Plant Secondary Metabolites PDF

Theory of plant defense  PSM not waste products, serve as defense compounds against herbivores (mechanical...

Theory of plant defense  PSM not waste products, serve as defense compounds against herbivores (mechanical defense - thorns, trichomes, thick epidermis, sticky or smooth surfaces - prevent eating or laying eggs; chemical defense - a series of compounds that are toxic, deter or cause the indigestibility of plant tissues - extensive groups: alkaloids, phenols, terpenoids, iridoid glycosides, cardenolides, cyanogenic glycosides; signaling molecules for pollinators or symbionts, additional role in plant development, membrane fluidity/integrity - sterols). PLANT SECONDARY METABOLITES  Compared to primary metabolites, evolutionary PSM not less important for the plant, Part I. primary and secondary metabolism are interwoven.  PSM in optimal or non-stress conditions (constitutive PSM), characteristic for species or higher taxonomic units (chemotaxonomic features of PSM - usually a single SM group dominates within a given taxon) (terpenoids: Apiaceae, Asteraceae, Lamiaceae, Rutaceae, Lauraceae; cyanogenic glycosides: Rosaceae; glucosinolates: Brassicaceae; alkaloids: Apocynaceae, Solanaceae...). SITUATION OF PHYTOPHARMAKA Plant secondary metabolites (PSM) TODAY AND IN THE FUTURE  Synthetic and herbal remedies: not mutually exclusive, but complementary Herbal remedies reduce the consumption of synthetic medicines/alleviate their side effects The use of phytopharmaceuticals makes sense: - treatment of slowly developing and chronic diseases - combinations of several active ingredients within the herbal preparation, which provide a better therapeutic effect regarding efficacy and acceptability (e.g. chamomile, yarrow) as a supplement of isolated active ingredients - if appropriate therapeutic effect is not found in the known isolated compound or if this is  few precursors (AA: precursors of alkaloids; activated isoprenes: precursors of uneconomical (e.g. phytopharmaceuticals based on hawthorn - Crataegus monogyna Jacq. - isoprenoids and their derivatives; shikimic acid: aromatic compounds, aromatic amino acids, - effective in heart insufficiency/feeling of pressure around the heart in an elderly heart that cinnamic acid and derivatives, polyphenolic compounds); does not yet need digitalization; even at high doses shows no toxicity - if the individual active ingredient is not yet known, but there is no doubt about the efficacy of  their abundance/diversity is caused by specific (substrate) construction enzymes, less the medicine (e.g. traditional medicines). specific degradative enzymes (β-glycosidases, esterases, hydrolases); Approval and enforcement of herbal remedies:  valuable tool in chemotaxonomy; guaranteed with pharmaceutical quality, efficiency and safety  determine the therapeutic effectiveness and quality of the herbal drug/preparation. Primary and secondary plant metabolites Connection between primary and secondary metabolism  Plant primary metabolites: Biosynthetic origin: division of PSM into three large groups: terpenoids (> 25,000 in all plant species (< meaning in chemotaxonomy); important role in plant metabolism; compounds, IPP derivatives: 5-C isopentenyl pyrophosphate, formed from acetyl - CoA), related to erg metabolism: essential for the synthesis of structural elements and the phenylpropanoids (~ 2,000) and related phenolic compounds (> 9,000 compounds, shikimi- growth and development of plants (organic acids, sugars - polysaccharides, AA - chorismate derivatives or mevalonate) and alkaloids (~ 12,000 compounds, derivatives of proteins, fatty acids - lipids, nucleic acids - nucleotides) amino acids).  Plant secondary metabolites (PSM): Precursor molecules: not a basis for distinguishing between primary and SM: between numerous and very diverse types of organic low-molecular compounds of complex diterpenes (C20) and triterpenes (C30) - both primary and secondary metabolites structure, produced by plants in small quantities; around 100,000 known structures, chemically only 5-15% of plants "studied“; most do not participate directly in growth and development (hence "secondary"), they make an important contribution to survival in evolution; Kaurenoic acid Abietic acid (intermediate in gibberellin biosynthesis) (component of resins in Fabaceae in Pinaceae) biosynthesis balanced with the basic biochemical and morphological processes of cell differentiation and development (the factors that trigger the synthesis of specific enzymes of secondary metabolism at a certain stage of ontogenetic development not known); appear in plants as a result of chemical conversions of precursors under the influence of cascade of enzymes. Proline Pipecolic acid (aminoacid – primary metabolite) (precursor in biosynthesis of alkaloids) 1  Diversity of PSM PSM: concentration/composition species/population/ developmental stage  High diversity of PSMs in many plant species - original thinking: a specific plant species encodes only a few enzymes that synthesize most PSMs; different PSMs  2 types of temporal changes: ontogenetic (genetically programmed developmental are formed from many metabolic substrates with a limited set of enzymes: a single changes during life period, i.e. maturation - seedling, adult plant, mature plant) and enzyme catalyzes the synthesis of multiple compounds from different substrates; in seasonal (result of temperature, lighting, water and nutrients - direct influence on the course of evolution: enzyme reactions adapt to available substrates - changed plant growth & development (GD) and induction of PSM synthesis) enzymes (changes in a limited number of amino acids) - change in gene expression;  Ontogenetic studies (constitutive PSM – plants not under stress, PSM not stress if a compound favorable to the plant was synthesized, the genetic changes were induced): concentration, regardless of the type of RSM, increases with the adapted. maturation of the seedling (herbaceous species, trees); with further ripening, the amount of PSM depends on species and the type of PSM  Populations with higher genetic variability - higher PSM variability, better adaptability and better utility.  Seasonal studies (constitutive PSM) (meta analysis - 42 studies, species from 20 families): at the same time as the leaves develop, the biosynthesis of RSM  Defensive role against herbivorous (insects and vertebrates) animals, bacteria and increases, the peak at the full development of the leaves, it begins to decline with fungi, viruses, other plants with which they compete for light, water and nutrients, aging (dependency on weather conditions) represent signaling molecules/compounds for animals/insects that enable pollination, for communication between plants and symbiotic organisms (mycorrhizal  The accumulation of any RSM in plant tissues is regulated by the ratio fungi,...), protection against environmental stresses biosynthesis/degradation. Some PSMs in the form of glycosides (heterosides) Biosynthesis of PSM  Glycosides (heterosides): many PSM are linked to sugars (hexoses: d- glucose, galactose, mannose; pentoses: arabinose, xylose, ribose; deoxy-2  metabolic pathways and genes involved in their synthesis are controlled, their sugars - digitoxose...); acids, alkalis, enzymes break them down into activation is linked to environmental, seasonal and external triggers; aglycone (genine) and glycone (sugar component)  PSM concentration reflects the synthesis/degradation ratio (balance) - daily  Number of sugar molecules: monoglycosides, diglycosides, oligoglycosides changes - monoterpenes, quinolizidine alkaloids; active transcription of the genes involved;  Glycone and aglycone can be linked by -O (mostly); -S (Brassicaceae senevole - glycosides, -C (Aloe anthra-glycosides)  plant genome: several thousand genes, only 15-25% of genes participate in secondary metabolism; these genes are regulated by various transcription  Solid crystalline substances, sometimes coloured (yellow flavonoids, red factors (TFs) – proteins that influence metabolism by influencing gene anthocyanosides, red or orange anthracenosides), often bitter in taste, their expression; solubility in water (glycon dependent)  PSM found throughout the plant, the initial site of synthesis is limited to 1 organ  Present in all organs: roots (gentiopicroside - Gentianaceae), bark (roots, shoots, leaves, flowers, fruit, seed); (esculoside - Aesculus hippocastanum, franguloside - Rhamnus frangula), leaves (cardiotonic - Digitalis spp., linamarin - Linaceae), fruits and seeds (prunasin, amygdalin, sambunigrin - Prunus spp , Sambucus nigra) PSM biosynthesis and storage Glycosides (heterosides) according to aglycone (genin) determines the effectiveness of the plant drug  most biosynthetic pathways in the cytoplasm (hydrophilic substances); in plastids heterosides of simple phenols (salicoside - salicin – white willow bark - Salicis (chloroplasts - some alkaloids - caffeine, coumarins, leucoplasts - monoterpenes,   22 22 cortex analgesic, antipyretic, arbutoside - arbutin - Uvae ursi folium: uroantiseptic), diterpenes, chromoplasts - carotenoids); in the endoplasmic reticulum (sesquiterpenes, sterols, post-synthetic changes - hydroxylation); mitochondria cyanogenic heterosides (capable of releasing hydrogen cyanide – HCN after (coniin, some amines) digestion) seeds, leaves Prunus spp.; obsolete antitussive: Aqua ammigdalae amarae - max. 1% HCN)  after formation, they can be transported through the phloem or xylem, via symplastic or apoplastic transport, and are stored in a variety of tissues (flowers, steroide heterosides (Foxglove leaf - Digitalis spp.- Scrophulariaceae, Nerium - fruits, seeds - annuals; bulbs, roots, rhizomes, bark - perennials) Apocynaceae, Lily of the Valley herb - Convallariae herba - Liliaceae, digitaloids: cardiotonics), anthraquinone heterosides (Aloe spp. - Liliaceae, Buckthorn bark - Rhamnus spp.- Rhamnaceae, Rhubarb root - Rheum spp.- Polygonaceae: laxatives), flavonoid heterosides (Asteraceae, Ginkoaceae, Rosaceae, Rutaceae antocianidini, flavanone - naringin, hesperetin, flavone - apigenin, luteolin, flavonol  stored in vacuoles (water-soluble compounds: alkaloids, flavonoids, – kemferol, kvercetin: capillary strengthening, venotonic, anti-inflammatory, glucosinolates, cyanogenic glycosides, saponins, tannins) in resin ducts, milk diuretic), ducts (isoquinoline alkaloids - Euphorbiaceae, Papaveraceae), glandular hairs (terpenoids - Asteraceae, Lamiaceae), hairs (amines - Urticaceae), epidermis coumarin heterosides(Meliloti herba – Fabaceae: antiedematous, anti- (alkaloids, flavonoids, anthocyanins, cyanogenic glycosides, coumarins) inflammatory, Ammeos visnagae fructus – Apiaceae: spasmolytic), saponosides(saponins, expectorants) 2  PSM – number of known copounds PSM – active ingredients of herbal drugs Nitrogen-containing compounds: Alkaloids 12.000 SACCHARIDES/ HETEROGENEOUS POLYSACCHARIDES Non-protein amino acids 700 heterogeneous or mixed polysaccharides consisting of various sugar monomers and uronic Amines 100 acids Cyanogenic glycosides 112 Plant drugs with heteroglycans that do not have mucus (slime) properties Glucosinolates 132 Purple coneflower herb (Echinaceae herba), (Echinaceae pallidae radix): Alkamides 150 Echinacea purpurea Moench., Echinacea angustifolia DC, Echinacea pallida Nutt. Nitrogen free compounds: branched heteroglycans (high molecular weights 35 - 450 kD), caffeic acid derivatives Monoterpenes 2.500 (cichoric, caftaric acid, echinacoside - AOX), isobutylamides/alkylamides: unsaturated Sesquiterpenes 5.000 aliphatic compounds with –NHCO- group (echinacein), polyins (polyacetylenes, linear, Diterpenes 2.500 partially cyclized; C-C triple bonds (analogous to polyenes with several C=C double bonds). Triterpenes (steroids, saponins) 5.000 Contraindication: Tetraterpenes 500 - Autoimmune diseases Flavonoids, fenolic compounds 9.000 Polyacetilenes, fatty acids 1.000 Polyketides 750 Carbohydrates > 200 PSM – active ingredients of herbal drugs Mucilaginous substances (Mucilaginosa) INORGANIC COMPOUNDS, ORGANIC ACIDS Herbal drugs with inorganic compounds (SiO2): A group of plant products (mucilages, gums, pectins) characterized by the - Horsetail herb (Equiseti herba); Equisetum arvense L., Equisetaceae: up to 6% common property of strong swelling in the presence of water and forming colloidal silicic acid (SiO2 + H2O H2SiO3) and flavonoids (kaempferol, quercetine solutions or gels glycosides): Mucilages (slimes) of higher plants: a physiological product of plants Recommended use: inflammation of the kidneys/bladder, weak diuretic effect Gums (sticky slimes): pathological protective product of the plant organism, formed only after wounding of plant tissues (biosynthesis from starch or cellulose) Pectins (polymers of galacturonic acid or rhamnose linked 1,4 - glycosidically; to – COOH Contraindications: can be attached – OCH3) in the fruits/seeds of apple (Malus domestica), pear (Pyrus - CVS disorders, kidney failure (diuresis - increased excretion of K+) communis), quince (Cydonia oblonga Mill.): due to the ability to reduce hypersensitivity to - children: high content of inorganic silicon and toxicity stimuli, reduce peristalsis, antidiarrheal activity Drug interactions: - cardiotonic glycosides and digitalis preparations: toxicity due to diuretic K+ loss - acceleration of the breakdown of thiamine (vitamin B1), due to the content of thiaminase PSM – active ingredients of herbal drugs Slimes of higher plants INORGANIC COMPOUNDS, ORGANIC ACIDS mixtures of hexoses (glucose, D- manose), pentoses (arabinose, xylose) and/or their oxidation products - uronic acids (glucuronic and galacturonic acids), which are obtained - Stinging nettle herb/leaf (Urticae herba/folium) after hydrolysis. (Ca2+, K+ - salts, silicic acid, flavonoids, triterpenes) Traditional herbal remedy: - urinary disorders, irrigation therapy - minor joint pains - seboroic skin alterations Contraindication: pregnancy (emenagogue, uterine-stimulating action) Interactions with drugs: increases the anti-inflammatory action of low doses of diclofenac depending on the plant species, they are distinguished by the monosaccharides they consist of Plant drugs with organic/fruit acids: in the plant: energy reserve function, water supply (water retention capacity) - Hibiscus flowers (Hibisci Flos) - Dog rose fruit (Rosa canina - Cynosbati fructus sine semine) vacuoles with slime accumulate along the cell walls in various organs - roots (Althaeae (refreshing teas) radix), tubers (Salep tuber), leaves (Althaeae folium, Malvae folium), flowers (Malvae flos, Verbasci flos), seeds (Cydoniae semen, Psylli semen, Lini semen, Foenugraeci semen). 3  The chemical structure of free and conjugated sterols. Slimes of higher plants R1 at C3 and R2 at C17 determine the diversity of sterols. Inflammatory diseases of the GIT and bronchial tract (they form a protective film on the inflamed stomach wall that protects the mucosa from the harmful attack of stomach acid and other chemical stimuli) Antidiarrheal action: the emollient property of slime is the basis for its use in diarrhea (protection of the intestinal mucosa can be achieved mechanically - coating the mucosa with slime - or chemically – slime adsorbs and buffers) Acute inflammation of the throat (adjuvant treatment) (resorbable slimes): (Althaeae radix) (slime covers the inflamed mucosa thus reducing its irritability) laxative effect: the ability of slime to bind water allows them to be used as a laxative: the increase in volume through water retention encourages the intestine to perform stronger peristalsis, which enables faster emptying of the intestinal contents (1) Tetracyclic skeleton of sterols; R1 determines the type of compound including, (2) free sterol, (3) steryl glucosides, (4) acylated steryl glucosides, (5) steryl esters, (6) steroidal glycoalkaloids. R2 determines the diversity of compounds. For example, the combination of (2) and (7) makes cholesterol (12), the combination of (2) and (8) makes campesterol (13), the combination of (2) and (9) makes β-sitosterol (14), the combination of (2) and (10) makes stigmasterol (15), the combination of (3) and (10) makes stigmasteryl glycose (16), the combination of (4) and (9) makes sitosteryl palmitoyl glucoside (17), the combination of (5) and (9) makes stigmasteryloleate (18), while the combination of (6) and (11) makes α-solanine (19). Plant SM = slime herbal drugs TERPENOIDS - Isoprenoids Slimes of higher plants - Marshmallow root and leaf (Althaeae radix, Althaeae folium) - Mallow leaf and flower (Malvae folium in flos) - Mullein flower (Verbasci flos: V. thapsus, densiflorum, phlomoides) - Plantain leaf/herb (Plantaginis lanceolatae folium/herba) - Black psyllium seed (Psylli semen) - Flaxseed (Lini semen) - Fenugreek seed (Foenugraeci semen) Cytoplasm: mevalonate pathway (MVA) Plastids: methylerythritol phosphate pathway (MEP) (acetyl Co-A condensation) (substrate: pyruvate and glyceraldehyde 3-phosphate) Isoprenoids PSM - Isoprenoids Compounds formed from activated isoprene (C-5 carbohydrate)  Terpenes: biosynthesis from activated isoprene. Monoterpenes: compounds formed from 2 isoprene molecules. According to no. of isoprene units (number of C atoms), terpenes are divided: Isoprenoids - starting compounds for a series of PSM in herbal drugs: terpenes - - monoterpenes (formed from 2 isoprene units; EO) compounds of essential oils (mono- and sesquitepenes); triterpene saponins (mostly 5 six-membered rings) formed from six isoprenoid units) - sesquiterpenes (formed from 3 isoprene units; bitters, EO) - diterpenes (formed from 4 isoprene units; bitters) - triterpenes (formed from 6 isoprene units; saponins, phytosterols) squalen - tetraterpenes (formed from 8 isoprene units; consisting of at least ten conjugated Sterols - cardiac glycosides (cardenolides/bufadienolides), steroid saponins, double unsaturated bonds sensitive to oxidation; carotenoids: α-carotene, phytosterols (β-sitosterol...): formed from mevalonic acid via triterpenoid biosynthetic β-carotene, β-cryptoxanthin (provitamine action), cardioprotective action pathway: triterpene squalene - starting compound of all biogenic steroids. (α-carotene, β-carotene, lycopene) cardenolid bufadienolide cyclopentane-perhydrophenanthrene – 4 - ring steroidal saponin structure 4  Plant drugs with triterpene saponins PSM - Isoprenoids Primrose root/flower (Primulae radix/flos) (Primulaceae)  Cardiotonic glycosides: aglycone backbone of all cardiac glycosides - sterane. - triterpene saponin aglycons (3 -10%) Primula veris L. Primula elatior (L.) Hill Contraindication: gastritis, stomach ulcer priverogenine A priverogenine B; protoprimulagenine A  Steroid aglycone (acting on a sick heart) - glycosidically linked to one to five monosaccharides (glucose, rhamnose, digitoxose) through the OH group. - phenolic glycosides (primverin, primulaverin) (2 - 3%)  Increase the force of contraction of the heart muscle and thus increase the capacity of the heart. Expectorant activity: local irritation of gastric mucosa, reflective increase in bronchial secretion, thinning of slime (decrease in sputum viscosity), decrease in surface tension in the back of the throat, accelerate movement of cilia/cilia of mucous epithelium; diluted slime coughed up easily Not recommended in pregnancy! Plant drugs with cardiotonic glycosides Plant drugs with triterpene saponins Licorice root (Liquiriziae radix) Foxglove leaf (Digitalis purpureae folium) Glycyrrhiza glabra L. (Fabaceae) Woolly foxglove leaf (Digitalis lanatae folium) 2 - 15 % triterpene saponins, flavonoids Adonis herb (Adonidis herba) glycyrrhizic acid glycyrrhetinic acid Lily of the Valley herb (Convallariae herba) Ginseng root (Panax ginseng) Panax ginseng C.A. Mayer (Araliaceae) Oleander leaf (Oleandri folium) 2 - 3 % triterpene saponins (ginsenosides) geriatric, revitalizing tonic, adaptogen, immunostimulant Scilla bulb (Scillae bulbus) Side effects: sleep disorders, AH, nervousness, diarrhea Plant drugs with triterpene saponins Plant drugs with saponins European goldenrod herb (Solidaginis herba) Solidago virgaurea L. (Asteraceae) Primrose root/flower (Primulae radix/flos) 2 % triterpene saponins, 1,5 % flavonoids, traces of tannins and EO Acute and chronic inflammation of the kidneys: diuretic, spasmolytic, anti-inflammatory Licorice root (Liquiriziae radix) Ginseng root (Panax ginseng) Horse - chestnut seed (Hippocastani semen) Aesculus hippocastanum L. (Hippocastanaceae) 3 - 6 % triterpene saponins (escine), flavonoids, coumarins European goldenrod herb (Solidaginis herba) Treatment of varicose veins (thickening of the vein wall, reduction of capillary permeability, reduction of edema) – anti-edematous and phlebotonic agent Horse - chestnut seed (Hippocastani semen) 5  Plant drugs with bitters Bitters: bitter herbal drugs reflexively increase the secretion of digestive juices, the appetite increases; amara pura – only bitters, amara aromatica – bitters and EOs, eterična olja, amara mucilaginosa - bitters and slimes Monoterpene bitters (Gentianaceae) Sesquiterpene bitters (Asteraceae) Centaury herb (Centaurii herba), Blessed Thistle herb (Cnici benedicti herba) Yellow gentian root (Gentianae radix) Artichoke leaf (Cynarae folium) Diterpene bitters (Lamiaceae) Horehound herb (Marrubii herba) Dandelion root with herb (Taraxaci radix cum herba) Wormwood herb (Absinthii herba) ESSENTIAL OILS  The volatile substances of plants and carriers of the scent (smell/aroma) of plant. They are mixtures of various biogenic substances, the main components of which are proven to be terpenes and phenylpropanes.  Resins must be distinguished from essential oils, as well as lipophilic mixtures of substances, which are solid and non-volatile. Resins dissolved in essential oils are called balms.  EOs formed in all plant parts in special cells and accumulate in special cavities, such as: 1. Oil glands such as glandular hairs or non-glandular trichomes. EO secreted from the secreting epidermal cells to the outside and accumulates between the cell wall and the cuticle (Lamiaceae, Asteraceae). 2. Oil cavities formed by the dissolution of parenchyma cells (Pinaceae, Rutaceae) 3. Schizogenically formed cavities, which are formed due to mutual spacing of tissue cells (Apiaceae) 4. Excretory cells that are isolated from the rest of the tissue by a suberin layer so that they can accumulate essential oil (Zingiberaceae). 6  Rastlinske droge z eteričnimi olji Absinthii herba (Wormwood herb) Matricariae flos Anethi fructus (Dill seed) Millefolii herba (Yarrow) Anisi fructus Piceae turiones recentes Arnicae flos (Fresh fir shoots) Carvi fructus Pini turiones (pine shoots) Cinammomi cortex Rosmarini folium Coriandri fructus Menthae folium Eucalypti folium Melissae folium Juniperi fructus Salviae folium Lavandulae flos Thymi herba Levistici radix Valerianae radix 7  Major branch pathways of (poly)phenol biosynthesis PSM of higher plants Part II Phenolics and Alkaloids PAL, phenylalanine ammonia-lyase; C4H, cinnamate-4-hydroxylase; 4CL, 4-coumaroyl:CoA-ligase; HCT, hydroxycinnamoyl transferase; C3H, p-coumarate-3-hydroxylase; CHS, chalcone synthase; CHI, chalcone isomerase; ANS, anthocyanidin synthase; DFR, dihydroflavonol reductase; FS, flavone synthase; FLS, flavonol synthase; F3H, flavanone 3-hydroxylase; IFS, isoflavone synthase; ANR, anthocyanidin reductase; LAR, leucoanthocyanidin reductase. PLANT PHENOLICS (PHENOLS, AROMATIC COMPOUNDS) PLANT PHENOLICS (PHENOLS, AROMATIC COMPOUNDS) - Phenols: aromatic metabolites with 1 or more "acidic" hydroxyl groups (polyphenols) attached to an aromatic ring: hydroquinone salycilic acid gallic acid - phenols with 1 aromatic ring: simple phenols (C6), phenol with 1 C- (C6-C1: salicylic, gallic acid), 2 - C (C6-C2: picein), 3 C – atoms (C6-C3: phenylpropanoids - caffeic, p- coumaric acid) picein caffeic acid p-coumaric acid - phenols with 2 aromatic rings, linked by 1 C (C6-C1-C6: xanthones) or 2 C (C6-C2-C6: stylbenes) xanthones stylbenes PLANT PHENOLICS (PHENOLS, AROMATIC COMPOUNDS) The shikimate – chorismate/ biosynthetic pathway of phenolics - phenols with 2 aromatic rings connected with 3 C (C6 – C3 – C6 : flavonoids) - originate from the phenyl - propanoid (shikimi - chorismin), less often/partly from the phenylalanine tiyosine polyketide biosynthetic pathway (condensation of acetyl - CoA: -CH3-CO-SCoA) main AA of shikimate/ chorismate pathway Phenylpropanoid structure Phenylpropanoid -acetate structure Conyferyl alcohol Quercetin (C6C3) (C6C3C6) 1  The acetate - malonate biosynthetic pathway of phenolics FUNCTIONS OF PHENOLIC COMPOUNDS IN PLANTS - participation in oxidation-reduction processes (photosynthesis, respiration, plant growth and development) - signaling role of flavonols in pollination and pollen maturation - protection against UV radiation (flavonoids absorb UV radiation in 280-320 nm range; accumulation in vacuoles of epidermal cells, limited penetration of UV rays into internal tissues); anthocyanins act as optical filter, scavenging radicals - adaptive-protective functions in pathogenesis; phenols present in plant tissues and organs: antimicrobial, fungicidal and insecticidal - signaling molecules in interaction of plants with soil microorganisms (MO): after infection with pathogens, activation of biosynthesis of species-specific polyphenols accompanied by an increase in the enzymatic activity of L-phenylalanine ammonia lyase, trans- cinnamic acid 4-hydroxylase and chalcone synthase; at the same time, the content of toxic phytoalexins in the covering tissues (phenolic compounds - Elongation of activated acetate units (acetyl-coenzyme A) via stepwise addition of activated stilbenes, flavans, coumarin, auron – toxic) malonyl unit (malonyl-SCoA) and decarboxylation; β - keto groups form polyketide - trigger oxidation-reduction processes in the soil - selective influence on the growth intermediates that cyclize: polyketide-resorcinol derivatives - THC, phloroglucine derivatives, and development of soil MOs anthracene derivatives (1 molecule of acetyl-coenzyme A and 7 molecules of malonyl-SCoA) PSM = active ingredients, useful substances Phenylpropanoid metabolism leading to the formation of p- PHENYLPROPANOIDS: coumaryl-, coniferyl - and sinapyl- alcohols and other phenolic - Coumarins (cis-o-coumarine lactones, coumarin derivative dicoumarol, melilotin), compounds; enzymes (and their flavonoids – Sweet Clover Herb (Meliloti herba) pains, heaviness, cramps in legs cofactors): 1. PAL; 2. PAL (or TAL); (chronic venous insuffiency), hemorrhoids; external bruises, bumps (anti- edematous by 3. cinnamate-4-hydroxylase (O2, increased venous reflux and improvement of lymphatic kinetics); cytochrome P450, NADPH); 4. hydroxylase (O2, cit. P450, NADPH); 5. CoA ligase, which participates in the binding of AMP and CoA (CoASH, ATP); 6. O- Uva ursi leaf: methyl transferase; 7. cinnamoyl- Uvae ursi folium Inflammatory disorders of the urinary tract (antibacterial effect: CoA: NADPH oxidoreductase (NADPH); 8. cinnamoyl alcohol 5-14 % hydroquinone derivatives (arbutin, methyl-arbutin - after ingestion: hydroquinone + dehydrogenase (NADPH); 9. glucose), tannins, flavonoids chalcone synthase; 10. chalcone Contraindication: pregnancy, lactation, children under 12 years isomerase; 11. stilbene synthase; Not more than 1 week; 5 –times per year 12. styrylpyrone synthase. Lignans (condensation of phenylpropane derivatives - coniferous, sinapic and p- coumaric alcohol), lignin (in addition to cellulose and hemicellulose main cell wall component, intended for mechanical support of higher plants, less important in phytotherapy PSM = active ingredients, useful substances PLANT PHENOLICS (PHENOLS, AROMATIC COMPOUNDS) POLYKETIDES: phloroglucine derivatives - Plants originate from an aquatic environment, their evolutionary adaptation was Hops strobile (Lupuli strobuli ) (Humulus lupulus L.) greatly contributed by the large-scale formation of phenolic compounds (structural α- and ß- bitter acids, 0,35 % essential oil and supporting role in the cell wall - lignin, defense compounds against pathogens, deterrents - tannins, at the same time antioxidants in flowers, seeds, seed shells, bark, leaves, roots - - lignans, protection against desiccation - suberinized tissues, antimicrobial agents, inhibit germination - coumarins, furanocoumarins, stilbenes, coloration of flowers - anthocyanins, signal molecules - isoflavonoids, specific smell and taste…) - Mood disturbaces (restlessness, anxiety, sleep disturbances): calming, sleep promoting St. John’s Wort herb (Hyperici herba ) (Hypericum perforatum L.) Hyperforine (phloroglucine derivative) hipericin (antracene derivative) - Psychovegetative disturbances, depressive moods, anxiety/nervous unrest - External: oil preparations: acute/contused injuries, myalgia, first degree burns 2  PSM = active ingredients, useful substances PSM - flavonoids POLYKETIDES: anthracene derivatives FLAVONOIDS Rhubarb root (Rhei radix): anthranoids (emodine, rheine)  belong to the group of natural compounds with C6C3C6 cyclic system (Rheum palmatum L., Rheum officinale Baillon)  formed by the condensation of phenyl-propanoids and 3 acetate building unit (malonyl- SCoA) Pharmacological effects: - strengthen the fragility of capillaries - have a tonic effect on the vascular walls Constipation (short term!) : laxative - inhibit vein permeability Contraindications: intestinal obstruction, acute intestinal inflammation, Chrohn’s - relieve inflammation and allergic phenomena disease, colitis ulcerosa, abdominal pain of unknown origin, pregnancy, children - act as a diuretic under 12 years  the role of flavonoids: involved in redox systems. Due to their distinct coloration used as Buckthorn bark (Frangulae cortex) (Rhamnus frangula L.) attractants for insects, they may also have a protective role against parasites (isoflavonoids); anticancer effects. Anthranoids (emodin, physcion, chrysophanol) Constipation (short term!) : laxative In plants (0.5% - 10%) mostly glycosidically bound, hydrophilic, dissolved in the cell sap. Contraindications: intestinal obstruction, acute intestinal inflammation, Chrohn’s  crystalline substances: yellow (flavones – apigenin; flavonols – quercetin) orange (chalcones), disease, colitis ulcerosa, abdominal pain of unknown origin, pregnancy, children red-blue (anthocyanidins), colorless (proanthocyanidins- flavan-3-ols) under 12 years PSM – tannins Diversity of flavonoids Polyphenolic, water-soluble compounds: astringent and bitter taste. Due to their "tanning properties“ properties in past used in the leather industry for tanning animal skins (they bind/coagulate proteins in the skin into insoluble compounds). Tannins of vegetable origin can be divided into 2 groups: - Sugar tannin esters (rarely glycosides) = tannins that can be hydrolyzed - gallotannins: polyphenolic acids (gallic, quinic, chlorgenic acid), their depsides (esters of 2 polyphenolic acids) or condensates (di- or polymers of polyphenolic acids via C-C bond, e.g. hexahydroxydiphenic acid) esterically (rarely glycosidically) linked with monosaccharides (or also with sugar alcohols) (defensive metabolites) Klasifikacija flavonoidov - Condensed tannins (tannins that cannot be hydrolyzed – catechin tannins or condensed proanthocyanidins) - polymers of flavan-3-ol units: potential beneficial effects on human health: immunomodulatory and anticancer activities, antioxidant and radical scavenging functions, anti-inflammatory activities, cardio-protective properties, vasodilating and antithrombotic effects, UV-protective functions PSM = active ingredients, useful substances Diversity of flavonoids TANNINS Tormentil root (Tormentillae rhizoma): tannins Potentilla tormentilla Necker (syn.: Potentilla erecta Raüschel), Rosaceae Uses: unspecified diarrhea; mild mucous membrane inflammations of the mouth/throat Side effects: stomach complaints in sensitive subjects Lady’s Mantle herb (Alchemillae herba): tannins, flavonoids Alchemilla vulgaris L., Rosaceae Uses: mild, unspecified diarrhea; adstringent activity Agimony herb (Agrimoniae herba): tannins, flavonoids Agrimonia eupatoria L., Rosaceae Uses: unspecified diarrhea; mild mucous membrane inflammations of the mouth/throat External: superficial inflammation of the skin; adstringent activity Oak bark (Quercus cortex): tannins Quercus robur L., Fagaceae Uses: unspecified diarrhea; mild inflammations of the mouth/throat, genital, anal area /rinses (20g/l) External: superficial inflammation of the skin; adstringent activity, virustatic 3  Herbal drugs with flavonoids Mechanisms of action: disruption of metabolic processes/enzyme chain reactions in signaling, initiation, promotion of cancer Hawthorn leaf with flower (Crataegi folium cum flore) (Crataegus monogyna Jaquin, C. laevigata DC)  inhibition of the formation of procarcinogens - antioxidant action/ ROS radical flavonoids: flavones – vitexin; flavonols- hyperoside, rutin; oligomeric procyanidins - catechins) scavengers (DNA lesions caused by reactive oxygen species (ROS) in mammalian Relieve symptoms of temporary nervous cardiac complaints - palpitations, extra heart beat due to anxiety, cells are considered an initial event that can lead to possible mutagenesis and/or increases coronary and myocardial circulatory perfusion; mild symptoms of mental stress/ to aid sleep. carcinogenesis and the observed incidence of cancer) 6 weeks minimum: if symptoms persist/in case of swelling of legs – physician consultation Traditionally: fresh juice, plant drug or extracts: a tonic and strenghtener of cardiac/circulatory functions  detoxification of carcinogens through induction of enzymes (superoxide dismutase, glutathione peroxidase, catalase and glutathione reductase), inhibition of DNA topoisomerase II, inhibition of intracellular tyrosine kinases Passionflower herb (Passiflorae herba) (Passiflora incarnata L.) flavonoids (vitexin) coumarine derivatives, small amount of essential oil  inhibition of tumor growth and spread (inhibition of nuclear factor (NF)-Kappa B in Relieve nervous restlessness (in animal experiments: motility-inhibiting effect) prostate cancer cells, inhibition of epidermal growth factor (EGF), inhibition of angiogenesis) Milk Tristle fruit (Cardui mariae fructus) (Silybum marianum (L.) Gaertner) Flavonoids (silibinin, silydianin, silychristin – silymarin)  apoptosis of cancer cells (condensation of chromatin, swelling of the plasma membrane, fragmentation of oligonucleosomal DNA, breakdown of the cell into Dyspeptic complaints (crude drug: 12-15 g), toxic liver damage, supportive smaller units - apoptotic bodies, finally phagocytosis of cell debris) therapy in chronic liver disease and hepatic cirrhosis (preparations 200-400 mg silymarin): stimulating regenerative ability of the liver and formation of new hepatocytes Side effect: mild laxative effect Herbal drugs with flavonoids Turmeric root Linden flower (Tiliae flos) (Tilia cordata Mill., Tilia platyphylos Scop.) (Curcuma longa L., Curcumae longae rhizoma) flavonoids, tannins, slime (mucilage) Colds, cold-related coughs – diaphoretic (increased sweeting lowering body T)  present in many spice mixes (curry, Worcestershire sauce) Meadowsweet flower (Spiraeae flos) (Filipendula ulmaria Max., syn. Spiraea ulmaria L.)  root and powder have a characteristic yellow color (dyeing of dishes, cloth) flavonoids, phenolic glycosides, essential oil Supportive therapy for colds curcumin Heart’s Ease herb (Violae tricoloris herba) (Viola tricolor L., Viola arvensis Gaudin) flavonoids, slime, essential oil, tannins Mild seborrheic skin diseases, anti - acne skin preparations  anti-inflammatory and antioxidant properties (inhibition of arachidonic acid metabolism and prostaglandin synthesis), dyspepsia (digestive problems, Calendula flower (Calendulae flos) (Calendula officinalis L.) abdominal pain) flavonoids, triterpene glycosides (alcohols), essential oil Inflammation of oral and throat mucosa (internal and topical use)  preventive properties in malignant diseases induced by a variety of chemical External: poorly healing wounds, ulcus cruris (shin ulcer) carcinogens. PHENOLIC COMPOUNDS IN SPICES AND HERBS Ginger root Pharmacological Perspectives (Zingiber officinale Roscoe) Epidemiological studies  known as a spice in cuisine for more than 2,500 years  Inverse association between consumption of foods rich in phenolic compounds and  oriental traditional medicine: relief of colds, fevers, rheumatic diseases, indigestion, nausea risk of CVD and cancer morbidity  modern phytotherapy: antiemetic for motion sickness, secretion of gastric juices, saliva  Protective effects of plant-based nutrition on cancer (initiation phase - kaempferol, genistein, galangin, diosmetin..., promotion - galangin, genistein..., progression - apigenin, naringenin, silymarin...)  Natural antioxidants in remedies and food supplements: protective agents, play important protective role in preventing destructive changes at the level of cellular gingerol paradol DNA and genotoxic effects in mammalian cells, even before tissue damage occurs.  ethanolic/water extracts of ginger and its components in lab experiments: antioxidant and topically anti-inflammatory properties, inhibitory properties on the growth of animal and SAR human cancer cells (spontaneous mouse breast cancer, many human cancer cell lines) and induced apoptosis of cultured cancer cell lines 4  Chinese cinnamon bark (Cinnamomum cassia Blume) Rosemary (Rosmarinus officinalis L.)  carnosol: induced apoptosis of many acute lymphoblastic leukemia (ALL) cell lines  aromatic component of foods: drinks, ice creams, sweets, chewing gums, and has traditionally been used as a remedy for digestive disorders, inflammation of the gastrointestinal tract, circulatory disorders and inflammatory diseases  Bioactive compound: cinnamic aldehyde  carnosic acid, also one of the main components of rosemary with a strong antioxidant effect, inhibited the proliferation of human myeloid leukemia cells without causing apoptosis or cell necrosis  exhibits many biological effects: loss of appetite, indigestion - GIT spasms,  rosmarinic acid: specifically induced apoptosis of cytotoxic T lymphocytes and killer cells in flatulence, antibacterial and antifungal; peripheral vasodilatation, antitumor freshly isolated human peripheral blood mononuclear cells (PBMCs), with activity activity, cytotoxic and antimutagenic activity (induction of apoptosis of human selectively targeting actively growing cells but not normal cells promyelocytic leukemic HL-60 cells, mechanism unclear). Paprika - Chili (Capsicum spp.) Sage (Salvia officinalis L.)  Capsaicin ("acria") exhibits gastroprotective, anti-inflammatory and antimutagenic  antioxidant activity is based on the content of phenolic compounds effects in moderate doses in various animal models and cell systems; locally hyperemic (external: muscle spasms of the limbs)  phenolic compounds with strong antioxidant potential: carnosic acid (known for its instability) and its degradation derivatives: carnosol, rosmanol, epirosmanol, 7-methyl-  Induction of apoptosis in malignant human and murine melanoma or transformed epirosmanol cells  Suppression of cancerogenesis.  rosmarinic acid contributes to the antioxidant activity of sage. Carnosol has been shown to have anti-inflammatory activity and tumor initiation inhibitory effects in mouse test systems Cloves Oregano (Syzygium aromaticum (L.) Merr.) (Origanum vulgare L.)  Essential oil as fragrance in food and cosmetic products  essential oil (added to the food of mice) induced glutathione-S-transferase activity in  Traditional Oriental Medicine: against intestinal parasites, antibacterial and toothache remedy various tissues (GST enzyme system involved in the detoxification of chemical carcinogens and plays an important role in the prevention of carcinogenesis)  Due to its ability to scavenge radicals, eugenol (4-allyl-2-methoxyphenol) strong inhibitor of lipid peroxidation, lowering low-density lipoproteins (LDL).  essential oil showed a high level of cytotoxicity (at dilutions up to 1:10000) against eukaryotic cancer cell lines (human squamous epidermoid cancer cells: Hep-2 and  effect on increased activity of glutathione-S-transferase in test systems in rats and mice, cervical epithelioid cells: HeLa) associated with their potential anticancer activity.  in vitro cytotoxic and/or anti-proliferative effects of extracts and isolated agents (carvacrol,  Participating in the inhibition of prostaglandin synthesis in macrophages of mice activated by thymol) lipopolysaccharide (LPS), eugenol directly reduces the expression of COX-2 genes in these cells, inhibits the proliferation of human colon cancer cells, has a cytotoxic effect and inhibits DNA synthesis - in human salivary cancer cell lines as in normal gingival fibroblasts  moderately suppressive activity of extracts of oregano (IC50 = 220 mg/ml) on human ovarian cancer cells (CaOv), of isolated carvacrol and thymol (IC50 = 120 μmol/l) on  eugenol and eugenol-rich essential oil inhibit papillomagenesis in rodent skin melanoma cells with high metastatic potential. 5  Alkaloids Alkaloidi  in plants dissolved in the cell sap of vacuoles in the form of salts (citrates, malates,  3000 years of use in human history (drinks, poisons, purgatives, antitussives, sedatives, tartrates, isobutyrates, benzoates...) or are linked to tannins snake bites, fever, insomnia, diseases - malaria)  they are contained in various plant organs, but most often they are localized in external  not only in plants (mostly in angiosperms), also in amphibians, reptiles, insects tissues (seed coat, outer part of the cortex of the stem or roots)  ecochemical function (defense due to physiological effects on herbivores - pyrrolizidine  alkaloids without oxygen are liquid, and those with oxygen atoms, which are the majority, alkaloids (Boraginaceae), antibiotic properties, toxic or deterrent to insects) are crystalline compounds  optically active (asymmetric chiral centers, 1 - nicotine or more - morphine)  as alkalis insoluble in water, they dissolve in concentrated solutions of HCl (a salt is formed) and non-polar or moderately polar organic solvents  basicity depends on the structural units directly next to the nitrogen (availability of a free Poison-hemlock Weed electron pair: if the free electron pair of nitrogen participates in aromatization, the (Conium maculatum L.) coniine compound is less basic (carbonyl group - amides - attracts electrons - neutral piperine, Jacques-Louis David (1787) pyrrole), if it does not participate, the compound is basic ( quinoline) nicotine caffeine morphine Physiologically active alkaloids used in medicine Biosynthesis of alkaloids  molecular weight between 100 and 900  complex chemical structure, molecules contain asymmetric centers (morphine - 5 asymmetric centers), formed from simple precursors with the participation of many specific enzymes  difficult detection of biosynthetic mechanisms/enzymes: slow generation and small amount of accessible metabolites/enzymes in biosynthetic stages; extraction of active enzymes is disturbed by tannins and other phenolic compounds; structures unknown before 1970 - development of callus cultures; plant cell cultures enable the development of higher concentrations of intermediate metabolites and included enzymes Alkaloids Biosynthesis of alkaloids  Hegnauer: - Berberine (antimicrobial activity): the first alkaloid (tetra-hydro-benzyl-isoquinoline), "Alkaloids - more or less toxic compounds that mainly affect the nervous for which all biosynthetic enzymes identified system. They are alcaline and contain heterocyclic nitrogen. In plants, they are formed from amino acids or their derivatives. In most cases, they have a limited distribution in plants."  Pelletier: “ Alkaloids - cyclic organic compounds that contain nitrogen in a negative oxidation state and have limited distribution in the living world."  Characteristic for families: Apocynaceae, Magnoliaceae, Solanaceae, Ranunculaceae, Berberis wilsoniae Loganiaceae, Papaveraceae, Rubiaceae cell suspension culture (high SM production) 6  Alkaloids Neurostimulants - drugs whose main action is stimulation of the central nervous system (not the same intensity on the entire nervous system) - stimulants in small doses, in large doses have a sedative effect (e.g. Erythroxylon coca Lamk. (Erythroxylaceae, coca leaves) (coca leaves, coca- alkaloid cocaine (methylbenzoyl -ecgonine) 0.5-2.5%) (longer use causes addiction!) (Indians Bolivia, Peru and Colombia: chewing the leaves reduces the feeling of hunger, thirst, fatigue, extract without cocaine - the smell of Coca cola - methylsalicylate) - local anesthetic (vasoconstrictor and local anesthetic action cause anesthesia and contraction of the mucous membrane) (1 - 4% solution) Biosynthesis of berberine from L-tyrosine. Biosynthesis of alkaloids Alkaloids Cola nitida A. Chev. (Sterculiaceae, kakavovke):  according to biosynthesis, alkaloids are divided into three groups: - fresh seeds, cotyledons (kola, kola nut, guru-guru) contain up to 3% caffeine (7.5 mg daily). In excessive amounts: cardiac arrest  protoalkaloids: simple derivatives of amino acids - modified amino acids (in the - medicinal dose (adults) for mental and physical fatigue: max. 250 mg of caffeine cytoplasm from 2 molecules of L-tyrosine: ephedrine - Ephedra sinica), capsaicin daily (3 cups of coffee); side effects: increased thermogenesis, increases overall (Capsicum frutescens.) or other AK: muscarine - Amanita muscaria); amines or energy expenditure, causes weight loss amides (the nitrogen is not part of the heterocyclic skeleton) - restriction necessary: pregnancy, gastric and duodenal ulcer, hypertension, heart disease. ephedrine muscarine capsaicin Biosynthesis of alkaloids Alkaloids  pseudoalkaloids usually have all the characteristics of true alkaloids but are not formed from amino acids; are of terpenoid origin (derivatives of monoterpenes: vinblastine, diterpenes and steroids), more rarely of polyketide origin Coffea arabica L. Rubiaceae (seed, 2% caffeine) - cardiostimulant, excites the heart muscle, increases the strength of myocardial contraction, also has a tonic-excitatory effect on the central nervous system, increases mental work. In high doses, it causes drowsiness, fatigue, confusion, muscle imbalance. quinine vinblastine  true alkaloids consist of a part derived from amino acids (one, two identical or two different, several identical) and a non-amino acid part whose C-atoms derive from other metabolic pathways: acetate and terpenoid 7 Camellia sinensis O. Kuntze (Theaceae): leaves (1-4 % caffeine) IIex paraguariensis St. Hill. (Aquifoliaceae) - maté, seed, leaves: 2 % caffeine Paulinia cupana L. (Sapindaceae), guarana: seed up to 3.5 % caffeine (8,75 mg dnevno), teobromine, teophyline (purine alkaloids), catechic tannins. Alkaloids Fumaria officinalis L. (Fumariaceae), Fumariae herba - isoquinoline alkaloids - fumarine, cryptokavin, corydalin, synactin, as well flavonoid glycosides, org. acids and slimes - spasmolytic properties, used for spasms of the bile ducts and the digestive tract - a daily dose of 6 g of the drug - stimulating and tonic if used for a short time (8-10 days in the form of an infusion - 5 g of the drug/100 ml of water, 3 times a day or tincture: 10 gtt before meals) - depressant when used for a longer time. 8