baicalein (IC50=28.1µM, 95% CI: 24.6–32.0µM) was weak. The isolated glycosides showed no inhibitory activity when tested at a concentration of 30µM. Quantification of the four active flavonoids in extracts and plant materials suggested that oroxylin A contributes to the NF-κB inhibitory activity of the stem barks of Oroxylum indicum to a greater extent than baicalein which was thought to be responsible for the anti-inflammatory activity of this plant.ConclusionsThe screening presented in this study identified the dichloromethane extracts of Chromolaena odorata and Oroxylum indicum as promising sources for NF-κB inhibitors. Hispidulin, baicalein, chrysin and oroxylin A, isolated from Oroxylum indicum, were identified as inhibitors of NF- κB activation