Antibiotics: Introduction to

Classification
Dr. Bhoj R Singh, Principal Scientist (VM)
Head of Division of Epidemiology
Indian Veterinary Research Institute, Izatnagar-243122, Bareilly, UP, India.
 WHY WE STUDY ANTIMICROBIALS?
IT IS TO UNDERSTAND :
A. SELECTIVITY
Antibiotics have selective toxicity toward the bacterium rather than the host.
Disinfectants lack selectivity.
Selectivity varies
Higher the selectivity lower the toxicity.
B. THERAPEUTIC INDEX
The ratio of the dose toxic to the host to the effective therapeutic dose. The higher the
therapeutic index the better the antibiotic.
C. CATEGORIES OF ANTIBIOTICS
Bactericidal . Bacteriostatic (reversible) Bactericidal antibiotics are preferred
Bacteriostatic antibiotic is used the duration of therapy must be sufficient to allow cellular and humoral defence
mechanisms to eradicate the bacteria.
Serious infections should be treated with bactericidal antibiotics for prompt eradication of the organisms.
D. ANTIBIOTIC SUSCEPTIBILITY TESTING
Minimum inhibitory concentration (MIC)
Minimum bactericidal concentration (MBC).
The MBC is the lowest concentration of the antibiotic that kills 99.9% of the inoculum in a given
time.
E. COMBINATION THERAPY
To prevent the emergence of resistant strains
• To treat emergency cases during the period when an etiological diagnosis is still in progress
• To take advantage of antibiotic synergism.
• Synergism: effects of a combination is greater than the sum of the effects of the individual
antibiotics.
• Antagonism : One interferes with the effects of another antibiotic.
F. ANTIBIOTICS AND CHEMOTHERAPEUTIC AGENTS
Antibiotic strictly refers to substances of biological origin.
Chemotherapeutic agent is a synthetic chemical.
CLASSIFICATION OF ANTIMICROBIALS
BASED ON CHEMICAL NATURE/
STRUCTURE
Β-Lactams
Penicillins
Cephalosporins
Monobactams
Carbapenems
Quinolones and fluroquinolones
Sulfonamides and trimethoprim
Glycopeptides
Phosphonic acids
Lipopetides
Peptide antibiotics
Ionophores
Β-LACTAM ANTIBIOTICS (HAVING LACTAM RING)
OTHER GROUPS OF CHEMICALLY DIFFERENT ANTIBIOTICS
CLASSIFICATION BASED ON MECHANISM
OF ACTION
• Affecting Protein Synthesis
• Affecting on Nucleic acid
synthesis
• Antimetabolite antimicrobials
• Acting on Cell wall
• Acting on Cell membrane
 PROTEIN SYNTHESIS AND SITE OF ACTION
OF ANTIMICROBIALS THAT INHIBIT PROTEIN
SYNTHESIS
A. Interfering with initiation of protein synthesis
Antimicrobials that bind to the 30s ribosomal subunit
– Irreversibly bind to the 30S ribosome and freeze the 30S initiation
complex (30S-mRNA-tRNA)-Aminoglycosides (Bactericidal)
– Reversibly bind to the 30S ribosome and inhibit binding of aminoacyl-t-
RNA to the acceptor site on the 70S ribosome. Tetracyclines
(Bacteriostatic).
– Reversibly interferes with mRNA interaction with the 30S ribosome
without causing misreading of mRNA unlike aminoglycosides.
Spectinomycin (Bacteriostatic)
Antimicrobials that bind to the 50s ribosomal subunit
– Bind to the 50S ribosome to inhibit peptidyl transferase activity.
Chloramphenicol, lincomycin, clindamycin (bacteriostatic)
– Inhibit translocation of the peptidyl tRNA from the A to the P site on the
ribosome by binding to the 50S ribosomal 23S RNA. Macrolides
(bacteriostatic)
B. Affecting peptide elongation
Binds to elongation factor G (EF-G) and inhibits release of EF-G from
the EF-G/GDP complex. Fusidic acid (bacteriostatic)
 INHIBITORS OF NUCLEIC ACID SYNTHESIS

A. Inhibitors of RNA synthesis: Bind to DNA-dependent RNA

polymerase and inhibit initiation of RNA synthesis. Rifampin,
rifamycin, rifampicin (bactericidal)
B. Inhibitors of DNA synthesis: Bind to the A subunit of DNA
gyrase (topoisomerase) and prevent supercoiling of DNA,
thereby inhibiting DNA synthesis. Quinolones,
fluoroquinolones, oxolinic acid (bactericidal).
C. Agents That Impair the Template Function of DNA. None of
them is therapeutically useful; however, chloroquine and
miracil D (lucanthone) inhibit plasmodia and schistosomes,
respectively by intercalating into the DNA and thereby to
inhibit further nucleic acid synthesis. Acridine dyes such as
proflavine act by intercalation mechanism, but because they
are toxicity and carcinogenicity in mammals they are not
used as antibacterial agents.
 ANTIMETABOLITE ANTIMICROBIALS

• Inhibitors of folic acid synthesis

– Analogues of para-aminobenzoic acid
competitively inhibiting formation of
dihydropteric acid. Sulfonamides, Sulfones,
Para-aminosalicylic acid (PSA),
Depsone (bacteriostatic).
– Bind to dihydrofolate reductase and inhibit
formation of tetrahydrofolic acid.
Trimethoprim, methotrexate,
pyrimethamine (bacteriostatic).
 ACTING ON CELL WALL
• Inhibit synthesis of mycolic acids. Isoniazid (bacteriostatic).
• Inhibition of Bacterial Cell Wall Synthesis: Peptidoglycan
synthesis occurs in three stages.
The first stage takes place in the cytoplasm, where the low-
molecular-weight precursors UDP-GlcAc and UDP-MurNAc-L-Ala-D-
Glu-meso-Dap-D-Ala-D-Ala are synthesized. Many antibiotics affect
this stage.
– UTP and N-acetylglucosamine α-1-P are converted to UDP-N-
acetylglucosamine, which is subsequently converted by the
enzyme phosphoenolpyruvate: UDP-GIcNAc-3-enol-
pyruvyltransferase. Fosfomycins block this transfer by a direct
nucleophilic attack on the enzyme.
– Three amino acids are added to the muramyl peptide to yield a
tripeptide. The dipeptide D-alanyl-D-alanine is synthesized from
two molecules of D-alanine by the enzyme D-alanyl-D-alanine
synthetase. D-Alanine is produced from L-alanine by an alanine
racemase. Cycloserine inhibits both alanine racemase and D-
alanyl-D-alanine synthetase, owing to the structural similarity
cycloserine binds to the enzymes better than the D-alanine.
 AFFECTING THE 2ND AND 3RD STAGE OF
CELL WALL SYNTHESIS
• 2nd stage is catalyzed by membrane-bound enzymes. The precursor
molecules are transferred sequentially to a carrier in the cytoplasmic
membrane. This carrier is a phosphorylated undecaprenyl alcohol. The lipid
carrier functions as a point of attachment to the membrane for the precursors
and allows for transport of the subunits across the hydrophobic interior of the
cytoplasmic membrane to the outside surface. Bacitracin, a peptide antibiotic,
specifically interacts with the pyrophosphate derivate of the undecaprenyl
alcohol, preventing further transfer of the muramylpentapeptide from the
precursor nucleotide to the nascent peptidoglycan.
• The third stage of cell wall synthesis involves polymerization and the
attachment of nascent peptidoglycan to the cell wall. Polymerization occurs by
transfer of the new peptidoglycan chain from its carrier in the membrane to the
nonreducing N-acetylglucosamine of the new saccharide-peptide that is
attached to the membrane. The new peptidoglycan is attached to preexisting
cell wall peptidoglycan by a transpeptidase reaction D-alanyl-D-alanine
terminus of two peptides. Transpeptidase enzyme cleaves the peptide bond
between two D-alanyl residues in the pentapeptide and become acylated via
the carbonyl group of the penultimate D-alanine residue. This final reaction can
be inhibited by β-lactam antibiotics. These antibiotics undergoes an acylation
reaction with the transpeptidases that cross-link the peptide polymers.
Penicillins (penams), Cephalosporins (oxacephems and cephamycins),
Penems, Thienamycins (carbapenems), and Aztreonam (monobactams)
 VANCOMYCIN, ALSO INTERFERES
WITH CELL WALL SYNTHESIS
• Vancomycin interrupts cell wall synthesis by forming a
complex with the C-terminal D-alanine residues of
peptidoglycan precursors.
• Complex formation at the outer surface of the cytoplasmic
membrane prevents the transfer of the precursors from a lipid
carrier to the growing peptidoglycan wall by transglycosidases.
• Biochemical reactions in the cell wall catalyzed by
transpeptidases and D,D-carboxypeptidases are also inhibited
by vancomycin and other glycopeptide antimicrobials.
• Because of its large size and complex structure, vancomycin
does not penetrate the outer membrane of gram-negative
organisms thus active only on GPBs.
 ACTING ON CELL MEMBRANE
• Permeabilizes cell membranes for sodium and potassium ions: Ionophore
antibiotics. Valinomycin permeabilizes membranes for K+ of both prokaryotic
and eukaryotic cells for potassium and therefore cannot be used for
antimicrobial chemotherapy. However, Monensin (in cattle) and salinomycin (in
pigs) are used exclusively in veterinary practice can inhibit bacteria, protozoa
(coccidia) and metazoan parasites.
• Binds to the cytoplasmic membrane and then forms oligomeric pores viz.,
permeabilization of liposomes by Lipopeptide antibiotics. Daptomycin
permeabilizes liposomes only when they contain phosphatidylglycerol (PG)
thus active on GPBs, outer membrane of GNBs lacking PG interferes its activity.
• Binding to LPS to disrupts outer membrane, Cyclopeptide antibiotics,
polymyxin B and E (colistin). LPS contains several negative charges interacting
with positively charged polymyxins, besides several hydrophobic interactions
between the two molecules also disrupts outer membranes. Amino groups in
polymyxin B pairs with the phosphates of lipid A in LPS.
• Quasi-ionophore antibiotics that include channel-forming agents such as
gramicidin and the polyene antibiotics. The polyene antibiotics, which act by
binding to membrane sterols, contain a rigid hydrophobic center and a flexible
hydrophilic section. They interact with fungal cells to produce a membrane-
polyene complex that alters the membrane permeability, resulting in internal
acidification of the fungus with exchange of K+ and sugars; loss of phosphate
esters, organic acids, nucleotides; and eventual leakage of cell protein.
• Interfering with the synthesis of lipid membranes. Imidazoles: miconazole,
ketoconazole, clotrimazole, and fluconazole. These compounds inhibit the
incorporation of subunits into ergosterol and may also directly damage the
fungal cell membrane.
 RESISTANCE
Clinical Resistance: The MIC of the drug for a particular strain of bacteria
exceeds threshold of safety in vivo. It is due to:
– By mutation in the gene that determines sensitivity/resistance to the agent
– By acquisition of extrachromosomal DNA (plasmid) carrying a resistance gene.
Cross Resistance: A single mechanism confers resistance to multiple
antimicrobial agents, commonly seen with closely related antimicrobial
agents.
Multiple Resistance: It implies to multiple mechanisms involved for resistance
to one or more antibiotics, seen with unrelated antimicrobial agents.
MECHANISMS OF RESISTANCE
1. Altered permeability of the antimicrobial agent
Altered permeability may be due to the inability of the antimicrobial agent to
enter the bacterial cell or alternatively to the active export of the agent from
the cell.
2. Inactivation of the antimicrobial agent
Resistance is often the result of the production of an enzyme that is capable
of inactivating the antimicrobial agent.
3. Altered target site
Resistance can arise due to alteration of the target site for the antimicrobial
agent.
4. Replacement of a sensitive pathway
Resistance can result from the acquisition of a new enzyme to replace the
sensitive one.