CEPHALOSPORINS

O
H
R C N H H
S

N OAc
O
CO2H

Assist.Prof.Dr. Mohammed Hassan

Lecture2
CEPHALOSPORINS
Naturally cephalosporin is cephalosporin C
The more useful semisynthetic modifications of the
basic 7-ACA nucleus have resulted from
-acylations of the 7-amino group with different acids
or nucleophilic substitution or reduction of the
acetoxyl group.
 Chemical Degradation
Cephalosporins undergo various hydrolytic degradation
reactions whose specific nature depends on the individual
structure .
Among 7-acylaminocephalosporanic acid derivatives, the 3-
acetoxylmethyl group is the most reactive site.
The acetoxyl function of this group readily undergoes
solvolysis in strongly acidic solutions to form the
desacetylcephalosporin derivatives. The latter lactonize to
form the desacetylcephalosporin lactones, which are virtually
inactive.
The 7-acylamino group of some
cephalosporins can also be
hydrolyzed under enzymatic
(acylases) and, possibly,
nonenzymatic conditions to give 7-
ACA (or 7-ADCA) derivatives.
Following hydrolysis or solvolysis of
the 3-acetoxymethyl group, 7-ACA
also lactonizes under acidic
conditions .
The reactive functionality common to all cephalosporins is the
β-lactam. Hydrolysis of the β-lactam of cephalosporins is
believed to give initially cephalosporoic acids (in which the R′
group is stable, [e.g., R′ = H or S heterocycle]) or possibly
anhydrodesacetylcephalosporoic acids (7-ADCA, for the 7-
acylaminocephalosporanic acids).
 SAR of Cephalosporins
H H H
R N S
1
7 6 2
O 8 5 3
N 4 O Me
O C

CO2H O

•Similar to penicillins
•The b-lactam ring is crucial to the mechanism
•The carboxylic acid at position 4 is important to binding
•The bicyclic system is important in increasing ring strain
•Stereochemistry is important
•The acetoxy substituent is important to the mechanism

Possible modifications
•7-Acylamino side chain
•3-Acetoxymethyl side chain
•Substitution at C-7
Variation of the 7-Acylamino Side Chain

•Not possible to generate analogues by fermentation

•Not possible to generate analogues by a full synthesis
•Restricted to semi-synthetic procedure

H H H H H
R N S
H2N S

RCOCl O
N O Me N O Me
O C
O C
O CO2H O
CO2H

7-ACA
Mechanism of Action

H H H
S
R N H H H
R N S
7
O -CH3CO2-
N O Me O
O C N
O
CO2H O
O
CO2H
OH Ser

Ser Enzyme
Enzyme

• The acetoxy group acts as a good leaving group and aids the
mechanism
 Oral Cephalosporins
The oral activity conferred by the phenylglycyl substituent is
attributed to increased acid stability of the lactam ring, resulting
from the presence of a protonated amino group on the 7-acylamino
portion of the molecule.
Also important for high acid stability is the absence of the
leaving group at the 3-position. Thus, despite the presence
of the phenylglycyl side chain in its structure, the
cephalosporanic acid derivative cephaloglycin is poorly
absorbed orally, because of solvolysis of the 3-acetoxyl
group in the low pH of the stomach. The resulting 3-
hydroxyl derivative undergoes lactonization under acidic
conditions. The 3-hydroxyl derivatives and, especially, the
corresponding lactones are considerably less active in vitro
than the parent cephalosporins.
 Spectrum of Activity

The cephalosporins are considered broad-spectrum antibiotics

with patterns of antibacterial effectiveness comparable to that
of ampicillin. Several significant differences exist, however.
Cephalosporins are much more resistant to inactivation by β-
lactamases, particularly those produced by Gram-positive
bacteria, than is ampicillin. Ampicillin however, is generally
more active against non-β-lactamase-producing strains of
Gram-positive and Gram-negative bacteria sensitive to both it
and the cephalosporins.
 β-Lactamase Resistance
The susceptibility of cephalosporins to various lactamases varies
considerably with the source and properties of these enzymes.
Cephalosporins are significantly less sensitive than all the β-
lactamase-resistant penicillins to hydrolysis by the enzymes from
S. aureus and Bacillus subtilis.
The “penicillinase” resistance of cephalosporins appears to be a
property of the bicyclic cephem ring system rather than of the
acyl group.
Despite natural resistance to staphylococcal β-lactamase, the
different cephalosporins exhibit considerable variation in rates of
hydrolysis by the enzyme. Thus, of several cephalosporins tested
in vitro, cephalothin and cefoxitin are the most resistant, and
cefazolin are the least resistant.
 H H H
S
N
7
S O 3
N OAc
O
CO2H

cephalothin cefoxitin

cefazolin
The introduction of polar substituents in the aminoacyl moiety of
cephalosporins appears to confer stability to some β-lactamases.
Two structural features confer broadly based resistance to β-
lactamases among the cephalosporins: (a) an alkoximino function
in the aminoacyl group and (b) a methoxyl substituent at the 7-
position of the cephem nucleus having α stereochemistry.
The structures of several β-lactamase-resistant cephalosporins,
including cefuroxime, cefotaxime, ceftizoxime, and ceftriaxone,
feature a methoximino acyl group.

cefotaxime
cefuroxime ceftriaxone
β-Lactamase resistance is enhanced modestly if the oximino
substituent also features a polar function, as in ceftazidime,
which has a 2-methylpropionic acid substituent on the
oximino group. Both steric and electronic properties of the
alkoximino group may contribute to the β-lactamase
resistance
Classification
Cephalosporins are divided into first-, second-,
third-, and fourth-generation agents, based roughly
on their time of discovery and their antimicrobial
properties (Table 1). In general, progression from
first to fourth generation is associated with
-a broadening of the Gram-negative antibacterial
spectrum.
-some reduction in activity against Gram-positive
organisms
-enhanced resistance to β-lactamases.
 (Table 1)

Generation Parenteral Oral Agents

Agents
First-generation Cefazolin Cefadroxil, cephalexin

Second-generation Cefotetan, cefoxitin, Cefaclor, cefprozil,

cefuroxime cefuroxime axetil,
loracarbef
Third-generation Cefotaxime, Cefdinir, cefditoren,
ceftazidime, cefpodoxime proxetil,
ceftizoxime, ceftibuten, cefixime
ceftriaxone
Fourth-generation Cefepime
 First Generation Cephalosporins
Cephalothin
H H H
S
N
7
S O 3
N OAc
O
CO2H

• First generation cephalosporin

• More active than penicillin G vs. some Gram -ve bacteria
• Less likely to cause allergic reactions
• Useful vs. penicillinase producing strains of S. aureus
• Not active vs. Pseudonomas aeruginosa
• Poorly absorbed from GIT
• Administered by injection
• Metabolised to give a free 3-hydroxymethyl group
(deacetylation)
• Metabolite is less active
Cephalothin - drug metabolism

H H H H H H
N S N S
7
S O 3 S O
N OAc N OH
O Metabolism O
CO2H CO2H

Less active
OH is a poorer leaving group
Cephaloridine
H H H
N S
7
S O 3
N N
O
CO2

• The pyridine ring is a good leaving group (neutralisation of

charge)
• Exists as a zwitterion and is soluble in water
• Poorly absorbed through the gut wall
• Administered by injection
Cefalexin
H2N H
H H H
S
N
7
O 3
N
O Me

CO2H

• The methyl group at position 3 is not a good leaving group

• The methyl group is bad for activity but aids oral absorption
• Cefalexin can be administered orally
• A hydrophilic amino group at the alpha-carbon of the side chain
helps to compensate for the loss of activity due to the methyl
group
 First Generation Cephalosporins

Cefazolin
Cefadroxil
Second Generation Cephalosporins
Oximinocephalosporins

Cefuroxime

• Much greater stability against some b-lactamases

• Resistant to esterases due to the urethane group
• Wide spectrum of activity
• Useful against organisms that have gained resistance to penicillin
• Not active against P. aeruginosa
• Used clinically against respiratory infections
• The second-generation cephalosporins have a
greater Gram-negative spectrum while retaining
some activity against Gram-positive cocci. They are
also more resistant to beta-lactamase.

• Cefaclor
 Cefonicid Cefprozil

Cefuroxime
 Forms of Cefuroxime
(2nd generation cephalosporin)

H OMe H
N S
7
S O 3
N O NH2
O C

CO2H O

Cefuroxime
Cefuroxime axetil
orally

The Second-generation cephalosporins include Cefotetan,

cefoxitin, and cefuroxime (all parenteral) as well as Cefaclor,
cefprozil, cefuroxime axetil, and loracarbef (all oral).
Third Generation Cephalosporins
Oximinocephalosporins
Aminothiazole
ring

• Aminothiazole ring enhances penetration of cephalosporins

across the outer membrane of Gram -ve bacteria
• May also increase affinity for the transpeptidase enzyme
• Good activity against Gram -ve bacteria
• Variable activity against Gram +ve cocci
• Variable activity vs. P. aeruginosa
 Ceftazidime

• Injectable cephalosporin
• Excellent activity vs. P. aeruginosa and other Gram -ve bacteria
• Can cross the blood brain barrier
• Used to treat meningitis
Cefotaxime ceftazidime

ceftriaxone
cefixime
 cefpodoxime proxetil

The Third-generation Cephalosporins include

Cefotaxime, ceftazidime, ceftizoxime, and
ceftriaxone (all parenteral) as well as Cefdinir,
cefditoren, cefpodoxime proxetil, ceftibuten, and
cefixime (all oral).
Fourth Generation Cephalosporins
Oximinocephalosporins

Cefepime

• Zwitterionic compounds
• Enhanced ability to cross the outer membrane of Gram negative
bacteria
• Good affinity for the transpeptidase enzyme
• Low affinity for some b-lactamases
• Active vs. Gram +ve cocci and a broad array of Gram -ve bacteria
• Active vs. P. aeruginosa
• parenterally effective.
 Newer beta-Lactam Antibiotics (CARBAPENEMS)
Thienamycin
Acylamino side
chain absent Opposite
stereochemistry
to penicillins
OH

H
Plays a role
H
in ß-lactamase
resistance H3C NH3
S
N

O
Double bond leading to
CO2 high ring strain and an increase
in -lactam ring reactivity
Carbapenam nucleus

• Potent and wide range of activity vs Gram +ve and Gram -ve
bacteria
• Active vs. Pseudomonas aeruginosa
• Low toxicity
• High resistance to b-lactamases
• Poor stability in solution (ten times less stable than Pen G)
Thienamycin analogues used in the clinic
NH
H OH HN
H
Me
S Imipenem
N
O
CO2

O
H
N C
H OH H N
Me
H
Me
Me Meropenem
S
N
O
CO2

O
H
N C
H OH Me N
H
H
Me
Ertapenem(2002)
S
N
O CO2
CO2