NATURAL PRODUCTS

Presented by:

Jennifer G. Valdez
NATURAL PRODUCTS
Definition
Chemical substances produced by living organisms i.e.
produced during metabolisms process.

Sources
 Plants
 Microorganisms (bacteria and fungi)
 Animals

 Marine organism/microorganisms
NATURAL PRODUCTS: TRADITIONAL VS
CONVENTIONAL MEDICINE
CONVENTIONAL TRADITIONAL
 Single bioactive metabolite for  Standardized crude extract
pharmaceutical formulation  Phytochemical analysis
 Physical analysis
 Safety assessment
 Tropical rainforest cover only  Dosage performance
12% of earth’s land area, but
constitute 50-70% of world
 80% world population depend on
species
alternative system of medicine
 Less than 1% of tropical  Phytochemical studies of
rainforests species tested, but at medicinal plant resulted isolation
least 25% of modern drug from of important drug
rainforests
 Cantharantus roseus originally
 Bioactive metabolites from plants investigated for hypoglycemic
from tropical rainforest formulate agents-yielded isolation important
as drug or precursor for drug anticancer drug (Vinca alkaloid)

(Kong et al., 2003) (Daniel, 2006; Newman et al., 2008)

 NATURAL PRODUCT: DRUG DISCOVERY

Lead Lead Optimization Lead Development

identification 1. Medicinal 1. Pharmacology
1. Isolation and chemistry 2. Toxicology DRUG
identification 2.Combinatorial CANDIDATES
3.
2. Target based chemistry Pharmacokinetics Clinical Trials
bioassay
4. ADME drug
3. In vivo and in delivery
vitro assays
INTERACTION OF MULTIDISCIPLINARY FIELD, INVESTIGATING
MEDICINAL AGENTS FROM HIGHER PLANTS

Plant SAR
selection
PHYTOCHEMIST/
MEDICINAL
SYSTEMIC PHARMACOGNOSIST
CHEMIST
BOTANIST Extraction, isolation,
Synthesis
structural elucidation

In vitro screening plant extracts

Bio-assay guided fractionation
In vivo screening of pure compounds

Pharmacologist AND/OR microbiologist and/or BIOCHEMIST

Pharmacological and/or chemotherapeutical and/or biochemical
evaluation
Determination of mode of action of active products

Pieters, L., & Vlietinck, A. J. (2005), Verpoorte, R. (1998)

PLANT
COLLECTION
AND
PROCESSING
 CRITERIA FOR PLANT
SELECTION

 Ethnopharmacology
based on traditional uses in folk
medicine

 Random screening
on site screening or large screening
program (dereplication)

 Chemotaxanomy
targeted for specific secondary
metabolites in certain taxa
 PLANT COLLECTION AND SELECTION
Plant collection and documentation

Alkaloid Phytochemical screening Flavonoid

Sample processing and extraction

Biological Activity Testing

Antioxidant
Antimicrobial
(DPPH)
PHYTOCHEMICAL ON-SITE SCREENING
FLAVONOID ALKALOID
TEST TEST
15g dried sample moistened
Methanolic Extract with 10% NH3
+
+ 15ml CHCl3 (Left for 10 min)
Mg ribbon + conc HCl
+
Equal amount 5% HCl + drops
Within 3 min Mayer’s reagent
Sample processing and extraction
Plant Material Dried ground plant material

Crude Extracts Extraction

 BIOLOGICAL ACTIVITY TESTING
(ANTIOXIDANT : DPPH METHOD)
A test samples in methanol will be added to 5 l of DPPH solution to give
final concentration of 250, 125, 62.5, 31.25, 15.625, 7.8125 and 3.906
μg/ml) in microtiter plate

Mixture is incubated for 30 min at room temperature

Absorbance at 517nm

The difference in absorbance between sample and control expressed as

percent inhibition
 EXPEDITION OUTCOMES
Eg: From two expeditions, 76 plant species
from 25 families were collected

Rubiaceae Rosaceae Lauraceae Rutaceae Commelinaceae

Annonaceae Moraceae Ebenaceae Simaroubaceae Burseraceae

Lecythidaceae Ulmaceae Verbanaceae Melastomaceae Convolvulaceae

Myristicaceae Menispermaceae Myrsinaceae Sterculiaceae Guttiferae

Tiliaceae Euphobiaceae Leeaceae Celastraceae Pandanaceae

20 species contain alkaloids
High content detected in the species in the table below
Psychotria sp Lasianthus Urophyllum hirsutum
(Rubiaceae) constrictus (Rubiaceae)
(Rubiaceae)
Chassalia cartaceae Polyalthia cauliflora Salacia grandiflora
(Rubiaceae) (Annonaceae) (Selastraceae)

Lasianthus constrictus
42 species contain flavonoid
High content detected in the species in the table below
Labisia Knema
Urophyllum sp
pumilla glauca
Baccaurea Leea Disoxylum
racemosa indica cauliflorum
Phyllagathis Myristica
rotundifolia cinnamomea

Baccaurea racemosa Leea

indica
 Antimicrobial assay:
140 crude extracts of plant parts tested against 12
gram-positive and gram negative bacteria.

Bacillus Enterococcus Eschericia Klebsiella

subtillis feacalis coli pneumonia
Proteus Pseudomonas Salmonella Serratia
mirabilis aeroginosa typhnimurium marcescens
Staphylococc Sterptococcus
S. epidermis S. pyogenes
us aureus pneumonia
 The antioxidant property was measured based radical
scavenging activity against DPPH radicals.
Out of 50 species, 16 species showed >80% inhibition at 250
μg/ml concentration.

Species Part Family IC50

Horsfieldia tomentosa Warb. Stem Myristicaceae 5.73
Microcos laurifolia (Hook. ex Mast.) Burret Stem Tiliaceae 13.6
Leaves 28.2
Prunus lamponga (Miq.) Kalkman Rosaceae
Stem 15.6
Gironniera parvifolia Planch. Stem Ulmaceae 14.6
Antidesma velutinosum Blume Stem Euphorbiaceae 5.9

Horsfiedlia polyspherula (Hook.f.) J. Sinclair var. Stem 1.6

Myristicaceae
sumatrana (Miq.) W.J. de Wilde Leaves 30.2
Knema glauca (Blume) Warb. Stem Myristicaceae 27.4
Leaves 22.4
Psydrax sp. Rubiaceae
Stem 30.2
Leaves 3.9
Psychotria malayana Jack Rubiaceae
Stem 29.7

Note: Strong < 30μg/ml, Medium 30-100μg/ml, Weak > 100μg/ml

Horsfiedlia polyspherula
(Hook.f.) J. Sinclair var. Psychotria malayana Jack
sumatrana (Miq.) W.J. de
Wilde
 FRACTIONATION AND PURIFICATION

PLANT MATERIAL

CRUDE EXTRACTS

FRACTIONS

PURE COMPOUNDS
OVERVIEW OF NATURAL PRODUCT RESEARCH

Single drugs
Spectroscopic
data on-line: Spectroscopic
- LC/UV synthesis Data off-line:
- LC/MS - UV,
- LCNMR PRODUCT - MS,
structure - NMR,
elucidation - IR

PLANT
EXTRACTS FRACTIONS PURE CONST’ toxicology
MATERIALS
extraction separation separation

BIOASSAY BIOASSAY structure

modification PRODUCT
Crude drugs
BIOASSAY
MEDICINAL PLANTS
Overview of Herbal Medicine Regulation in the World

• WHO recognizes the important and historical role of

herbal medicine in health. Herbal medicine is a major
form of treatment for more than 70% of the world's
population.
• The Guidelines for the Assessment
of Herbal Medicines states that a
substance’s historical use is a valid
way to document safety and efficacy
in the absence of scientific evidence
to the contrary.
• Appropriate use
• Quality control methods
• Recommended labeling
http://www.holisticonline.com/Herbal-Med/hol_herbalmed-
drugreg.htm
WHAT IS “MEDICINAL PLANT”?
 A plant that contains a substance or substances which
modulate beneficially the physiology of sick
mammals, and that it has been used by man for that
purpose.

What is a “weed”?
 A plant whose virtues have not yet been discovered
 Ralph Waldo Emerson
POTENTIAL OF TROPICAL RAIN FORESTS AS
SOURCE OF NEW THERAPEUTIC AGENTS
 Indonesia is the second biodiverse nation in the world

 Estimated no. of plant species in selected countries

 Market value of drugs/medicinal produced from plants is US 43 billion in
1989.

Country No. of Species

Malaysia 12 000
Indonesia 20 000
Philippines 10 000
USA 9 000
Brazil 55 000
Britain 1 550
Germany 2 600
PHARMACEUTICALS OF PLANT/
MICROORGANISM ORIGIN
Pharmaceuticals Sources
Codein Papaver somniferum
Reserpine Rauwolfia serpentine
Digoxin Digitalis lanata
Digitoxin Digitalis purpurea
Scopolamine Datura metel
Vincristine/vinblastine Catharanthus roseus
Cephalosporins Cephalosphorium sp.
Ephedrine Ephedra sinica
Quinidine Cinchona ledgerina
Morinda elliptica
 Common names:
“Mengkudu kecil”
 Medicinal uses:
1.Loss of appetite, headache, cholera,
diarrhea and fever
2.Enlarged spleen and wounds
3.Hemorrhoid and after childbirth
O OH
15 O OCH3
8
CHO
1
9
7
12 13
CHO
2

6 3
11 10 14

5 4
OH
O
O

2-formyl-2- Damnacanthal
hydroxyAQ (new) *strong anti-HIV
*anticancer

Ismail, N. H., Ali, A. M., Aimi, N., Kitajima, M., Takayama, H., & Lajis, N.
H. (1997). Anthraquinones from Morinda elliptica. Phytochemistry,
45(8), 1723-1725.
 O

Cassia alata
 Common names:
Gelenggang besar
OH O OH

 Medicinal uses:
Chrysophanol
Laxative, ringworm, constipation,
intestinal worm
O
OH O OH

OH

OH
OH O OH
O

*Aloe-emodin
Emodin

*Anticancer,
antimicrobial
 Andrographis paniculata
 Common names:
sambiloto, hempedu bumi
 Medicinal uses:
liver disease, high blood pressure
O O

O O
HO

HO HO
H
OH OH

Andrographolide 1,4-deoxyandrographolide

Reddy, M. K., Reddy, M. V. B., Gunasekar, D., Murthy, M. M., Caux, C., & Bodo, B. (2003). A
flavone and an unusual 23-carbon terpenoid from Andrographis paniculata.
Phytochemistry, 62(8), 1271-1275.
 Centella asiatica
 Common names:
pegagan
 Medicinal uses:
liver disease, high blood pressure

Asiaticoside

Kim, W.-J., Kim, J., Veriansyah, B., Kim, J.-D., Lee, Y.-W., Oh, S.-G., et al.
(2009). Extraction of bioactive components from Centella asiatica using
subcritical water. The Journal of Supercritical Fluids, 48(3), 211-216.
Eurycoma longifolia Jack
 Common names:
pasak bumi, tongkat ali
 Medicinal uses:
aphrodisiac

1-eurycomanone
2-14,15β-Dihydroxyklaineanone
3-13,21-Dihydroeurycomanone
Chan, K.-L., Choo, C.-Y., Abdullah, N. R., & Ismail, Z. (2004). 4-13,21-Epoxyeurycomanone
Antiplasmodial studies of Eurycoma longifolia Jack using the 5-Eurycomanol
lactate dehydrogenase assay of Plasmodium falciparum. 6-Eurycomalactone
Journal of Ethnopharmacology, 92(2-3), 223-227. 7-9-Methoxycanthin-6-one
 "And the earth brought forth grass, and herb
yielding seed after his kind, and the tree yielding
fruit after his kind, whose seed was in itself, and
after his kind: and God saw that it was good."
-Genesis 1:12
THANK YOU 

33
PLANTS SECONDARY
METABOLITES

JANICE D. DARISAN
 PLANT SECONDARY METABOLITES
Secondary metabolites are those metabolites which are often
produced in a phases of subsequent to growth, have no function in
growth (although they may have survival function), are produced
by certain restricted taxonomic groups of microorganisms, have
unusual chemical structures, and are often formed as mixture of
closely related members of a chemical family.

They are not generally included in standard metabolic charts

 A metabolic intermediate or product, found as a differentiation

product in restricted taxonomic groups, not essential to growth
and the life of the producing organism, and biosynthesis from one
or more general metabolism.
 PLANT SECONDARY METABOLITES
They have a wide range of chemical structures and biological
activities. They are derived by unique biosynthetic pathways from
primary metabolites and intermediates.

 Plants produce as amazing diversity of low molecular weight

compounds.

Of the estimated 400,000-500,000 plant species around the globe,

only a small percentage has been investigated phytochemically
and the fraction subjected to biological or pharmacological
screening is lower.
 PLANT SECONDARY METABOLITES

The ability to synthesize secondary metabolites has been

selected through the course of evolution in different plant
lineage when such compounds address specific needs.
- Floral scent volatiles and pigments have evolved to attract
pollination and thus enhance fertilization.
- To synthesize toxic chemicals has evolved to ward off
pathogens and herbivores or to suppress the growth of
neighboring plants.
- Chemicals found in fruits prevent spoilage and act as signals
(in the form of color, aroma, and flavor) the presence of
potential rewards (sugars, vitamins, and flavor) for animals
that eat the fruit and thereby help to disperse the seed.
 Primary and Secondary Metabolism

 The pathways for generally modifying and synthesizing

carbohydrates, proteins, fats and nucleic acids are found
to be essentially the same in all organisms, apart from
minor variations.
- These processes demonstrate the fundamental unity of
all living matter, and are described as primary
metabolites.
 Primary Metabolisms
 Degradation of carbohydrates and sugars generally proceeds
via the well characterized pathways, known as glycolysis and
the krebs/citric acid/tricarboxylic acid cycle, which release
energy from the organic compounds by oxidative reactions.

 Aerobic organisms are able to optimize these processed by

adding on a further process, oxidative phosphorylation. This
improves the efficiency of oxidation by incorporating a more
general process applicable to oxidation of a wide variety of
substrates rather then having to provide specific process for
each individual substrate.
 Secondary Metabolisms
 The compounds which synthesized from the secondary
metabolisms are so-called secondary metabolites.

 Secondary metabolites are formed in only specific organisms,

or group of organisms, are expression of the individuality of
species.

 Secondary metabolites are not necessarily produced under all

conditions, and in the vast majority of cases the function of
these compounds and their benefit to the organism is not yet
known.

 It is this area of secondary metabolism that provides most of

the pharmacologically active natural products.
 Secondary Metabolisms
 To make such compounds as sugars, waxes, lignin,
starch, pigments or alkaloids, plants utilize very specific
enzymes, each of which catalyzes a specific metabolic
reaction.
- The enzymes are proteins called organic catalysts.
- These enzymes are coded by specific genes in the
plants DNA and are made via processed we call
transcription and translation.
- When there is a series of enzymatically catalyzed
reaction in a well defined sequence of step, we have what
is termed a metabolic pathway.
 Biosynthetic Pathway
 Nucleoside diphosphate sugar pathway: cellulose,
hemicellulose, glycosides.

 Shikimate-cinnamete pathway: lignin, lignans,

hydrolized tannins.

 Mixed pathways of 3 and 4: some prenylflavonoids,

quinones and stilbenes.
 Classes of Secondary Metabolites
 The majority of secondary metabolites belong to one of a
number of families, each of which have particular
structural characteristics arising from the way in which
they are built up in nature (biosynthesis).

 The classes of secondary metabolites are:

- Polyketides and fatty acids
- Terpenoids and steroids
- Phenylpropanoids
-Alkaloids
- Others (specialize amino acids and carbohydrates
 Polyketide and Fatty acids
 Polyketides are formed by the linear combination of
acetate units derived from the “building block” acetyl co-
enzyme A.
 The acetate origin of these compounds leads to a
preponderance of even-numbered carbon chains.
 Many plant oils and animal fats contain long-chain
monocarboxylic acids known as fatty acids.
 The common fatty acids have an even number of carbon
atoms, typically C12 – C20, linked together in a straight
chain with up four double bonds.
 In plants, the fatty acids and the corresponding alcohol
are found in leave waxes and seed coating:
- Myristic acid (C14) is found in nutmeg seeds.
- Palmitic acid (C16) found in almost oil plants.
- Stearic acid (C18) occurs in long amounts in
animal fat.
 Polyketide and Fatty acids
 Unsaturated fatty acids
are important to us in
food.
- Oleic acid is the most widely
distributed, and a major
constituent of olive oil.
- Linoleic acid and linolenic
acids are most highly
unsaturated and found in
linseed oil.
- Linolenic acid is easily
oxidized by air, and is one of
the “drying oil” used in paint
and varnishes.
 Polyketide and Fatty acids

- Linolenic acid is oxidized by

plants to jasmonic acid,
which is a signaling
substances that stimulates
plant defense mechanisms.

- Arachidonic acid ( C20) is a

precursors of the
prostaglandin hormones
 Polyketide and Fatty acids

• Polyacetylenes
- They are a group of naturally
occurring hydrocarbon
derivatives characterized by
one or more acetylenic groups
in their structures

- Araliaceae, Campanulaceae,
Asteraceae, Pittosporaceae
and some fungi
 Terpenes
• The most widespread and chemically diverse groups of natural
products.

• Unique group of hydrocarbon-based natural products whose

structures may be derived from isoprene.

• Classified by the number of 5- carbon units.

• Its function in plants is generally considered to be both ecological

and physiological.
- Allelopathy
-Insecticidal
-Insect pollinators
-Plant Hormones (Abscisic acid, gibberlellin)
Myrcene

Myrcene is found in the essential oil bay leaves as

well as hops. It is used as an intermediate in the
manufacture of perfumes.
Geraniol and Linalool

Geraniol is isometric with linalool, constituents the

major part of the oil of roses and is also found in
essential oils of citronella, lemon grass, and others
Menthol

Menthol is well-known monoterpene which is found in

the essential oil of peppermint and other members of
the mint family
Sesquiterpenoids
• More than 10000 identified, representing a wide variety
compounds of different skeletal types from acyclic to
tetra cyclic systems.
Diterpenoids
• Constitute a major part of
oleoresin.
• This group can be divided
into acyclic, bicyclic,
tricyclic, tetracyclic, and
macrocyclic structural
types.
• Present either
hydrocarbons or as
derivatives with hydroxyl,
carbinyl, or carboxyl
groups
 Phenolic Constituents
• Heartwood and bark contain a large variety of complex aromatic
extractives. Most of them are phenolic compounds, and many are
derived from the phenylpropanoid structure.
Phenolic Compounds
 Lignans
• Large amount of research devoted to investigating medicinal
properties of lignans.
- Particularly from tropical hardwoods and grasses
-Antiviral
- Antitumor
- Biocidal
- Bioactive Agents
• Can be isolated from the bark, fruit, heartwood, leaves, roots and
resin of plants.
 Flavonoids/Tannins
• Concentrated in certain parts of plants
• Plant sources
- Pulp of fruits
-Broccoli, green peppers, onions, etc.
-Green tea, red wine
- Herbs
- Tree bark
Condensed Tannins
• Sources hardwoods but present in softwoods
- Wattle (Acacia)
-Quebarcho (Schnopsis)
- Mangrove (Rhizophora)
- Hemlock
 Alkaloids

• Derived from plants

• With a basis character (hence the term alkaloid from alkali)
• Contained a nitrogen based heterocyclic ring within their
molecules
• Subcategories:
- Monocyclic alkaloids
- Bicyclic alkaloids
- Polycyclic alkaloids
Physiologically active alkaloids used in modern medicine
THE FUNCTIONS OF SECONDARY
METABOLITES IN PLANTS
Secondary Metabolites Function Mechanism
Defensive Deterring herbivores due to their
Alkaloids agents bitter taste

Volatile oils Pheromones Attracting insect for pollination and

protect the plants from microbes.
When present in resin, perform
Diterpenes and Antimicrobial wound healing and antimicrobial
triterpenes
function.
Limonoids and Insect Protect the plants especially wood
quassinoids repellents from insect attack.
Protect cellular membranes and
Phenolics Antioxidants tissues containing lipids from
oxidation.
Anthocyanins and Pollinator Pigments which make flowers visible
flavanoids guides to ‘colour blind’ insects

Pigments, Intermediates in cellular respiration

Quinones Respiration and photosynthesis

(Daniel, 2006;
Thank you!!!