Concise &Conceptual Series

-Lactam Antibiotics

-LACTAM ANTIBIOTICS
-lactam antibiotics are a broad class of antibiotics that include penicillin derivatives, cephalosporins , monobactams, and carbapenems, that is, any antibiotic agent that contains a -lactam nucleus in its molecular structure. A beta-lactam ring (-lactam) is a lactam with a heteroatomic ring structure, consisting of three carbon atoms and one nitrogen atom. A lactam is a cyclic amide.

-LACTAM SKELETONS
The agents having current or potential therapeutic use can be represented by following principal chemical skeletons: 1) PENAMS Penicillins are bicyclic structures where the p-lactam ring is fused with a fivemembered thiazolidine ring. 2) PENEMS These differ from penams by the presence of a double bond between the C2 and C3 positions. No natural penems have so far been described, but many have been synthesized in the expectation that they would combine desirable properties of penicillins and carbapenems. 3) CARBAPENAMS AND CARBAPENEMS These compounds differ from penams and penems by the presence of a carbon atom at position 1. Many natural and synthetic members of the group have been described, and some of them are currently used in the clinics.

Concise &Conceptual Series Penam Cephem

-Lactam Antibiotics Carbapenem

Penem

Oxacephem

Oxapenam

Monobactam

Carbacephem

Carbapenam

4) CEPHEMS, OXACEPHEMS, AND CARBACEPHEMS

Concise &Conceptual Series

-Lactam Antibiotics

These compounds are characterized by the presence of a P-lactam ring fused with a sixmembered unsaturated ring, having at position 1 a sulfur, an oxygen, or a carbon atom, respectively. In particular, the Cephem class has been very prolific in generating good antibiotics, which have found extensive application in the treatment of bacterial infections. 5) OXAPENAMS This class is generally characterized by a very weak antibacterial activity but has found therapeutic applications as inhibitors of bacterial p-lactamases. Its skeleton differs from that of penams by the presence of an oxygen atom at position 1. 6) MONOBACTAMS The monocyclic p-lactam is the simplest structure still retaining antibacterial activity.

-LACTAM REACTIVITY AND BIOLOGICAL ACTIVITY

The remarkable properties of penicillin molecule, including its antibacterial activity could possibly be explained due to the fusion of the -lactam ring with the thiazolidine ring. The strain developed the fusion of the five-membered thiazolidine ring with the four-membered b-lactam ring leads to nonplanarity of the molecule, which results in a large angle and torsional rotation. These factors make it enormously labile to any kind of nucleophilic attack as well as in the presence of acid, alkali or even neutral molecules like water and alcohol. Carbapenems like thienamycin and meropenem, panipenem have shown exceptionally high broad-spectrum activity as well as ability to inactivate lactamases, since they combine the functional feature of the best of b-lactams as well as -lactamase inhibitors.

-LACTAM ANTIBIOTIC ALLERGY

-Lactam antibiotics (penicillin in particular) are known to cause allergic reaction in 3 to 5% of patients who take the drug by parenteral route. All -lactam antibiotics ranging from benzyl penicillin to the more recently introduced - lactams such as aztreonam or even the related b-lactamase inhibitors, like clavulanic acid76 may induce allergic reactions to some extent depending on their degradation products. The allergic reactions usually appear within a maximum interval of 1 h after drug intake and are mediated by IgE antibodies, while the symptoms are 3

Concise &Conceptual Series

-Lactam Antibiotics

produced by rapid release of histamine and other vasoactive inflammatory mediators immediately after haptenantibody interaction. Another characteristic reaction of penicillin is that it acts as acylating (penicilloylating) agent with reactivity comparable to carboxylic acid anhydrides. This factor leads to many difficulties in isolation, manipulation as well as polymer formation causing allergenicity of the penicillins. EXAMPLES OF SOME NON CLASSICAL -LACTAM ANTIBIOTICS
OH CH3 H H H3C O N NH H3C O HOOC H H N O N CH3 SO3H H2N O N OH CH3 H H

S COOH

N H

O S NH N CH3 CH3

Imipenem [9] O H3C CH3 HOOC C O N C C S N NH3 Aztreonam[11]

Figure 3.

COOH Meropenem [10] O N H OH O H

OH N

Norcardicin [12] COOH

Some non -classical -lactam antibiotics.

DEGRADATION OF -LACTAMS IN THE PRESENCE OF METAL IONS

Degradation of penicillins and cephalosporins has been studied extensively in the presence of various metal ions, viz. mercury, zinc, cadmium, cobalt and copper. It was found that these ions catalysed the rate of inactivation or hydrolytic opening of -lactams. It has been established that the metal ions catalyse penicillin degradation following two routes, i.e. first by interacting with sulphur atom of the penicillenic acid intermediate to form metal mercaptide (36) and second by complexing the penicillin with metal ion to form chelate (37). It has been observed that the former pathway plays a major role during penicillin degradation by metal catalysis.

Concise &Conceptual Series

M N R C O H CH S CH3 CH3 COOH

-Lactam Antibiotics
R C O O M [37] H N H H N S CH3 CH3 COOH

N O [3 6]

DEGRADATION OF PENICILLIN
Penicillin is a unique molecule containing unstable, highly strained and reactive b-lactam amide bond. The degradation of penicillin takes place in various conditions, viz. alkaline or acidic, in the presence of enzyme -lactamase or treatment of weak nucleophiles like water and metal ions. The strained bicyclic system in penicillin is supposed to remain in equilibrium with pseudopenicillin (14) under neutral condition, which contains an oxazolone structure leading to various degradation products under a variety of conditions as detailed herein (Figure 4). It undergoes further isomerization on aging to yield penicillenic acid (17), as characterized by its strong UV absorption at 322 nm and has been regarded as haptenic component for penicillin allergenicity. Hence most of the oral and parenteral -lactam preparations are generally kept below 1% moisture level during their shelf life. The highly strained -lactam ring and its amide bond break open in the presence of acid giving an array of complex products, including penilloic acid, penicillamine and penilloaldehyde through the highly unstable intermediate, viz. penillic acid, penicilloic acid and penicillenic acid. Penicilloic acid (16) exists in its isomeric form, i.e. penamaldic acid (19) by opening of the thiazolidine ring which has been characterized by its UV peak around 320 nm under strong acid condition. It further degrades to give the ultimate decomposed products, viz. penilloaldehyde (22) through its parent acid i.e. penaldic acid (20), and penicillamine (21); possibly through its N-formyl penicillamine. It may also undergo ready decarboxylation to give penilloic acid (18). In strong acidic medium (pH 2 or less), it undergoes rearrangement through oxazoline formation giving rise to penillic acid (15). It was thought worthwhile to protect the degradation of penicillins under acidic conditions by putting some electron-withdrawing groups in the -position of the N-acyl side chain, which would decrease the electron density on the side chain 5

Concise &Conceptual Series

-Lactam Antibiotics

carbonyl group and reduce its tendency to act as nucleophile and thus protect these penicillins. Such modifications resulted in the introduction of some acid-resistant penicillins, viz. ampicillin, amoxicillin, carbenicillin, ticarcillin, etc. in the market. Penicillins undergo degradation rapidly in alkaline conditions (pH 7.59.0), wherein the amide bond gets open to give penicilloic acid (16). The carboxyl group present on penicilloic acid after bond opening, undergoes decarboxylation giving rise to penilloic acid (18). The formation of these degradation products of penicillin leads to the total loss of activity. The extensive degradation of ampicillin was studied under alkaline conditions. The first degradation product in alkaline condition was 5R-penicilloic acid, which subsequently undergoes epimerization at C-5 to form the 5S-isomer via the imine tautomer. Penicillin undergoes degradation by an enzyme -lactamase or penicillinase, which is produced by many penicillin- resistant bacteria. It is also reported that cleavage of - lactam amide bond takes place in water when it is heated, but this cleavage is slower than -lactamase inhibitors.

Concise &Conceptual Series

-Lactam Antibiotics
[19] COOH H Strong HC acid N

C O

H N

H CH HN O H

CH3 CH3 COOH

CH3 CH3 + [19] COOH

C O

H H H N S N H

CH3

idi Ac

H cp

-H H S CH3 CH3

R Penillic acid [15] HgCl2 [20] + [ 21] N-Penicilloyl Derivative

CH3

Neutral pH R

C O

CH HN

O Penicillin [13]

COOH
H / se k .p n a A l ic i l l i n Pe

C O

H N

H CH2 HN

CH3

CH3

R -CO2

Penilloic acid [18] Acidic pH [20] + [21]

COOH

H COOH O Pseudopenicillin [14] O Isomeri2 SH H sation H H HS CH3 N CH3 N S R C CH H2O CH3 CH3 HN O O C HN COOH O O COOH OH O] Penicillenic acid [17] [H 2 Penicilloic acid [16] HS +

Isomerisation CH3 HS Strong Acidic R C C CH3 pH O C HN O COOH OH Penamaldic acid [19] H N R= C6H5CH2 - or -CH2OC6H5

C O

NH

CH

CHO

CH3 CH3

COOH O -CO2

Penaldic acid [20]

H2N COOH Penicillamine [21]

C NH CH2 CHO Penilloaldehyde[22]

Figure 4. Pathways of degradation of penicillin in acidic and alkaline conditions.

Concise &Conceptual Series

R C O H N H H N S CH3 CH3

-Lactam Antibiotics
R C OH N H H N S CH3 CH3

O COOK Potassium Penicillin-G or V[1] R= C6H5CH2 - or -CH2OC6H5

i) Trimethylchlorosilane ii) Pyridine [Chemical process]

CO2SiMe3 Silyl ester (enolform) i) PCl5 ,Pyridine ii) ROH N H H N S CH3 CH3 CO2SiMe3

H2N

H N

Penicillin amidase/acylase (Enzymatic process) CH3 S CH3 COOH R COOR [H2O]

C OR

6-APA[2]
F igure 1.

By-product [4]

Imino ether [3] (R= alkyl)

6-A prod ction b chem andenzym d rad PA u y ical atic eg ation of penicillin.

DEGRADATION OF CEPHALOSPORINS
Although cephalosporins are more stable to hydrolytic degradation reactions than penicillins, they experience a variety of chemical and enzymatic transformations, whose specific nature depends on the side chain at C-7 and the substituent on C-3 atom. The presence of a good leaving group at C-3 facilitates spontaneous expulsion of the 3-substituent by concerted event due to hydrolysis of CN bond of lactam nucleus by any general nucleophile or b-lactamase. Thus, desacetyl cefotoxime is more stable to hydrolysis in comparison to cefotoxime. The absence of a leaving group at position 3 of cephalosporins makes them more acid-stable, thus rendering them suitable for oral consumption. Hence cephalexin posses sing methyl group at position 3 is much better absorbed than cephaloglycin having acetoxymethyl group at position 3, while both have identical phenylglycyl side chain at the position 7. The nature of substituents at C-7 of cephalosporins plays an important role in determining the facility with which the reactive -lactam bond is hydro lysed or broken either by chemical or enzymatic means of degradation of cephalosporin C. Isomerization of D3-cephalosporin to its D2-isomer occurs with great loss of antibacterial activity, but it leads to more stability. Cephalosporins are significantly less sensitive to hydrolysis by various lactamases of different origin than penicillins, which appears due to the intrinsic property of bicyclic ring system as discussed earlier in b-lactam reactivity. 8

Concise &Conceptual Series

-Lactam Antibiotics

However, the rate of hydrolysis by b-lacatmase varies considerably with different cephalosporins due to the nature of substituents at C-7 and C-3 as well as steric factor involved therein; although no empirical rule for the substituents exists. Like penicillins, cephalosporins are comparatively less soluble and unstable in aqueous solution and therefore are used generally as sodium salts. On enzymatic degradation in the presence of acylase cephalosporin C (23) gives 7-amino cephalosporanic acid (24), which in the presence of acid undergoes lactonization to give des acetyl-7-amino cephalosporanic acid lactone (25). In the presence of esterase, cephalosporin-C gave desacetyl cephalosporin (26) and then desacetyl cephalosporin lactone (27). The enzyme b-lactamase or cephalosporanase degraded cephalosporin C into cephalosporoic acid (28), anhydrodesacetyl cephalosporoic acid (29) and desacetyl cephalosporoic acid (30). Further breakdown of these acidic products leads to many other fragmented and rearranged products.
H2N HOOC O H H N H H H N O H N

NOCl

N CH2OAc O Cephalosporin C[5] COOH

i) Beta lactamase ii) Acetyl esterase

HOOC

S CH2OAc COOH

i) D-amino acid oxidase/H2O2 ii) Glutaryl-7-ACA amidase H2N CH3 O H H N S HOOC [H2O] CH2OAc O H H N O H N S CH2OAc

H2N O

H N OH

COOH Desacetylcephalosporanic acid [8]

F igure 2.

COOH 7-ACA [7]

COOH Imino ether [6]

7-A Aprod ction b enzym andchem d rad C u y atic ical eg ation of cephalosporin C .

Concise &Conceptual Series

-Lactam Antibiotics
O

H2N O

H N

H H N O

H N

O O Desacetyl-7-aminocephalosporanic acid lactone [25] H H N O H / -H2O S CH2R H N R

O O Desacetylcephalosporin lactone [27] O C H H N O S H N H / -H2O S CH2OH

H2N

COOH O 7-Aminocephalosporanic acid [24] H H (R=Ac) R C N [H2O] [H 2O]/Acylase 370C O S

COOH Desacetylcephalosporin [26] Esterase CH2R

COOH Cepahlosporin-C[23] R N COOH -Lactamase (Cephalosporanase) Thiazole-4-carboxylic acid(31) OH or H3O O O H H H H H H R C N S R C N S O HN OH CH2R O N OH CH2

COOH COOH Cephalosporoic acid [28] Anhydrodesacetyl cephalosporoic acid [29] (R=OAc) O Acetyl H H H R C N S esterase Fragmented and rearranged products NH3 HN CH3 CH O OH R = (CH2)3 CH COOH , R= (CH2)3 CO2 COOH Desacetyl cephalosporanic acid [30] NH2
F igure 6. Possib pathw of d rad le ays eg ation of cephalosporin C .

REFERENCES: Burger Asutoshkar Degradation of -lactam antibiotics, A. D. Deshpande, K. G. Baheti and N. R. Chatterjee*, Padmashree Dr D. Y. Patil Institute of, Pharmaceutical Sciences and Research, Pimpri, Pune 411 018, India, http://www.ias.ac.in.com www.wikipedia.com

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