Antibiotics

An antibiotic is a chemical compound derived from or produced by a living organism, which is

capable, in small concentrations, of inhibiting the life processes of micro-organisms.
Classification Based on Chemical Structure
1. β-lactam antibiotics
2. Aminoglycoside antibiotics
3. Tetracycline antibiotics
4. Polypeptide antibiotics
5. Macrolide antibiotics
6. Lincomycins
7. Other antibiotics
β-lactam antibiotics
The β-lactam antibiotics may be further sub-divided into two categories, namely Penicillins and
Cephalosporins.
Penicillins
Penicillin was the first antibiotic developed and used clinically. It was discovered accidentally by
Alexander Fleming. The source of penicillin is high-yielding penicillium chrysogenum.

Mechanism of Action:
Penicillins inhibit bacterial cell wall synthesis by binding to penicillin-binding proteins (PBPs).
This binding blocks the cross-linking of peptidoglycan chains, weakening the cell wall and causing
the bacterial cell to lyse and die. This mechanism is especially effective against actively dividing
bacteria. Penicillins are particularly effective against Gram-positive bacteria. Resistance can occur
through β-lactamase production, PBP alterations, or membrane permeability changes.

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Therapeutic Applications

 Respiratory Tract Infections: Used to treat streptococcal pharyngitis, pneumonia, and

acute exacerbations of chronic bronchitis.
 Skin and Soft Tissue Infections: Effective against cellulitis, impetigo, and erysipelas
caused by streptococci and staphylococci.
 Sexually Transmitted Infections: Penicillin G is the treatment of choice for syphilis.
 Central Nervous System Infections: Used for bacterial meningitis caused by susceptible
organisms such as Neisseria meningitidis and Streptococcus pneumoniae.
 Endocarditis: Often used in combination with other antibiotics to treat infective
endocarditis caused by streptococci and enterococci.
 Intra-abdominal Infections: Ampicillin/sulbactam and piperacillin/tazobactam are used
for peritonitis and other abdominal infections.
 Urinary Tract Infections: Amoxicillin and ampicillin are used for UTIs caused by
susceptible enterococci and E. coli.
 Bone and Joint Infections: Penicillinase-resistant penicillins like nafcillin and oxacillin
are used for osteomyelitis and septic arthritis caused by Staphylococcus aureus.

Synthesis

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Methicillin

Oxacillin, Cloxacillin, Dicloxacillin, Floxacillin

Ampicillin

Structure-Activity of Penicillins

6-Acyl side chain

1. The substitution of R on the primary amine with an
electron withdrawing group decreases the electron density
on the side chain and protects from acid degradation.

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 2. Substituents on the α-carbon of the side chain, such as amino (ampicillin), chloro, and
guanidine exerts good resistance to inactivation by acids.
3. Substitution of α-aryl of the alkyl group in the side chain gives increased stability and
oral absorption.
4. Substitution of bulky groups on α-carbon of the side chain confers β-lactamase resistance.
Examples: methicillin, nafcillin, oxacillin, etc. The size of the ring systems play an
important role in determining the ability of the ortho substitutent to confer penicillinase
resistance.
5. The isomeric forms of penicillins differs in their activity. Example: D-isomer is 2–8 times
more active than L-isomer of amoxicillin.
6. The introduction of polar group or ionized molecule into the α-position of the side chain
in the benzyl carbon atom of penicillin-G confers against the gram-negative bacilli.
Amino, hydroxyl, carboxyl, and sulphonyl increases gram-negative activity. Example:
ampicillin and carbenicillin.
7. Replacement of acyl side chain with hydroxymethyl groups shows improved gram-
negative activity and introduction of C-6 α-methoxy group produces greater stability
against β-lactamase.
Substituents at sulphur: Sulphur is the only atom at position 1 of the penicillin in order to
retain appreciable antibacterial activity.
C-2 substituents: The geminal dimethyl group at C-2 is characteristic of the penicillin.
C-3 substituents: In general, derivatization of the C-3 carboxylic acid functionality is not
tolerated unless the free penicillin carboxylic acid can be generated Antibiotics. Doubly activated
penicillin esters such as alkanoyloxyalkyl congeners undergo rapid cleavage Antibiotics to
generate active penicillin—e.g., pivampicillin and becampicillin.
Variation at N-4: The nitrogen atom at the ring junction is vital for antibacterial activity; the
nitrogen atom contributes to the reactivity of the β-lactam carbonyl centre

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 Cephalosporins
The cephalosporins are β-lactam antibiotics isolated from Cephalosporium species and/or prepared
semisynthetically. This comes under the class of 7-amino cephalosporonic acid (7-ACA)
derivatives and are much more acid-stable than the corresponding 6-APA compounds. The
cephalosporin nucleus, 7-aminocephalosporanic acid (7-ACA), was derived from cephalosporin C
and proved to be analogous to the penicillin nucleus 6-aminopenicillanic acid, but it was not
sufficiently potent for clinical use. Modification of the 7-ACA side-chains resulted in the
development of useful antibiotic agents, and the first agent cephalothin (cefalothin) was launched
by Eli Lilly in 1964.
Cephalosporins can be divided into three classes:
 Cephalosporin N: It has a penicillin-like structure being a derivative of 6-
aminopenicillanic acid.
 Cephalosporin P: An acidic antibiotic, which is steroidal in nature.
 Cephalosporin-C: It is a true cephalosporin and it is a derivative of 7 amino-
cephalosporanic acid
Classification

Mechanism of Action
Cephalosporins inhibit bacterial cell wall synthesis by binding to penicillin-binding proteins
(PBPs), which disrupts peptidoglycan cross-linking by inhibiting the transpeptidase, leading to a
weakened cell wall, cell lysis, and bacterial death. They are bactericidal and their effectiveness
depends on their ability to reach PBPs, their affinity for PBPs, and their stability against beta-
lactamases.
Spectrum
First-generation cephalosporins have good activity against Gram-positive bacteria (e.g.,
Staphylococcus aureus, Streptococcus species) and limited activity against Gram-negative
bacteria.
Second-generation cephalosporins have expanded activity against Gram-negative bacteria while
retaining good Gram-positive coverage. They are effective against organisms such as Haemophilus
influenzae, Enterobacter species, and some Neisseria species.
Third-generation cephalosporins have further expanded Gram-negative coverage and are more
resistant to beta-lactamases. They are effective against a broader range of Gram-negative bacteria,

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including Pseudomonas aeruginosa (some agents), and have diminished activity against Gram-
positive bacteria compared to first-generation cephalosporins
Fourth-generation cephalosporins have a broad spectrum of activity, including enhanced Gram-
negative coverage and better beta-lactamase stability. They are effective against both Gram-
positive and Gram-negative bacteria, including Pseudomonas aeruginosa.
Fifth-generation cephalosporins are effective against a wide range of Gram-positive and Gram-
negative bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). They have
enhanced binding to PBPs of resistant organisms.
Therapeutic Applications
First-generation cephalosporins are commonly used to treat skin and soft tissue infections, such
as cellulitis and impetigo, as well as for prophylaxis in surgical procedures to prevent infections.
Second-generation cephalosporins are used for respiratory tract infections, including sinusitis,
otitis media, and community-acquired pneumonia. They are also effective for urinary tract
infections (UTIs) and some intra-abdominal infections.
Third-generation cephalosporins have applications in more severe and hospital-acquired
infections. They are used for bacterial meningitis, gonorrhea, severe pneumonia, sepsis, and
complicated UTIs. Ceftriaxone is also frequently used for its convenience in outpatient parenteral
antibiotic therapy (OPAT).
Fourth-generation cephalosporins are reserved for serious infections caused by multi-drug
resistant Gram-negative bacteria, including Pseudomonas aeruginosa. They are used for severe
nosocomial infections, febrile neutropenia, and complicated intra-abdominal and urinary tract
infections.
Fifth-generation cephalosporins are used for treating infections caused by methicillin-resistant
Staphylococcus aureus (MRSA) and are also effective against acute bacterial skin and skin
structure infections (ABSSSI) and community-acquired bacterial pneumonia (CABP).

Structures:

First Generation cephalosporins

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Second Generation cephalosporins

Third Generation Cephalosporins

Synthesis
Synthesis of 7-aminocephalosporonic acid from cephalosporin C

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Cephalexin

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Amino group of phenylglycine is protected with t-BOC, and activated carboxylic acid reacts with
7-ACA to give amide derivative. t-BOC is deprotected by treatment with trifl uoro acetic acid.
Reducing this product with hydrogen using a palladium on barium sulphate catalyst results in the
deacetoxylation at the third position of 7-aminocephalosporanic acid, making the desired
cephalexin

Cefadroxil

7-ACA is deacetylated by hydrogenolysis; carboxylate is protected with silyl derivative; and

acylation with t-BOC-protected tyrosine and deprotection afford cefadroxil. It is an analogue of
cephalexin and differs only in the presence of a hydroxyl group in the fourth position of the
phenyl ring of phenylglycine.

Cephradine

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Ceftizoxime

Acylation of 6-ACA derivative with thiazole derivative in presence of DCC, followed by

deprotection of trityl group, affords ceftizoxim.

Structure-Activity Reltionship of Cephalosporins

1. 7-Acylamino substituents:
i. Acylation of amino group generally increases the potency against gram-positive bacteria,
but it is accompanied by a decrease in gram-negative potency.
ii. High antibacterial activity is observed only when the new acyl groups are derived from
carboxylic acids for gram-positive bacteria.
iii. Substituents on the aromatic ring that increases lipophilicity provide higher gram-positive
activity and generally lower gram-negative activity.
iv. The phenyl ring in the side-chain can be replaced with other heterocycles with improved
spectrum of activity and pharmacokinetic properties, and these include thiophene,
tetrazole, furan, pyridine, and amino thiazoles.
2. C-3 substituents:
The nature of C-3 substituents influences pharmacokinetic and pharmacological properties as
well as antibacterial activity. Modification at C-3 position has been made to reduce the
degradation (lactone of desacetyl cephalosporin) of cephalosporins
i. The benzoyl ester displaces improved gram-positive activity but lower gram-negative
activity

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 ii. Pyridine and imidazole-replaced acetoxy groups show improved activity against P.
aeruginosa. Displacement of acetoxy group by azide ion yields derivatives with relatively
low gram-negative activity.
iii. Displacement with aromatic thiols of 3-acetoxy group results in an enhancement of activity
against gram-negative bacteria with improved pharmacokinetic properties.
iv. Replacement of acetoxy group at C-3 position with —CH3, Cl has resulted in orally active
compounds
3. Cephamycins: Introduction of C-7 α-methoxy group shows higher resistance to hydrolysis by
β-lactamases
4. Oxidation of ring sulphur to sulphoxide or sulphone greatly diminishes or destroys the
antibacterial activity.
5. Replacement of sulphur with oxygen leads to oxacepam (latamoxef) with increased
antibacterial activity, because of its enhanced acylating power. Similarly, replacement of sulphur
with methylene group (loracarbef) has greater chemical stability and a longer half-life.
6. Olefinic linkage at C 3-4 is essential for antibacterial activity. Isomerization of the double
bond to 2-3 position leads to great losses in antibacterial activity

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 Aminoglycosides
The aminoglycoside antibiotics contain one or more amino sugars linked to an aminocytitol ring
by glycosidic bonds. These are broad-spectrum antibiotics; in general, they have greater activity
against gram-negative than gram-positive bacteria. The development of streptomycin, the fi rst
antibiotic of this group, was a well-planned work of Waksman (1944) and his associates, who
isolated it from a strain of Streptomyces griseus.
The aminoglycoside can produce severe adverse effects, which include nephrotoxicity,
ototoxicity, and neuro effects. These properties have limited the use of aminoglycoside
chemotherapy to serious systemic indications. Some aminoglycosides can be administered for
ophthalmic and topical purposes.

Examples:

Mechanism of Action:
Aminoglycosides work by binding to the 30S subunit of bacterial ribosomes. This binding
interferes with the initiation complex of protein synthesis and causes misreading of mRNA. As a
result, incorrect amino acids are incorporated into the growing polypeptide chain, leading to the
production of nonfunctional or toxic proteins. This disruption of protein synthesis ultimately
results in bacterial cell death. Additionally, aminoglycosides can also cause the breakup of
polysomes into nonfunctional monosomes. Their uptake into bacterial cells is oxygen-dependent,
meaning they are more effective against aerobic bacteria.

Streptomycin
 Streptomycin is chiefly employed in the treatment of tuberculosis in conjunction with other
drugs such as isoniazid and rifampicin.

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  Streptomycin and penicillin exert a synergistic action against bacteria and are usually
employed together in the treatment of subacute bacterial endocarditis caused by
Streptococcus faecalis.
 It exerts bacteriostatic action in low concentrations and bactericidal in high concentrations
against a plethora of Gram-negative and Gram-positive organisms.
 It is a triacidic base and has an aldo sugar.
 It consist of three parts; Streptidine (1,3-diguano ring), Streptose sugar (hexose), Amino
sugar (N-methyl L-glucosamine

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SAR of Streptomycin

.
Neomycin
Neomycin is mostly used in a wide variety of local infection such as burns, ulcers, wounds,
impetigo, infected dermatoses, furunculosis, conjunctivitis, etc. It is also employed as an adjuvant
in topical steroid preparations to control secondary infections in the case of inflammatory
disorders.
Neomycin is a mixture of closely related epimers, neomycin B, and C. Neomycin B differ from
neomycin C by the nature of the sugar attached terminally to D-ribose, this sugar called neosamine.
B1 differs from neosamine C in its stereochemistry. In neomycin B1 , the neobiosamine moiety
contains. β-L-iodopyranosyl, whereas in neomycin C the confi guration is inverted and it is 2-D-
glucopyranosyl.

SAR of Neomycin
The structures of neomycin A(neamine), neomycin B and neomycin C have been established ;
besides, the absolute configurational structures of neomycin and neamine have been reported. It
has been demonstrated that neamine could be obtained by the methanolysis of neomycin B and C
respectively, whereby the glycosidic linkage existing between D-ribose and deoxystreptamine
undergoes cessation.

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Gentamycin
Gentamycin is a mixture of C1 , C2, and C1 A compounds, obtained commercially from
Micromonospora purpurea. Gentamycin sulphate exists as white hygroscopic powder, soluble in
water, and practically insoluble in alcohol, although it is a broad-spectrum antibiotic. It is used in
the treatment of infections caused by gram-negative bacteria of particular interest and has a high
degree of activity against P. aeruginosa, where the important causative factor is burned skin. It is
used topically in the treatment of infected bed sores, pyodermata, burns, and in the eye infection

Kenamycin
The mixture consists of three related structures, that is
Kanamycin A, B, and C. The kanamycins do not possess
D-ribose molecule that is present in neomycins and
paramomycins. The use of kanamycins is restricted to
infections of the intestinal tract and to systemic infections.

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Amikacin
Amikacin is a semisynthetic drug derived form kanamycin A.
It retains 50% of the original activity of kanamycin A.
L-Isomer is more active than D-isomer. It resists attack
by most bacterial inactivating enzyme. Therefore,
it is very effective and less ototoxic than other aminoglycosides.

Tobramycin
Its activity is similar to gentamycin. The superior activity of tobramycin against P. aeruginosa
may make it useful in the treatment of bacterial oesteromyelitis, and pneumonia caused by P.
species.

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 Chloramphenicol
Chloramphenicol (chloromycetin) is a levorotatory broadspectrum antibiotic originally produced
from several streptomycetes, namely : S. venezualae, S. omiyamensis and S. phacochromogenes
var. chloromyceticus.
 The molecular formula of chloramphenicol is C11H12O5N2Cl2.

Spectrum

Chloramphenicol has a broad-spectrum antibacterial activity, meaning it is effective against a wide

variety of microorganisms. It is active against many gram-positive and gram-negative bacteria,
including anaerobes. Specifically, chloramphenicol is effective against bacteria such as
Streptococcus pneumoniae, Haemophilus influenzae, Neisseria meningitidis, Salmonella species,
and certain strains of Escherichia coli. It also works against Rickettsia, Chlamydia, and
Mycoplasma species, making it useful in treating atypical bacterial infections.

However, due to its potential severe side effects, including bone marrow suppression and the risk
of aplastic anemia, the use of chloramphenicol is limited to serious infections for which no safer
alternatives are available. It is often used as a last resort for treating infections such as typhoid
fever, bacterial meningitis, and rickettsial diseases when other antibiotics are ineffective or
contraindicated.

Mechanism of Action
Chloramphenicol is a broad-spectrum antibiotic that exerts its bacteriostatic effect by inhibiting
bacterial protein synthesis. It binds reversibly to the 50S ribosomal subunit of the bacterial
ribosome. This binding interferes with the peptidyl transferase activity of the ribosome, (an
enzyme crucial for the formation of peptide bonds between adjacent amino acids during protein
elongation). By blocking the peptidyl transferase, chloramphenicol prevents the transfer of the
growing peptide chain from the aminoacyl-tRNA to the peptidyl-tRNA, effectively halting the
elongation process of protein synthesis. This inhibition disrupts the production of essential proteins
required for bacterial growth and replication.

Therapeutic Applications
 Typhoid Fever and Paratyphoid Fever: Chloramphenicol is particularly effective against
Salmonella typhi and Salmonella paratyphi, the causative agents of these infections.

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  Bacterial Meningitis: It is used in cases where the causative organisms are susceptible to
chloramphenicol and other more commonly used antibiotics, like beta-lactams, are not an
option.
 Rickettsial Infections: Diseases such as Rocky Mountain spotted fever, caused by
Rickettsia rickettsii, can be treated with chloramphenicol, especially in patients who cannot
tolerate tetracyclines.
 Anaerobic Infections: Chloramphenicol is effective against a range of anaerobic bacteria,
making it useful for treating infections in anaerobic environments, such as intra-abdominal
and central nervous system infections.
 Eye Infections: Topical chloramphenicol (in the form of eye drops or ointments) is used
to treat bacterial conjunctivitis and keratitis due to its broad-spectrum activity and good
penetration into ocular tissues.

Synthesis

Continue………

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para-Nitroacetophenone( on bromination gives the corresponding bromo derivative which on
treatment with hexamine followed by acidic ethanol yields α-amino-p-nitroacetophenone
hydrochloride. This on acetylation gives the acetamido derivative which on treatment with
formaldehyde followed by aqueous sodium carbonate affords the corresponding hydroxy methyl
analogue. Reduction of the keto moiety is effected by treatment with aluminium iso-propoxide to
give the product in DL-form which on reaction with HCl removes the acetyl function to yield the
chloramphenicol base in its DL-form. The resulting product is first subjected to resolution with α-
camphoric acid and secondly with dichloromethyl acetate to afford the addition of the side chain
to yield chloramphenicol.

Structure-Activity Relationship

1. Modification of p-nitrophenyl group:

The p-nitrophenyl group may be modified through the following ways:
 Replacement of the nitro group by other substituents leads to a reduction in activity.
 Shifting of the nitro group from the para position also reduces the antibacterial activity.
 Replacement of phenyl group by the alicyclic moieties results in less potent
compounds.

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 2. Modification of dichloroacetamido side chain: Other dihalo derivatives of the side chain
are less potent although major activities are retained,
3. Modification of 1,3-propanediol: If the primary alcoholic group on C-1 atom is modified,
it results in a decrease in activity; hence, the alcoholic group seems to be essential for
activity.
4. The propanediol moiety should be in D threo-isomer. Other isomers are inactive.
5. Replacement of OH, and extension or suppression of terminal CH2OH abolishes the
activity.
6. Replacement of nitro group by other electron withdrawing groups gives active compounds
as:
 CH3SO2 (Thiamphenicol) or
 CH3CO (Cetophenicol)

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 Tetracyclines
Tetracyclines have a ring system of four linear annelated six-membered rings and are
characterized by a common octahydronaphthacenes skeleton. They are potent, broad-spectrum
antibacterial agents effective against gram-positive and gram-negative aerobic and anaerobic
bacteria. As a result, the tetracyclines are drugs of choice or well-accepted alternatives for a
variety of infectious diseases.

Mechanism of Action
Tetracyclines exert their antibacterial effect by inhibiting protein synthesis in bacteria. They
achieve this by binding to the 30S ribosomal subunit of the bacterial ribosome. This binding
blocks the attachment of aminoacyl-tRNA to the mRNA-ribosome complex, preventing the
addition of new amino acids to the growing peptide chain. By interfering with the proper
alignment of tRNA and mRNA, tetracyclines disrupt the elongation phase of protein synthesis.
This results in the inhibition of bacterial growth and replication, making tetracyclines
bacteriostatic rather than bactericidal.

Spectrum

1. Gram-positive bacteria: Streptococcus species, including Streptococcus pneumonia,

Staphylococcus aureus (including MRSA), Bacillus anthracis (anthrax), Listeria
monocytogenes.
2. Gram-negative bacteria: Haemophilus influenza, Escherichia coli, Klebsiella species,
Enterobacter species, Neisseria gonorrhoeae, Brucella species (brucellosis), Vibrio
cholerae (cholera), Yersinia pestis (plague)
3. Atypical bacteria: Chlamydia trachomatis (chlamydial infections), Mycoplasma
pneumoniae (atypical pneumonia), Rickettsia species (Rocky Mountain spotted fever,
typhus), Coxiella burnetii (Q fever)
4. Anaerobes: Clostridium species, including Clostridium perfringens and Clostridium
tetani
5. Other organisms: Treponema pallidum (syphilis), Borrelia burgdorferi (Lyme
disease),Propionibacterium acnes (acne).
Tetracyclines are also effective against some protozoa, including Plasmodium species (malaria)
and certain amoebas.
Therapeutic Applications
Respiratory Tract Infections:
 Atypical pneumonia caused by Mycoplasma pneumoniae.
 Chlamydial pneumonia caused by Chlamydia pneumoniae.
 Respiratory infections caused by Haemophilus influenza.
Sexually Transmitted Infections (STIs):
 Chlamydia trachomatis infections, including urethritis, cervicitis, and pelvic
inflammatory disease
 Syphilis (Treponema pallidum), especially in patients allergic to penicillin
Rickettsial Infections:
 Rocky Mountain spotted fever

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  Typhus
 Q fever (Coxiella burnetii)
Acne:
 Moderate to severe acne vulgaris due to its activity against Propionibacterium acnes
Zoonotic Infections:
 Lyme disease (Borrelia burgdorferi)
 Brucellosis (Brucella species)
 Plague (Yersinia pestis)
 Tularemia (Francisella tularensis)
Gastrointestinal Infections:
 Cholera (Vibrio cholerae)
 Helicobacter pylori infections, often as part of a combination therapy
Malaria
Anthrax:

Structures and Classification

Classes of tetracyclines: 1) Natural tetracyclines (biosynthetic) 2) Semisynthetic tetracyclines
3) Protetracyclines.
1. Natural tetracyclines

2. Semisynthetic tetracyclines

3. Protetracyclines.

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Synthesis
Methacycline

It is synthesised from oxytetracycline, which is reacted with a sulphur trioxide-pyridine

complex, resulting in an oxidation reaction. Simultaneous sulphonation gives a naphthacen
sulphotetrahydrofuran derivative intermediate, which when reacted with hydrofluoric acid forms
methacycline.

Doxycycline (Vibramycin)

Doxycycline is synthesised in two different ways; one of the ways suggests dehydrating
oxytetracycline at C6 by reducing the tertiary hydroxyl group with hydrogen using a rhodium on
carbon catalyst. It is also synthesised by reducing methylene group of methacycline using
rhodium on carbon catalyst

Minocycline

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Hydrogenolysis of demethylchlortetracycline removes both the chlorine and the benzylic
hydroxyl group. The product, 6-demethyl-6-deoxytetracycline, on nitration gives mixture of 7
nitro 6-demethyl-6-deoxytetracycline; is reductively methylated by catalytic hydrogenation in
the presence of formaldehyde to give minocycline.

Rolitetracycline

It is prepared by reacting tetracycline, formaldehyde, and pyrrolidine; amino methylation takes

place in amide group (Mannich reaction).
Structure-Activity Relationship of Tetracyclines

The key structural feature is a linearly fused tetracyclic nucleus and each ring needs to be six
membered and purely carbocyclic. A tetracyclic backbone skeleton is essential for activity.
 The D-ring needs to be aromatic and the A-ring must be appropriately substituted at
each of its carbon atoms for notable activity.
 The B-ring and the C-ring tolerate certain substituent changes as long as the keto-
enol systems (at C-11, 12, 12a) remain intact and conjugated to the phenolic D-ring.
 The D, C, B-ring phenol, keto-enol system is imperative and the A-ring must also
contain a conjugated keto enol system.
1. C-1 substituents: The keto-enol system of the Α-ring is indispensable for antibacterial
activity. No variation at the C-1 position has been successful.
2. C-2 substituents: The carboxamide moiety is present in all naturally occurring
tetracyclines and this group is crucial for antibacterial activity. The amide is best left
unsubstituted, or mono-substitution is acceptable in the form of activated
alkylaminomethyl amide (Mannich bases). An example is rolitetracycline. Large alkyl
group on the carboxamide may alter the normal keto-enol equilibrium of the C-1, 2, and 3
conjugated system, and diminishes inherent antibacterial activity. The replacement of
carboxamide group or dehydration of carboxamide to the corresponding nitrile results in
the loss of activity.
3. C-3 substituents: In conjugation with the C-1 position, the keto-enol conjugated system
is imperative for antibacterial activity.

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 4. Modification of C-4 position: The keto-enolic character of the A-ring is due to the α-C-4
dimethyl amino substituent. Loss of activity is exerted when dimethyl amino group is
replaced with hydrazone oxime or hydroxyl group.
5. Modification of C-4a position: The α-hydrogen at C-4a position of tetracyclines is
necessary for useful antibacterial activity.
6. C-5 substituents: Many naturally occurring antibacterial tetracyclines have an
unsubstituted methylene moiety at the C-5 position. However, oxytetracycline contains C-
5 α-hydroxyl group and was found to be a potent compound, and has been modified
chemically to some semi-synthetic tetracyclines. Alkylation of the C-5 hydroxyl group
results in a loss of activity. Ester formation is only acceptable if the free oxytetracycline
can be liberated in vivo; only small alkyl esters are useful.
7. C-5a substituents: The configuration of the naturally occurring tetracyclines places the C-
5a hydrogen atom in an α-configuration. Epimerization is detrimental to antibacterial
activity.
8. Modification at the C-6 position: The C-6 methyl group contributes little to the activity
of tetracycline. The C-6 position is tolerant to a variety of substituents. The majority of
tetracyclines have α-methyl group and α β-hydroxyl group at this position. Demeclocycline
is a naturally occurring C-6 demethylated chlortetracycline with an excellent activity.
Removal of C-6 hydroxyl group affords doxycycline, which exerts good antibacterial
activity.
9. C-7 and C-9 substituents: The nature of the aromatic D-ring predisposes the C-7 position
to electrophilic substitution, and nitro and halogen groups have been introduced. Some C-
7 nitro tetracyclines are among the most potent of all tetracyclines in vitro, but these
compounds were potentially toxic/carcinogenic. Halogenated derivatives are less active.
The C-7 acetoxy, azido, and hydroxyl tetracyclines are inferior in terms of antibacterial
activity.
10. C-10 substituents: The C-10 phenolic moiety is absolutely necessary for antibacterial
activity.
11. C-11 substituents: The C-11 carbonyl moiety is part of one of the conjugated keto-enol
systems required for antibacterial activity.
12. C-11a substituents: In general, few modifications at the C-11a position of tetracycline
have been tolerated. This is probably due to the detrimental effects exerted upon the keto-
enol system, which is vital for magnesium cation binding and subsequent tetracycline
uptake by the bacterial cell.
13. C-12 substituents: As with the C-11 position, the C-12 position is part of the keto-enol
system vital for drug uptake, binding, and observed antibacterial activity.
14. C-12a substituents: The C-12a hydroxyl group is needed for antibacterial activity,
although this moiety can be esterified to provide tetracycline with increased lipophilicity.
Antibacterial properties are retained if the alkyl ester is small in size, and readily undergoes
hydrolysis to liberate free tetracycline.

M. Awais Fareed Pharm D IUB (2019-24)

References
 Lectures by Dr. Mohsin Abbas Khan
 Textbook of Medicinal Chemistry—V. Alagarsamy.
 Medicinal Chemistry— Ashutosh Kar.
 Medicinal Chemistry—D. Sri Ram

M. Awais Fareed Pharm D IUB (2019-24)