Isolation Of Bioactive Natural Products Research Articles

Recent bioanalytical methods for the isolation of bioactive natural products from genus Codonopsis.

Chromatography and spectroscopy are nowadays well-validated techniques allowing to isolate and purify different class of natural products from the genus Codonopsis. Several categories of phytochemicals with drug like properties have been selectively extracted, isolated, characterised by this methodology. The present review aims to provide up-to-date and comprehensive information on the chromatography, phytochemistry and pharmacology of natural products of Codonopsis with an emphasis on the search for natural products having various biological activities and the semi-synthetic derivatives of bioactive ones and to highlight current gaps in knowledge. A literature search was performed in the SciFinder Scholar, PubMed, Medline, and Scopus databases. During the period covered in this review, several classes of compounds have been reported from genus Codonopsis. Codonopsis pilosula and Codonopsis lanceolata are the most popular in the genus especially as per phytochemical and bioactive studies. Phytochemical investigation demonstrates that Codonopsis species contain mainly xanthones, flavonoids, alkaloids, polyacetylenes, phenylpropanoids, triterpenoids and polysaccharides, which contribute to numerous bioactivities. The major bioactive compounds isolated were used for semi-synthetic modification to increase the chance to discover lead compound. It can be concluded that genus Codonopsis has been used as traditional medicines and food materials around the world over years due to chemical constituents with diverse structural types, exhibiting extensive pharmacological activities in immune system, blood system, cardiovascular system, central nervous system, digestive system, and so forth, with almost no obvious toxicity and side effect. Therefore, Codonopsis can be used as a promising ethnopharmacological plant source.

Isolation of bioactive natural products from Rumex dentatus and their antimicrobial evaluation: A comparative study against various pathogenic bacteria

Rumex dentatus is traditionally used to treat different ailments including bacterial and fungal infections. The presented study disclosed isolation of five bioactive molecules from Rumex dentatus. All the isolates were evaluated for antimicrobial activities. In this protocol the comparative study has been made on various extracts and isolated compounds in order to identify the lead molecule. Thus the activity carried, resulted, compound X2 with strong anti-microbial potential and exhibiting IC50 value between 14–32 μM. While rest of isolates showed moderate antibacterial potential. Further, the investigation of antimicrobial activity on various plant extracts against six human pathogens (five bacteria and a fungal pathogen), most of the extracts showed strong to moderate antimicrobial activity with IC50 ranging from 24–35 μM. Out of all the extracts which were evaluated for antimicrobial activities, the hexane and methanol: DCM (1:1) extracts were found to have good antibacterial potential in microtiter plate assay. The IC50 value of compound X2 was found to be favorable and presented a strong antimicrobial potential. Thus the results inferred that the plant has strong antibacterial value and hence can be used in medication for the treatment of various bacterial infections.

A New Citrinin Derivative from the Indonesian Marine Sponge-Associated Fungus Penicillium citrinum.

Sponge-associated fungi are attractive targets for the isolation of bioactive natural products with different pharmaceutical purposes. In this investigation, 20 fungi were isolated from 10 different sponge specimens. One isolate, the fungus Penicillium citrinum strain WK-P9, showed activity against Bacillus subtilis JH642 when cultivated in malt extract medium. One new and three known citrinin derivatives were isolated from the extract of this fungus. The structures were elucidated by 1D and 2D NMR spectroscopy, as well as LC-HRMS. Their antibacterial activity against a set of common human pathogenic bacteria and fungi was tested. Compound 2 showed moderate activity against Mycobacterium smegmatis ATCC607 with a minimum inhibitory concentration (MIC) of 32 µg/mL. Compound 4 exhibited moderate growth inhibition against Bacillus subtilis JH642, B. megaterium DSM32, and M. smegmatis ATCC607 with MICs of 16, 16, and 32 µg/mL, respectively. Furthermore, weak activities of 64 µg/mL against B. subtilis DSM10 and S. aureus ATCC25923 were observed for compound 4.

Mass spectrometry-based metabolomics approach in the isolation of bioactive natural products

Metabolomics is a powerful tool in the analysis and identification of metabolites responsible for biological properties. Regarding natural product chemistry, it constitutes a potential strategy to streamline the classic and laborious process of isolating natural products, which often involves the re-isolation and identification of known compounds. In this contribution, we establish a mass spectrometry-based metabolomics strategy to discover compounds with larvicidal activity against Aedes aegypti. We analyse the Brazilian plant Annona crassiflora using different platforms to annotate the active compounds in different extracts/fractions of various plant parts. The MetaboAnalyst and GNPS platforms, which consider LC-MS and LC-MS/MS data, respectively, were chosen to identify compounds that differentiate active and inactive samples. Bio-guided isolation was subsequently performed to confirm compound activity. Results proved the capacity of metabolomics to predict metabolite differences between active and inactive samples using LC-MS and LC-MS/MS data. Moreover, we discuss the limitations, possibilities, and strategies to have a broad view of vast data.

NCI Program for Natural Product Discovery: A Publicly-Accessible Library of Natural Product Fractions for High-Throughput Screening.

The US National Cancer Institute's (NCI) Natural Product Repository is one of the world's largest, most diverse collections of natural products containing over 230,000 unique extracts derived from plant, marine, and microbial organisms that have been collected from biodiverse regions throughout the world. Importantly, this national resource is available to the research community for the screening of extracts and the isolation of bioactive natural products. However, despite the success of natural products in drug discovery, compatibility issues that make extracts challenging for liquid handling systems, extended timelines that complicate natural product-based drug discovery efforts and the presence of pan-assay interfering compounds have reduced enthusiasm for the high-throughput screening (HTS) of crude natural product extract libraries in targeted assay systems. To address these limitations, the NCI Program for Natural Product Discovery (NPNPD), a newly launched, national program to advance natural product discovery technologies and facilitate the discovery of structurally defined, validated lead molecules ready for translation will create a prefractionated library from over 125,000 natural product extracts with the aim of producing a publicly-accessible, HTS-amenable library of >1,000,000 fractions. This library, representing perhaps the largest accumulation of natural-product based fractions in the world, will be made available free of charge in 384-well plates for screening against all disease states in an effort to reinvigorate natural product-based drug discovery.

Bioactive Natural Products Prioritization Using Massive Multi-informational Molecular Networks.

Natural products represent an inexhaustible source of novel therapeutic agents. Their complex and constrained three-dimensional structures endow these molecules with exceptional biological properties, thereby giving them a major role in drug discovery programs. However, the search for new bioactive metabolites is hampered by the chemical complexity of the biological matrices in which they are found. The purification of single constituents from such matrices requires such a significant amount of work that it should be ideally performed only on molecules of high potential value (i.e., chemical novelty and biological activity). Recent bioinformatics approaches based on mass spectrometry metabolite profiling methods are beginning to address the complex task of compound identification within complex mixtures. However, in parallel to these developments, methods providing information on the bioactivity potential of natural products prior to their isolation are still lacking and are of key interest to target the isolation of valuable natural products only. In the present investigation, we propose an integrated analysis strategy for bioactive natural products prioritization. Our approach uses massive molecular networks embedding various informational layers (bioactivity and taxonomical data) to highlight potentially bioactive scaffolds within the chemical diversity of crude extracts collections. We exemplify this workflow by targeting the isolation of predicted active and nonactive metabolites from two botanical sources (Bocquillonia nervosa and Neoguillauminia cleopatra) against two biological targets (Wnt signaling pathway and chikungunya virus replication). Eventually, the detection and isolation processes of a daphnane diterpene orthoester and four 12-deoxyphorbols inhibiting the Wnt signaling pathway and exhibiting potent antiviral activities against the CHIKV virus are detailed. Combined with efficient metabolite annotation tools, this bioactive natural products prioritization pipeline proves to be efficient. Implementation of this approach in drug discovery programs based on natural extract screening should speed up and rationalize the isolation of bioactive natural products.

Metabolomic-Guided Isolation of Bioactive Natural Products from Curvularia sp., an Endophytic Fungus of Terminalia laxiflora.

Endophytic fungi associated with medicinal plants are a potential source of novel chemistry and biology. Metabolomic tools were successfully employed to compare the metabolite fingerprints of solid and liquid culture extracts of endophyte Curvularia sp. isolated from the leaves of Terminalia laxiflora. Natural product databases were used to dereplicate metabolites in order to determine known compounds and the presence of new natural products. Multivariate analysis highlighted the putative metabolites responsible for the bioactivity of the fungal extract and its fractions on NF-κB and the myelogenous leukemia cell line K562. Metabolomic tools and dereplication studies using high-resolution electrospray ionization mass spectrometry directed the fractionation and isolation of the bioactive components from the fungal extracts. This resulted in the isolation of N-acetylphenylalanine (1: ) and two linear peptide congeners of 1: : dipeptide N-acetylphenylalanyl-L-phenylalanine (2: ) and tripeptide N-acetylphenylalanyl-L-phenylalanyl-L-leucine (3: ).

Concentrical coils counter-current chromatography for natural products isolation: Salvia miltiorrhiza Bunge as example

Countercurrent chromatography (CCC) is an efficient separation technique without the solid support matrix, largely depending on the partition of two-immiscible liquid phases in the separation column. Since the helical coil planet centrifuge was invented in early 1970s by Yoichiro Ito, a series of coils columns, including spiral coils and conical coils columns have been developed for CCC separation. In this work, we introduced a new simple and efficient concentrical coils column for CCC separation, which was prepared by winding the whole polytetrafluoroethylene (PTFE) tube into the circular grooves from the rotation axis in the same direction. Once the PTFE tube filled in all space of one round of the circular groove, it was jumped into the nearby outer circular groove through the gap and until the whole groove was filled. The three same concentrical coils distributed on three disc-shaped holders were connected by the same PTFE tube to form concentrical coils separation column. The separation capacity was further investigated using ten tanshinones of the extracts of a Traditional Chinese Medicine Salvia miltiorrhiza Bunge as a model natural product. All results indicated that the concentrical coils column could hold satisfactory retention of the stationary phase, higher theoretical plate number and better resolution for CCC separation of more than ten tanshinones. It may be an alternative CCC column for non-targeted and targeted isolation of bioactive natural products.

Schinus terebinthifolius scale-up countercurrent chromatography (Part I): High performance countercurrent chromatography fractionation of triterpene acids with off-line detection using atmospheric pressure chemical ionization mass spectrometry

‘Countercurrent chromatography’ (CCC) is an ideal technique for the recovery, purification and isolation of bioactive natural products, due to the liquid nature of the stationary phase, process predictability and the possibility of scale-up from analytical to preparative scale. In this work, a method developed for the fractionation of Schinus terebinthifolius Raddi berries dichloromethane extract was thoroughly optimized to achieve maximal throughput with minimal solvent and time consumption per gram of processed crude extract, using analytical, semi-preparative and preparative ‘high performance countercurrent chromatography’ (HPCCC) instruments. The method using the biphasic solvent system composed of n-heptane–ethyl acetate–methanol–water (6:1:6:1, v/v/v/v) was volumetrically scaled up to increase sample throughput up to 120 times, while maintaining separation efficiency and time. As a fast and specific detection alternative, the fractions collected from the CCC-separations were injected to an ‘atmospheric pressure chemical ionization mass-spectrometer’ (APCI-MS/MS) and reconstituted molecular weight MS-chromatograms of the APCI-ionizable compounds from S. terebinthifolius were obtained. This procedure led to the direct isolation of tirucallane type triterpenes such as masticadienonic and 3β-masticadienolic acids. Also oleanonic and moronic acids have been identified for the first time in the species. In summary, this approach can be used for other CCC scale-up processes, enabling MS-target-guided isolation procedures.

Tracking antiangiogenic components fromGlycyrrhiza uralensisFisch. based on zebrafish assays using high‐speed countercurrent chromatography

Natural products are some of the most important sources of lead compounds for drug discovery. The advanced isolation technique of lead compounds of natural origin using therapeutically relevant bioassays is capable of enhancing work efficiency from complex multiconstituent extracts. In the present study, a bioassay-guided isolation strategy combined with bioactivity screening was used to identify novel angiogenesis inhibitors from licorice (Glycyrrhiza uralensis Fisch.) based on the zebrafish model and rapid preparative separation by high-speed countercurrent chromatography. Zebrafish embryos at 24 h postfertilization were chosen as the angiogenesis inhibition model for bioactivity screening. A solvent system (n-hexane-ethyl acetate-methanol-water) with different ratios was optimized and applied in the high-speed countercurrent chromatography separation of two fractions, Fr5 and Fr6, from the ethyl acetate extract of licorice. Blood circulation and vascular outgrowth in intersegmental vessels were found to be simultaneously inhibited by isoliquiritigenin and isolicoflavonol in a dose-dependent manner. Thus, these two compounds were identified and considered as active inhibitors against angiogenesis. These experimental results indicate that zebrafish bioassays combined with high-speed countercurrent chromatography may provide an alternative pathway for the rapid isolation of bioactive natural products.

Valorization of Vegetable Waste: Identification of Bioactive Compounds and Their Chemo-Enzymatic Optimization

A growing demand – above all in industrialised countries – for safer foods and cleaner production processes has been observed in recent times. This has led to a drive, by the agro-alimentary industries, for waste reduction and upgrading as a strategy to reduce costs and achieve new sources of income. In fact, waste may be a source of high-added value products potentially useful as beneficial food constituents, food flavours and antioxidants, cosmetics, chemopreventive agents, drugs or drug adjuvants. In this regard, we have recently examined some vegetable wastes available in the Mediterranean basin as bioresources of useful compounds and we report here a short account of our work as an example of integrated approach between the query of agro-food industry for waste upgrading and the search of some sectors of chemical industry for bioactive products obtainable from renewable resources. We employed two different strategies aimed to the exploitation of vegetable by-products: 1) Identification or isolation of bioactive natural products in vegetable waste as possible source of lead compounds or enriched fractions. 2) Chemical and enzymatic modification of lead compounds available from vegetable waste to obtain optimized analogues, food additives, drugs or cosmetics. The results here summarised are focused on: a) isolation of antiproliferative constituents from almond (Prunus dulcis) hulls and grape (Vitis vinifera) stems; b) chemical analysis of extracts / fractions with antioxidant properties obtained from grape pomace and grape stems; c) chemical / enzymatic modification of hydroxytyrosol (26) and trans-resveratrol (11), two bioactive polyphenols available from vegetable waste.

Biodiversity conservation and drug discovery: Can they be combined? The Suriname and Madagascar experiences

The approach to new drugs through natural products has proved to be the single most successful strategy for the discovery of new drugs, but in recent years its use has been deemphasized by many pharmaceutical companies in favor of approaches based on combinatorial chemistry and genomics, among others. Drug discovery from natural sources requires continued access to plant, marine, and microbial biomass, and so the preservation of tropical rainforests is an important part of our drug discovery program. Sadly, many of the tropical forests of the world are under severe environmental pressure, and deforestation is a serious problem in most tropical countries. One way to combat this loss is to demonstrate their value as potential sources of new pharmaceutical or agrochemical products. As part of an effort to integrate biodiversity conservation and drug discovery with economic development, we initiated an International Cooperative Biodiversity Group (ICBG) to discover potential pharmaceuticals from the plant biodiversity of Suriname and Madagascar. The Group, established with funding from agencies of the United States government, involved participants from the USA, Suriname, and Madagascar. The basic approach was to search for bioactive plants in the Suriname and Malagasy flora, and to isolate their bioactive constituents by the best available methods, but the work included capacity building as well as research. Progress on this project is reported, drawing on results obtained from the isolation of bioactive natural products from Suriname and Madagascar. The benefits of this general approach to biodiversity and drug discovery are also discussed.

Extraction of microalgal toxins by large-scale pumping of seawater in Spain and Norway, and isolation of okadaic acid and dinophysistoxin-2

Marine biotoxins from microalgae can accumulate in shellfish and lead to poisoning of human consumers as well as fish, marine mammals and sea birds. Toxicological assessment of the toxins and development of analytical methods require large amounts of high-purity toxins and their metabolites. Although these toxins can be obtained in limited amounts from contaminated shellfish or from microalgal cultures, difficulties arise when the toxin-producing microalga is difficult to culture or its identity is not known. To circumvent this problem, we have developed a large-scale method for solid-phase extraction (SPE) of lipophilic biotoxins from natural microalgal blooms in seawater. To enhance subsequent purification of toxins adsorbed on the column, we included a filtration step to release the toxins from the cells while removing insoluble compounds and cellular debris. The efficacy of the method was illustrated by extraction and purification of okadaic acid and dinophysistoxin-2 from a high-density Dinophysis acuta bloom in Spain and from a mixed bloom containing low densities of D. acuta in Norway. Isolation of the toxins adsorbed on the SPE column was simple and efficient, and results obtained so far indicate that the method is potentially applicable to a wide range of microalgal toxins such as azaspiracids, pectenotoxins, spirolides and microcystins. The method should also be useful for harvesting toxins from large-scale microalgal cultures, and for bioprospecting for and isolation of bioactive natural products from marine and freshwater environments.

Burning the Hay to Find the Needle – Data Mining Strategies in Natural Product Dereplication

The acquisition and use of data from the LC/MS-ELSD analysis of extracts is described. The methodology requires MS spectra to be recorded in the positive/negative ESI mode, as well as the determination of retention time and peak area from ELSD. Subsequent calculation of molecular weight, referenced retention time, and normalized peak area, results in the creation of a peak library, which can be used for different data mining strategies: i) the dereplication of previously isolated natural products; ii) clustering/ranking of extracts for the creation of highly diverse natural product libraries; iii) a selection tool for the focused isolation of bioactive natural products and iv) to search for alternative sources of a target natural product.

Isolation of Bioactive Natural Products from Myxomycetes

The Myxomycetes (true slime molds) are an unusual group of primitive organisms that may be assigned to one of the lowest classes of eukaryotes. As their fruit bodies are very small and it is very difficult to collect much quantity of slime molds, few studies have been made on the chemistry of myxomycetes. Cultivation of the plasmodium of myxomycetes in a practical scale for natural products chemistry studies is known only for very limited species. Here is described a review on the recent results on isolation of bioactive natural products from myxomycetes obtained in these two years in the laboratories. Spore germination experiments were studied of hundreds of field-collected myxomycetes collected in Japan and succeeded in laboratory culture of plasmodia of several myxomycetes in a practical scale for natural products chemistry studies. As a result, pyrroloiminoquinones, polyene yellow pigments, and a peptide lactone from cultured plasmodia of Didymium iridis, Physarum rigidum and P. melleum, respectively were isolated. New naphthoquinone pigments, cycloanthranilylprolines, tyrosine-kinase inhibitory bisindoles, and a cytotoxic triterpenoid aldehyde lactone were also isolated from field-collected fruit bodies of Cribraria purpurea, Fuligo candida, Tubifera casparyi, and Tubifera dimorphotheca, respectively.

Recent applications of counter-current chromatography to the isolation of bioactive natural products

Recent applications of counter-current chromatography to the isolation of bioactive natural products

The application of high‐speed countercurrent chromatography to the semipreparative separation of vincamine and vincine

AbstractHigh‐speed countercurrent chromatography (HSCCC) has gained increasing popularity in the isolation of bioactive natural products. This paper reports its application to the separation of the alkaloids vincamine and vincine which differ only by a methoxy group. It is demonstrated in this study that the resolution and speed of HSCCC is comparable to high‐performance liquid chromatography (HPLC) and its capability in semi‐preparative purification via continuous sample injections can be a very useful application in phytochemical research·alkaloids; vincamine; vincine; high‐speed countercurrent chromatography

ChemInform Abstract: The Application of True Countercurrent Chromatography in the Isolation of Bioactive Natural Products.

ChemInform Abstract: The Application of True Countercurrent Chromatography in the Isolation of Bioactive Natural Products.

The Application of True Countercurrent Chromatography in the Isolation of Bioactive Natural Products

ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTThe Application of True Countercurrent Chromatography in the Isolation of Bioactive Natural ProductsY. W. Lee, Q. C. Fang, Y. Ito, and C. E. CookCite this: J. Nat. Prod. 1989, 52, 4, 706–710Publication Date (Print):July 1, 1989Publication History Published online1 July 2004Published inissue 1 July 1989https://doi.org/10.1021/np50064a006RIGHTS & PERMISSIONSArticle Views155Altmetric-Citations18LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit PDF (296 KB) Get e-Alerts Get e-Alerts

Application of analytical high-speed counter-current chromatography to the isolation of bioactive natural products

Application of analytical high-speed counter-current chromatography to the isolation of bioactive natural products