BIOSYNTHESIS OF

FLAVONOIDS

Anna Blzovics Dr. D.Sc.

OCCURRENCE

flavonoids are widely distributed in the flora they can be found in petals, crops, seeds, leaves, stems, roots and barks it is the reddish braunish - yellowish colour of heartwood flavonoids of ferns are C-methyl derivatives glycoflavonoids can be isolated from see-weeds inferior plants and mushrooms have them only very rarely

microorganisms and algae species do not synthetize

STRUCTURE OF FLAVONOIDS
Polyphenols, namely favonoids are low molecular weight substances The skeleton can be represented as the C6 - C3 - C6 system Three main groups can be distinguished on the places of phenyl ring.

The chemical structure of flavonoids are based on a C15 skeleton with a chromane ring bearing a second aromatic ring B in position 2, 3 or 4.

The oxidation state of propane chain means the difference. Oxidation degree rises from 0 to 6. (Tetrahedron, 8, 336, 1960.)

SYNTHESIS OF FLAVONOIDS

Flavonoids are synthesized by the phenylpropanoid metabolic pathway in which the amino acid phenylalanine is used to produce 4-coumarate-CoA.
This can be combined with malonyl-CoA to yield the true backbone of flavonoids, a group of compounds called chalcones, which contain two phenyl rings.

Conjugate ring-closure of chalcones results in the familiar form of flavonoids, the three-ringed structure of a flavone.

The metabolic pathway continues through a series of enzymatic modifications to yield flavanones dihydroflavonols anthocyanins.

Along this pathway, many products can be formed, including the flavonols, flavan3-ols, proanthocyanidins (tannins) and a host of other various polyphenolics.

ENZIMATIC CONVERSION OF NARINGENIN TO APIGENIN

EC 5.5.1.6 chalcone isomerase EC 1.14.11.22 flavone synthase

ENZIMATIC CONVERSION OF TAXIFOLIN TO QUERCETIN

EC 1.14.11.23 flavonol synthase

ENZYMATIC CONVERSION FROM DIHYDROKAEMPHEROL TO LEUCOPELARGONIDIN

EC 1.1.1.219 dihydrokaempferol 4-reductase

SYNTHESIS OF FLAVONOIDS
EC 1.1.1.219 dihydrokaempferol 4-reductase EC 1.14.11.9 flavanone 3-dioxygenase EC 1.14.11.22 flavone synthase EC 1.14.11.23 flavonol synthase EC 1.14.13.21 flavonoid 3'-monooxygenase EC 1.14.13.88 flavonoid 3',5'-hydroxylase EC 1.17.1.3 leucoanthocyanidin reductase EC 5.5.1.6 chalcone isomerase
http://www.chem.qmul.ac.uk/iubmb/enzyme/reaction/phenol/flavonoid.html

FLAVONOIDS ARE DIVIDED INTO GROUPS

According to the IUPAC nomenclature, they can be classified into:
flavone: flavonol: 2-phenylchromen-4-one (3-hydroxyflavone) 3-hydroxy-2-phenylchromen-4-one

flavanone: 2,3-dihydro-2-phenylchromen-4-one
flavanonol: (3-hydroxyflavanone) 3-hydroxy-2,3-dihydro-2-phenylchromen-4-one isoflavone anthocyanidin

proantocianidin

other flavonoids

catechin chalcone dihydrochalcone aurone

Both the oxidation state of the heterocyclic ring and the position of ring B are important in the classification.

ANTHOXANTHINES ARE THE REAL FLAVONOIDS

Natural derivatives from benzo--pyron stucture are considered as flavonoids by certain researchers. These molecules are anthoxantines.
benzo--pyron chromon

-pyron

SPECIAL FLAVON DERIVATIVES

cumaroflavones

BIOCHEMICAL PATHWAY OF THE FORMATION OF ROTENONE

isocumaroflavon
furanoflavonoids biflavones

coumestan derivatives
rotenoids

blue: carbons derived from methionine red: carbons derived from prenyl (isoprenoid).

REASON FOR THEIR VARIETY

OH groups number and position of O-methyl, O-alkil, O-glycosyl-groups variance of glycosyl groups, acylation grade with or without conjugated double bonds

The most often occuring substituents can bond to C3 -, C5 , C7 -, C3 -, C4 atoms. Sugars can bond: through oxygen atom (O-glycosides) directly to C-atom (C-glycosides)

Antocyanides are always in their glycoside forms in vacuoles (in cell fluids) catechines and procyanidines can be stored in their aglycone forms (tannic acid holders, dissolved in essential volatile oils)

THE ROLE OF FLAVONOIDS IN PLANTS

Flavonoids are the secondary metabolites of plants, and they are providing the colour and flavour materials. protect against UV-radiation fungus- , insect- and snail pests they are signal for N-bound bacteria they modify enzyme reactions 75% of Angiospermae contain kaempferol and quercetin and 10% of them myricetin.

kaempferol

quercetin

myricetin

THE MOST OFTEN OCCURING GLYCOSIDE

rutin

Rutine can be found in 50% of Angiospermae. Quercetin is the aglycone form of a number of other flavonoid glycosides, such as rutin and quercitrin, found in citrus fruits, buckwheat and onions. Quercetin forms the glycosides quercitrin and rutin together with rhamnose and rutinose, respectively.

BIOFLAVONOIDS HAVE BIOACTIVITY

flavone, flavonol, flavanone, flavanonol 2-phenyl-benzo--pyron

catechin

High concentration of quercetin in in vitro isoflavone experiments causes 3-phenyl-benzo--pyron DNA mutations in spite of that epidemiological studies declare that this flavonoid consuming prevents atherosclerosis, infarct and others [Dunnick anthocyanidin
1998, Williams 2004].

Recent in vitro examinations strenghten the antiproliferative effects of prenylated naringenin (Humulus lupulus L., hops) on prostate tumour cell line culture.
(Phytomedicine. 13 (9-10), 732-4. 2006.)

 Oxygen of pyrane ring is quaterner.

Oxoniumbases with acids form salines.

20 dkg soylent contains ca. 300 mg izoflavones.

CONDITIONS OF ANTIOXIDANT PROPERTY

Fenolic antioxidants can function free radical scavengers and metal chelators. Flavonoids are highly effective scavengers of all types of oxidizing radicals. Rate constant of quercetin with superoxide anion is 0.9x105 at pH 7.5. The arised fenoxy radical is a relatively stable molecule. This molecule limits the metal catalysed degradation of hydroperoxides.

CHELATING ABILITY OF FLAVONS AND FLAVANONS

3, 4- dihydroxy configuration
carbonyl group at the position 4 free OH-groups at 3- and 5 positions

Conditions of chelating property:

MESOMER STRUCTURE OF QUERCETIN AROXYL RADICAL

PHYSIOLOGICAL ACTIVITY OF FLAVONOIDS

Flavonoids have been referred to as "nature's biological response modifiers" because of strong experimental evidence of their inherent ability to modify the body's reaction
antioxidant and /or scavenger activity

immunomodulating and antiinflammatory effects anti-asthmatic and anti-allergic effects anti-viral and anti-bacterial and anti-fungal effects oestrogen activity (isoflavonoids) influencing effect on mutagenicity and carcinogenicity hepatoprotective effect effects on circulation, modification of permeability

SILIBININ INCREASES SYNTHESIS OF RNA POLIMERASE (Flavanolignane)

BANNED!
Catechol-type antioxidant was used in cases of alcoholic liver diseases.

CATERGEN

It was also available in Hungary, but became banned because it caused haemolytic anaemia. Depending on concentration, catergen inhibits the activities of Mg++ ATP-ase and Na+K+ ATP-ase.

METABOLISM OF FLAVONOIDS DEPENDS ON SEVERAL FACTORS Only 1 % of the original form of flavonoids can get into the circulation.
stucture, oxidation state rate of glycosylation /acylation conjugation property

rate of polymerisation
antioxidant property rate of absorption

bacterial enzyme activity

effects of their derivatives are often different from natural molecules
(Hertog 1993, Hollman 1995, Breinholt 1999.)

WHAT HAPPENS WITH FLAVONOIDS IN OUR SYSTEM?

Flavonoids are extensively metabolised, metabolites arise on intestinal absorption. and their

Flavonoids are also transformed into phenolic acids by the enzymes of colonised gut microbes. These phenolic acids are also absorbed and metabolised in the liver. Generally, flavonoids may undergo conjugation, oxidation and P450-related transformation intracellularly. Glucuronide and sulfate conjugates are formed with the majority of flavonoids in the small intestine and in the liver.

BIOTRANSFORMATION OF FLAVONOIDS IN MICROSOMES

The liver is the main detoxicating tissue, where the elimination of exogenic toxic agents happen. Mixed functioning oxidases are localised in the endoplasmic reticulum and they can catalyse a lot of oxidation reactions. They too have the main role of decompositioning lipid peroxidation endproducts.

TRANSFORMATION OF LUTEOLIN BY BACTERIA AND MICROSOME

Flavonoid glycosides can unergo changes because of the effects of bacterial enzymes colonised in the bowel in the terminal ileum and colon.

Mixed functioned monooxigenases are able to modify flavonoids in the liver, bowel, kidneys and skin.

PLASMA LEVELS OF FLAVONOIDS

(mg/ttg)

(ug/ml)

quercetin (+) - catechin

65 32

<0,1 0,064

Gungler (1975) Lee (1995)

3-methoxycatechin
diosmin rutin

30
10 1,4

11
0,4 0,09

Hackett (1985)
Cova (1992) Hollman (1997)

REGULATION OF CELL CYCLE DEPENDS ON

REDOX HOMEOSTASIS
Flavonoids can influence the redox homeostasis in several steps:
in PG-biosynthesis
in leukotriene biosynthesis in NF-B translocation to the nucleus in PKC activity and in PI3-kinase activity in signal
We can simply say that free radicals in small concentrations may cause apoptosis, and in large quantities necrosis.

transduction in MMP-ase activity in extracellular space in tumour

BIOSYNTHESIS OF PROSTAGLANDINS

Kempferol inhibits COX2-enzyme

BIOSYNTHESIS OF LEUKOTRIENES

Quercetin inhibits the activities of lipoxygenases, therefore they have anti-inflammatory properties.
Prenylated flavonoids can inhibit the 15lipoxygenase activity.

FLAVONOIDS AND SIGNAL TRANSDUCTION

The polyphenol like compounds inhibit signal transduction pathways, the TNFmediated activation of NF-B pathway, partly through the inhibition of IB kinase and IL-1 activated NF-B pathway, which is a partly distinct route. NF-B activation by IL-1 requires an IL-1 receptor-associated protein kinase activity [Croson 1995].

LOCATION OF FLAVONOIDS IN THE ACTIVE CENTRE OF PI3-KINASE

Flavonoids are the potential inhibitors of protein kinases. Glycosylation of aglicons moderate the inhibitory effect. Small molecular alterations can strenghten the inhibitory activity and selectivity. It is hard to predict between structure and function any better.

FLAVONOIDS IN THE SIGNAL TRANSDUCTION

PROLIFERATION AND SPREDING OF TUMOUR DEPENDS ON ACTIVITIES OF MATRIX-METALLOPROTEINASES

In the formation of extracellular the matrix, collagenes take part in the delimitation of individual cell populations. Zn-containned matrix metalloproteinases do the continous renewal of the extracellular matrix. Tissue inhibitors and alpha 2- macroglobuline inhibit Flavonoids could also induce mechanisms the MMP ases.

that help kill cancer cells and inhibit tumor invasion.

BONDING OF FLAVONOIDS TO THE MATRIX METALLOPROTEINASE-9

Flavonoids are potent inhibitors of MMP-ases. There is small difference concerning the inhibitory effect in the case of flavonoid aglicons. Glycosylation increases the inhibitory effect. The connection between structure and function can be predicted well.

FLAVONOIDS AND SIGNAL TRANSDUCTION

Flavonoids can modulate the activity of protein kinase and lipid kinase signaling cascades. Direct and indirect inhibitory actions of flavonoids within ERK1/2 and Akt/PKB pathways could initiate cell death, whereas inhibitory actions within JNK pathways are more likely to be protective.
[Agullo 1997, Gamet-Payrastre 1999,]

NF-B, JNK and p53 signaling proteins play an important role in apoptotic death and free radicals and metals are very important mediators in the apoptotic process as well.

DATA CONCERNING OF FLAVONOIDS

Consuming vegetables and fruits containing one gram of flavonoid and polyphenol per day is advised.

According to surveys in different countries, people consume flavonoids in the quantity of 20-50 mgs per day.
According to Dr. Lugasi and colleagues studies, the average flavonoid intake in our country is: children: adults: 19,5 26,6 mg/person 18,8 28,9 mg/person

Personal consumption
children: adults: 0-179,3 mg, 0,5-309,7 mg

CONCLUSION

Flavonoids are required substances from plant kingdom through animals to human, but their type and concentrations are very determined for healthy life.

THANK YOU FOR YOUR ATTENTION!