Transcript Alkaloids * Natural nitrogenous secondary metabolites from plants

Some important classes of alkaloids
Class
Piperidine alkaloids
Pyrrolidine/tropane
alkaloids
Precursors
L-lysine (C5N)
Pyridine alkaloids
L-Trp or L-Asp
Examples
Piperine
_____________
L-ornithine (C4N) Cocaine, scopolamine
Niacin (Vit. B3), nicotine
Catecholamines
L-Tyr (C6C2N)
and tetrahydroisoquinolines
Opiates
2 L-Tyr units
Dopamine, adrenaline, mescaline
anhalamine
Morphine, tubocurarine
Phenylalanine-derived
L-Phe (C6C3N)
Capsaicin, ephedrine
Indole alkaloids
Quinoline alkaloids
Purine alkaloids
L-Trp
“
L-Gly, L-Gln, L-Asp
Serotonin, ergotamine, LSD, vinblastine
quinine, camptothecin
Caffeine, theobromine
Building blocks from the acetate, shikimate, or deoxyxylulose phosphate pathways are also
frequently incorporated into the alkaloid structures.
Many alkaloids acquire their N via transamination reactions (catalyzed by Vitamin B6).
Most alkaloids are quite toxic and produced by the plant as a defense against herbivores.
Opiates: morphine and related compounds
Focus:
• pieces/precursors
• key assembly steps
• structure-activity
• physiological interactions
First methylation
Precursor is formed by combining two
C6C2 units, both originating from tyrosine
Two stereoisomers lead to different
classes of alkaloids
From (S)-N-methyl
coclaurine
Further methylation
is common
Berberine is found in many members of the
Berberidaceae, the Ranunculaceae, and other
families used in traditional medicine.
Berberine has antiamoebic, antibacterial, and
anti-inflammatory properties.
Curare (Chondrodendron tomentosum)
Arrow poison prepared in the rain forests by native South American tribes from bark
and stems may contain >30 different plants, C. tomentosum is the main one.
Radical dimerization of the two coclaurine isomers produces a dimer.
Tubocurarine is an ACh agonist, once it gets into the bloodstream, it relaxes voluntary
muscles to paralysis by blocking nerve impulses at neuromuscular junction.
Synthetic derivatives used during surgery as muscle relaxant.
Oh oh…I think I just
stuck myself… better
get out my
cholinesterase
inhibitor pen!
Making the morphine
skeleton
Key steps:
1) oxidative coupling to
make the 4th ring
2) reduction and acetylation
promotes formation of
the furan ring
3) Demethylation then
reduction of the ketone
“antitussive”
Opium
• 80% of the world’s illegal opium is grown in
Afghanistan, supported by the Taliban.
• $ 4 billion/year industry
• Northern alliance initiative to destroy poppy fields pits
farmers and corrupt local officials against military
• U.S. troops are supporting efforts to provide farmers
with crop alternatives like pomegranates or grapes
• However, many farmers told in 2007 that they will
receive assistance in establishing legal crops are still
waiting for help
R. Draper, “Opium Wars”, National Geographic, Feb. 2011.
Properties of opiates
Brain has opioid receptors (d, k and m) that mediate adenylcyclase, lower cAMP levels,
increase K+ conductance, close Ca2+ channels, decrease transmission of pain signals
Morphine’s use as analgesic involves overall CNS depression so side effects include
drowsiness, lethargy, constipation, unconsciousness
Structural basis for bioactivity of opiates
Enkephalins and endorphins are
endogenous opioid peptide ligands produced
during labor, other times of stress or shock
“Runner’s high”
ACh inhibitors, rapidly degraded
“Morphine rule”: structure must contain an aromatic ring attached to a
quarternary C (Cq) with a 2-C linker to a tertiary (3o) amine
Drugs derived from morphine semi-synthetically
Similar activity, can be
produced from morphine
by methylation
Synthetic analogues
Effective
analgesic and
less addictive
Heroin was introduced into cough syrup in 1898
Acetylation decreases polarity, so it
crosses blood-brain barrier quickly.
More addictive though.
Used in cough medicine 
Antitussive but not analgesic
“Demerol”
Fast, but short-acting analgesia
50-100X more active than
morphine due to lipophilicity
Orally active, produces less
euphoria, less withdrawal
From L-Tryptophan:
Indole alkaloids
Produced from L-tryptophan
plus an isoprene unit,
the indole alkaloids have
polycyclic ring structures
Fungi from Claviceps genus are
best-known producers because
they infect some grain crops,
but the indoles are produced
by other fungi including
Aspergillus and Penicillium
serotonin
skeleton
Lysergic acid is probably the
most well-known, as it is the
precursor for hallucinogen LSD
(lysergic acid diethylamide) and
the ergot alkaloids – mixed
agonist/antagonist effects on
5-HT (serotonin) receptors
leads to hallucinations
Ergot – not a fun guy
Ergot is the dried sclerotium of the fungus Claviceps purpurea
that develops on the ovary of rye and other grasses consumed
by humans or animals.
The poisonous properties of ergots are caused by a group of
indole alkaloids, the ergot alkaloids or ergolines.
Consumption of ergot-infected rye produces a disease called
ergotism. Ergot poisoning affects two types of receptors:
1) The a-adrenergic receptors for norepinephrine which causes
• GI upsets, e.g. diarrhea, abdominal pains, and vomiting.
• Circulatory changes, e.g. coldness of hands and feet due to
vasoconstriction of the blood vessels to the extremities
Ergot – not a fun guy
2) Ergot also acts on the serotonin (5-HT) receptors and the
a-dopaminergic receptors causing neurological symptoms:
Headache, vertigo, convulsions, psychotic disturbances,
hallucinations
Vasoconstriction can cause restricted blood flow in small
terminal arteries, death of the tissue, gangrene, and even the
loss of hands, feet, or limbs.
Gangrenous ergotism was known as St. Anthony’s Fire because
the Order of St. Anthony cared for the sufferers during the
Middle Ages in Europe when outbreaks of the disease in
humans and animals were relatively frequent.
Indole-isoprenoid is modified by attachment of a small peptide (Phe & Pro usually)
Ergotamine’s vasoconstrictive activity has led to its use in treatment of migraines
For more information:
A Trip Through LSD and the Ergot
Alkaloids
Melissa Medeiros
Natural Products
May 2006
Amaryllidaceae
alkaloids
narciclasine
(abundant)
synthetic derivative
trans-dihydronarciclasine
pancratistatin
Compounds 1 – 6 are found in Narcissus pseudonarcissus (daffodils) and other plants from the
Amaryllidaceae family.
Pancratistatin (1) selectively induces apoptosis in cancer cells through a mitochondrial pathway,
with little effect on normal cells. However, it’s not very abundant.
Narciclasine (3) disrupts organization of the actin cytoskeleton of cancer cells at 30-90 nM.
Compound (5) also has potent anticancer activity while its cis-isomer has none.
Problems: low water-solubility, low natural abundance of (1) and (5), and toxicity of (3) ...
Narciclasine inhibits CYP3A4 (major liver detoxifying cytochrome), which can damage the liver.
SAR shows this
part of structure
is key...
Goal of study: prepare trans-dihydro
derivative of narciclasine that has the anticancer activity without the liver toxicity.
Protection of the OH groups
followed by hydrogenation
usually produces a mixture of cisand trans-isomers.
Since the cis-isomer (7) has no
anticancer activity, reaction
conditions were modified until
maximum yield of trans isomer
was achieved.
Compound 5 did not inhibit
any of the cytochromes tested.
Result: an effective way to
prepare a less toxic drug!
More indole alkaloids...from Uncaria tomentosa
Complex polycyclic indole alkaloids are prevalent in species
such as Uncaria tomentosa (Cat’s claw)
see: Heitzman, M. E.; Neto, C. C.; Winiarz, E.; Vaisberg, A. J.,
Hammond, G. B.* Review: Ethnobotany, phytochemistry and
pharmacology of Uncaria (Rubiaceae), Phytochemistry, 66: 5-29
(2005).
Traditional uses of U. tomentosa
Aqueous extracts of bark or roots used by Ashaninka in
Peruvia rainforest:
• Allergies, asthma
• Wound healing, viral infections, fevers
• Arthritis, gastric ulcers, rheumatism, inflammation
• Contraception, menstrual irregularities, childbirth
• Some of these activities are associated with alkaloids
• Uncaria also contains flavonoids, catechins,
cinchonains, caffeic acid and ursolic acid – these may
play a role in some of the bioactivities.
Pentacyclic oxindole
alkaloids have immuneboosting effect, upregulate
B & T cell production
Tetracyclic oxindole alkaloids
block immune factor, may also
inhibit central dopamine
release, depressing movement
Weaker dopamine receptor
antagonist – some locomotor
depression observed
More indole alkaloids...terpene indole
alkaloids with multicyclic structures
Coupling of tryptamine
(decarboxylated Trp)
with the secoiridoid
secologanin, followed
by a variety of cyclizations
forms terpene indole
alkaloid skeletons with
lots of structural diversity
Some interesting terpene indole
alkaloids
Yohimbine
• From Pausinystalia yohimbe
• Folk use: Aphrodisiac,
weight loss
• Pharmacology: dilates blood
vessels, binds a-adrenergic
receptors
Reserpine, deserpidine
• From Rauwolfia serpentina
• Folk use: treat snake bite,
insanity
• Pharmacology: lowers blood
pressure, tranquilizer, depletes
stored catecholamines
Skeletal rearrangements and dimerization lead
to vincristine/vinblastine structure
• Intermediates form through
rearrangement of
isoprenoid moieties
Anti-cancer agent, used to treat
leukemia and lymphoma.
Strychnine
• From dried seeds of Strychnos
nux-vomica
• Biosynthesis: Rearrangements in
strictosidine
• Highly toxic convulsant
• Binds to glycine binding sites in
spinal cord, causes asphyxiation
• Has been used as a rat poison
Quinine – Antimalarial agent from bark of Cinchona trees
Discovered in 1600’s , cured a Peruvian countess
Depolymerizes toxic byproducts of Plasmodium
Quinine was originally added to tonic water as a
prophylactic against malaria – gin
was later added to mask the
bitter taste 
Some important classes of alkaloids
Class
Piperidine alkaloids
Pyrrolidine/tropane
alkaloids
Precursors
L-lysine (C5N)
Pyridine alkaloids
L-Trp or L-Asp
Examples
Piperine
_____________
L-ornithine (C4N) Cocaine, scopolamine
Niacin (Vit. B3), nicotine
Catecholamines
L-Tyr (C6C2N)
and tetrahydroisoquinolines
Opiates
2 L-Tyr units
Dopamine, adrenaline, mescaline
anhalamine
Morphine, tubocurarine
Phenylalanine-derived
L-Phe (C6C3N)
Capsaicin, ephedrine
Indole alkaloids
Quinoline alkaloids
Purine alkaloids
L-Trp
“
L-Gly, L-Gln, L-Asp
Serotonin, ergotamine, LSD, vinblastine
quinine, camptothecin
Caffeine, theobromine
Building blocks from the acetate, shikimate, or deoxyxylulose phosphate pathways are also
frequently incorporated into the alkaloid structures.
Many alkaloids acquire their N via transamination reactions (catalyzed by Vitamin B6).
Most alkaloids are quite toxic and produced by the plant as a defense against herbivores.
From amino acids to purines to caffeine
and xanthine alkaloids
Purine heterocyclic
ring system is derived
from amino acids and
various one C donors
muscle relaxant
The xanthines
Caffeine, Theobromine, and Theophylline
•
•
•
•
The purine alkaloids caffeine, theobromine, and theophylline are all methyl
xanthines that commonly co-occur in plants. Major sources are stimulant
beverages and foods such as tea, coffee, cocoa, and cola.
Xanthines competitively inhibit phosphodiesterase, causing an increase in cyclic
AMP and adrenaline release. This leads to CNS stimulation, relaxation of bronchial
smooth muscle, and induction of diuresis. Inhibition of TNF-a and leukotriene
synthesis is thought to occur, reducing inflammation and innate immunity.
The effects vary among the three compounds. Caffeine is the best CNS stimulant.
As a vasoconstrictor it can be combined with a therapeutic agent to increase
effectiveness (e.g. compound analgesics). It has weaker diuretic action.
Theobromine has little stimulant action, but has more diuretic activity and also
muscle relaxant properties. Theophylline also has low stimulant action and is an
effective diuretic, but it relaxes smooth muscle better than caffeine or
theobromine and is frequently used in slow-release formulations.
Caffeine and adenosine
• Caffeine readily crosses the blood-brain barrier, and once in the brain, the
principal mode of action is as a nonselective antagonist of adenosine
receptors (competitive inhibition)
• Adenosine is found in every part of the body, but it has special functions in
the brain. Concentrations of brain adenosine are thought to be increased
by metabolic stresses such as anoxia or ischemia. It also may have a
specific role in control of the sleep-wake cycle.
• Brain adenosine may also protect the brain by suppressing neural activity
and increasing blood flow through A2A and A2B receptors located on
vascular smooth muscle. By counteracting adenosine, caffeine reduces
resting cerebral blood flow – it’s a vasoconstrictor.
• Adenosine is released in the brain through a complex mechanism. It is not
likely that adenosine is the primary neurotransmitter for any group of
neurons, but rather is released together with other transmitters by a
number of neuron types.
So you can’t start the day
without Joe?
You’re not alone!
• Is caffeine addictive? Several classes of adenosine receptors
are known, and there is evidence that A 2A receptors interact
with the dopamine system, which is involved in reward and
arousal.
• Tolerance: because caffeine is primarily an antagonist of
adenosine receptors, regular caffeine consumers may adapt
to its continuous presence by increasing the number of
adenosine receptors.
• This reduces the stimulatory effects (tolerance adaptation)
and makes one much more sensitive to adenosine, so that
reducing caffeine intake results in withdrawal symptoms.
From Drugs and the Human Body, 6th edition, K. Liska
Chocolate and pet poisoning
• Dogs are most often affected, due to their ability to find
chocolate and the common 'sweet tooth' they develop, but
cats and other mammals are susceptible to the toxic effects
of chocolate, too.
• Theobromine is the major stimulant in chocolate. Its effects:
– CNS and cardiovascular stimulant
– Increases blood pressure (mild)
– Nausea and vomiting
• Are some chocolates more toxic than others?
Yes. Unsweetened chocolate contains 8-10 times the amount of
Theobromine as milk chocolate. Semi-sweet or dark chocolate
falls roughly in between the two.
• The toxic dose of Theobromine (or caffeine) for pets is 100200mg/kg body weight. However, reports by the ASPCA have
noted problems at doses much lower than this - i.e. 20mg/kg.
• Using the 20mg/kg as a measure of "problems can be seen” a
50 lb (23 kg) dog would have to consume 9 oz of milk chocolate.
Some dogs won't see problems at this rate but others may.
http://vetmedicine.about.com/cs/nutritiondogs/a/chocolatetoxici.htm
Background
• A Dutch cohort study in 2002 and others found that
higher coffee consumption correlates with lower risk of
Type-2 diabetes.
• Coffee constituents that have been previously shown
to affect glucose metabolism include caffeine,
chlorogenic acid, lignans, trigonelline.
• Caffeine also reported to induce energy expenditure by
boosting thermogenesis.
• However some cohort studies suggest that decaf also
reduces risk.
• Is caffeine the active constituent against diabetes?
Mouse model
• KK-A3’ mice spontaneously develop type-2 diabetes by
6 weeks old
• Obesity, hyperglycemia, high triglycerides and fatty
liver also develop
• Hyperinsulinemia is observed, suggesting that insulin
resistance occurs in peripheral tissues.
• Altered signaling in adipose tissues: decreased
adiponectin (a adipocytokine) and increased MCP-1
(monocyte chemoattractant protein) and TNF-a which
induce insulin resistance.
• Study looked at mechanism of coffee’s action against
type-2 diabetes and the effect of caffeine alone on
development of hyperglycemia.
Experiments
• Expt 1: Diabetic mice were given diluted coffee vs.
plain drinking water for 5 weeks ad libitum
• Blood drawn and blood glucose measured weekly
• Insulin tolerance test given at 5 weeks
• At study end, mice were sacrificed and tested for
serum insulin, triglycerides, total cholesterol, and proinflammatory markers MCP-1 and IL-6.
• Liver and adipose tissues were removed and examined
for cholesterol, TG and phospholipid content.
• Expt 2: Same study was repeated with caffeine solution
(250ug/mL) vs. plain drinking water
Coffee treatment lowered blood glucose and
insulin tolerance in the mouse model
Coffee treatment significantly reduced liver weight, most types of body
fat, liver lipid content, insulin and TAG levels, and inflammatory markers
Conclusions
• Coffee improved insulin sensitivity and
decreased hyperglycemia (high blood sugar)
• Expression of inflammatory cytokines such as
MCP-1, IL-6 and TNFa in adipose tissue was
lower in the coffee treatment group.
• Coffee decreased adipose tissue inflammation,
thus improving insulin resistance.
• Caffeine alone also reduced hyperglycemia,
inflammation in some but not all fat tissues.
• Both coffee and caffeine improved fatty liver.