Fumarprotocetraric acid

Efficiency and selectivity of ionic liquids in microwave-assisted extraction of major lichen phenolic compounds: a scalable process with recycling of ionic liquids.[Pubmed:33150689]

Phytochem Anal. 2020 Nov 4.

INTRODUCTION: Pseudevernia furfuracea, a lichen used classically for cosmetic applications, contains interesting metabolites possessing antimicrobial and anti-inflammatory or antioxidant properties. OBJECTIVES: Ionic liquid combined to microwave-assisted extraction (IL-MAE) was successfully applied for metabolites extraction from Pseudevernia furfuracea. MATERIALS AND METHODS: Three imidazolium and pyridinium-based ionic liquids (ILs): 1,3-dimethylimidazolium methylsulphate [C1 C1 Im][MeSO4 ], 1-ethyl-3-methylimidazolium ethylsulphate [C2 C1 Im][EtSO4 ], and N-ethylpyridinium ethylsulphate [C2 Py][EtSO4 ] were assessed for this process. The efficiency of the extraction method was evaluated using thin-layer chromatography (TLC) coupled to a Camag(R) spectrophotodensitometer and using high-performance liquid chromatography (HPLC) analysis. RESULTS: ILs under MAE showed extraction time efficiency (15 min vs. 24 h for conventional heating) and high selectivity in extracting the targeted metabolites: atranorin (AT), methyl-beta-orcinol carboxylate (MOC), Fumarprotocetraric acid (Fum. Ac.), and physodic acid (Phys. Ac.) despite the increased degradation of AT under MAE. We showed a tunable selectivity of ILs towards extracting metabolites by changing anion or cation due to the modification of the interaction between the IL and the metabolites. While [C2 Py][EtSO4 ] was the most efficient IL and could extract all the targeted metabolites, [C2 C1 Im][EtSO4 ] was the most selective. It fully extracted AT and partially Fum. Ac. Moreover, the lichen prepared by mixing procedure provided AT and Fum. Ac. more than the milled one. A 100 times scale-up extraction was successfully performed on mixed samples with full IL recycling after back extraction. CONCLUSION: IL-MAE is reliable for lichen metabolites extraction. The method is reproducible, scalable, with possible IL recycling, opening the door for potential industrial applications.

Tissue-specific localization of polyketide synthase and other associated genes in the lichen, Cladonia rangiferina, using laser microdissection.[Pubmed:30296707]

Phytochemistry. 2018 Dec;156:142-150.

The biosynthesis of two polyketides, atranorin and Fumarprotocetraric acid, produced from a lichen-forming fungus, Cladonia rangiferina (L.) F. H. Wigg. was correlated with the expression of eight fungal genes (CrPKS1, CrPKS3, CrPKS16, Catalase (CAT), Sugar Transporter (MFsug), Dioxygenase (YQE1), C2H2 Transcription factor (C2H2), Transcription Factor PacC (PacC), which are thought to be involved in polyketide biosynthesis, and one algal gene, NAD-dependent deacetylase sirtuin 2 (AsNAD)), using laser microdissection (LMD). The differential gene expression levels within the thallus tissue layers demonstrate that the most active region for potential polyketide biosynthesis within the lichen is the outer apical region proximal to the photobiont but some expression also occurs in reproductive tissue. This is the first study using laser microdissection to explore gene expression of these nine genes and their location of expression; it provides a proof-of-concept for future experiments exploring tissue-specific gene expression within lichens; and it highlights the utility of LMD for use in lichen systems.

Amelioration of anti-hepatotoxic effect by Lichen rangiferinus against alcohol induced liver damage in rats.[Pubmed:29395895]

J Ayurveda Integr Med. 2019 Jul - Sep;10(3):171-177.

BACKGROUND: Reindeer lichen, Lichen rangiferinus syn. or Cladonia rangiferina (L.) F. H. Wigg. (Cladoniaceae) has been traditionally reported as a remedy to treat fever, colds, arthritis as well as convulsions, liver infections, coughs, constipation, and tuberculosis. The current study is aimed at rectification of alcohol induced liver damage by the use of L. rangiferinus extract. OBJECTIVES: The aim of the study was to compare some biochemical markers for liver injury and hematological indices in normal untreated rats and treated rats. MATERIAL AND METHODS: The study was performed using male Wistar rats. Animals were categorized into five groups, negative control group (normal diet only), treated groups (2 groups were lichen treated along with 10% ethanol & 1 group was only ethanol treated) and positive control group (Silymarin+10% ethanol) of six animals in each group. Biochemical markers for liver injury and hematological indices of all animals were measured using standard diagnostic tools. The animals were then sacrificed and livers were sent to the pathology lab for histopathological analysis. RESULTS: Lichen extract showed a significant restoration of altered biochemical parameters towards normal in both in vitro and in vivo conditions. The total phenolic and flavonoid content of the LRE was found to be 21.78 mug PE/mg of extract and 5.13 mug RE/mg of extract respectively. The IC50 values for atranorin and Fumarprotocetraric acid were found to be 128.48 and 218.46 mg/mL respectively. Reducing power of the extract was found to be quite significant. After administration of lichen extract, endothelial cells were less injured around central vein and number of fat vacuoles was also lesser in hepatocytes. CONCLUSION: Conclusively, treatment with lichen extract assuages alcohol-related damage and guards hepatic tissue from alcohol-induced toxicity.

In vitro neuroprotective potential of lichen metabolite fumarprotocetraric acid via intracellular redox modulation.[Pubmed:28041784]

Toxicol Appl Pharmacol. 2017 Feb 1;316:83-94.

The lichen-forming fungi Cetraria islandica has been largely used in folk medicines, and it has recently showed promising in vitro antioxidant effects in glial-like cells. Current work aimed at investigating the neuroprotective potential of its major isolated secondary metabolite: the depsidone Fumarprotocetraric acid (FUM). H2O2 was used herein to induce oxidative stress (OS)-mediated cytotoxicity in two models of neurons and astrocytes cells (SH-SY5Y and U373-MG cell lines). We found that a pre-treatment with FUM significantly enhanced cell viability compared to H2O2-treated cells, and we selected the optimal concentrations in each model (1 and 25mug/ml, respectively) for assessing its cytoprotective mechanisms. FUM, which exerted effective peroxyl radical scavenging effect in the chemical oxygen radical antioxidant capacity (ORAC) assay, alleviated the alterations in OS markers provoked by H2O2. It attenuated intracellular ROS formation, lipid peroxidation and GSH depletion. At mitochondrial level, FUM prevented from the dissipation of mitochondrial membrane potential and the increase in mitochondrial calcium, implying a protective role against oxidative damage in mitochondrial membrane. Similarly, FUM pre-treatment diminished H2O2-induced apoptosis, as evidenced by the reduction in caspase-3 activity and expression; inmunoblot analysis also revealed a decrease in Bax and an increase in Bcl-2 proteins levels. Furthermore, FUM up-regulated the expression of the antioxidant enzymes catalase, superoxide dismutase-1, and hemeoxigenase-1. These findings and the activation of Nrf2 binding activity in nuclear extracts suggest a plausible involvement of Nrf2 signaling pathway in the cytoprotection by FUM. In conclusion, FUM emerges as a potential drug candidate in the therapy of OS-related diseases, such as the neurodegenerative disorders.

Algal carbohydrates affect polyketide synthesis of the lichen-forming fungus Cladonia rangiferina.[Pubmed:27091386]

Mycologia. 2016 Jul-Aug;108(4):646-56.

Lichen secondary metabolites (polyketides) are produced by the fungal partner, but the role of algal carbohydrates in polyketide biosynthesis is not clear. This study examined whether the type and concentration of algal carbohydrate explained differences in polyketide production and gene transcription by a lichen fungus (Cladonia rangiferina). The carbohydrates identified from a free-living cyanobacterium (Spirulina platensis; glucose), a lichen-forming alga (Diplosphaera chodatii; sorbitol) and the lichen alga that associates with C. rangiferina (Asterochloris sp.; ribitol) were used in each of 1%, 5% and 10% concentrations to enrich malt yeast extract media for culturing the mycobiont. Polyketides were determined by high performance liquid chromatography (HPLC), and polyketide synthase (PKS) gene transcription was measured by quantitative PCR of the ketosynthase domain of four PKS genes. The lower concentrations of carbohydrates induced the PKS gene expression where ribitol up-regulated CrPKS1 and CrPKS16 gene transcription and sorbitol up-regulated CrPKS3 and CrPKS7 gene transcription. The HPLC results revealed that lower concentrations of carbon sources increased polyketide production for three carbohydrates. One polyketide from the natural lichen thallus (Fumarprotocetraric acid) also was produced by the fungal culture in ribitol supplemented media only. This study provides a better understanding of the role of the type and concentration of the carbon source in fungal polyketide biosynthesis in the lichen Cladonia rangiferina.

Neuroprotective activity and cytotoxic potential of two Parmeliaceae lichens: Identification of active compounds.[Pubmed:26220632]

Phytomedicine. 2015 Aug 15;22(9):847-55.

BACKGROUND: Lichens are symbiotic organisms capable of producing unique secondary metabolites, whose pharmacological activities are attracting much interest. PURPOSE: The present study aimed to investigate the in vitro neuroprotective effects and anticancer potential of methanol extracts of two Parmeliaceae lichens: Cetraria islandica and Vulpicida canadensis. The chemical composition of the two lichens was also determined. METHODS: Neuroprotective activity was studied with respect to the antioxidant properties of the extracts; radical scavenging tests (ORAC and DPPH assays) were performed and oxidative stress markers (intracellular ROS production, caspase-3 activity, MDA and glutathione levels) were assessed in a hydrogen peroxide-induced oxidative stress model in astrocytes. Cytotoxic activity was tested against human HepG2 (hepatocellular carcinoma) and MCF-7 (breast adenocarcinoma) cell lines. RESULTS: Cell viability studies identified a single concentration for each extract that was subsequently used to measure oxidative stress markers. Lichen extracts were able to reverse the oxidative damage caused by hydrogen peroxide, thus promoting astrocyte survival. Both lichen extracts also had anticancer activity in the cell lines, with IC50 values of 19.51-181.05 microg/ml. The extracts had a high total phenolic content, and the main constituents identified by HPLC were Fumarprotocetraric acid in Cetraria islandica, and usnic, pinastric and vulpinic acids in Vulpicida canadensis. The biological activities of the lichen extracts can be attributed to these secondary metabolites. CONCLUSION: The lichen species studied are promising sources of natural compounds with neuroprotective activity and cytotoxic potential, and warrant further research.

Antitrypanosomal activity & docking studies of isolated constituents from the lichen Cetraria islandica: possibly multifunctional scaffolds.[Pubmed:24660683]

Curr Top Med Chem. 2014;14(8):1014-21.

Chemical investigation of the lichen Cetraria islandica has led to the isolation of four compounds identified as protolichesterinic acid, lichesterinic acid, protocetraric acid and Fumarprotocetraric acid. Their structures were characterized using their physical and spectroscopic data. Using an Alamarblue 96 well microplate assay, these compounds were tested to evaluate their trypanocidal activity against Trypanosoma brucei brucei. Protolichesterinic acid (MIC = 6.30 microM) and lichesterinic acid (MIC = 12.5 microM) showed very significant activity against the test organism. Docking studies (GRIP technique) of these molecules revealed their strong affinity towards possible targets of Trypanosoma brucei such as riboflavin kinase, sterol-14alpha-demethylase (CYP51), rohedsain and glutathione synthetase. Hydrophobicity played a significant role in their antitrypanosomal activity.

Expectorant and antioxidant activities of purified fumarprotocetraric acid from Cladonia verticillaris lichen in mice.[Pubmed:23872116]

Pulm Pharmacol Ther. 2014 Apr;27(2):139-43.

UNLABELLED: The lichen Cladonia verticillaris produces bioactive secondary metabolites, such as fumarprotocetraric (FUM) and protocetraric acids. Species of the genus Cladonia demonstrate anti-tumor, anti-inflammatory and antipyretic activities and have been used in folk medicine to treat respiratory diseases (throat irritation, cough, asthma and tuberculosis). The aim of the present study was to evaluate the expectorant and mucolytic activities of Fumarprotocetraric acid in albino Swiss mice. FUM was extracted and purified from an acetone extract of C. verticillaris. The phenol red quantification method was used on the bronchoalveolar lavage fluid following the administration of FUM (25, 50 or 100 mg/kg orally or intraduodenally and 12.5, 25 or 50 mg/kg, intraperitoneally) for the evaluation of expectorant activity. Control groups received either saline solution (7.5 mL/kg) or ambroxol (1 mg/kg) through the same administration routes. Antioxidant activity was evaluated using the thiobarbituric acid reactive species assay in mouse lung tissue treated with the FUM at 25, 50 or 100 mg/kg orally, followed by a lipopolysaccharide solution at 1 mg/kg intrapleurally. The same protocol was used for the control groups using either saline solution (7.5 mL/kg, orally) or N-acetylcysteine (20 mg/kg, orally). RESULTS: Orally administered FUM at doses of 25 and 50 mg/kg promoted significantly greater dose-dependent phenol red activity in the bronchoalveolar lavage and expectorant activity in comparison to the controls (p < 0.05). Lipid peroxidation (malondialdehyde equivalent) was reduced by 50% in the lung tissue. CONCLUSION: The results confirm the expectorant and antioxidant properties of Fumarprotocetraric acid produced by the lichen C. verticillaris.

Antimicrobial and antibiofilm activity of secondary metabolites of lichens against methicillin-resistant Staphylococcus aureus strains from cystic fibrosis patients.[Pubmed:23374132]

Future Microbiol. 2013 Feb;8(2):281-92.

AIM: Three secondary metabolites of lichens - usnic acid, atranorin and Fumarprotocetraric acid - were evaluated for their in vitro antibacterial and antibiofilm activities against three strains each of methicillin-susceptible and methicillin-resistant Staphylococcus aureus (MRSA) from cystic fibrosis patients. MATERIALS & METHODS: Antibacterial activity was assessed by broth microdilution, while antibiofilm activity was evaluated by spectrophotometry or viable count. RESULTS: Usnic acid was significantly more active than atranorin against planktonic cells, while Fumarprotocetraric acid exhibited no activity. Atranorin was the most effective in counteracting adhesion to polystyrene, although usnic acid was more active against MRSA. Usnic acid and atranorin showed comparable activity against biofilm formation, although atranorin was more active against MRSA. Usnic acid was significantly more active than atranorin against preformed biofilms. CONCLUSION: Secondary metabolites of lichens may be considered to be 'lead compounds' for the development of novel molecules for the treatment of S. aureus infections in cystic fibrosis patients.

Antibacterial activity of selected metabolites from Chilean lichen species against methicillin-resistant staphylococci.[Pubmed:23030591]

Nat Prod Res. 2013;27(17):1528-31.

The in vitro antibacterial activities of eight compounds isolated from lichens, collected in several Southern regions of Chile (including Antarctica), were evaluated against methicillin-resistant clinical isolates strains of Staphylococcus aureus, Staphylococcus haemolyticus and Staphylococcus warneri. The minimum inhibitory concentrations, calculated in microdilution, were ranging from 8 microg mL(-1) for sphaerophorin to 1024 microg mL(-1) for Fumarprotocetraric acid. These findings suggest, however, that the natural compounds from lichens are good candidates for the individuation of novel templates for the development of new antimicrobial agents or combinations of drugs for chemotherapy.

Antioxidant and antimicrobial properties of some lichens and their constituents.[Pubmed:21861720]

J Med Food. 2011 Dec;14(12):1624-30.

The aim of this study is to examine the in vitro antioxidant and antimicrobial activities of the acetone, methanol, and aqueous extracts of the lichens Cladonia furcata (many-forked Cladonia), Hypogymnia physodes (puffed lichen), and Umbilicaria polyphylla (manyleaf navel lichen) and the antimicrobial activity of Fumarprotocetraric acid, gyrophoric acid, and physodic acid isolated from the respective lichen species. Antioxidant activity was evaluated by five different methods: free radical scavenging, superoxide anion radical scavenging, reducing power, determination of total phenolic compounds, and determination of total flavonoid content. U. polyphyla exhibited more powerful antioxidant activities than the other lichen species examined: methanol extracts of U. polyphyla showed 90.08% free radical scavenging activities (the ratio was greater than those of standard antioxidants). Moreover, the extracts tested had effective reducing power and superoxide anion radical scavenging. Total contents of phenol and flavonoid in extracts were determined as pyrocatechol equivalents and as rutin equivalents, respectively. A significant strong relationship between total phenolic and flavonoid contents and their antioxidative activities was observed. The antimicrobial activity was estimated by determination of the minimal inhibitory concentration by the broth microdilution method against six species of bacteria and 10 species of fungi. In general, the lichen extracts tested and their respective acids had relatively strong antimicrobial activity against the microorganisms tested. The present study shows that the lichen species tested demonstrated strong antioxidant and antimicrobial activities. The results suggest that lichens may be used as good sources of natural antioxidants and for pharmaceutical purposes in treating various diseases.

High acidity tolerance in lichens with fumarprotocetraric, perlatolic or thamnolic acids is correlated with low pKa1 values of these lichen substances.[Pubmed:19464777]

Environ Pollut. 2009 Oct;157(10):2776-80.

The depsidone Fumarprotocetraric acid as well as the depsides perlatolic and thamnolic acids are lichen secondary metabolites. Their first dissociation constants (pK(a1)) in methanol were determined to be 2.7 for perlatolic acid and 2.8 for fumarprotocetraric and thamnolic acids by UV spectroscopy. Lower pK(a1) values are, so far, not known from lichen substances. Several lichens producing at least one of these compounds are known for their outstanding tolerance to acidic air pollution. This is demonstrated by evaluating published pH preferences for central European lichens. The low pK(a1) values suggest that strong dissociation of the studied lichen substances is a prerequisite for the occurrence of lichens with these compounds on very acidic substrata, as protonated lichen substances of different chemical groups, but not their conjugated bases, are known to shuttle protons into the cytoplasm and thereby apparently damage lichens.

Cladonia peziziformis (Lichenized Ascomycota, Cladoniaceae) New to Korea.[Pubmed:23997624]

Mycobiology. 2008 Sep;36(3):193-4.

Cladonia peziziformis (With.) J.R. Laundon was collected from Baega mountain, Jeonnam Province, Korea in 2008. It is characterized by short and slender podetia with verruculose surface, split along the sides. Apothecia large, pale brown, always growing on the top of the podetia. Primary squamules shell-like, thick, and convex. Fumarprotocetraric acid contained in thallus. This is the first record of this species in Korea.

Susceptibility to acidic precipitation contributes to the decline of the terricolous lichens Cetraria aculeata and Cetraria islandica in central Europe.[Pubmed:18053625]

Environ Pollut. 2008 Apr;152(3):731-5.

The effective quantum yield of photochemical energy conversion in photosystem II (Phi2) was shown to be reduced in the terricolous lichens Cetraria aculeata and Cetraria islandica by short-term exposure to aqueous SO2 at pH values occurring in the precipitation of areas with high SO2 pollution. Significant reduction of Phi2 was found at pHFumarprotocetraric acid, a major lichen substance of C. islandica, increases the pollution tolerance in lichens.