scholarly journals The Feasibility of Establishing a Morphological Cell Profiling Assay to Assess the Bioactivity of Natural Products

2021 ◽  
Author(s):  
◽  
Lauren Taylor
Keyword(s):  
Flow Cytometry ◽  
Natural Products ◽  
Morphological Analysis ◽  
Accurate Information ◽  
Biologically Relevant ◽  
Multiple Components ◽  
Selection Protocol ◽  
Cellular Components ◽  
Compound Concentration ◽  
Product Compound

<p><b>Morphological cell profiling (MCP) is an assay which quantifies the morphology of cells and cellular components. Changes in the morphology of cells following compound treatment has been shown to assess the bioactivity, and even propose a target of said compound. It is a powerful assay that can be used to assess novel compounds for drug candidature. However, it is currently not transferable between institutions. Thus, this project evaluated the feasibility of establishing an MCP assay at Victoria University of Wellington - Te Herenga Waka to assess the bioactivity of novel natural products from the natural products laboratory.</b></p> <p>First, a new individualistic approach to select compound concentration for MCP was assessed by flow cytometry with six exemplary drugs. Results concluded this flow cytometry approach can realise an ideal concentration for individual compounds in which the compound treatment was bioactive but not cytotoxic. This approach provides more biologically relevant and accurate information about a compound's bioactivity than previous MCP methods.</p> <p>Second, the feasibility of establishing the assay was assessed by testing the same six drugs through protocol common to MCP assays. Various stains and microscopes were tested for suitability for an MCP; image quantitative software CellProfiler was evaluated for MCP analysis ease; and quality control protocol was attempted. Ultimately, an MCP is currently not feasible at Te Herenga Waka as multiple components to establish the assay are too difficult and time intensive to complete.</p> <p>Third, discorhabdin E, a pyrroloiminoquinone alkaloid, was isolated from the New Zealand marine sponge, Latrunculia kaakaariki. The compound was tested as if it were a novel natural product compound through the new flow cytometry concentration selection protocol to test its effectivity, and was subsequently assessed by morphological analysis.</p>

scholarly journals The Feasibility of Establishing a Morphological Cell Profiling Assay to Assess the Bioactivity of Natural Products

2021 ◽  
Author(s):  
◽  
Lauren Taylor
Keyword(s):  
Flow Cytometry ◽  
Natural Products ◽  
Morphological Analysis ◽  
Accurate Information ◽  
Biologically Relevant ◽  
Multiple Components ◽  
Selection Protocol ◽  
Cellular Components ◽  
Compound Concentration ◽  
Product Compound

<p><b>Morphological cell profiling (MCP) is an assay which quantifies the morphology of cells and cellular components. Changes in the morphology of cells following compound treatment has been shown to assess the bioactivity, and even propose a target of said compound. It is a powerful assay that can be used to assess novel compounds for drug candidature. However, it is currently not transferable between institutions. Thus, this project evaluated the feasibility of establishing an MCP assay at Victoria University of Wellington - Te Herenga Waka to assess the bioactivity of novel natural products from the natural products laboratory.</b></p> <p>First, a new individualistic approach to select compound concentration for MCP was assessed by flow cytometry with six exemplary drugs. Results concluded this flow cytometry approach can realise an ideal concentration for individual compounds in which the compound treatment was bioactive but not cytotoxic. This approach provides more biologically relevant and accurate information about a compound's bioactivity than previous MCP methods.</p> <p>Second, the feasibility of establishing the assay was assessed by testing the same six drugs through protocol common to MCP assays. Various stains and microscopes were tested for suitability for an MCP; image quantitative software CellProfiler was evaluated for MCP analysis ease; and quality control protocol was attempted. Ultimately, an MCP is currently not feasible at Te Herenga Waka as multiple components to establish the assay are too difficult and time intensive to complete.</p> <p>Third, discorhabdin E, a pyrroloiminoquinone alkaloid, was isolated from the New Zealand marine sponge, Latrunculia kaakaariki. The compound was tested as if it were a novel natural product compound through the new flow cytometry concentration selection protocol to test its effectivity, and was subsequently assessed by morphological analysis.</p>

scholarly journals Natural product fragment combination to performance-diverse pseudo-natural products

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Michael Grigalunas ◽  
Annina Burhop ◽  
Sarah Zinken ◽  
Axel Pahl ◽  
José-Manuel Gally ◽  
...  
Keyword(s):  
Natural Products ◽  
Product Design ◽  
Natural Product ◽  
Chemical Space ◽  
Biological Evaluation ◽  
Product Structure ◽  
Biologically Relevant ◽  
Biologically Relevant Chemical Space

AbstractNatural product structure and fragment-based compound development inspire pseudo-natural product design through different combinations of a given natural product fragment set to compound classes expected to be chemically and biologically diverse. We describe the synthetic combination of the fragment-sized natural products quinine, quinidine, sinomenine, and griseofulvin with chromanone or indole-containing fragments to provide a 244-member pseudo-natural product collection. Cheminformatic analyses reveal that the resulting eight pseudo-natural product classes are chemically diverse and share both drug- and natural product-like properties. Unbiased biological evaluation by cell painting demonstrates that bioactivity of pseudo-natural products, guiding natural products, and fragments differ and that combination of different fragments dominates establishment of unique bioactivity. Identification of phenotypic fragment dominance enables design of compound classes with correctly predicted bioactivity. The results demonstrate that fusion of natural product fragments in different combinations and arrangements can provide chemically and biologically diverse pseudo-natural product classes for wider exploration of biologically relevant chemical space.

scholarly journals Optimization of Extraction or Purification Process of Multiple Components from Natural Products: Entropy Weight Method Combined with Plackett–Burman Design and Central Composite Design

Molecules ◽  
2021 ◽  
Vol 26 (18) ◽  
pp. 5572
Author(s):  
Yu Du ◽  
Pengcheng Huang ◽  
Weifeng Jin ◽  
Chang Li ◽  
Jiehong Yang ◽  
...  
Keyword(s):  
Natural Products ◽  
Purification Process ◽  
Entropy Weight ◽  
Multiple Components ◽  
Entropy Weight Method ◽  
Weight Method ◽  
Burman Design ◽  
Plackett Burman Design ◽  
Central Composite ◽  
Comprehensive Score

In this paper, the optimization of the extraction/purification process of multiple components was performed by the entropy weight method (EWM) combined with Plackett–Burman design (PBD) and central composite design (CCD). We took the macroporous resin purification of Astragalus saponins as an example to discuss the practicability of this method. Firstly, the weight of each component was given by EWM and the sum of the product between the componential content and its weight was defined as the comprehensive score, which was taken as the evaluation index. Then, the single factor method was adopted for determining the value range of each factor. PBD was applied for screening the significant factors. Important variables were further optimized by CCD to determine the optimal process parameters. After the combination of EWM, PBD and CCD, the resulting optimal purification conditions were as follows: pH value of 6.0, the extraction solvent concentration of 0.15 g/mL, and the ethanol volume fraction of 75%. Under the optimal conditions, the practical comprehensive score of recoveries of saponins was close to the predicted value (n = 3). Therefore, the present study provided a convenient and efficient method for extraction and purification optimization technology of multiple components from natural products.

Rearranged ergostane-type natural products: chemistry, biology, and medicinal aspects

2019 ◽  
Vol 17 (7) ◽  
pp. 1624-1633 ◽  
Author(s):  
Fenja L. Duecker ◽  
Franziska Reuß ◽  
Philipp Heretsch
Keyword(s):  
Natural Products ◽  
Biologically Relevant ◽  
Natural Products Chemistry

Rearranged ergostane-type natural products are structurally intriguing and exhibit biologically relevant properties.

scholarly journals N-(1-Phenylethyl)aziridine-2-carboxylate esters in the synthesis of biologically relevant compounds

2019 ◽  
Vol 15 ◽  
pp. 1722-1757
Author(s):  
Iwona E Głowacka ◽  
Aleksandra Trocha ◽  
Andrzej E Wróblewski ◽  
Dorota G Piotrowska
Keyword(s):  
Amino Acids ◽  
Natural Products ◽  
Natural Compounds ◽  
Review Article ◽  
Chiral Auxiliary ◽  
Biologically Relevant ◽  
Alternative Approaches ◽  
Approved Drugs ◽  
Enantioselective Syntheses ◽  
New Procedures

Since Garner’s aldehyde has several drawbacks, first of all is prone to racemization, alternative three-carbon chirons would be of great value in enantioselective syntheses of natural compounds and/or drugs. This review article summarizes applications of N-(1-phenylethyl)aziridine-2-carboxylates, -carbaldehydes and -methanols in syntheses of approved drugs and potential medications as well as of natural products mostly alkaloids but also sphingoids and ceramides and their 1- and 3-deoxy analogues and several hydroxy amino acids and their precursors. Designed strategies provided new procedures to several drugs and alternative approaches to natural products and proved efficiency of a 2-substituted N-(1-phenylethyl)aziridine framework as chiron bearing a chiral auxiliary.

scholarly journals Studies on the Identification of Constituents in Ethanol Extract of Radix Glycyrrhizae and Their Anti-Primary Hepatoma Cell Susceptibility

2014 ◽  
Vol 2014 ◽  
pp. 1-5
Author(s):  
Jie Liu
Keyword(s):  
Flow Cytometry ◽  
Natural Products ◽  
Cell Cycle Progression ◽  
Chemical Constituents ◽  
Hepatoma Cell ◽  
Ethanol Extract ◽  
Hepatoma Cells ◽  
Primary Hepatoma ◽  
Isolation And Purification ◽  
Radix Glycyrrhizae

The objective of this paper is to study the chemical constituents of Radix Glycyrrhizae and to apply the resulting natural products in the study of drug susceptibility of hepatoma cells so as to provide a scientific basis for quality standards and clinical application of medicinal Radix Glycyrrhizae. Chromatographic materials were used for isolation and purification; structural identification was performed based on physicochemical properties and spectral data. MTT colorimetry was used to detect the proliferation inhibition rate against primary hepatoma cells by natural products, and flow cytometry was used to detect the changes in cell cycle progression. Five compounds were isolated and identified, namely, liquiritigenin (1), liquiritin (2), isoliquiritigenin (3), betulinic acid (4), and oleanolic acid (5). In the study, 5-FU (5-fluorouracil) is used as a positive control to the hepatoma cells. Primary hepatoma cells were highly susceptible to 5-FU and liquiritigenin, both of which markedly inhibited the proliferation of hepatoma cells; flow cytometry results showed an increase in G0/G1 phase cells, a decrease in S phase cells, and a relative increase in G2/M phase cells. Primary hepatoma cells are highly susceptible to liquiritigenin, a natural product; the testing of tumor cell susceptibility is of important significance to the improvement of therapeutic effect of cancer.

scholarly journals Cheminformatics Analysis of Natural Product Scaffolds: Comparison of Scaffolds Produced by Animals, Plants, Fungi and Bacteria

2020 ◽  
Author(s):  
Peter Ertl ◽  
Tim Schuhmann
Keyword(s):  
Natural Products ◽  
Natural Selection ◽  
Natural Product ◽  
Selection Process ◽  
Structural Features ◽  
New Drugs ◽  
Large Database ◽  
Biologically Relevant ◽  
Different Types ◽  
Selection Of

AbstractNatural products (NPs) have evolved over a very long natural selection process to form optimal interactions with biologically relevant macromolecules. NPs are therefore an extremely useful source of inspiration for the design of new drugs. In the present study we report the results of a cheminformatics analysis of a large database of NP structures focusing on their scaffolds. First, general differences between NP scaffolds and scaffolds from synthetic molecules are discussed, followed by a comparison of the properties of scaffolds produced by different types of organisms. Scaffolds produced by plants are the most complex and those produced by bacteria differ in many structural features from scaffolds produced by other organisms. The results presented here may be used as a guidance in selection of scaffolds for the design of novel NP-like bioactive structures or NP-inspired libraries.

scholarly journals Validation of immunophenotyping by flow cytometry in the investigation of diagnostic and prognostic markers for breast cancer

Mastology ◽  
2020 ◽  
Vol 30 (Suppl 1) ◽  
Author(s):  
Daniella Serafin Couto Vieira ◽  
Sandro Wopereis ◽  
Laura Otto Walter ◽  
Maria Claudia Santos da Silva
Keyword(s):  
Breast Cancer ◽  
Flow Cytometry ◽  
Predictive Value ◽  
Morphological Analysis ◽  
Prognostic Markers ◽  
Molecular Subtype ◽  
Histological Grade ◽  
Tumor Development ◽  
Progesterone Receptors ◽  
The Individual

Introduction: Given its high prevalence, breast cancer has a great financial impact on health systems. Currently, the diagnosis is made by morphological analysis and immunohistochemistry (IHC). However, this methodology has some limitations. Therefore, new methods should be developed to assist those in use, based on the fast and safe detection of tumor cells. Objective: To validate immunophenotyping by flow cytometry (FC) in the investigation of diagnostic and prognostic markers for breast cancer and study lymphocyte subtypes infiltrating the tumor and their relationship with tumor development. Method: A total of 52 samples of breast tumors were sectioned, macerated in phosphate-buffered saline, stained with antibodies against estradiol receptors (ER), progesterone receptors (PR), HER2, Ki67, CD3, CD4, CD8, and CD45, and analyzed by FC. All results were compared with IHC (standard method) as to sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV), except for Ki67, whose analysis involved a comparison of bias between the methods and correlation between lymphocyte subtypes and tumor characteristics. Results: The comparison between FC and IHC for each marker presented the ER analysis (sensitivity: 75%, specificity: 90%, PPV: 96.7%, NPV: 47.4%); PR analysis (sensitivity: 72%, specificity: 70%, PPV: 79.3%, NPV: 60.8%); HER2 analysis (sensitivity: 80%, specificity: 90.2%, PPV: 66.7%, NPV: 94.9%). The FC Ki67 analysis proved to be equivalent to that of IHC, with the advantage of not having observational bias. No correlations were identified between the profile of the population of intratumoral lymphocytes and the molecular subtype or the histological grade of the tumor. Conclusion: The results show the ability of FC in safely and promptly detecting breast cancer markers used in clinical practice. The use of FC, together with morphological analysis and IHC, might overcome the individual limitations of each methodology, efficiently providing reliable and rapid results, which would lead to faster diagnosis and more accurate prognosis, directly benefiting the patients.

scholarly journals Chalkophores

2018 ◽  
Vol 87 (1) ◽  
pp. 645-676 ◽  
Author(s):  
Grace E. Kenney ◽  
Amy C. Rosenzweig
Keyword(s):  
Natural Products ◽  
Iron Homeostasis ◽  
Copper Binding ◽  
Copper Uptake ◽  
Biochemical Evidence ◽  
Biologically Relevant ◽  
Current State ◽  
Methane Oxidizing Bacteria ◽  
Copper Chelators ◽  
Key Aspects

Copper-binding metallophores, or chalkophores, play a role in microbial copper homeostasis that is analogous to that of siderophores in iron homeostasis. The best-studied chalkophores are members of the methanobactin (Mbn) family—ribosomally produced, posttranslationally modified natural products first identified as copper chelators responsible for copper uptake in methane-oxidizing bacteria. To date, Mbns have been characterized exclusively in those species, but there is genomic evidence for their production in a much wider range of bacteria. This review addresses the current state of knowledge regarding the function, biosynthesis, transport, and regulation of Mbns. While the roles of several proteins in these processes are supported by substantial genetic and biochemical evidence, key aspects of Mbn manufacture, handling, and regulation remain unclear. In addition, other natural products that have been proposed to mediate copper uptake as well as metallophores that have biologically relevant roles involving copper binding, but not copper uptake, are discussed.

scholarly journals Type S Non Ribosomal Peptide Synthetases for the rapid generation of tailor-made peptide libraries

2021 ◽  
Author(s):  
Nadya Abbood ◽  
Tien Duy Vo ◽  
Jonas Watzel ◽  
Kenan A. J. Bozhueyuek ◽  
Helge B. Bode
Keyword(s):  
Natural Products ◽  
Natural Product ◽  
De Novo ◽  
Chemical Space ◽  
Biologically Relevant ◽  
Peptide Synthetases ◽  
Medical Needs ◽  
Natural Product Research ◽  
Rapid Generation ◽  
Traditional Natural

Bacterial natural products in general, and non-ribosomally synthesized peptides in particular, are structurally diverse and provide us with a broad range of pharmaceutically relevant bioactivities. Yet, traditional natural product research suffers from rediscovering the same scaffolds and has been stigmatised as inefficient, time-, labour-, and cost-intensive. Combinatorial chemistry, on the other hand, can produce new molecules in greater numbers, cheaper and in less time than traditional natural product discovery, but also fails to meet current medical needs due to the limited biologically relevant chemical space that can be addressed. Consequently, methods for the high throughput generation of new-to-nature natural products would offer a new approach to identifying novel bioactive chemical entities for the hit to lead phase of drug discovery programms. As a follow-up to our previously published proof-of-principle study on generating bipartite type S non-ribosomal peptide synthetases (NRPSs), we now envisaged the de novo generation of non-ribosomal peptides (NRPs) on an unreached scale. Using synthetic zippers, we split NRPS in up to three subunits and rapidly generated different bi- and tripartite NRPS libraries to produce 49 peptides, peptide derivatives, and de novo peptides at good titres up to 145 mgL-1. A further advantage of type S NRPSs not only is the possibility to easily expand the created libraries by re-using previously created type S NRPS, but that functions of individual domains as well as domain-domain interactions can be studied and assigned rapidly.

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