Brigitte Kopp

Discovery and resupply of pharmacologically active plant-derived natural products: A review.

Medicinal plants have historically proven their value as a source of molecules with therapeutic potential, and nowadays still represent an important pool for the identification of novel drug leads. In the past decades, pharmaceutical industry focused mainly on libraries of synthetic compounds as drug discovery source. They are comparably easy to produce and resupply, and demonstrate good compatibility with established high throughput screening (HTS) platforms. However, at the same time there has been a declining trend in the number of new drugs reaching the market, raising renewed scientific interest in drug discovery from natural sources, despite of its known challenges. In this survey, a brief outline of historical development is provided together with a comprehensive overview of used approaches and recent developments relevant to plant-derived natural product drug discovery. Associated challenges and major strengths of natural product-based drug discovery are critically discussed. A snapshot of the advanced plant-derived natural products that are currently in actively recruiting clinical trials is also presented. Importantly, the transition of a natural compound from a “screening hit” through a “drug lead” to a “marketed drug” is associated with increasingly challenging demands for compound amount, which often cannot be met by re-isolation from the respective plant sources. In this regard, existing alternatives for resupply are also discussed, including different biotechnology approaches and total organic synthesis. While the intrinsic complexity of natural product-based drug discovery necessitates highly integrated interdisciplinary approaches, the reviewed scientific developments, recent technological advances, and research trends clearly indicate that natural products will be among the most important sources of new drugs also in the future.

Aconitum in Traditional Chinese Medicine—A valuable drug or an unpredictable risk?

Aconitum species have been used in China as an essential drug in Traditional Chinese Medicine (TCM) for 2000 years. Reviewing the clinical application of Aconitum, their pharmacological effects, toxicity and detoxifying measures, herb-herb interactions, clinical taboos, famous herbal formulas, traditional and current herbal processing methods based upon a wide range of literature investigations serve as a case study to explore the multidisciplinary implications of botanicals used in TCM. The toxicological risk of improper usage of Aconitum remains very high, especially in countries like China, India and Japan. The toxicity of Aconitum mainly derives from the diester diterpene alkaloids (DDAs) including aconitine (AC), mesaconitine (MA) and hypaconitine (HA). They can be decomposed into less or non-toxic derivatives through Chinese traditional processing methods (Paozhi), which play an essential role in detoxification. Using Paozhi, the three main forms of processed aconite -- yanfuzi, heishunpian and baifupian -- can be obtained (CPCommission, 2005). Moreover, some new processing techniques have been developed in China such as pressure-steaming. The current development of fingerprint assays, in particular HPLC, has set a good basis to conduct an appropriate quality control for TCM crude herbs and their ready-made products. Therefore, a stipulation for a maximum level of DDA content of Aconitum is highly desirable in order to guarantee the clinical safety and its low toxicity in decoctions. Newly developed HPLC methods have made the accurate and simultaneous determination and quantification of DDA content interesting.

Natural product agonists of peroxisome proliferator-activated receptor gamma (PPARγ): a review.

Graphical abstract

Bufadienolides from animal and plant sources

Abstract Naturally occurring bufadienolides, which were isolated from both animal and plant sources and structurally elucidated in the period from 1967–1995, are reviewed and compiled.

Ethnopharmacological in vitro studies on Austria's folk medicine—An unexplored lore in vitro anti-inflammatory activities of 71 Austrian traditional herbal drugs

Ethnopharmacological relevance In Austria, like in most Western countries, knowledge about traditional medicinal plants is becoming scarce. Searching the literature concerning Austrias ethnomedicine reveals its scant scientific exploration. Aiming to substantiate the potential of medicinal plants traditionally used in Austria, 63 plant species or genera with claimed anti-inflammatory properties listed in the VOLKSMED database were assessed for their in vitro anti-inflammatory activity. Material and methods 71 herbal drugs from 63 plant species or genera were extracted using solvents of varying polarities and subsequently depleted from the bulk constituents, chlorophylls and tannins to avoid possible interferences with the assays. The obtained 257 extracts were assessed for their in vitro anti-inflammatory activity. The expression of the inflammatory mediators E-selectin and interleukin-8 (IL-8), induced by the inflammatory stimuli tumor necrosis factor alpha (TNF-α) and the bacterial product lipopolysaccharide (LPS) was measured in endothelial cells. The potential of the extracts to activate the nuclear factors PPARα and PPARγ and to inhibit TNF-α-induced activation of the nuclear factor-kappa B (NF-κB) in HEK293 cells was determined by luciferase reporter gene assays. Results In total, extracts from 67 of the 71 assessed herbal drugs revealed anti-inflammatory activity in the applied in vitro test systems. Thereby, 30 could downregulate E-selectin or IL-8 gene expression, 28 were strong activators of PPARα or PPARγ (inducing activation of more than 2-fold at a concentration of 10 µg/mL) and 21 evoked a strong inhibition of NF-κB (inhibition of more than 80% at 10 µg/mL). Conclusion Our research supports the efficacy of herbal drugs reported in Austrian folk medicine used for ailments associated with inflammatory processes. Hence, an ethnopharmacological screening approach is a useful tool for the discovery of new drug leads.

Valerenic acid potentiates and inhibits GABAA receptors: Molecular mechanism and subunit specificity

Valerian is a commonly used herbal medicinal product for the treatment of anxiety and insomnia. Here we report the stimulation of chloride currents through GABA(A) receptors (I(GABA)) by valerenic acid (VA), a constituent of Valerian. To analyse the molecular basis of VA action, we expressed GABA(A) receptors with 13 different subunit compositions in Xenopus oocytes and measured I(GABA) using the two-microelectrode voltage-clamp technique. We report a subtype-dependent stimulation of I(GABA) by VA. Only channels incorporating beta(2) or beta(3) subunits were stimulated by VA. Replacing beta(2/3) by beta(1) drastically reduced the sensitivity of the resulting GABA(A) channels. The stimulatory effect of VA on alpha(1)beta(2) receptors was substantially reduced by the point mutation beta(2N265S) (known to inhibit loreclezole action). Mutating the corresponding residue of beta(1) (beta(1S290N)) induced VA sensitivity in alpha(1)beta(1S290N) comparable to alpha(1)beta(2) receptors. Modulation of I(GABA) was not significantly dependent on incorporation of alpha(1), alpha(2), alpha(3) or alpha(5) subunits. VA displayed a significantly lower efficiency on channels incorporating alpha(4) subunits. I(GABA) modulation by VA was not gamma subunit dependent and not inhibited by flumazenil (1 microM). VA shifted the GABA concentration-effect curve towards lower GABA concentrations and elicited substantial currents through GABA(A) channels at > or = 30 microM. At higher concentrations (> or = 100 microM), VA and acetoxy-VA inhibit I(GABA). A possible open channel block mechanism is discussed. In summary, VA was identified as a subunit specific allosteric modulator of GABA(A) receptors that is likely to interact with the loreclezole binding pocket.

Bufadienolides and their antitumor activity

Covering: 1998 to July 2010 Since 1998, a great number of bufadienolides have been reported from plants, animals and plant cell suspensions, as well as from fungi and bacteria cultures. This review summarizes both new bufadienolides and those obtained from new sources, together with data regarding the antitumor activity of bufadienolide derivatives.

Achillea millefolium L. s.l. revisited: recent findings confirm the traditional use.

ZusammenfassungDie Schafgarbe (Achillea millefolium L. s.l.) wird traditionell bei krampfartigen und entzündlichen Magen-Darm-Erkrankungen, als Gallenmittel und äußerlich bei Entzündungen eingesetzt. In neuesten Untersuchungen konnte gezeigt werden, dass die Flavonoide aus der Schafgarbe spasmolytisch wirken, während die Dicaffeoylchinasäuren choleretisch aktiv sind. Außerdem wurde eine in vitro-Hemmung der humanen neutrophilen Elastase, einer am Entzündungsgeschehen beteiligten Protease, durch Extrakte und Fraktionen aus der Schafgarbe beobachtet, was einen zusätzlichen Hinweis zur antiphlogistischen Wirkweise von Achillea millefolium L. s.l. darstellt. Somit bestätigen diese Ergebnisse die volksmedizinische Anwendung der Droge.SummaryYarrow (Achillea millefolium L. s.l.) is traditionally used in the treatment of inflammatory and spasmodic gastro-intestinal disorders, hepato-biliary complaints and inflammation. Now we could show that the flavonoids mediated the antispasmodic properties of yarrow, whereas the dicaffeoylquinic acids caused the choleretic effects. Moreover, we observed an in vitro-inhibition of human neutrophil elastase, a protease involved in the inflammatory process, by extracts and fractions from yarrow, which suggests additional mechanisms of antiphlogistic action. The presented results confirm the traditional use of yarrow.

The genus Rhododendron: an ethnopharmacological and toxicological review.

ETHNOPHARMACOLOGICAL RELEVANCE The vast genus Rhododendron includes species that have been used in traditional medicine for the treatment of inflammatory conditions, pain, gastro-intestinal disorders, common cold, asthma, skin disease, etc. Rhododendrons are also well known for their toxicity and some species have been traditionally used as poison. AIM OF THE REVIEW The work reviews and analyses the traditional use, biological activities with the corresponding chemical constituents, and toxicological data on Rhododendron species. The review aims at characterizing the ethnopharmacology of the genus in relation to its toxicity in order to identify the therapeutic potential of Rhododendron species and future directions for research. METHODS Data regarding Rhododendron spp. was collected using electronic databases (SciFinder, PubMed, Google Scholar) and library search for selected peer-reviewed articles. Plant taxonomy was validated by the databases The Plant List, Tropicos, eFloras, Flora Iberica and Flora Europaea (RBGE). Additional information on traditional use and botany was obtained from published books. The review encompasses literature, mainly regarding biological activity and toxicological data, from 1898 to the end of December 2012. RESULTS Rhododendrons have been used in Asian, North American and European traditional medicine mainly against inflammation, pain, skin ailments, common cold and gastro-intestinal disorders. In vivo and in vitro testing of plant extracts and isolated compounds determined diverse biological activities including anti-inflammatory, analgesic, anti-microbial, anti-diabetic, insecticidal and cytotoxic activity. Rhododendron spp. can cause intoxications in humans following intake of rhododendron honey or medicinal preparations. The toxicity is due to grayanotoxins, diterpenes which activate voltage-gated sodium channels and lead to gastro-intestinal, cardiac and central nervous system symptoms. CONCLUSION Rhododendron species are useful traditional remedies for the treatment of inflammation, pain, skin ailments, common cold and gastro-intestinal disorders. Pharmacological data has validated most indications of rhododendrons in ethnomedicine and toxicology studies have confirmed the toxicity observed by traditional use. Ethnopharmacological data point to the therapeutic potential of the genus Rhododendron for the treatment of inflammatory conditions and pain and, thus, research should focus on identification of active compounds and related mechanistic studies. Prolonged and high dose intake of traditional formulations containing rhododendrons should be avoided until more in depth toxicity studies become available.

Stimulation of nitric oxide synthesis by the aqueous extract of Panax ginseng root in RAW 264.7 cells

In this study, we investigated the effect of Panax ginseng root aqueous extracts upon inducible nitric oxide synthesis in RAW 264.7 cells. Panax ginseng root extract has been used in the Asian world for centuries as a traditional herb to enhance physical strength and resistance and is becoming more and more popular in Europe and North America. Incubation of murine macrophages (RAW 264.7 cells) with increasing amounts of aqueous extracts of Panax ginseng (0.05 – 0.8 μg μl−1) showed a dose dependent stimulation of inducible nitric oxide synthesis. Polysaccharides isolated from Panax ginseng showed strong stimulation of inducible nitric oxide synthesis, whereas a triterpene‐enriched fraction from an aqueous extract of Panax ginseng did not show any stimulation. Inducible nitric oxide synthase protein expression was enhanced in a dose dependent manner as revealed by immunoblotting when cells were incubated with increasing amounts of Panax ginseng extract. This was associated with an incline in inducible nitric oxide synthase mRNA‐levels as determined by semiquantitative polymerase chain reaction and electromobility shift assay studies indicated enhanced nuclear factor‐κB DNA binding activity. As nitric oxide plays an important role in immune function, Panax ginseng treatment could modulate several aspects of host defense mechanisms due to stimulation of the inducible nitric oxide synthase.