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Planta Med 2021; 87(04): 294-304
DOI: 10.1055/a-1289-9569
Biological and Pharmacological Activity
Original Papers

Biochemometric Analysis of Fatty Acid Amide Hydrolase Inhibition by Echinacea Root Extracts

Rui Liu
1   Department of Biology, University of Ottawa, Ottawa, ON, Canada
,
Kelly Burkett
2   Department of Mathematics and Statistics, University of Ottawa, Ottawa, ON, Canada
,
Michel Rapinski
3   Institut de recheche en biologie végétale (IRBV), University of Montreal, Montreal, QC, Canada
,
John T. Arnason
1   Department of Biology, University of Ottawa, Ottawa, ON, Canada
,
Franklin Johnson
4   Trout Lake Farm, LLC, Trout Lake, WA, USA
,
Phil Hintz
4   Trout Lake Farm, LLC, Trout Lake, WA, USA
,
John Baker
5   CBD Baker Inc., Stirling, ON, Canada
,
Cory S. Harris
1   Department of Biology, University of Ottawa, Ottawa, ON, Canada
› Author AffiliationsSupported by: Mitacs IT09228
Supported by: Natural Sciences and Engineering Research Council of Canada 2015-05453
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Abstract

Recent research demonstrates that Echinacea possesses cannabimimetic activity with potential applications beyond common contemporary uses for relief of cold and flu symptoms. In this study, we investigated the in vitro inhibitory effect of root extracts of Echinacea purpurea and Echinacea angustifolia on fatty acid amide hydrolase, the main enzyme that degrades the endocannabinoid anandamide. The objective was to relate variation in bioactivity between commercial Echinacea genotypes to their phytochemical profiles and to identify determinants of activity using biochemometric analysis. Forty root extracts of each of species were tested for inhibition of fatty acid amide hydrolase and analyzed by HPLC-DAD/MS to identify and quantitate alkylamides and caffeic acid derivatives. Fatty acid amide hydrolase inhibition ranged from 34 – 80% among E. angustifolia genotypes and from 33 – 87% among E. purpurea genotypes. Simple linear regression revealed the caffeic acid derivatives caftaric acid and cichoric acid, and the alkylamide dodeca-2E,4Z-diene-8,10-diynioc acid 2-methylbutylamide, as the strongest determinants of inhibition in E. purpurea (r* = 0.53, 0.45, and 0.20, respectively) while in E. angustifolia, only CADs were significantly associated with activity, most notably echinacoside (r* = 0.26). Regression analysis using compound groups generated by hierarchical clustering similarly indicated that caffeic acid derivatives contributed more than alkylamides to in vitro activity. Testing pure compounds identified as determinants of activity revealed cichoric acid (IC50 = 45 ± 4 µM) and dodeca-2E,4E,8Z,10E-tetraenoic acid isobutylamide (IC50 = 54 ± 2 µM) as the most active. The results suggest that several phytochemicals may contribute to Echinaceaʼs cannabimimetic activity and that ample variation in genotypes exists for selection of high-activity germplasm in breeding programs.

Key words

Echinacea - Asteraceae - coneflowers - alkamides - phenolics - endocannabinoid system

Supporting Information

    Enzyme inhibition activity and CAD and AKA content of tested E. angustifolia and E. purpurea root extracts as well as the concentration-dependent inhibition of FAAH activity by pure Echinacea CADs and AKAs are available as Supporting Information.

  • Supporting Information


Publication History

Received: 30 May 2020

Accepted after revision: 02 October 2020

Article published online:
09 December 2020

© 2020. Thieme. All rights reserved.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

  • References

  • 1 Shemluck M. Medicinal and other uses of the compositae by Indians in the United States and Canada. J Ethnopharmacol 1982; 5: 303-358
    CrossrefPubMedGoogle Scholar
  • 2 Smith T, Guillespie M, Eckl V, Knepper J, Morton-Reynolds C. US Retail Sales of Herbal Supplements Increase by 9.4 % in 2018. HerbalGram 2018; 123: 62-73
    PubMedGoogle Scholar
  • 3 Blumenthal M, Hall T, Goldberg A, Kunz T, Dinda K. eds. The ABC Guide to clinical Herbs. Austin, Texas: American Botanical Council; 2003: 235-246
    Google Scholar
  • 4 Binns SE, Livesey JF, Arnason JT, Baum BR. Phytochemical variation in echinacea from roots and flowerheads of wild and cultivated populations. J Agric Food Chem 2002; 50: 3673-3687
    CrossrefPubMedGoogle Scholar
  • 5 Binns SE, Livesey JF, Arnason JT, Baum BR. Phytochemical variation within populations of Echinacea angustifolia (Asteraceae). Biochem Syst Ecol 2002; 30: 837-854
    CrossrefPubMedGoogle Scholar
  • 6 Barrett B. Medicinal properties of Echinacea: a critical review. Phytomedicine 2003; 10: 66-86
    CrossrefPubMedGoogle Scholar
  • 7 Haller J, Hohmann J, Freund TF. The effect of Echinacea preparations in three laboratory tests of anxiety: comparison with chlordiazepoxide. Phytother Res 2010; 24: 1605-1613
    CrossrefPubMedGoogle Scholar
  • 8 Haller J, Freund TF, Pelczer KG, Füredi J, Krecsak L, Zámbori J. The anxiolytic potential and psychotropic side effects of an Echinacea preparation in laboratory animals and healthy volunteers. Phytother Res 2013; 27: 54-61
    CrossrefPubMedGoogle Scholar
  • 9 Chicca A, Raduner S, Pellati F, Strompen T, Altmann KH, Schoop R, Gertsch J. Synergistic immunomopharmacological effects of N-alkylamides in Echinacea purpurea herbal extracts. Int Immunopharmacol 2009; 9: 850-858
    CrossrefPubMedGoogle Scholar
  • 10 Di Marzo V. New approaches and challenges to targeting the endocannabinoid system. Nat Rev Drug Discov 2018; 17: 623-639
    CrossrefPubMedGoogle Scholar
  • 11 Di Marzo V, Bisogno T, Melck D, Ross R, Brockie H, Stevenson L, Pertwee R, De Petrocellis L. Interactions between synthetic vanilloids and the endogenous cannabinoid system. FEBS Lett 1998; 436: 449-454
    CrossrefPubMedGoogle Scholar
  • 12 Guindon J, Hohmann AG. Cannabinoid CB2 receptors: a therapeutic target for the treatment of inflammatory and neuropathic pain. Br J Pharmacol 2008; 153: 319-334
    CrossrefPubMedGoogle Scholar
  • 13 Nackley AG, Makriyannis A, Hohmann AG. Selective activation of cannabinoid CB2 receptors suppresses spinal fos protein expression and pain behavior in a rat model of inflammation. Neuroscience 2003; 119: 747-757
    CrossrefPubMedGoogle Scholar
  • 14 Hohmann J, Rédei D, Forgo P, Szabó P, Freund TF, Haller J, Bojnik E, Benyhe S. Alkamides and a neolignan from Echinacea purpurea roots and the interaction of alkamides with G-protein-coupled cannabinoid receptors. Phytochemistry 2011; 72: 1848-1853
    CrossrefPubMedGoogle Scholar
  • 15 Woelkart K, Xu W, Pei Y, Makriyannis A, Picone RP, Bauer R. The endocannabinoid system as a target for alkamides from Echinacea angustifolia roots. Planta Med 2005; 71: 701-705
    Article in Thieme ConnectPubMedGoogle Scholar
  • 16 Thors L, Belghiti M, Fowler CJ. Inhibition of fatty acid amide hydrolase by kaempferol and related naturally occurring flavonoids. Br J Pharmacol 2008; 155: 244-252
    CrossrefPubMedGoogle Scholar
  • 17 Raduner S, Majewska A, Chen JZ, Xie XQ, Hamon J, Faller B, Altmann KH, Gertsch J. Alkylamides from Echinacea are a new class of cannabinomimetics. Cannabinoid type 2 receptor-dependent and -independent immunomodulatory effects. J Biol Chem 2006; 281: 14192-14206
    CrossrefPubMedGoogle Scholar
  • 18 Oboh G, Agunloye OM, Adefegha SA, Akinyemi AJ, Ademiluyi AO. Caffeic and chlorogenic acids inhibit key enzymes linked to type 2 diabetes (in vitro): a comparative study. J Basic Clin Physiol Pharmacol 2015; 26: 165-170
    CrossrefPubMedGoogle Scholar
  • 19 Chiou SY, Sung JM, Huang PW, Lin SD. Antioxidant, antidiabetic, and antihypertensive properties of Echinacea purpurea flower extract and caffeic acid derivatives using In Vitro models. J Med Food 2017; 20: 171-179
    CrossrefPubMedGoogle Scholar
  • 20 Rastogi H, Jana S. Evaluation of inhibitory effects of caffeic acid and quercetin on human liver cytochrome p450 activities. Phytother Res 2014; 28: 1873-1878
    CrossrefPubMedGoogle Scholar
  • 21 Baell JB. Feeling natureʼs PAINS: Natural products, natural product drugs, and Pan Assay Interference Compounds (PAINS). Nat Prod 2016; 79: 616-628
    CrossrefPubMedGoogle Scholar
  • 22 Woelkart K, Kollroser M, Magnes C, Sinner F, Frye RF, Derendorf H, Bauer R, Butterweck V. Absolute/relative bioavailability and metabolism of dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamides (tetraenes) after intravenous and oral single doses to rats. Planta Med 2011; 77: 1794-1799
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Bel-Rhlid R, Pagé-Zoerkler N, Fumeaux R, Ho-Dac T, Chuat JY, Sauvageat JL, Raab T. Hydrolysis of chicoric and caftaric acids with esterases and Lactobacillus johnsonii in vitro and in a gastrointestinal model. J Agric Food Chem 2012; 60: 9236-9241
    CrossrefPubMedGoogle Scholar
  • 24 Wang SJ, Zeng J, Yang BK, Zhong YM. Bioavailability of caffeic acid in rats and its absorption properties in the Caco-2 cell model. Pharm Biol 2014; 52: 1150-1157
    CrossrefPubMedGoogle Scholar
  • 25 Bergeron C, Livesey JF, Awang DV, Arnason JT, Rana J, Baum BR, Letchamo W. A quantitative HPLC method for the quality assurance of Echinacea products on the North American market. Phytochem Anal 2000; 11: 207-215
    CrossrefPubMedGoogle Scholar
  • 26 Liu R. Pharmacology and Toxiclogy of Echinacea, Souroubea and Platanus spp. [PhD Thesis]. Ottawa: University of Ottawa; 2019: 1-178
    PubMedGoogle Scholar
  • 27 Anderson MJ, Legendre P. An empirical comparison of permutation methods for tests of partial regression coefficients in a linear model. J Stat Comput Sim 1999; 62: 271-303
    CrossrefPubMedGoogle Scholar
  • 28 Legendre P, Anderson MJ. Distance-based redundancy analysis: testing multispecies responses in multifactorial ecological experiments. Ecol Monogr 1999; 69: 1-24
    CrossrefPubMedGoogle Scholar
  • 29 Legendre P, Legendre L. Numerical Ecology. 3rd ed.. ed. Amsterdam: Elsevier; 2012
    Google Scholar
  • 30 R Core Team. R: A Language and Environment for Statistical Computing. Vienna, Austria: R Foundation for Statistical Computing; 2019. Accessed February 2, 2020 at: https://www.r-project.org/
    Google Scholar