Antibiotics and

Chemiotherapeutics
Antibiotics - Still Miracle
Drugs

Paul Ehrlichs Magic Bullets

Salvarsan No. 606

Selective Toxicity
Drugs that specifically target microbial processes, and
not the human host cellular processes

Antibiotics

Antibiotics - antimicrobials of microbial origins, most of the

produced by fungi or bacteria (Streptomyces)
Antimicrobial, antimicrobic- any suspastance with
sufficient antimicrobial activity, used in treatment of
inf.dis
Bactericidal that kill bacteria
Bacteriostatic inhibit growth, not kill bacteria
Chemotherapeutics encompasses antibiotics,
antimicrobial, drugs used in the teratment of cancer; not
natural, created in lab.
Semisynthetic drugs antibiotics that are chemicaly
modified
Narrow spectrum / broad spectrum

Selective Toxicity
Mechanisms and sites
Mechanism of action
1. Bacterial cell wall
2. Nucleic acid synthesis
3. Protein synthesis
4. Cell membrane
5. Folic acid synthesisc

tigecycline

daptomycin

Most antibiotics have 2 names, the trade or brand name,

Svaki
lek poseduje dva imena:
created by the drug company that manufactures the drug, and a
Generiko
univerzalno
u svetu
(pr.amoxicillin)
generic name, based
on the antibiotic's
chemical
structure or
Zatieno,
fabrika
chemical class.koje
Tradedaje
names
such askoja
Keflexpravi
and Zithromax are
capitalized. Generics such as cephalexin and azithromycin are
not capitalized.

1.

Inhibition of Cell wall synthesis

Bacteria cell wall unique in
construction
Contains peptidoglycan
Antimicrobials that interfere with the
synthesis of cell wall do not interfere
with eukaryotic cell
Due to the lack of cell wall in
animal cells and differences in cell
wall in plant cells
These drugs have very high therapeutic
index
Low toxicity with high effectiveness
Antimicrobials of this class include
lactam drugs
Vancomycin
Bacitracin
Isoniazide, ethambutol - TBC

Target- Cell Wall Synthesis

The bacterial cell wall is a cross linked polymer called
peptidoglycan which allows a bacteria to maintain its
shape despite the internal turgor pressure caused by
osmotic pressure differences.
If the peptidoglycan fails to crosslink the cell wall will lose
its strength which results in cell lysis.
All -lactams disrupt the synthesis of the bacterial cell
wall by interfering with the transpeptidase which
catalyzes the cross linking process.

Peptidoglycan
Peptidoglycan is a carbohydrate composed of alternating
units of NAMA and NAGA.
The NAMA units have a peptide side chain which can be
cross linked from the L-Lys residue to the terminal DAla-D-Ala link on a neighboring NAMA unit.
This is done directly in Gram (-) bacteria and via a
pentaglycine bridge on the L-lysine residue in Gram (+)
bacteria.

1.

Inhibition of Cell wall synthesis

Penicillins and cephalosporins
Part of group of drugs called
lactams
Have shared chemical
structure called -lactam ring

Competitively inhibits function

of penicillin-binding proteins
Inhibits peptide bridge
formation between glycan
molecules
This causes the cell wall to
develop weak points at the
growth sites and become
fragile.

1.

MECHANISMS OF ACTION OF
inhibitors of cell wall synthesis

The weakness in the cell wall

causes the cell to lyze.

1.

Beta lactam antibiotics

1.

Antibiotics that inhibit cell wall

synthesis
Beta lactam antibiotics (BLA) - 4 groups:
Penicillins
Cephalosporins
Carbapenems
Monobactams

1. Antibiotics that inhibit cell wall synthesis

Beta lactam antibiotics - Penicillins
Natural (penicillin G and V)
Semisynthetic (ampicillin, carbenicillin,
antistaphylococcal ..)
Structure
Thiazolidine ring
Beta-lactam ring
Variable side chain)

Antibiotics that inhibit cell wall synthesis

Chemical structure of penicillins

The R group is responsible for

the activity of the drug, and
cleavage of the beta-lactam
ring will render the drug
inactive.

Beta lactam ring

Beta lactam antibiotics 1.

cephasloposrins
The cephalosporins broad spectrum
Chemical structures make them resistant to
inactivation by certain -lactamases
Tend to have low affinity to penicillin-binding proteins
of Gram + bacteria, therefore, are most effective
against Gram bacteria.
Chemically modified to produce family of related
compounds
First, second, third, fourth and fifth generation
cephalosporins

1.

Antibiotics that inhibit cell wall synthesis

Beta lactam antibiotics -Carbapenems

Imipenem and others

Activity broad spectrum
Resistant to most beta lactamasis
Indication therapy of very serious infetions caused by
multiploresistant bacteria

1.

Bacteria can produce enzyme

penicillinases (beta Lactamases) that
can destroy BLA

1.

Antibiotics that inhibit cell wall

Vancomycin
Inhibits formation of glycan chains
Inhibits formation of peptidoglycans and cell wall construction
Does not cross lipid membrane of Gram Gram - organisms innately resistant

Important in treating infections caused by penicillin resistant Gram +

organisms
Must be given intravenously due to poor absorption from intestinal
tract

1.

Antibiotics that inhibit cell wall

Bacitracin inhibits first phases in cell
wall synthesis bactericidal
Toxicity limits use to topical applications
active against Gram +
Isoniazid for TBC treatment
Ethambutol - for TBC treatment

2. Injury to the Plasma Membrane

Polymyxin B, Colistin (Gram negatives)
Topical
Combined with bacitracin and neomycin
(broad spectrum) in over-the-counter
preparation

New - daptomycin

2. Polymyxin: Antibacterial activity

However, the polymyxins are only active against

gram negative bacteria (P. aeruginosa, E. coli, K.
pneumoniae), while daptomycin is used to treat
gram positive bacteria
The polymyxins are highly nephrotoxic and are
thus only used topically

2. Polymyxins: Mechanism of action

Bind the the lipopolysaccharide in the outer

membrane, thus destroying OM integrity.
Bind to the cytoplasmic membrane (to the
phosphatidylethanolamine) and make the membrane
more permeable.

3. Inhibition
of protein
synthesis

3. Inhibition of protein synthesis

Inhibition of protein synthesis
Structure of prokaryotic ribosome acts as target for many
antimicrobials of this class
Differences in prokaryotic and eukaryotic ribosomes responsible for
selective toxicity

Drugs of this class include

Aminoglycosides
Tetracyclins
Macrolids
Chloramphenicol
Oxazolidines - lincosamide

3. Sites of inhibition on the procaryotic ribosome

3. Inhibition of Protein synthesis

Aminoglycosides

Aminoglycosides

Irreversibly binds to 30S

ribosomal subunit

Causes distortion and

malfunction of ribosome
Blocks initiation translation
Causes misreading of
mRNA

Not effective against

anaerobes, enterococci and
streptococci
Often used in synergistic
combination with -lactam
drugs
Allows aminoglycosides to
enter cells that are often
resistant

3. Inhibition of Protein synthesis

Aminoglycosides

Examples of aminoglycosides
include
Gentamicin, streptomycin
and tobramycin

Side effects with extended

use include
Otto toxicity
Nephrotoxicity

3. Inhibition of Protein synthesis

Tetracyclins

Tetracyclins

Reversibly bind 30S ribosomal subunit

Blocks attachment of tRNA to ribosome
Prevents continuation of protein synthesis
Effective against certain Gram + and Gram Newer tetracyclines such as doxycycline have longer half-life
Allows for less frequent dosing
Resistance due to decreased accumulation by bacterial cells
Can cause discoloration of teeth if taken as young child

3. Inhibition of Protein synthesis

Tetracycline
Broad spectrum and low cost
Commonly used to treat sexually transmitted
diseases and intracellular bacteria
Minor side effect gastrointestinal disruption
Doxycicline

Tigecycline new tetracycline

3. Inhibition of Protein synthesis

Macrolides - Erythromycin
Macrolids
Reversibly binds to 50S
ribosome
Prevents continuation
of protein synthesis
Effective against variety of
Gram + organisms and
those responsible for
atypical pneumonia
Often drug of choice for
patients allergic to
penicillin
Macrolids include
Erythromycin,
clarithromycin and
azithromycin

3. Inhibition of Protein synthesis

Oxazolidines - Linezolid

Active against
resistant Gram
positive
bacteria

3. Inhibition of Protein synthesis

Chloramphenicol

Binds to 50S ribosomal subunit

Prevents peptide bonds from
forming and blocking proteins
synthesis
Bacteriostatic agent
Effective against a wide variety of
organisms
Generally used as drug of last
resort for life-threatening
infections
Rare but lethal side effect is
aplastic anemia

3. Mupirocin (90% Pseudomonic acid A)

Isolated from Pseudomonas fluorescens

Mupirocin works against Gram-positive bacteria only
Can be used to treat MRSA (although resistance is
rising)
Mupirocin inhibits bacterial isoleucyl-tRNA synthetase.

Products containing Mupirocin

4. Inhibitors of Nucleic Acid

Synthesis
Inhibition of nucleic acid synthesis
These include
Fluoroquinolones
Rifamycins

4. Inhibitors of Nucleic Acid Synthesis

Fluoroquinolones
Inhibit action of topoisomerase DNA gyrase
Topoisomerase maintains supercoiling of
DNA
Effective against Gram -; newer also against
Gram +
Examples include
Ciprofloxacin and ofloxacin
There are 4 generation of fluoroquinolones

Fluorohinoloni
Fluoroquinolones - Inhibits DNA gyrase

Inhibiraju enzimsku aktivnost dva predstavnika topoizomeraza:

DNK giraze (topoizomeraze II) i topoizomeraze IV (dovode do superuvrtanja DNK
lanca u hromozmu i plazmidima bakterija i odravaju DNK u stabilnoj formi)
Superuvrnutost DNK utie na njenu replikaciju i prepis gena
Posledica - inhibicija replikacije bakt. DNK i ev. degradacija DNK
DNK giraza konvertuje relaksiranu
dvolananu formu DNK u negativno
superuvrnutu (obrnuti smer
dvostrukog heliksa)
Aktivnost je u fazama:
1. Pozitivno superuvrtanje
2. See lance DNK
3. Provlai DNK kroz prolazni dvolanani
prekid i vri negativno superuvrtanje
energetski stabilna forma

4. Inhibitors of Nucleic Acid

Synthesis
Rifamycins
Block prokaryotic RNA polymerase
Block initiation of transcription

Rifampin most widely used rifamycins

Effective against many Gram + and some Gram - as well as members
of genus Mycobacterium
Primarily used to treat tuberculosis and Hansens disease (lepra) as
well as preventing meningitis after exposure to N. meningitidis

5. Inhibition of metabolic pathways folic

acid synthesis
Inhibition of metabolic pathways
Relatively few
Most useful are folate
inhibitors
Mode of actions to inhibit
the production of folic
acid
Antimicrobials in this class
include
Sulfonamides
Trimethoprim

Sulfonamides compete with PABA for the active

site on the enzyme.

The sulfonamide Sulfamethoxazole is commonly used

in combination with trimethoprim

Drugs used for treatment of TBC are

Antitubercular drugs (tuberculostatics)
Several compaunds with different mechanism of action
Treatment must be administered over a prolonged
period; and the use of several anti-TB drugs is required.

First line tuberculostatic drug (bactericidal, except

etanbutol)

Isoniazid, rifampicin, streptomycin, ethambutol, pyrazinamid

Very effective with low toxicity
Second line tuberculostatic drug
PAS, ethionamide, amikacin, kanamycin, fluorochinolones
Less effective and toxic

What is antibiotic resistance?

Antimicrobial Resistance
Intrinsic
Acquired - relative or complete
lack of effect of antimicrobial
against a previously susceptible
microbe

Mechanisms of Antibiotic
Resistance
Enzymatic destruction of drug
Alteration of drug's target site
Prevention of penetration of drug
Rapid ejection of the drug (efflux)

Mechanism of bacterial resistance

Mechanisms of bacterial resistance

1. Enzymatic destruction of drug:
Beta lactamasis inactivate beta lactam
antibiotics
Esterasis inactivates macrolides
Acetyltransferase, adenyltransferase
inactivate aminoglycosides

Produkcija beta

Beta Lactamases
Beta lactam
antibiotics

Porin
Peptidoglikan

PBP

Periplazmatski prostor

-lactamase

2. Mechanisms of bacterial resistance

Alteration of targets:
Alteration of penicilllin binding proteins
(PBPs) resistance to BLA
Alteration of ribosomes resistance to
macrolides, tetracyclines
Alteration of DNA gyrases resistance to
fluorochinolones

Binding to targets (PBP) (PVP)

PBP

 Lactam antibiotic

PVP

Vezivanje BLA za

High affinity
for PBP

Inactive PBP
Inhibition of peptidoglycan synthesis

High affinity to PBP

Cell wall destruction

Bacterial death

 Lactam antibiotic

Vezivanje BLA za ciljna mesta (PVP)

Low affinity to PBP

Active PBP
Synthesis of PG

3. Mechanisms of bacterial resistance

Decreaseses uptake in bacterial cells
A. Decreased infflux (Gram negative
bacteria, due to porins)
B. Increased efflux

Porins in Gram negative bacteria

Efflux pump protein transport both in Gram positive and Gram
negative bacteria i kod Gram negativnih
Outside

Protein
chanell

Efflux pump
Inside

Porini kod Gram negativnih bakterija i proteinski transportni sistem kod efluks
pumpe
antibiotic

Porins

outside

Efflux pump
inside

Antibiotic
Excreted
antibiotic

porins

outside

Efflux pump
inside

Mechanisms of Resistance in
Beta Lactam antibiotics
Altered target (Gram negative/positive)
Altered permeability (Gram negative)
Production of inactivating enzymes (Gram
negative/positive)

Beta lactamases
Over than 340 different beta laktamazses
They cleave beta lactam ring and destroy BLA

Gram negative bacteria

are main producers ; they
store these anzymes in
periplasmatic space

Bakterije iz
Enterobacteriaceae
(E.coli,
Klebsiella...)
i Pseudomonas
Bacteria
infamilije
Enterobacteriacea
family
(E.coli,
Klebsiella)
and
stvaraju brojne beta laktamaze
Pseudomonas are main
beta lactamases producers

Beta lactamases
Depending on group of beta lactam antibiotics that is their main
target beta lactamses colud be
penicilinases

cephalosporinases

carbapenemases

Due to emergence of new beta-lactamases, there was a need for

the synthesis of new drugs
Rusult: after a short time, bacteria sucseed to develop new, more
powerful beta lactamases
Eg.
Wide Spectrum Beta Lactamses (mid 1970s)
Extended spectrum beta lactamses (ESBLs) 1990
Metallo betalactamses
Carbapenemases

Antibiotic Resistance
Intermicrobial transfer of plasmids, transposons and chromosiomal DNA containing
resistance genes (R factors) occurs by conjugation, transformation,and transduction

Figure 20.20

Transfer of genetic material spreeding

bacterial resistance

What Factors Promote

Antimicrobial Resistance?
Exposure to sub-optimal levels of
antimicrobial
Inappropriate use
Exposure to microbes carrying
resistance genes

Inappropriate Antimicrobial Use

Prescription not taken correctly
Antibiotics for viral infections
Antibiotics sold without
medical supervision
Spread of resistant microbes
in hospitals due to lack of hygiene

Inappropriate Antimicrobial Use

Inadequate surveillance or defective
susceptibility assays
Poverty or war
Use of antibiotics in foods
Lack of quality control in manufacture or
outdated antimicrobial

Antibiotics in animal feeds

Antibiotics in Foods
Antibiotics are used in animal feeds and
sprayed on plants to prevent infection and
promote growth
Multi drug-resistant Salmonella typhi has
been found in 4 states in 18 people who
ate beef fed antibiotics

Antibiotic Drug and Host Interaction

Toxicity to organs
Allergic reactions
Suppress/alter microflora
Effective drugs

Tetracycline treatments
can cause teeth discoloration.

Disrupting the normal flora in the intestine can result in

superinfections.

Finding an effective drug for

treatment
Identify infectious agent
Perform sensitivity testing
Often the Minimum Inhibitory
Concentration (MIC) is determined

Proposals to Combat Antimicrobial

Resistance

Speed development of new antibiotics

Track resistance data nationwide
Restrict antimicrobial use
Direct observed dosing (TB)

Proposals to Combat Antimicrobial

Resistance
Use more narrow spectrum antibiotics
Use antimicrobial cocktails

MRSA nosocomial pathogen

till ~1996, since than it became community
acquired