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ABiosyntheticApproachof

NaturalProductsto
Phytopharmaceutics

Definitions
Biosynthesisisthetermfortheinvivo
synthesisofmetabolites/naturalproducts.

Theformationofachemicalcompoundbya
livingorganism.
Thepathwaysforgenerallymodifyingand
synthesizingcarbohydrates,proteins,fats,and
nucleicacidsarefoundtobeessentiallythesamein
allorganisms,apartfromminorvariations.

Theseprocessesarecollectivelydescribedas
primarymetabolism,withthecompoundsinvolved
inthepathwaysbeingtermedprimarymetabolites.

PrimaryMetabolites
Primarymetabolitesarecompoundsthatare
commonlyproducedbyallplantsandthatare
directlyusedinplantgrowthanddevelopment.
Moleculesthatareessentialforgrowthand
developmentofanorganism.
Themainprimarymetabolitesare:carbohydrates,
proteins,nucleicacids,andlipids.

Primarymetabolism( Biochemistry)
SecondaryMetabolism
Secondarymetabolites :metabolicpathwaysthatare
notessentialforgrowth,developmentor
reproduction,butthatusuallyhaveecological
function.
Secondarymetabolites arethosechemicalcompounds
inorganismsthatarenotdirectlyinvolvedinthenormal
growth,developmentorreproductionofanorganism.
Inthissensetheyare"secondary".

Secondarymetabolites,arefoundinonlyspecific
organisms,orgroupsoforganisms,andarean
expressionoftheindividualityofspecies.

Secondarymetabolitesarenotnecessarilyproduced
underallconditions,andinthevastmajorityof
casesthefunctionofthesecompoundsandtheir
benefittotheorganismisnotyetknown.
Somesecondarymetabolitesareproducedforeasily
appreciatedreasons,e.g.
1. Astoxicmaterialprovidingdefenseagainst
predators.
2. Asvolatileattractantstowardsthesameorother
species.
3. Ascoloringagentstoattractorwarnotherspecies.

Secondarymetabolism( Naturalproducts
chemistry).

ClassificationofNaturalProducts
Secondarycompoundsaregroupedintoclassesbasedonsimilarstructures,
biosyntheticpathways,orthekindsofplantsthatmakethem.
Naturalproductsaregenerallygroupedintofivemainclasses
Secondary metabolites are derived
from primary metabolites

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secondarymetabolites
Over300,000secondarymetabolitesexist
Organismsvarywidelyintheircapacitytosynthesize
andtransformchemicals.

Forinstance,plantsareveryefficientatsynthesizing
organiccompoundsviaphotosynthesisfrominorganic
materialsfoundintheenvironment,whilstother
organismssuchasanimalsandmicroorganismsrelyon
obtainingtheirrawmaterialsintheirdiet,e.g.by
consumingplants.

Thebuildingblocks
Thebuildingblocksforsecondarymetabolites
arederivedfromprimarymetabolism.

Thenumberofbuildingblocksneededis
surprisinglyfew.
Themostimportantbuildingblocks
employedinthebiosynthesisofsecondary
metabolitesarederivedfrom:

1. AcetylcoenzymeA(acetylCoA)
2. Shikimicacid
3. Mevalonicacid
4. 1deoxyxylulose5phosphate
5. Aminoacids

1.Acetatepathway
Theforminwhichacetateisusedinmostofits
importantbiochemicalreactionsisacetylcoenzymeA
(acetylCoA).
AcetylCoA isformedbyoxidativedecarboxylationof
theglycolyticpathwayproductpyruvicacid.
Importantsecondarymetabolitesformedfromthe
acetatepathwayincludes:

1. Phenols
2. Prostaglandins
3. Macrolideantibiotics
CoenzymeA: presentinalllivingcellsthat
functionsasanacylgroupcarrier.
NH2
N
N
O O H3C CH3 N
OH OH N
RS O O O
N N P P
H H O
OH O O
HO
HO O OH
P
O

CoenzymeAactsasanacyltransfer/
carbonactivationreagentbyforming
reactiveacylthioesters

2.Shikimatepathway
Shikimicacidisproducedfromacombinationof
phosphoenolpyruvate,aglycolyticpathway
intermediate,anderythrose4phosphatefromthe
pentosephosphatepathway.
Theshikimatepathwayleadstoavarietyof:
1. Phenols
2. Cinnamicacidderivatives
3. Lignans
4. Alkaloids
3.Mevalonatepathway
Mevalonicacidisitselfformedfromthree
moleculesofacetylCoA,butthemevalonate
pathwaychannelsacetateintoadifferentseries
ofcompoundsthandoestheacetatepathway.

4.Deoxyxylulosephosphatepathway
Deoxyxylulosephosphatearisesfromacombination
oftwoglycolyticpathwayintermediates,namely
pyruvicacidandglyceraldehyde3phosphate.
Themevalonateanddeoxyxylulosephosphate
pathwaysaretogetherresponsibleforthe
biosynthesisofavastarrayofterpenoidandsteroid
metabolites.
5.Aminoacidspathway
Peptides,proteins,alkaloidsandmanyantibiotics
arederivedfromaminoacids.

Intermediatesfromtheglycolyticpathwayandthe
Krebscycleareusedinconstructingmanyofthem.

Thearomaticaminoacidsphenylalanine,tyrosine,
andtryptophanarethemselvesproductsfromthe
shikimatepathway.

Secondarymetabolitescanbesynthesizedbycombining
severalbuildingblocksofthesametype,orbyusinga
mixtureofdifferentbuildingblocks.
Manyofsecondarymetabolitesalsocontainoneor
moresugarunitsintheirstructure.
Toappreciatehowanaturalproductiselaborated,itis
ofvaluetobeable:
1. Todissectitsstructureintothebasicbuildingblocks
fromwhichitismadeup.
2. Toproposehowthesearemechanisticallyjoined
together.
Relativelyfewbuildingblocksareroutinely
employed,andthefollowinglistincludesthose
mostfrequentlyencounteredinproducingthe
carbonandnitrogenskeletonofanatural
product.

C1:thesimplestofthebuildingblocksiscomposedofa
singlecarbonatom,usuallyintheformofamethyl
group,andmostfrequentlyitisattachedtooxygenor
nitrogen,butoccasionallytocarbon.Itisderivedfrom
theSmethylofLmethionine.

C2:AtwocarbonunitmaybesuppliedbyacetylCoA.
AcetylCoAisfirstconvertedintothemorereactive
malonylCoA beforeitsincorporation.
C5:thebranchedchainC5 isoprene unitisafeatureof
compoundsformedfrommevalonate ordeoxyxylulose
phosphate.

C6C3:thisreferstoaphenylpropylunitandisobtained
fromthecarbonskeletonofeitherLphenylalanine orL
tyrosine.

C6C2N:again,thisbuildingblockisformedfromeither
Lphenylalanine orLtyrosine.

Indole.C2N: thethirdofthearomaticaminoacidsis
Ltryptophan.

C4N: theC4Nunitisusuallyfoundasaheterocyclic
pyrrolidinesystemandisproducedfromLornithine
(nonproteinaminoacid).

C5N: itisproducedbyusingLlysine andtheunittends


tobefoundasapiperidineringsystem.
Theconstructionmechanisms
Naturalproductmoleculesarebiosynthesizedbya
sequenceofreactionswhicharecatalyzedbyenzymes.
Mostproteinsinlivingcellsareenzymes.
Enzymeshavethepowertoeffectthese
transformations:
1. Moreefficientlyandmorerapidlythanthechemical
analogy.
2. Underverymuchmilderconditions.
3. Carryoutreactionsinastereospecificmanner.

BiochemicalReactions
1. Alkylationreactions
2. WagnerMeerweinrearrangements
3. AldolandClaisenreactions
4. SchiffbaseformationandtheMannichreaction
5. Transamination
6. Decarboxylationreactions
7. Oxidationandreductionreactions
8. Phenolicoxidativecoupling
9. Glycosylationreactions
Why secondary metabolites?

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Secondary metabolites

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Fatty acid-derived substances and
polyketides

Morethan10,000known

Biosynthesizedfromsimpleacylprecursorssuchasacetyl
CoA,propionylCoA,andmethylmalonylCoA

Naturalproductsderivedfromfattyacids,suchas
prostaglandinE1,generallyhavemostoftheoxygenatoms
removed

Alkaloids

34
Phenolics

36
Terpenoids

37
Taxol

39

Where does taxol come from?


Taxol is found in the bark of the tree.

Taxus brevifolia Nutt. What can we do?


Pacific Yew

40
FromGGPtoTaxotere

42
Where does camptothecin come from?

Camptotheca acuminata

Happy tree

43

zaad

wildgroei teelt GAP

oogsten

wassen,drogen
uitgangsmateriaal

verkleinen
decontamineren
GLP
transporteren GMP
opslaan

extraherenplantenmateriaal kwaliteitscontrole
identiteit
tussenproduct zuiverheid
standaardiserenextract gehalte

concentrerenextract
drogenextract GMP

formuleren
eindproduct
fytotherapeuticum
Repercolation

Repercolation
CounterCurrentextractie

Whatisanextract?

3000to5000constituents
Analyticalcomplex
Mainconstituents
minorconstituents
Saponins
Sugars
Tannins
Lignins
Solventhasinfluenceonextractionpattern

Chamomille, Matricaria recutita, Asteraceae

Ethanolfreeextracts

Extractionwith
Sorbitol/water
Polyethylenglykol400(PEG)/water
PEG400/Propylenglykol(PG)/water

Examples:
Prospan(Sorbitol/Water)
Tevoninforte(PEG/Water)
Ethanolfreeextracts
Ethanolismostappropiatesolvent,but
problemsfor
Babies,children
Pregnantwomen
Alcoholproblems,Parkinson,Epilepsiepatienten

Qualitycontrol
EuropeanPharmacopea.
Identity,purity.
Fingerprints:DLC,HPLC,GC.
Toxiccontaminantions:pathogenicbacteria,
fungi;yeast,endo andexotoxines;heavy
metals;pesticids,herbicids;volatile
decontaminants;radioactiveisotops.
Fraudoruseofwrongspecies.
Preparationofherbalmedicinal
products
SolventshaseffectonqualityofHMP
HighqualitybyGoodManufacturingPractice
(GACP,GMP,GLP)
Formulation:tablet,capsule,liqid,crme,
ointment.
Informationonbox
Stability

Standardisationofherbalmedicinal
products
Oneormoredefiningconstitutents
Biologic active constitutent is responsible for
activityofHMP
Markercompounds,characterisitcforspecific
plantspeciesandvarietieswhichdonotshow
specialbiological/pharmacologicalactivity
Essentialforpharmacological,toxicologicaland
clinicalevaluationforrationaluseand
registration
Standardisation

Fytotherapeutica:characterisitics

Complexmixtureisthe Mostlydifferentplant
drug varietiesandspecies
Mild,sometimesstrong Pharmacologicalactive
Synergism,Antagonism
activities
Safe,goodandbraod
therapeuticuse
Sideeffectsand
interactionspossible
Sideeffects

Allergicreactions(Arnicamontana,Calendulaflos)
Phototoxicity(Ammivisnaga,Hypericum
perforatum)
Hepatotoxiceffects(KavaKava,Tussilagofarfara)
Nephrotoxicity(Brassicanigra)
Cardiovascularreactions(Glycyrrhizaglabra)
DrugInteractions(Hypericumperforatum)

HMPsinEurope
Fytotherapeuticsmostlydonotfullfillall
criteriaasrequestedbyEU
Regulationsmostlydependonnational
jurisdictionandwehavemajordifferences
betweenEUmemberstates
EMEAissettingupcommonguidelinesfor
marketharmonisation
Guidelinesfortraditionaluse

Simplifiedregistrationprocessfortradional
use(e.g.homeopathy,TCM):adaptionto
guidelinesformanufactering,butmoreliberal
regardingefficacytesting
Europeanstandardshavetobemetfor
qualitycontrol,safetyandproduction