Current Bioactive Compounds - Volume 15, Issue 2, 2019

The interest in Cannabis sativa L. phytocomplex as a medicinal tool is a recently-emerging topic. Neurodegenerative diseases represent a promising field of application for cannabis and its preparations, as most of this pathologic conditions relies on an inflammatory etiology. Several cannabis constituents display anti-inflammatory effects targeting multiple pathways. In this review, a comprehensive overview of the available literature on C. sativa constituents activities in neuroinflammation is given. On the basis that the anti-inflammatory activity of cannabis is not attributable to only a single constituent, we discuss the possible advantages of administering the whole phytocomplex in order to fully exploit the “entourage effect” in neuroinflammatory-related conditions.

Endocannabinoid System (ES) has gained over the years a leading position in scientific research thanks to its involvement in numerous patho/physiological conditions. Accordingly, its main components, such as receptors, enzymes and mediators, have become important drug targets for the management of diseases where it is dysregulated. Within the manuscript, several classes of cannabinergic ligands are examined, emphasizing molecules coming from the natural world, unique source of active compounds. Firstly, the endogenous lipid ES modulators are described, starting from the major endocannabinoids to the plethora of endocannabinoid congeners. Afterwards, Cannabis-derived cannabinoids, namely well-known phytocannabinoids and new constituents from different varieties of Cannabis, are reviewed also mentioning the huge effort of pharmaceutical research in obtaining synthetic analogues. Finally, an overview of peptides and miscellaneous natural products points out new opportunities to modulate ES, offering an enormous chemical heterogeneity. Accordingly, hemopressin and related peptides, plant-derived alkylamides, terpenoid derivatives, neolignans and examples from the marine world can provide interesting hints and original ideas to develop new cannabinergic compounds.

In recent years, the Cannabis plant (Cannabis sativa L.) has been rediscovered as a source of new medicines around the world. Despite the fact that a number of registered medicines have been developed on the basis of purified cannabis components, there is a rapid increasing acceptance and use of cannabis in its herbal form. Licensed producers of high quality cannabis plants now operate in various countries including The Netherlands, Canada, Israel, and Australia, and in many US states. The legal availability of cannabis flowers allows to prescribe and prepare different cannabis galenic preparations by pharmacists. It is believed that synergy between cannabis components, known as “entourage effect”, may be responsible for the superior effects of using herbal cannabis versus isolated compounds. So far, only a few cannabis components have been properly characterized for their therapeutic potential, making it unclear which of the isolated compounds should be further developed into registered medicines. Until such products become available, simple and accessible galenic preparations from the cannabis plant could play an important role. In cannabis, phytochemical and pharmacological attention has been attributed mainly to four major cannabinoids (Δ9- tetrahydrocannabinol, cannabidiol, cannabigerol and cannabichromene) and to terpene components. This means a basic knowledge of these compounds and their bioavailability in different administration forms is useful for producers as well as prescribers of galenic preparations. This work will outline the most important aspects of cannabinoids and terpenes, and their behaviors during preparation and use of various administration forms including vaporizing, cannabis oils and extracts, tea, and skin creams.

Background: The medicinal properties of cannabis are now recognized. Indeed, in many countries, lawmakers have introduced specific laws and programmes to allow patients to use cannabis preparations in various forms; nevertheless, controversial situations can still be found. In Europe, medicinal- grade cannabis is mainly available as standardized dried flower tops of the cannabis plant used in magistral formulas. These are pharmaceutical preparations compounded in a pharmacy – following Good Compounding Practices - in accordance with a physician’s prescription for an individual patient. Aim of this work is to discuss the available Cannabis based preparations and regulation for prescription and distribution, with reference to Italy. Methods: We undertook a structured search of Italian laws, ministerial decree and circulars related to therapeutic use of Cannabis, interpreted in light of Italian Society of Compounding Pharmacists’ (SIFAP) position. Results: Seven documents were analysed. Cannabis can be orally administered to patients - as a dried drug dosed in filter or capsules for infusion or as an extract in olive oil prepared in a pharmacy - or inhaled with a specific vaporizer. Considering the costs and limited availability of Dutch medicinal-grade cannabis, the Italian Government, with the latest Decree of November 9th, 2015, has authorized the production of cannabis in the Military Pharmaceutical Chemical Institute on Florence, describing also the therapeutic use of cannabis and the requirements to be adhered to by physicians and pharmacists. Conclusion: A well-established national regulation guarantees the proper therapeutic use of Cannabis. Description of preparation method in monograph Pharmacopoeia will be the way to assure quality and therefore the efficacy of these products.

Background: Fibre-type Cannabis sativa L. (hemp) usually contains cannabidiolic acid and cannabidiol as the main non-psychoactive cannabinoids. Even though there is evidence of the neuroprotective activity of pure cannabidiol, no in vitro studies have reported so far the role of hemp extracts on neuroprotection. The objective of this study was to evaluate the neuroprotective effect of hemp extracts in in vitro cellular models of neurotoxicity. Methods: One extract was obtained from raw hemp inflorescences, while the other was prepared from the same plant material submitted to a decarboxylation process. The composition of both these extracts was evaluated by HPLC-UV/DAD. Human neuroblastoma SH-SY5Y and microglial BV-2 cell lines treated with rotenone were selected as the model of neurodegeneration. The neuroprotection of hemp extracts was assessed also in serum-free conditions both in the presence and in the absence of rotenone as the toxic agent by using the same cell lines. The neuroprotective potential of cannabidiol was tested in parallel. Results: The decarboxylated hemp extract possesses a mild neuroprotective activity on BV-2 cells treated with rotenone, higher than that of pure cannabidiol. As regards serum-free experiments, the nondecarboxylated hemp extract was the most effective neuroprotective agent toward SH-SY5Y cells, while BV-2 cells were better protected from the toxic insult by the decarboxylated extract and cannabidiol. Conclusion: Both hemp extracts and pure cannabidiol displayed a moderate neuroprotective activity in the neurotoxicity models considered in this study; in addition, they showed a trophic effect on SHSY5Y cells.

Background: Exploration of antibiotics from microorganisms became widespread in the academia and the industry with the serendipitous discovery of Penicillin from Penicillium notatum by Sir Alexander Fleming. This embarked the golden era of antibiotics which lasted for over 60 years. However, the traditional phenotypic screening was replaced with more rational and smarter methods of exploration of bioactive compounds from fungi and microorganisms. Fungi have been responsible for providing a variety of bioactive compounds with diverse activities which have been developed into blockbuster drugs such as Cyclosporine, Caspofungin, Lovastatin and Fingolimod etc. It has been reported that ca. 40% of the 1453 New Chemical Entities (NCE’s) approved by USFDA are natural products, natural product inspired or mimics many of which have their origins from fungi. Hence fungal compounds are playing a very important role in drug discovery and development in the pharmaceutical industry. Methods: We undertook structured searches of bibliographic databases of peer-reviewed research literature which pertained to natural products, medicinal chemistry of natural products and drug discovery from fungi. With the strategic improvement in screening and identification methods, fungi are still a potential resource for novel chemistries. Thus the searches also comprised of bioactive agents from fungi isolated or derived from special ecological groups and lineages. To find different molecules derived or isolated from fungi under clinical studies, clinical trial data from the NIH as well as from pharmaceutical companies were also explored. This comprised of data wherein the pharmaceutical industries have acquired or licensed a fungal bioactive compound for clinical study or a trial. Results: Natural product chemistry and medicinal chemistry continue to play an important role in converting a bioactive compound into therapeutic moieties or pharmacophores for new drug development. Conclusion: Thus one can say fungal bioactive compounds are alive and well for development into new drugs as novel ecological groups of fungi as well as novel chemistries are being uncovered. This review further emphasizes the collaboration of fungal biologists with chemists, pharmacologists and biochemists towards the development of newer drugs for taking them into the drug development pipeline.

Background: Endophytic fungi are microorganisms that colonize inside the plant tissues without causing any disease symptoms. Endophytic fungi isolated from medicinal plants have emerged as an interesting source for the isolation of bioactive compounds. In this study, we selected Calotropis procera, a member of the Asclepiadaceae family commonly called ‘Aak’ to evaluate the antioxidant potential of isolated endophytic fungi. Objective: The aim of this study was to assess the total phenolic content (TPC), antioxidant capacity by using different assay and phytochemical screening of endophytic fungi isolated from Calotropis procera (leaves, stem and root). Method: Crude ethyl acetate extracts of 20 different endophytic fungi isolated from Calotropis procera were tested for their preliminary phytoconstituents presence, TPC estimation (by Folin–Ciocalteu colorimetric assay) and antioxidant potential [1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, hydrogen peroxide (H2O2) free radical scavenging assay, β-carotene-linoleic free radical scavenging assay, metal chelating assay and xanthine oxidase inhibitory assay]. Results: Phenols, carbohydrates, saponins, tannins, alkaloids and flavanoids were the main phytoconstituents present in the endophytic fungi. Among the endophytes, Aspergillus nomius showed the highest TPC [72.71±1.67 μg GAE (gallic acid equivalent) /mg dry weight of fungi] and antioxidant activity for DPPH free radical scavenging assay (68.86±0.19%). A high positive linear correlation was found between TPC and xanthine oxidase inhibitory assay (R2-0.890) and between TPC and DPPH free radical scavenging assay (R2-0.839). Aspergillus and Fusarium genus species showed significant antioxidant activity by the different assay. Conclusion: The present study revealed some endophytic fungi from Calotropis procera could be a potential source of novel natural antioxidant compounds.

Background: Recently, many carbohydrates and their derivatives are being investigated for bioactivity. In the present study, we aimed at developing a novel antioxidant and prebiotic component by radiation processing of sodium alginate. Method: Gamma irradiated (0.5 to 40 kGy) aqueous solution of sodium alginate was characterised by UV-visible, fluorescence, Fourier transform infrared (FT-IR) spectroscopy and thin layer chromatography (TLC) analysis. Antioxidant potential of processed alginate was determined using different in vitro assays and prebiotic activity was evaluated by co-culturing of E. coli and Lactobacillus plantarum. Observation: Approximately, 50% of superoxide radicals and 75% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals were scavenged by oligomers at concentrations of 1 mg/ml and 5 mg/ml, respectively. The ability to chelate iron and to inhibit the oxidation of β-carotene was not affected. Radiation processing improved the prebiotic activities as seen by enhanced cell number of L. plantarum by one log and reduction in E. coli count. The efficiency in reducing E. coli depended directly on the absorbed dose. Conclusion: Results suggest that radiation processing of alginate is a practical method for improving its antioxidant and prebiotic activity and thus improving the applicability of alginate in nutraceutical industries.

Background: Extraction or sample preparation is the first step in any research related to natural products and the objectives of the research could be seriously jeopardized if the extract prepared is not the true representative of the original crude material. The work reports a microwave assisted extraction model for the rapid extraction of phenolics and flavonoid bioactives from the leaves of Taraxacum officinale which is an edible plant with profound ethobotanical significance. The work aims to showcase new technological ventures to the nutraceutical industries for large scale production of such components which are greatly used as dietary supplements. Methods: The process optimization of the proposed method has been carried out using Taguchi L9 orthogonal array approach and total phenolics content (μg/gm dried extract in terms of chlorogenic acid equivalent) was used as the performance evaluation parameter. SEM reports and test of integrity of biological activity has also been reported. Results: Eight phenolic/flavonoid bioactives were identified using HPTLC. The final optimum conditions for rapid microwave assisted extraction of phenolics were reported as 160 W microwave power, 6 min irradiation time, 50 °C temperature and 2 min soak time. Results in terms of yield of total phenolics were very superior compared to 36 hours of conventional Soxhlet and maceration extraction. SEM images clearly indicated cellular ruptures, thus facilitating easy exit of bioactives from inside the cell to the external bulk solvent. Conclusions: The work basically attempts to encourage researchers in adapting to green technologies so that science, industry and environment can progress in tandem.

Background: Cancer being a deadly disease, many reports of new chemical entities are available. Pyranopyrazole (PPZ) compounds have also been disclosed as bioactive molecules but mainly as antimicrobial agents. Based on one previous report and our interest in anticancer drug design, we decided to explore PPZs as anticancer agents. To the best of our knowledge, we found that a comprehensive study, involving synthesis, in-vitro biological activity determination, exploration of the mechanism of inhibition and finally in-silico docking studies, was missing in earlier reports. This is what the present study intends to accomplish. Methods: Ten spiro and eleven non-spiro PPZ molecules were synthesized by environment-friendly multicomponent reaction (MCR) strategy. After subjecting each of the newly synthesized molecules to Hep3b hepatocellular carcinoma cell lines assay, we selectively measured the Optical Density (OD) of the most active ones. Then, the compound exhibiting the best activity was docked against human CHK- 1 protein to get an insight into the binding affinities and a quick structure activity relationship (SAR) of the PPZs. Results: The two series of spiro and non-spiro PPZs were easily synthesized in high yields using microwave assisted synthesis and other methods. Among the synthesized compounds, most compounds showed moderate to good anticancer activity against the MTT assay. After performing the absorbance studies we found that the non-spiro molecules showed better apoptosis results and appeared to bind to DNA causing disruption in their structures. Finally, the docking results of compound 5h (having N,Ndimethylamino substituted moiety) clearly showed good binding affinities as predicted by our experimental findings. Conclusion: The paper describes a comprehensive synthesis, in-vitro and docking studies done on new PPZs. The newly synthesized series of spiro and non-spiro PPZs were found to possess antineoplasmic activity as evinced by the studies on hep3b cells. Also, the UV visible absorbance study gave clues to the possible binding of these molecules to the DNA. Docking studies corroborated well with the experimental results. Thus, these new molecules appear to be potential anticancer agents, but further studies are required to substantiate and elaborate on these findings.