Antimicrobial agents and their mode of action

Bereket Dessalegn (DVM, MSc)

Assistance professor of Veterinary Microbiology

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 Objectives

• After completing this lecture you should able to:

– Explain how each group of antimicrobial agents targets key
structures or metabolic pathways in bacteria
– List the mechanisms of resistance in bacteria

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Antimicrobial
• The term ‘antimicrobial’ refers to a
substance that inhibits or destroys
bacteria, parasites, viruses or fungi
• it also refers to medicines used to treat
or prevent infections caused by
bacteria, parasites, viruses or fungi.
Antimicrobials fall into four categories:
• Antibacterials (antibiotics)
• Antifungals
• Antivirals
• Antiparasitics.
.

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 Based on effects on bacterial populations, Antimicrobial drugs are divided into two classes:

 Bacteriostatic drugs Prevents the growth of bacteria (keeps the bacteria in the
stationary phase of growth) without directly causing bacterial death.

 They depend upon the immune system to kill and remove the bacteria.
o Examples: Tetracyclines, Sulfonamides and Chloramphenicol

 Bactericidal drugs  Kills the bacteria

 Important in animals with impaired immune systems

 Examples: Penicillin, Streptomycin, Cephalosporins, Polymyxin and

Neomycin. In high concentrations, erythromycin may be bactericidal

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 Mechanism of action of antimicrobial drugs

 Mechanism of action can be placed under the followings:

1. Inhibition of growth by anti-metabolites

2. Inhibition of cell wall synthesis
3. Inhibition of cell membrane function

4. Inhibition of protein synthesis

5. Inhibition of nucleic acid synthesis

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 Target sites of some antimicrobial agents
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 Interference with cell wall synthesis

• Antimicrobial agents that interfere with cell wall synthesis block

peptidoglycan synthesis and thus are active against growing bacteria

• Antimicrobial agents that interfere with cell wall synthesis are

bactericidal.
– Example, β-lactam antibiotics (beta-lactam antibiotics)
– They are a class of broad-spectrum antibiotics, consisting of all
antibiotic agents that contain a beta-lactam ring in their molecular
structures. This includes penicillin derivatives (penams),
cephalosporins (cephems), monobactams, and carbapenems.
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 Interference with cell wall synthesis
• Activity of β-lactam on Gram-negative bacteria
– In Gram-negative bacteria, β-lactam antimicrobials enter the cell through porin channels in
the outer membrane
– In susceptible cells, β-lactam molecules bind to penicillin binding proteins (PBPs) that are
enzymes required for cell wall synthesis
– The attachment of the β-lactam molecules to the PBPs, located on the surface of the
cytoplasmic membrane, blocks their function. This causes weakened or defective cell walls
and leads to cell lysis and death
• Activity of β-lactam on Gram-positive bacteria

– Since Gram-positive bacteria do not possess an outer membrane, beta-lactam

antimicrobials diffuse through the cell wall. The next steps are similar to those in Gram-
9
negative bacteria
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 Interference with cytoplasmic membrane

• Polymyxin molecules diffuse through the outer membrane and cell

wall of susceptible cells to the cytoplasmic membrane

• They bind to the cytoplasmic membrane and disrupt and destabilize

it

• This causes the cytoplasm to leak out of the cell resulting in cell
death

• Antimicrobial agents that interfere with the cytoplasmic membrane

are bactericidal 10

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 Interference with Protein synthesis by binding to the 30S
ribosomal subunit

• Tetracyclines (e.g. tetracycline, minocycline and doxycycline) bind to the 30S

subunit of the ribosome and block the attachment of tRNA

• Since new amino acids cannot be added to the growing protein chain,
synthesis of protein is inhibited

• The action of tetracyclines is bacteriostatic

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 Interference with Protein synthesis by binding to the 30S
ribosomal subunit

• Aminoglycosides (e.g. gentamicin, tobramycin, amikacin, and streptomycin) also bind

to the 30S ribosomal subunit and can block protein synthesis in two different ways
– First they can attach to the 30S subunit of the ribosome and prevent the 30S
subunit from attaching to the mRNA
– Second, the presence of the aminoglycoside on the ribosome may cause misreading
of the mRNA. This leads to the insertion of the wrong amino acid into the protein or
interference with the ability of amino acids to connect with one another

• These activities often occur simultaneously and the overall effect is bactericidal

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 Inhibition of Protein synthesis by binding to the
50S ribosomal subunit

• Macrolides (e.g. erythromycin, azithromycin and clarithromycin) and

lincosamides (e.g. clindamycin) attach to the 50S ribosomal subunit causing
termination of the growing protein chain and inhibition of protein synthesis
– They are primarily bacteriostatic

• Chloramphenicol also binds to the 50S subunit of the ribosome and interferes
with binding of amino acids to the growing protein
– Antimicrobial agents that inhibit protein synthesis in this manner are
bacteriostatic
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 Inhibition of Protein synthesis by Inhibition of
the 70S initiation complex

• Linezolid (an oxazolidinone) is a new potent inhibitor of protein

synthesis

• It binds to a site on the bacterial 23S ribosomal RNA of the 50S

subunit and prevents the formation of a functional 70S initiation
complex, which is necessary for bacterial protein synthesis

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 Interference with nucleic acid synthesis
• It is caused by two classes of drugs

• Fluoroquinolones (e.g. nalidixic acid, ciprofloxacin, levofloxacin and gemifloxacin)

interfere with DNA synthesis by blocking the enzyme DNA gyrase
– DNA gyrase helps to wind and unwind DNA during DNA replication

– The enzyme binds to the DNA and introduces double stranded breaks that
allow the DNA to unwind

• Fluoroquinolones bind to the DNA gyrase-DNA complex and allow the broken DNA
strands to be released into the cell, which leads to cell death

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 Interference with nucleic acid synthesis
• Trimethoprim acts on the folic acid synthesis pathway at a point after the sulfonamides

– Interfere with the synthesis of folic acid at different levels and prevent/inhibit the
growth of microorganisms.
– It inhibits the enzyme dihyrofolate reductase

• necessary for the synthesis of amino acids, hence necessary for the bacterial
protein synthesis

• Trimethoprim and sulfonamides can be used separately or together

– When used together they produce a sequential blocking of the folic acid synthesis
pathway and have a synergistic effect
– Both trimethoprim and the sulfonamides are bacteriostatic
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Antimicrobial resistance (AMR)?
• Antimicrobial resistance (AMR) occurs when bacteria, parasites, viruses or fungi
change to protect themselves from the effects of antimicrobial drugs designed to
destroy them
• This means previously effective antimicrobial drugs (e.g. antibiotics) used to treat or
prevent infections may no longer work

 Resistant microorganisms are able to withstand attack by antimicrobial drugs.

 Standard treatments become ineffective

 Infections persist
 The risk of spreading resistant microorganisms to others increases

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  Origin of antimicrobial resistance
 It can be:
I. Non-genetic
II. Genetic origin
I. Non - genetic origin
 Microorganisms may lose the specific target
structure for a drug for several generations and
thus become resistant.
 For example:  Penicillin susceptible
organisms may change to cell wall deficient
L forms during penicillin administration.
 Lacking cell walls leads to resistant to
cell wall-inhibitor drugs.

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 II. Genetic origin
 Most drug resistant microbes emerge as a result of genetic change
Two types:

a. Chromosomal resistance

This develops as a result of spontaneous mutation

 It occurs rarely, thus is an infrequent cause of drug resistance.

b. Extra chromosomal resistance

 Bacteria often contain extra chromosomal genetic elements called
plasmids. This genetic elements are frequently carries resistant
genes
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 Mechanisms of drug resistance
 Pathogens often become resistant to a certain drug by the following
ways:
I. Preventing the entrance of the drug

II. Pumping the drug out of the cell after it has entered ( efflux pump)
III. Enzymatic destruction of drug

IV. Alteration of target site

V. Using of alternative pathway to bypass the sequence inhibited by
the drug

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 Figure: Mechanisms of Antimicrobial resistance

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 I. Preventing the entrance of the drug

 Many Gram-negative bacteria are unaffected by penicillin

 M. tuberculosis can resist many drugs because of the high content
of mycolic acid in a complex lipid layer out side their peptidoglycan

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 II. By pumping the drug out of the cell after it has entered

 Some pathogens have plasma membrane translocases, often called

efflux pumps that expel the entered drugs (multidrug resistant pumps).

 Efflux pumps are transport proteins involved in the extrusion of toxic

substrates (including virtually all classes of clinically relevant
antibiotics) from within cells into the external environment

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 III. By production of an enzyme that destroys the drug

 For example, the hydrolysis of the β-lactam ring of penicillins by the enzyme
penicillinase (β-lactamase)

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 IV. By alteration of target site

 Resistance can arise when the target enzyme or cellular structure is modified so
that it is no longer a susceptible drug

For example:  The affinity of ribosomes for erythromycin and

chloramphenicol can be decreased by a change in the rRNA to which they
bind

 Enterococci become resistant by masking the terminal D-alanine-D-

alanine target site of vancomycin. This drastically reduces antibiotic
binding

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 V. By using an alternative pathway to bypass the sequence
inhibited by the agent

 For example, those bacteria resistant to sulfonamides adapt to

using preformed folic acid from their surroundings are
resistant to sulfonamides. However, other strains increase
their rate of folic acid production and thus counteract
sulphonamide inhibition
 Folic acid is an essential nutrient necessary for protein and
nucleic acid synthesis (DNA and RNA). It is synthesized by bacteria
from the substrate para-amino-benzoic acid (PABA)

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 Mechanism of drug resistance transfer

 Bacteria can share genes with each other in a process called horizontal gene transfer

 This can occur both between bacteria of the same species and between different
species

 Gene transfer results in genetic variation and is a large problem when it comes
to the spread of antibiotic resistance genes

 Antibiotic resistance genes can be transferred by the following mechanisms :

I. Transformation

II. Transduction

III. Conjugation- Most common mechanism

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 THANK YOU

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