ANTIBIOTICS

The term antibiotic was derived from the word antibiosis coined in 1889 by Paul Vuillemin
which means against life. (Anti = against bios = life)
Waksman defined antibiotics as chemical substances produced by various species of
microorganisms such as bacteria & fungi, which has the capacity to inhibit or destroy the other
microorganisms in low concentrations.
An antibiotic is a naturally occurring (produced by microorganisms), semi synthetic or
synthetic type of agent (a structural analogue of naturally occurring antibiotic) that destroys or
inhibits growth of other microorganisms used in the treatment of external & internal infections.
Historical background:
Centuries earlier, humans had learned to use crude preparations empirically for the topical
treatment of infections. In 500 to 600 BC, molded curd of soybean was used in Chinese folk
medicine to treat boils. Moldy cheese had also been used by Chinese and Ukrainian peasants
to treat infected wounds.
In 1877, Pasteur and Joubert discovered that anthrax bacilli were killed when grown in culture
in the presence of certain bacteria, along with similar observations by other microbiologists,
led Vuillemin to define antibiosis (literally “against life”).
In 1928 SIR ALEXANDER FLEMING, a British bacteriologist discovered penicillin from the
fungus, Penicillium notatum. He observed that colonies of S. aureus failed to grow in the areas
contaminated by Penicillium notatum. He isolated the mould, grew it in a fluid medium and
found that it produced a substance capable of killing many of the common bacteria that infect
humans. He coined the term PENICILLIN. It was unstable and he was unable to purify.
In June 1938 FLOREY & CHAIN introduced penicillin into therapy by adapting Freeze drying
& Chromatography technique to isolate penicillin & shared the Noble prize with Fleming.
In his Nobel Prize acceptance speech, Fleming warned the world of the dangers of misusing
antibiotics. He had already noted bacteria in his lab becoming resistant to penicillin, just a few
years after its discovery. After decades of antibiotic misuse, today we find ourselves facing
bacteria which has become resistant to most, if not all antibiotics.
Based on the year of discovery some of the natural antibiotics are listed below
1928- Penicillin
1940-Streptomycin
1947-Chloramphenicol
1948-Chlortetracycline
1952- Erythromycin
1956- Vancomycin
1960-Gentamicin
Ideal characteristics of antibiotics:
1. Kill or inhibit the growth of different species of pathogens
2. Selectively toxic to the microbe but nontoxic to host cell
3. Cause no allergic reaction to the host
4. Should be chemically stable to isolate, processing and storage for reasonable period of
time.
5. Remain in specific tissues in the body long enough to be effective i.e the rate of
biotransformation and elimination should be slow.
6. Kill the pathogens before they mutate and become resistant to it
7. Relatively soluble, function even when highly diluted in body fluid.
8. Reasonably priced.
Classification of Antibiotics:
Antibiotics are classified based on structure, spectrum, source & pharmacological activity.
I Classification based on structure:
1. Beta lactam Antibiotics: Penicillin, Cephalosporin, Beta lactamase inhibitors &
Monolactams
2. Amino glycosides: Streptomycin, Neomycin and Kanamycin.
3. Tetracyclines: Chlortetracycline, Tetracycline, Oxytetracycline, doxycycline,
minocycline
4. Macrolides: Erythromycin, Clarithromycin and Azithromycin
5. Miscellaneous: Chloramphenicol* & Clindamycin
II Classification based on spectrum:
1. Narrow spectrum antibiotics: The term refers to an antibiotic that acts against a
narrow range of disease-causing bacteria. Ex: Benzyl penicillin, Bacitracin, Cloxacillin,
Vancomycin, Fluxacillin
2. Broad spectrum antibiotics: The term broad-spectrum antibiotic refers to an antibiotic
that acts against a wide range of disease-causing bacteria. Ex: Chloramphenicol,
Streptomycin, Amoxicillin, Tetracycline, Cephalosporin, Erythromycin
III Classification based on Mode of action:
1. Bacteriostatic: A bacteriostatic agent is a biological or chemical agent that stops
bacteria from reproducing, while not necessarily killing them otherwise. Ex:
Tetracycline, Chloramphenicol, Lincomycin.
2. Bactericidal: is a substance which kills bacteria Ex: Penicillin, Cephalosporins,
Erythromycin, Monobactams.
IV Classification on the basis of mechanism of action:
1) Cell Wall Synthesis inhibitors: Penicillins, Cephalosporins, Vancomycin, Beta-
lactamase Inhibitors, Polymycin & Bacitracin.
 2) Agents acting on Cell membrane permeability: Amphotericin B, Nystatin,
Polymixins
3) Protein Synthesis Inhibitors

a) Inhibit 30s Subunit Aminoglycosides (gentamycin), Tetracyclines

b) Inhibit 50s Subunit Macrolides, Chloramphenicol, Clindamycin
4) Agents that interfere with the nucleic acid biosynthesis: Actinomycin,
Griseofulvin, Rifamycin
V. Classification on the basis of source:
1. Natural Antibiotics: they are obtained from the fermentation of microorganisms.
Ex: Bacitracin, Polymixin B
2. Semisynthetic Antibiotics: These antibiotics are commercially synthesized from
natural antibiotics.
Ex: Ampicillin, Methicillin, Cloxacillin, Oxacillin

3. Synthetic Antibiotics: These antibiotics are synthesized commercially.

Ex: Chloramphenicol
Commercial production:
The commercial production of antibiotics for medicinal use follows a general pattern. The
general scheme may be divided into six steps:
(a) Preparation of a pure culture of the desired organism.
(b) Fermentation, during which the antibiotic is formed.
(c) Isolation of the antibiotic from the culture medium.
(d) Purification.
(e) Assays for potency, sterility, absence of pyrogens, and other necessary data.
(f) Formulation into acceptable and stable dosage forms
Antibiotic resistance
● Antibiotic resistance is form of drug resistance where bacteria are able to survive after
exposure to one or more antibiotics.
● Cross-resistance - Cross-resistance to a particular antibiotic that often results in
resistance to other antibiotic, usually from a similar chemical class, to which the
bacteria may not have been exposed. For-example Clindamycin and lincomycin.
How does antibiotic resistance occur?
Bacteria mutate at a constant rate from mistakes being made during cell replication. These
mistakes can be good, bad or neutral. If they are good, we say that they confer a fitness
advantage and the phenotype gives the cell an advantage to survive in that particular
 environment. If they are neutral, they have no effect on the bacterium. However, if they are
bad the cell is likely to die and that particular mutation will not survive in future
generations.
Antibiotic Resistance Mechanisms:
A. Bacteria develop ability to hydrolyze these drugs using β lactamase confers resistance
to penicillin e.g. E. coli, Staph epidermidis, Pseudomonas aeruginosa, Klebsiella
pneumonia. Hence, add β lactamase inhibitor e.g. Clavulanic acid in amoxicillin-
Clavulanate (Augmentin).
B. Genetic mutation.
C. Misuse of antibiotics: Antibiotic misuse, sometimes called antibiotic abuse or
antibiotic overuse. The misuse or overuse of antibiotics, may produce serious effects
on health. It is a contributing factor to the creation of multidrug-resistant bacteria,
informally called "super bugs" relatively harmless bacteria can develop resistance to
multiple antibiotics and cause life-threatening infection.
Preventing antibiotic resistance

✔ Take antibiotics exactly as doctor prescribes.

✔ Complete the prescribed course of treatment.

✔ Do not save the antibiotics for next illness.

✔ Only take antibiotics prescribed for you; do not share or use leftover antibiotics.

✔ Do not ask for antibiotics when your doctor thinks you do not need them.
Mechanism of action of antibiotics:

Site of Action Name of the Antibiotic Process interrupted Type of activity

 1. CELL WALL Penicillins, 1.Mucopolypeptide Bactericidal
Cephalosporins, Synthesis
Bacitracin, Vancomycin
2.Cell wall cross linking
2. CELL Polymyxin Cell membrane integrity Bactericidal
MEMBRANE

3. Ribosomes Chloramphenicol Protein synthesis Bactriostatic

50s ribosomal sub Macrolides,Clindamyci Protein synthesis Bactriostatic

unit n
30s ribosomal sub Tetracyclines, Protein synthesis Bactericidal
unit Aminoglycosides

Β-Lactam Antibiotics:

Beta-lactam (β-lactam) ring is a four membered cyclic amide. They are the dominant class of
agents currently used for the chemotherapy of bacterial infections.
Beta lactam antibiotics are further subdivided into
1. Penicillins- Penicilln G, Ampicillin, Amoxycillin.
2. Cephalosporins- Cephalexin, Cephadroxil.
3. Carbapenams – Thienamycin
4. Oxopenams- Clavulanic acid
Mechanism of Action:
Beta lactam antibiotics act by inhibiting the bacterial cell wall synthesis. Specifically, they
inhibit the biosynthesis of the peptidoglycan that provides strength and rigidity to the cell wall.
The bacterial cell consists of cell membrane and peptidoglycan strands. The peptidoglycan
layer is a polymer made up of N-acetyl muramic acid alternating with N-acetyl glucosamine.
In Gram +ve bacteria peptidoglycan strand is very thick and in Gram -ve bacteria it is thin.
Proteins called Transpeptidases, also called as Penicillin binding proteins (PBPs) helps in
crosslinking the peptide glycan strands, which is essential for stability and rigidity of cellwall.
The β-lactam antibiotics containing β-lactam ring acylate a specific bacterial D-transpeptidase,
thereby rendering it inactive for its role in forming peptide cross-links of two linear
peptidoglycan strands. Hence they are bactericidal in nature.

1. Penicillins:
Penicillin is derived from the Penicillium notatum and Penicillium chrysogenum. It acts on
bacteria by inhibiting the enzymes responsible for the formation of the cell wall in the bacterial
cells. Penicillin is used to treat different types of grams positive and gram-negative bacterial
infections.
Nomenclature:

In the Chemical Abstracts system penicillins are named as 4-thia-l-azabicyclo[3.2.0]heptanes.

Three simplified forms of penicillin nomenclature have been adopted for general use.
• As derivatives of Penam: Use the name “penam” for the unsubstituted bicyclic system
(a fused thiazolidine and beta lactam ring), including the amide carbonyl group. Thus,
penicillins are generally designated as 5- acylamino-2,2-dimethylpenam-3-carboxylic
acids.

• As derivatives of Penicillanic acid: Use the name “penicillanic acid” to describe the
ring system with substituents that are generally present (i.e., 2,2-dimethyl and 3-
carboxyl).
 • As derivatives of penicillin: (on the basis of “R” group) - TRIVAL SYSTEM 6-
carbonyl amino penicillanic acid portion of the molecule is named as penicillin.

Structure of Penicillins:

Stereochemistry:

1. The penicillin molecule contains three chiral carbons at C-3, C-5 & C-6.
 2. All natural, semi synthetic and synthetic penicillin’s have the same absolute
configuration about these three centres.
3. The carbon atom bearing the acylamino group (C-6) has L configuration. Whereas, the
carbon to which the carboxyl group is attached has the D configuration ie: the
hydrogens on C6 & C3 are trans to each other.
4. The acylamino group and carboxyl group are trans to each other, with the former alpha
and later in the beta orientation relative to penam ring.
5. The absolute stereochemistry of the penicillins is designated 3S:5R:6R, as shown
below.
Structure Activity Relationship:

Position 1 – When the sulphur atom of the Thiazolidine ring is oxidized to a sulfone or
sulfoxide, it improves acid stability, but decreases the activity of the agent. The sulphur of
thiazolidine ring if replaced by other atom like oxygen or carbon increased broad spectrum
activity. Ex – Clavulanic acid and thienamycin.
Position 2 – No substitutions allowed at this position. Any change will lower activity. The
methyl groups are necessary.
Position 3 – The carboxylic acid of the Thiazolidine cannot be reduced to alcohol. However,
esterified resulting in the prodrug, longer duration of action Ex - Bacampicillin.
Position 4 – The nitrogen is a must.
Position 5 – No substitutions allowed.
Position 6 – Substitutions are allowed on the side chain of the amide. An electron withdrawing
group added at this position will give the compound better acid stability because this
substitution will make the amide oxygen less nucleophillic. A bulky group added close to the
ring will make the compound more resistant to Beta-lactamases. Steric hindrance also provides
protect to the Beta-lactam ring.
Three chiral centres are required for Penicillins bioactivity.
Side chain can be replaced with different R group to obtain different compounds with broad
antibacterial spectrum. Substituent on alpha carbon of side chain like-amino, guanidino and
chloro - resistant against acid. Ex – Ampicillin. Amino group - confers more activity to pass
through cell wall barriers.
Aromatic ring attached to the carbonyl group and has some substitution on the aryl ring at the
position ortho to the point of attachment- resistant to acid hydrolysis, making this analogue of
pencillin more resistant against ß- lactam ring and protect the lactam ring from degradation
Ex- oxacillin , cloxacillin . Cloxacillin better absorbed than oxacillin Ex Methicillin, Naficillin.
Position 7 – The carbonyl group on the Beta-lactam ring is a must.
The thiazolidine ring can be removed in penicillins resulting in the monobactams which are
monocyclic ß- lactam. Ex – Azetreonam.
Pka of penicillin is 2.5-3.0 Penicillin contain basic group – zwitter ion – pka 7.4 and these free
acids are sparingly soluble in water. Active against gram positive organism.
Chemical Degradation:
• The main cause of deterioration of penicillin is hydrolysis. The β-lactam carbonyl group
of penicillin readily undergoes nucleophilic attack by water or especially, hydroxide
ion to form the inactive penicilloic acid, which is reasonably stable in neutral to alkaline
solutions but readily undergoes decarboxylation and further hydrolytic reactions in
acidic solutions.
• Other nucleophiles, such as hydroxylamines, alkylamines, and alcohols, open the β-
lactam ring to form the corresponding hydroxamic acids, amides, and esters.
• In strongly acidic solutions (pH 3), penicillin undergoes a complex series of reactions
leading to various inactive degradation products. The first step appears to involve
rearrangement to the penicillanic acid. This process is initiated by protonation of the β-
lactam nitrogen, followed by nucleophilic attack of the acyl oxygen atom on the β-
lactam carbonyl carbon.
• The subsequent opening of the β-lactam ring destabilizes the thiazoline ring, which then
also suffers acid-catalyzed ring opening to form the penicillanic acid. The latter is very
unstable and experiences two major degradation pathways. The most easily understood
path involves hydrolysis of the oxazolone ring to form the unstable penamaldic acid.
Because it is an enamine, penamaldic acid easily hydrolyzes to penicillamine (a major
degradation product) and penaldic acid.
• The second path involves a complex rearrangement of penicillanic acid to a penillic
acid through a series of intramolecular processes.
• Penillic acid (an imidazoline-2-carboxylic acid) readily decarboxylates and suffers
hydrolytic ring opening under acidic conditions to form a penilloic acid.
• Penicilloic acid exists in equilibrium with penamaldic acid and undergoes
decarboxylation in acid to form penilloic acid.
• The third major product of the degradation is penicilloaldehyde, formed by
decarboxylation of penaldic acid.
• By controlling the pH of aqueous solutions within a range of 6.0 to 6.8 and refrigerating
the solutions, aqueous preparations of the soluble penicillins may be stored for up to
several weeks.
• Oxidizing agents also inactivate penicillins, but reducing agents have little effect on
them. Temperature affects the rate of deterioration; although the dry salts are stable at
room temperature and do not require refrigeration, prolonged heating inactivates the
penicillins.
Bacterial Resistance:
• The most species of Gram-negative bacilli, are naturally resistant to the action of
penicillins. Other normally sensitive species can also develop penicillin resistance.
• The most important biochemical mechanism of penicillin resistance is by the bacterial
enzymes that inactivate penicillin called penicillinases, are of two types: β- lactamases
and acylases.
• The β-lactamases, catalyze the hydrolytic opening of the β-lactam ring of penicillins to
produce inactive penicilloic acids.
• Another important resistance mechanism, especially in Gram-negative bacteria, is
decreased permeability to penicillins. The cell envelope in most Gram-negative bacteria
is more complex, which creates a physical barrier to the penetration of antibiotics.
• Bacterial resistance can result from changes in the affinity of PBPs for penicillins.
• Genetic mutation.
• Certain strains of bacteria are resistant to the lytic properties of penicillins but remain
susceptible to their growth inhibiting effects. Thus, the action of the antibiotic has been
converted from bactericidal to bacteriostatic. This mechanism of resistance is termed
tolerance.
 Preventing antibiotic resistance

✔ Take antibiotics exactly as doctor prescribes.

✔ Complete the prescribed course of treatment.

✔ Do not save the antibiotics for next illness.

✔ Only take antibiotics prescribed for you; do not share or use leftover antibiotics.

✔ Do not ask for antibiotics when your doctor thinks you do not need them.
Benzyl Penicillin:

It is also known as penicillin G, is an antibiotic used to treat a number

of bacterial infections. Benzylpenicillin possess effectiveness mainly against Gram-
positive organisms. Some Gram-negative organisms such as Neisseria
gonorrhoeae and Leptospira weilii are also reported to be susceptible to benzylpenicillin.
This includes pneumonia, strep throat, syphilis, enterocolitis, diphtheria, gas
gangrene, leptospirosis, cellulitis, and tetanus. Benzylpenicillin is given by injection into a
vein or muscle. Two long-acting forms benzathine benzylpenicillin and procaine
benzylpenicillin are available for use by injection into a muscle.
Side effects include diarrhea, seizures and allergic reactions including anaphylaxis. When
used to treat syphilis a reaction known as Jarisch–Herxheimer may occur. It is not
recommended in those with a history of penicillin allergy. Use during pregnancy is generally
safe.
Derivatives of Penicillin G:
Procaine Penicillin G: It is a mixture of procaine hydrochloride with penicillin G. The salt is
less soluble, hence used as suspension for intramuscular use for prolonged duration of action.
Procaine acts as local anaesthetic, which reduces pain during & after injection.
Benzathine Penicillin G: It is a mixture of two molecules of penicillin G with N,N-Dibenzyl
ethylene diamine. It is tasteless compound almost insoluble in water, administered only by IM
route for sustained action in the treatment of streptococci infection.
Phenoxymethylpenicillin:
It is also known as penicillin V , is an antibiotic useful for the treatment of strep throat, otitis
media and cellulitis. It is also used to prevent rheumatic fever and to prevent infections
following removal of the spleen. It is given by mouth.
Side effects include diarrhoea, nausea, and allergic reactions including anaphylaxis. It is not
recommended in those with a history of penicillin allergy. It is relatively safe for use
during pregnancy. Infections caused by Streptococcus pyogenes, Tonsillitis, Pharyngitis,
Skin infections, Anthrax (mild uncomplicated infections), Rheumatic fever (primary and
secondary prophylaxis), Streptococcal skin infections.
Ampicillin:

It was the first "broad spectrum" penicillin with activity against Gram-positive and Gram-
negative bacteria. It is an antibiotic used to prevent and treat a number of bacterial infections,
such as respiratory tract infections, urinary tract infections, meningitis, salmonellosis, and
endocarditis. It may also be used to prevent streptococcal infection in newborns. It is given by
mouth, intramuscularly or intravenously. Its spectrum of activity is enhanced by co-
administration of sulbactam, a drug that inhibits beta lactamase enzyme.
Common side effects include rash, nausea, and diarrhoea.
Bacampicillin Hydrochloride: Bacampicillin hydrochloride, is the hydrochloride salt of the
1-ethoxycarbonyloxyethyl ester of ampicillin. It is a prodrug of ampicillin with no antibacterial
activity. After oral absorption, bacampicillin is hydrolyzed rapidly by esterases in the plasma
to form ampicillin.
Amoxicillin:

It is an antibiotic used to treat a number of bacterial infections. These include middle ear
infection, strep throat, pneumonia, skin infections, urinary tract infections
Salmonella infections, Lyme disease, and chlamydia infections. It is taken by mouth or less
commonly by injection. Common adverse effects include nausea and rash. It should not be used
in those who are allergic to penicillin.
Amoxicillin is susceptible to degradation by β-lactamase-producing bacteria. So, it may be
combined with clavulanic acid, a β-lactamase inhibitor. This drug combination is commonly
called co-amoxiclav.
Methicillin:

It is a synthetic analog of penicillin. It is a narrow-spectrum β-lactam antibiotic of

the penicillin class. Methicillin sodium is particularly resistant to inactivation by the
penicillinase found in staphylococci and Bacillus cereus. It is mainly used in the treatment of
staphylococcal infections caused by strains resistant to other penicillins. It is recommended
that it not be used in general therapy, to avoid the possible widespread development of
organisms resistant to it.
Nafcillin sodium:

It is another semisynthetic penicillin and is a narrow-spectrum beta-lactam antibiotic. As

a beta-lactamase-resistant penicillin, it is used to treat infections caused by Gram
positive bacteria, in particular, species of staphylococci that are resistant to other penicillins.
Oxacillin:

It is 5-methyl-3-phenyl-4-isoxazolyl derivative of penicillin. It is penicillinase resistant

and narrow-spectrum beta-lactam antibiotic of the penicillin class.
The use of oxacillin is restricted to the treatment of infections caused by staphylococci resistant
to penicillin G.
Cloxacillin Sodium:

The chlorine atom ortho to the position of attachment of the phenyl ring to the isoxazole ring
enhances the activity of cloxacillin sodium. In almost all other respects, it resembles oxacillin,
with better oral absorption, leading to higher plasma levels.