Bratton, Heather Bui, Steven

Bautista, Candie Chen, John

Chem 444
Chan, Hiu Fai Doan, Chad Ho, Joyce Karimbabai Massihi, Anna Krasner, Danielle Loi, William Chida, Odette Cueva, Carla Golanbar, Gelareh

Biol 444

Kashiwabara, Claudine Kopilec, Jaimisyn Lau, Jensen

Malette, Jacqueline
Nguyen, Trang Robles, Marisa Tallorin, Lorillee Truong, Daniel C Udeh, Francis Villalvazo, Adrianne Yeh, Enrenn

Le, Giang-Tuong
Lee, Margaret Mata, Francisca Nguyen, Thuy Nhi Phillips, Irving Romero, Marcelo Wang, Tony Wittig, Michelle

Primary (1) Metabolism

- Construct common biological macromolecules from simple building blocks found within every cell - Typically a process of polymerization, stringing monomers together into a macromolecule that performs a cellular function sugars amino acids fats polysaccharides proteins phospholipid bilayers

- Block production: cell dies (primary metabolites are essential)

Secondary (2) Metabolism

- Synthesize compounds that are unique to a particular species or genus (unlike common proteins, lipids, etc.) - Molecules may have extremely complex structures - These molecules typically have no effect on the producing organism, but are often highly biologically active against other organisms (competitors, pathogens, predators) - Often present at extraordinary concentrations, >10% of the dry weight of the organism inference is, they must do something

Natural Products: Folk to Modern Medicines

- Humans have long used chemicals in plant and animal extracts for many purposes: medicines, poisons, recreational stimulants - Since the 1800s, chemists have characterized and synthesized such natural products, uncovering the basis for folk remedies

- Identifying the mechanism of action of natural products was the genesis of modern drug discovery - The active ingredients in many ancient curatives are still used medicinally today

Natural Products 1: Toxins

Rotenone

- Natural fish + insect poison

Curare (Tubocurarine chloride) - Derived from 2 South American plant genera

- Used as arrow-tip poison by native peoples - Blocks transmission of acetylcholine signal to muscles, causing instant paralysis - 1st drug used as muscle relaxant in surgery

Natural Products 1: Toxins

Hyoscine (= scopolamine)

- Derived from plants: deadly nightshade (belladonna), mandrake

- In ancient Greece, mandrake was used as an anaesthetic (or poison, in higher doses)

- Witches smeared extracts on their armpits: avoided toxic oral route, got maximum hallucinogenic effect = felt like they were flying - Blocks certain acetylcholine receptors - Modern use: prevention of motion sickness (patches behind the ear)

Natural Products 2: Medicines

COCH3

Salicin

Acetyl-salicilic acid: aspirin

- From Willow tree bark, which was used in folk remedies for treating fevers
- Led to synthetic analogue, modern aspirin

Natural Products 2: Medicines

Ventolin Ephedrine
- From Ephedra plants, basis of ancient Chinese herbal remedy Ma Huang (100 AD) for treating respiratory illness - Used clinically since 1926 as bronchodilator to treat asthma

- Similar to adrenaline, but also stimulates heart (not good in a drug) - Stimulated research resulting in the non-stimulant drug Ventolin

Natural Products 2: Medicines

Quinine

- Found exclusively in the bark of the Chinona tree - Used as a malaria treatment since 1600s - Now synthetic derivatives used, due to widespread resistance

Natural Products 3: Stimulants

Caffeine

Nicotine

- Caffeine-containing plant leaves + seeds have long been brewed to produce stimulant drinks - Such stimulants naturally act as feeding deterrents to repel insect herbivores that consume plants leaves - Tobacco plant can up its production of nicotine 4-fold when under attack by insects; nicotine by-products are used as insecticides

Natural Products 3: Stimulants

Cocaine
- Coca leaves have been used as a source of cocaine for > 2,000 yrs - Used by Incas in religious ceremonies - Introduced to Europe by conquistadores - Leaves chewed daily by >8 million native peoples in the Andes, to alleviate feelings of hunger and fatigue - Inhibits re-uptake of excitatory neurotransmitter dopamine

Natural Products 4: Halucinogens

Morphine (= opium)

Heroin (synthetic derivative)

- Found in only 2 species of poppy flower (Papaver)

- Milky exudate of seed capsules is 25% opiate - Used as a baby calming treatment in ancient Egypt - Binds to brain receptors for short peptides called enkephalins, derived from endorphins

Natural Products 4: Halucinogens

Similar spatial relationships of (*)-marked atoms responsible for similar pharmacological effects

Enkephalin Morphine

* *
*

Natural Products 4: Halucinogens

Tetrahydrocannabinol

Lysergic acid

- Lysergic acid is the parent compound from which ergot alkaloids are derived, such as LSD - Produced by fungus; often affected stored grain in Middle Ages - Such compounds found in Aztec magical preparation ololuiqui - Structural mimics of human neurotransmitter 5-hydroxytryptamine

Lysergic acid Diethylamine (LSD, synthetic)

Secondary Metabolites
- Produced from a small number of key intermediates, often generated as by-products of primary metabolism acetate (in the form of acetyl coA) mevalonate 4-carbon sugars - Perhaps arose as a means of dealing with excess metabolic intermediary compounds -2o metabolites then took on ecological roles as toxins, etc.

Secondary Metabolites
- Typically play ecological roles in nature, deterring would-be pests, predators or pathogens - Affect humans due to structural resemblance to innate neurotransmitters, or by binding to proteins in a way that disrupts normal cellular function - Natural products are the basis for a big % of pharmaceutical drugs currently on the market -

Simple building blocks serve as the basis for each major pathway of secondary metabolism: (1) Shikimate Aromatics (ring - C3 chain)

(2)

Amino acids

Alkaloids, Penicillins (N-containing) Terpenes, Steroids

(3)

Mevalonate

(4)

Acetate

Polyketides (aromatics, macrocycles)

(photosynthesis)

Polysaccharides Glycosides Nucleic Acids

phosphoenol pyruvate

Shikimate pathway
Shikimate Aromatic Compounds Lignans

(1)

pyruvate

aromatic amino acids aliphatic amino acids

CITRIC ACID CYCLE
CH3COSCoA

Alkaloids (2)
Peptides Penicillins Cyclic Peptides

acetyl CoA CH3COSCoA

-O

2CCH2COSCoA

CH3COCH2COSCoA
CH3COSCoA

CH3COSCoA

Isoprenoids (terpenes, steroids,

mevalonate

(3)

carotenoids)

(4)

Polyketides, Fatty Acids

Prostaglandins, Macrocyclic Antibiotics

(1) Shikimate pathway

from the Japanese flower shikimi (, Illicium anisatum) (A) Biosynthesis of aromatic amino acids (tyrosine, phenylalanine, and tryptophan) lead to alkaloids (B) Build aromatic secondary metabolites

Shikimate
Podophyllotoxin

Shikimate biosynthesis
Shikimate + PEP Chorismate

Prephenate

Other shikimate metabolites: ring-C3 ...ring-C2 ...ring-C1 Cinnamic acid - NH3

+ NH3

Tyrosine, Phenylalanine

Biosynthesis of phenyl compounds

ring-C1 type

vanillin

vanillic acid

salicin

salicylic acid

- In plants, many shikimate metabolites are allelopathic: they inhibit growth of competitors - Insoluble forms often linked to sugars in plant tissue - Soluble acidic forms leach out into surrounding soil in rain

phenylalanine

Lignans & Lignins

Cinnamic acid

Polymerization: complex lignins

Enzymatic coupling: dimeric lignans Cinnamyl alcohol

Podophyllotoxin

Large % of woody plant biomass

Podophyllotoxin
- An important shikimate compound - Used by native Americans to cure warts - Powerful inhibitor of mitosis; found to block enzyme tubulin polymerase
-

Flavanoid Biosynthesis
Shikimate + 3 acetates = flavanones

Cause bitter tastes in plants, especially polymerized tannins; deter feeding by herbivores

Derived compounds are responsible for much of plant color

(2) Alkaloids
(2) Amino acids Alkaloids, Peptides, Penicillins

Penicillin

Strychnine (alkaloid) Cyclosporin A (cyclic peptide)

Alkaloid characteristics
- Non-(normal)-peptide, non-nucleic acid compounds that contain nitrogen - Common in fungi, plants, insects + amphibians

- Derived from amino acid precursors

Tyramine

Tyrosine Dopamine

Mescaline - potent hallucinogen from the peyote cactus - competitively binds to dopamine receptors

Penicillin Biosynthesis
a-adipate + cysteine + valine

- start with peptide made of 3 amino acids (including a non-standard a.a., a-adipate)

a-adipate + cysteine + valine

isopenicillin

synthase

penicillin N

epimerase

isopenicillin N

cephalosporins

penicillins

Penicillins: Mechanism of Action

This class of antibiotics interferes with synthesis of the cell wall of Gram-positive bacteria (Staphylococci, Streptococci) Cell wall is a repeating polymer of disaccharide, tetrapeptide repeats cross-linked into a 3D matrix (1) cleave here (transpeptidase) sugar - sugar - phospholipid
L-ala

D-glu L-lys D-ala D-ala (glycine)5 (2) cross-link here

Penicillins: Mechanism of Action

Penicillins inhibit the bacterial transpeptidase enzyme by mimicking its natural substrate, the terminal D-alaD-ala Transpeptidase attacks the b-lactam ring of penicillin, forms a covalent bond; enzyme is now out of business

(1) NO CLASS next Monday (2) For NEXT WEDNESDAY - bring to class a small amount of some spice, seeds, leaves, fruit, or medicinal herb - pick something with a strong taste/smell, or that you know is used in a folk remedy - at Wednesdays lecture, you will put your material in a tube and label it; I will then add solvent and extract the natural products from it over the weekend - in the next lab, you will bioassay the extracts of your material for antibiotic activity and cytotoxicity