Liquids that kill bacteria

e.g. phenol based too toxic for skin surfaces

 Topical (e.g. skin) e.g. iodine or 70% alcohol reduce bacterial load

 Selectively toxic for bacteria bactericidal (killing) bacteriostatic (growth inhibition)

minimize harm to patient

4

destroy structures
present in bacteria not present in host

Mechanism of action include:

Inhibition of cell wall synthesis Inhibition of protein synthesis Inhibition of nucleic acid synthesis Inhibition of metabolic pathways Interference with cell membrane integrity


vancomycin

Amphotericin

Aminoglycosides

Tetracyclines

Inhibitors of cell wall synthesis. Penicillins, cephalosporin, bacitracin, carbapenems and vancomycin. Inhibitors of Cell Membrane. Polyenes - Amphotericin B, nystatin, and condicidin. Imidazole - Miconazole, ketoconazole and clotrimazole. Polymixin E and B. Inhibitors of Protein Synthesis. Aminoglycosides - Streptomycin, gentamicin, neomycin and kanamycin. Tetracyclines - Chlortetracycline, oxytetracycline, doxycycline and minocycline. Erythromycin, lincomycin, chloramphenicol and clindamycin.
BC Yang

For lecture only

Inhibitors of metabolites

(Antimetabolites).
Sulfonamides - Sulfanilamide, sulfadiazine silver and sulfamethoxazole. Trimethoprim, ethambutol, isoniazid.

Inhibitors of nucleic acids (DNA/RNA

polymerase).
Quinolones - Nalidixic acid, norfloxacin and

ciprofloxacin.
Rifamycin and flucytosine. rifamycin

For lecture only

BC Yang

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

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.

The weakness in the cell wall causes the cell to lyze.

The weakness in the cell wall causes the cell to lyze. Penicillins and cephalosporins are considered bactericidal. Penicillins are more effective against Gram+ bacteria. This is because Gram + bacteria have penicillin binding proteins on their walls.

The cephalosporins

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 and fourth generation

cephalosporins

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 Acquired resistance most often due to alterations in side chain of NAM molecule
Prevents binding of vancomycin to NAM component of glycan

Bacitracin

Interferes with transport of peptidoglycan precursors across cytoplasmic membrane Toxicity limits use to topical applications Common ingredient in non-prescription first-aid ointments

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

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

Examples of aminoglycosides include

Gentamicin, streptomycin and tobramycin

Side effects with extended use include

Otto toxicity Nephrotoxicity

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

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

Resistance due to decreased accumulation by bacterial cells Can cause discoloration of teeth if taken as young child

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

Resistance can occur via modification of RNA target

Other mechanisms of resistance include production of enzyme that

chemically modifies drug as well as alterations that result in decreased uptake of drug

Chloramphenicol

Binds to 50S ribosomal subunit

Prevents peptide bonds from forming and blocking

proteins synthesis

Effective against a wide variety of organisms Generally used as drug of last resort for lifethreatening infections Rare but lethal side effect is aplastic anemia

Chloramphenicol

Binds to 50S ribosomal subunit

Prevents peptide bonds from forming and blocking proteins synthesis

Effective against a wide variety of organisms Generally used as drug of last resort for lifethreatening infections Rare but lethal side effect is aplastic anemia

Inhibition of nucleic acid synthesis

These include
Fluoroquinolones Rifamycins

Fluoroquinolones

Inhibit action of topoisomerase DNA gyrase

Topoisomerase maintains supercoiling of DNA

Effective against Gram + and Gram Examples include

Ciprofloxacin and ofloxacin

Resistance due to alteration of DNA gyrase

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 as well as preventing meningitis after exposure to N. meningitidis Resistance due to mutation coding RNA polymerase
Resistance develops rapidly

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

Group of related compounds

Collectively called sulfa drugs

Inhibit growth of Gram + and Gram - organisms

Through competitive inhibition of enzyme that aids in production of folic acid

Structurally similar to para-aminobenzoic acid

Substrate in folic acid pathway

Human cells lack specific enzyme in folic acid pathway

Basis for selective toxicity

Resistance due to plasmid

Plasmid codes for enzyme that has lower affinity to drug

Trimethoprim

Inhibits folic acid production

Interferes with activity of enzyme following enzyme

inhibited by sulfonamides

Often used synergistically with sulfonamide Most common mechanism of resistance is plasmid encoded alternative enzyme
Genes encoding resistant to sulfonamide and

trimethoprim are often carried on same plasmid

Interference with cell membrane integrity

Few damage cell membrane

Polymixn B most common
Common ingredient in first-aid skin ointments

Binds membrane of Gram cells

Alters permeability
Leads to leakage of cell and cell death

Also bind eukaryotic cells but to lesser extent

Limits use to topical application

Mechanisms of resistance

Drug inactivating enzymes

Some organisms produce enzymes that chemically modify drug
Penicillinase breaks -lactam ring of penicillin antibiotics

Alteration of target molecule

Minor structural changes in antibiotic target can prevent binding
Changes in ribosomal RNA prevent macrolids from binding to ribosomal subunits

Mechanisms of resistance

Decreased uptake of the drug

Alterations in porin proteins decrease permeability of cells
Prevents certain drugs from entering

Increased elimination of the drug

Some organisms produce efflux pumps
Increases overall capacity of organism to eliminate drug Enables organism to resist higher concentrations of drug Tetracycline resistance

Probably the most widely used testing method is the diskdiffusion method, also known as the KirbyBauer test.

Conventional disc diffusion method

Kirby-Bauer disc diffusion routinely used to qualitatively determine susceptibility Standard concentration of strain uniformly spread of standard media Discs impregnated with specific concentration of antibiotic placed on plate and incubated

Clear zone of inhibition around disc reflects susceptibility

Based on size of zone organism can be described as susceptible or resistant

Sometimes the chemotherapeutic effects of two drugs given simultaneously is greater than the effect of either given alone. This is called synergism. For example, penicillin and streptomycin in the treatment of bacterial endocarditis. Damage to bacterial cell walls by penicillin makes it easier for streptomycin to enter.

Other combinations of drugs can be antagonistic. For example, the simultaneous use of penicillin and tetracycline is often less effective than when wither drugs is used alone. By stopping the growth of the bacteria, the bacteriostatic drug tetracycline interferes with the action of penicillin, which requires bacterial growth.

Combinations of antimicrobial drugs should be used only for:

1. 2. 3.

To prevent or minimize the emergence of resistant strains. To take advantage of the synergistic effect. To lessen the toxicity of individual drugs.