_ BeeHARM D APP acum ANTIBACTERIAL 1 ANTIBIOTICS 1, INTRODUCTION Antibiotic is a chemical compound that inhibits or abolishes the growth of microorganisms, such as bacteria, fungi or protozoans. It also includes any agent with biological activity against living organisms; however, the term is commonly used to refer to substances with anti-bacterial, anti-fungal or anti-parasitical activity. History: Although potent antibiotic compounds for treatment of human diseases caused by bacteria such as tuberculosis, plague or leprosy were not isolated and identified until the twentieth century. The first known use of antibiotics was by the ancient Chinese over 2,500 years ago. Many other ancient cultures, including the ancient Egyptians and ancient Greeks already used molds and plants to treat infections, owing to the production of antibiotic substances by these organisms. At that time, however, the compounds having antibiotic activity present in molds or plants were unknown The antibiotic properties of Penicillium spp. were first described in France by Ernest Duchense in 1897. However, his work went by without much notice from the scientific community until Alexander Fleming’s discovery of Penicillin. Modern research on antibiotic therapy began in Germany with the development of the narrow-spectrum antibiotic Salvarsan by Paul Ehrlich in 1909, for the first time allowing 4n efficient treatment of the then-widespread problem of syphilis. The drug, which was also effective against other spirochaetal infections, is no longer in use in modern medicine. Antibiotics were further developed in Britain following the re-discovery of Penicillin im 1928 by Alexander Fleming. More than ten years later, Est Chain and Howard Florey interested in his work and came up with the purified form of Penicillin. The three the 1945 Nobel Prize in Medicine. "Antibiotic" was originally used to refer only to extracted from a fungus or other microorganism, but has come to include also ‘synthetic and semi-synthetic drugs that have antibacterial effects. 143Chapter 10 Antibacterial Antibiotics 2. CLASSIFICATION Antibiotics are classified in many ways based on chemical structure, its spectrum of activity, pharmacological activity. I. Based on Chemical Structure a) b) 8) B - Inctam antibiotics: eg. Penicillins, Cephalosporins, Monobactams, Carbapenems, Aminoglycoside antibiotics: eg. Streptomycin, Neomycin, Paromomycin, Kanamycin, Amikacin, Gentamicin, Tobramycin, Netilmicin, Sisomicin, Spectinomycin. Tetracyclines: eg. Tetracycline, Chlortetracycline, Rolitetracycline, Oxytetracycline, Methacycline, Demeclocycline, Meclocycline, Doxycycline, Minocycline. Macrolide antibiotic: eg. Erythromycin, Clarithromycin, Azithromycin, Dirithromycin, Troleandomycin, Lincomycins: eg. Lincomycin, Clindamycin. ics: eg. Vancomycin, Teicoplanin, Bacitracin, Polymyxin B, Colistin, Gramicidin. Polypeptide anti Miscellaneous: eg. Chloramphenicol, Novobiocin, Mupirocin, Quinupristin, Fusidic acid. IL. Based on Pharmacological Activity a) » Antifungal antibiotics. (@ __ Polyene: eg. Amphotericin B, Nystatin, Natamycin, Gi) Others: eg. Griseofulvin. Anticancer antibiotics: eg. actinomycin, Daunorubicin, — Doxorubic' Bleomycin, Idarubicin, Mitomycin, Plicamycin, Streptozocin, Valrubicin. Antityphoid antibiotic: eg. Chloramphenicol. 144Chapter 10 Antibacterial Antibiotics 4) Antidiarrheal antibiotic: eg. Colistin. ¢) Antituberculor antibiotics: eg. Rifampicin, Rifabutin, Cycloserine, Capreomyc' 1 B- Lactam Antibiotics B-lactam antibiotics are a broad class of antibiotics which include Penicillin derivatives, Cephalosporins, Monobactams, Carbapenems and f-lactamase inhibitors; basically any antibiotic agent which contains a i-lactam nucleus in its molecular structure. ‘They are the most widely used group of antibiotics available. Clinical Uses: P-lactam antibiotics’ are indicated for the prophylaxis and treatment of bacterial infections caused by susceptible organisms. Whilst traditionally f-lactam antibiotics were mainly active only against Gram-positive bacteria, the development of broad-spectrum f-lactam antibiotics active against various Gram-negative organisms has increased their usefulness, Mode of Action: \-lactam antibiotics are bactericidal and act by inhibiting the synthesis of the peptidoglycan layer of bacterial cell walls. The peptidoglycan layer is important for cell wall structural integrity, especially in Gram-positive organisms. The final transpeptidation step in the synthesis of the peptidoglycan is facilitated by transpeptidases known as penicillin binding proteins. 2.1.1 Penicillins Penicillin refers to a group of -lactam antibiotics used in the treatment of bacterial infections caused by susceptible, usually Gram- positive orga ms. History: The discovery of penicillin is usually attributed to Scottish scientist Sir Alexander Fleming in 1928, though others had earlier noted the antibacterial effects of Penicillium. The development of penicillin for use as a medicine is attributed to the Adelaide bom Nobel Laureate Howard Walter Florey. In March 2000, doctors of the San Juan de Dios Hospital in San Jose published manuscripts belonging to the Costa Rican scientist and medical doctor Clodomiro Picado Twight (1887-1944). The manusctipts explained Picado’s experiences between 1915 and 1927 about the inhibitory actions of the fungi of genera penic. Apparently Clorito Picado had reported his discovery to the Paris Academy of Sciences in Paris, yet did not patent it, even though his investigation had started years before Fleming’s. Fleming, at his-laboratory in St. Mary’s Hospital in London, noticed a halo of inhibition of bacterial growth around a contaminant blue-green mold Staphylococcus plate culture. 145Chapter 10 Antibacterial Antibiotics Fleming concluded that the mold was releasing a substance that was inhibiting bacterial growth and lysing the bacteria, He grew a pure culture of the mold and discovered that it was a Penicillium mold, now known to be Penicillium notatum. Fleming coined the term “Pencicillin” to describe the filtrate of a broth culture of the Penicillium mold. Even in these early stages, penicillin was found to be most effective against Gram-positive bacteria and ineffective against Gram-negative organisms and fingi. The chemical structure of penicillin was determined by Dorothy Crowfoot Hodgkin in the early 1940s, enabling synthetic production. A team of Oxford research scientists led by Australian Howard Walter Florey and including Emst Boris Chain and Norman Heatley discovered a method of mass producing the drug. Chemist John Shechan at MIT completed the first total synthesis of penicillin and some of its analogues in the early 1950s, but his methods were not efficient of mass production. Florey and Chain shared the 1945 Nobel Prize in medicine with Fleming for this work. Penicillin has since become the most widely used antibiotic to date and is still used for many Gram- positive bacterial infections. Development from Penicillin: The narrow spectrum activity of penicillins, along with the poor activity of the orally-active Phenoxymethyl penicillin, led to the search for derivatives of penicillin which could treat a wider range of infections. The first major development was Ampicillin, which offered a broader spectrum of activity than either of the original Penicillins. Further development yielded beta-lactamase- resistant penicillins including Flucloxacillin, Dicloxacillin and Methicillin. These are ineffective against the Methicillin-resistant Staphylococcus aureus strains that subsequently emerged. The line of true penicillins was the antipseudomonal penicillins, such as Ticarcillin and Piperacillin, useful for their activity against Gram-negative bacteria. However, the usefulness of the beta-lactam ring was such that related antibiotics, including the Mecillinams, the Carbapenems and most importantly, the Cephalosporins, have this at the center of their structures. Mechanism of Action: B-lactam antibiotics work by inhibiting the formation of peptidoglycan crosses links in the bacterial cell wall. The B-lactam moiety of penicillin binds to the enzyme (transpeptidase) that links the peptidoglycan molecules in bacteria and this weakens the cell wall of the bacterium. In addition, the build-up of peptidoglycan precursors triggers the activation of bacterial cell wall hydrolysis which further digests the 146Chapter 10 Antibacterial Antibiotics bacteria’s existing peptidoglycan. When the bacterial lose their cell walls they are then called spheroplasts. 2.1.1.1 Classification Penicillins are also classified on many ways depending upon the nature and spectrum of activity. I. a) Natural (Biosynthetic) Penicillins: eg. Benzyl penicillin (Penicillin - G), 2-Pentenyl penicillin (Penicillin -.F), 3-Pentenyl penicillin, n-Pentyl penicillin, n-Heptyl penicillin (Penicillin - K), p-Hydroxy benzyl penicillin, (Penicillin- X), Phenoxymethyl penicillin. (Penicillin-V) and Zero allergy penicillin (Penicillin-O). > b) Semi-synthetic Penicillins: eg. Ampicillin, Amoxicillin, Methicillin, Nafacillin, Oxacillin, Cloxacillin, Dicloxacillin, Flucloxacillin, Carbenicillin, Ticarcillin, Piperacillin, Mezlocillin, Cyclacillin. : ©) Penicillin Combination: eg. Benzathine penicillin, Procaine penicillin. Il. Based on Spectrum of Activity a) Effective against Gram-positive bacteria: eg. Benzyl penicillin, Ampicillin, Oxacillin. b) _Efffective against Gram-negative bacteria: eg. Temocillin. ©) _ Broad spectrum penicillin: eg. Ampicillin, Amoxicillin, Carbenicillin. 4) Penicillinase resistant penicillin: eg. Methicillin, Nafacillin, Oxacillin, Cloxacillin, Dicloxacillin, Flucloxacillin, ©) Acid resistant penicillin: eg. Benzathine penicillin, Phenoxymethyl penicillin, Phenoxy ethyl penicillin, Phenoxy propyl penicillin, 2.1.1.2 Chemistry ‘The structure penicillin composed of fused 4- member B-lactam ring, thiazolidine nucleus and a side chain, It contains three chiral carbon atoms (Cs, Cs and Cs). 6-Amino penicillanic acid (6-APA) is the basic skeleton structure of all penicillins. 147Antibacterial Antibiotics Chapter 10 cu, H iH S, 7 mt fs eo =H-CooH ° 6 - Amino penicillanic acid (6 - APA) Table 10, 1 Structure of Pei oO CH; Il HoH US. R.CNH-C—c~ SC | Sc, CH- COOH fe} Basic skeletan structure S.No. | Generic Chemical Name R Group Name 1. | Penicillin-G | Benzyl penicillin a — 2. | Penicillin- V Phenoxymethyl o-om— : penicillin 3. | Penicittin-x | P-Hydroxy benzyl Ho. on, penicillin 4. "| Penicillin-F | 2-Pentenyl penicillin | CHy— CHa CH = CH Ci= 5. | Penicillin-K | —n-Heptyl penicillin Cis (Cay — 6. = 3-Pentenyl penicillin | CH. CH= CH.CH; CH= 7. 7 1Pentyl penicillin CH(CHa)s— 148Antibacterial Antibiotics Chapter 10 ——— 8 | Phenethicilin | Phenoxy ethyl penictin CS... 7 be | Propiciliin Phenoxy propyl one = Penicillin oor Me) aticnin | 25Dimetiorypheny ' penicillin oot, Nafacillin penicillin 2-Ethoxy-I-naphthyl SS A C. Oxacillin 5-Methyl-3-phenyl-4- isoxazolyl penicillin 5-Methyl-3-(2- chlorophenyl)-4- isoxazoly penicillin 5-Methyl-3-(2,6- dichloropheny!)-4- isoxazolyl penicilliny Chapter 10 Antibacterial Antibiotics . . D-c-Amino benzyl- EX Is. Ampicillin tie i penicillin —/ hy, 16. | Amoxicillin | P-%-Amino-p-hydroxy Ho ( ca benzyl penicillin —/ |, -Ami 1 17. | Cyclaciltin | !Aminocyclohexy penicillin 18. | Carbeniciliin | %-Carboxy benzyl- penicillin 19. | Ticarcitlin | %Carboxy-3-thienyl- penicillin a-(4-Ethyl-2,3-dioxo-I- 20. | Piperaciliin | piperazinyl carbonyl- amino) benzyl penicillin 2.1.1.3 Structure Activity Relationship Penicillins are a group of antibiotics that contain 6-amino penicillanic acid with side chain attached at 6 amino group. Basic skeleton essential to show bactericidal activity are the bicyclic ring, free carboxylate group, Cis stereochemistry at C-5 and C-6 and acylamino side chain. (Fig. 10.1) 150Chapter 10 Antibacterial Antibiotics ‘Amide essential a Cis Stereochemistry essential — ~ 7 HoH R’ “Eps. Me Wi Ss. ? _ 7 ~2/ He B-Lactam essential Carboxylic acid essential ‘CooH=— Bicyclic system essential Fig. 10.1 Essential Structural Features of Penic ins ‘* Penicillin's core structure consists of a fused f-lactam ring and a thiazolidine ring. © The strained ® - lactum ring is essential. The greater the strain, the greater the activity. B- lactam ring binds with the pencillinase transpeptidase is responsible for cross linking of linear peptidoglycan layer. , * B- lactam ring is a good acylating agent for active penicillin binding protein serine. «Sulfur is not essential. Activity decreases if the sulphur is oxidised to sulphone or sulfoxide. + The N at position 4 is must for antibacterial activity. © The free carboxylate group is essential. The negative charge of carboxlate group directs the drug to positively charged active binding pocket at neutral pH. ‘© The carboxylate ion binds to the charged ammonium ion of lysine residue in the binding site * The acylamino side chain is a necessary feature. The modification at acylamino side chain is aimed for strengthening of activity, stability, resistance, absorption and distribution. © Enhancement of Gram-negative activity is increased introducing hydrophilic group(NH2, OH, -COOH group) to the carbon a to the carbonyl group on the side chain.eg Ampicillin. 151_ Chapter: os Antibacterial Antibiotics ‘The broad spectrum activity was further extended by introducing strong acidic Broup to the carbon a to the carbonyl group on the side chain.eg. Carbenicillin, Ticarcillin, Substitution of a side chain R on the acylamino group with an electron withdrawing group increases the electron density on the side chain carbonyl group and protect these -pencillin from acid degradation result in greater stability and oral availability. eg. Ampicillin. Since the amino group and the carboxylic group are ionisable, these compounds are poorly absorbed through the gut. Modifications such as, changing the carboxylic acid to esters were made at the carboxylic group to overcome this problem. —_-Pivampicillin and Bacampicillin are examples of ester prodrugs of Ampicillin. By placing a bulky group on the acylamino side chain, degradation of the drug by B-lactamases would be minimized. However, if a steric shield is too bulky, the penicillin would not be able to bind is an example of penicillin with a bulky group. to transpeptidase. Methicil This semi-synthetic penicillin possesses a dimethoxy benzene R group. Both the methoxy groups of the benzene are at the ortho position. 7 Nafacillin possess a naphthalene ring in its acylamino side chain which acts as @ steric shield. Flucloxacillin contains a bulky and electron-withdrawing _ heterocyclic acylamino side chain. Thus Flucloxacillin is an acid-resistant, narrow-spectrum, B-lactamase-resistant penicillin. Presence of urea group at carbon a to the carbonyl group (ureido penicillins) result in broad spectrum antibiotic. They are used parenterally and are active against Pseudomonas aeruginosa. Presence of urea group gives improved penetration into Gram-negative species. They are more active than carboxy pencillins against Streptococci and Haemophilus species.’ Azlocillin and Piperacillin are examples of ureido penicillins. 152 \ ~Chapter 10 Antibacterial Antibiotics © In vitro degradation of Pencillins can be retarded by keeping the pH of the solution between 6 to 6.8 and refrigerating them. © Cis stereochemistry of Hs and He is essential. Penicillin molecule has 3 chiral carbon, C-3, C-5 and C-6. Modification of these special arrangement results in loss of activity. « Natural Penicillins have narrow spectrum of activity and more susceptible to penicillinase. Development of semi-synthetic pencillins overcome these disadvantages. Semi-synthetic penicillins are made in the laboratory by adding different side chains onto the B-lactam ring after it is synthesized by a fungus. Semi- synthetic penicillins have various specific properties such as resistance to stomach acids so that they can be taken orally, Y a‘degree of resistance to penicillinase (a penicillin-destroying enzyme produced by some bacteria) Y extended range of activity against some Gram-negative bacteria. A. Benzyl Penicillin Potassium ° M1 . CH : crema TT i, o® cod -—W: K o It is a white crystalline powder, very soluble in water, practically insoluble in chloroform, ether, fixed oils and liquid paraffin. It contains not less than 96 percent and not more than 100.5 percent of penicillins. It is stored in tightly — closed container in a cool, dry place. It is available as intramuscular or intravenous injection or infusion in divided doses. | B. Benzathine Penicillin | { ° f] S. cH, @ | (CH,CNH TT cH prsctaca | }— [fe CHiN, CAs | ° 2 ® \ | 153Chapter 10 Antibacterial Antibiotics It is a combination of two molecule of Benzyl penicillin with one molecule of Benzathine (N, N’- dibenzyl ethylene diamine). It is a white crystalline powder, odorless, freely soluble in DMF, slightly soluble in ethanol (95%), very slightly soluble in chloroform and water, practically insoluble in ether. In should be stored in a tightly-closed container, in acool and dry place. It is given orally, intramuscularly or intravenously in a divided dose. A single injection is found to be effective for a week. C. Procaine Peni CO0(CH):N (GHD s, cuLCNH oad cy -—w- ‘coo It is a combination of equimolar mixture of one molecule of Benzyl penicillin with one molecule of Procaine. It is a white crystalline powder, slightly soluble in water, sparingly soluble in ethanol (95%). It should be stored in a tightly closed container in a cool and dry place. It is given as intramuscular injection. It is a popular respiratory product which reduces the injection frequency. NH, 2.1.1.4 Chemical Degradation . ‘The main cause of deterioration of penicillin is the reactivity of the strained lactam ring. The hydrolysis and the nature of degradation products are influenced by pH of the solution. Thus the B-lactam carbonyl group of penicillin readily undergoes nucleophilic attack by water or hydroxide ion to form inactive penicilloic acid, which on decarboxylation produces penilloic acid. On treatment with strong acid the ultimate product formed is penicillamine and penilloaldehyde. On subject to pH 2.0 and 4.0 give rise to penillic acid and penicillenic acid respectively. 154 -155 Antibacterial Antibiotics cH re ae nent —o—F \ (ee eer § ——-coon noc ° Penilloicacid Te te REN — CHC NZ? Penile acid %y, Penicilinase [ee % x—— hs % orNsOH " COOH on, Penilloie ac E ee wo ANY oie acid nou 7% eHow pew —o— \ ae ee anil a Mech 0 | on, PO H-COOH — wooo} 4 ee ar eee loate 7 ue 4 ab Met pencio |p K+ Reon 5 Mech, —ch—coon mN-cH-coon Petilloadehyde M cH : aon Penicillamine I aris 1c, covet, cits 1N-CH-COOIL Methyl penaldate 1,0 Pencillamine ° R.ENILG = CH. NH.CH. COOH ; é ° Hoo s—C— CH i 1 cay R.CNHC=.CH..NH.CH.COOH 1 Penaldie acid coon ns—C-cH, 1 Penamaldic acid Cs ° °° ome | 1 1 a + RENHCHCHO —Y» — RCNHCHCHO | cx, 1 CO; Penilloaldehyde HN-cH-Coont coon Penicillamine Penaldic acid Fig. 10.2 Degradation of Penicillin | | |Chapter 10 Antibacterial Antibiotics 2.1.1.5 Synthesis of Phenoxy methyl Penicillin Step -I: Preparation of DL Penicillamine (cuse030 ernercncoot + crcricoa BAe HnCHSHCOoM — Fo cm -HCL 7 NH, Chloroacetyl NHCOCH,CI chloric DL- Valine ie owe (CH). — C—cH COOH ii) Pyridine Bet fey e—ent-coot H,0 =} est, 2 [et emme i Boil DL-Pencilamine smu wreocr, | Fo 1 on, 2, 5, 5'= Trimethyl - 2 - thiazoline - 4 - carboxylic acid Step - Il: Preparation of t- Butylphthalimido malonaldehyde ° ° cg ° f . i ‘cca i NK + CHyCOKCH, Nee ocicr, < Br No NS Potassium phthalimide 9 ce cHo Nel eH, ° t~ Butylphthalimido malonaldehyde 156Chapter 10 Step - III: Condensation of Step I and Step I Products Antibacterial Antibiotics ° cro | JO COE 4 (CH= |—er4 COOH — ba, Cemenstin | 140 DL-Penicillamine t- Burylphthalimido malonaldehyde ore -CH.-COOH Hy) -coo8 Sat a 1NHNHy HCL PX ¢ ow Hydrine 8 NH a \ Nou Not t foN i — coocici), q coocice, sa |(-oake Phenoxy sey cre macy H-COOH Ci —c. ‘CH= COOH a Cie) | x a a Se? Ng SH-coocicasy, GH cooH mweaio sien ° Cyetstion 10 Dicyeloheny carbodiimide # fae | fo CH -COOH Phenoxymethy! penicillin 157Antibacterial Antibiotics Chapter 10 2.1.1.6 Synthesis of Benzyl Penicillin “= Oh=O$ ‘coon Phibalimide pic nie 140 NS Phihaic acid KOH /Fesion e | BrcH:co0CCCH), Noe CODEC, ss 7 “Kr 7 < f wcooen, % a one, Potassium pbaimise + eure prom Ls Penicilamine ony anct c¢cy, 5 (Calyx Hee ecerty, | 0/1 eo ia a a eavond mip — 2 eran, SHEED a He bd - L—n. CH -COOH ©” 7 Amino penicillanic acid ’ Benzyl penicillin 2.1.2 Cephalosporins Cephalosporin compounds were first isolated from cultures of Cephalosporium acremonium from a sewer in Sardinia in 1948 by Italian scientist Giuseppe Brotzu. He noticed that these cultures produced substances that were effective against Salmonella typhi, the cause of typhoid fever. Researchers at the Sir William Dunn School of Pathology at the University of Oxford isolated Cephalosporin C, which had stability to B-lactamases but was not sufficiently potent for clinical use. The cephalosporin nucleus, 7-Amino 158Chapter 10 Antibacterial Antibiotics cephalosporanic acid (7-ACA), was derived from Cephalosporin C and proved to be analogous to the penicillin nucleus 6-Amino penicillanic acid. Modification of the 7-ACA side-chains resulted in the development of useful antibiotic agents and the first agent Cephalothin was launched by Eli Lilly in 1964. Mode of Action: Cephalosporins are bactericidal and have the same mode of action as other B-lactam antibiotics (such as Penicillins). Cephalosporins disrupt the synthesis of the peptodoglycan layer of bacterial cell walls. The peptidoglycan layer is important for cell wall structural integrity, especially in Gram-positive organisms. The final transpeptidation step’ in the synthesis of the peptidoglycan is facilitated by transpeptidases known as penicillin binding proteins. Clinical Uses: Cephalosporins are indicated for the prophylaxis and treatment of bacterial infections caused by susceptible organisms. First-generation cephalosporins are predominantly active against Gram-positive bacteria and successive generations have increased activity against Gram-negative bacteria. Chemistry ‘The nomenclature of Cephalosporin is more complex than penicillins because of the presence of a double bond in dihydrothiazine ring. Basic nucleus is made up of 7-Amino cephalosporanic acid (7-ACA). 5 nut nn 8 8S Glial eet sf 'CH,OCCHy ° Coon +7 Amino cephalosporanic acid oy Ao coon Cepham : as Cephalosporanic acid ee Semi synthetic modification of the basic 7-ACA nucleus has resulted from acylation of the 7-amino group with different acids or nucleophilic substitution or reduction of the acetoxyl group. 159Chapter 10 Antibacterial Antibiotics The semi-synthetic Cephalosporins has got the following advantages over natural Caphalosporins . increased acid stability J © better oral absorption © broad antimicrobial spectrum © increased activity against resistant microorganisms. © decreased allergenicity © increased tolerance. Za 33 2.1.2.1 Classification 33 £i i The classification of Cephalosporin into generations is commonly practiced, E J} atthough the exact categorization of Cephalosporins is often in precise. \) _ I. Based on Generations '~ 4) First Generation: eg. Cefacetrile, Cefalexin, Cefadroxil, Cefaloglycin, Cagnatoini Cefalonium, Cefaloridine, Cefalotin, Cefapirin, Cefatrizine, Cefazaflur Cefazedone. Cefazolin, Cefradine, Cefroxadine, Ceftezole. b) Second Generation: eg. Cefonicid, Cefprozil, Cefuroxime, Cefuzonam, Cefaclor, Cefamandole, Ceforanide, Cefotiam, Carbacephems, > Loracarbef, Cephamycins, Cefouperazone, Cefmetazole, % eS Cefminox, Cefotetan, Cefoxitin. a > c) Third Generation: eg. Cefeapene, Cefdaloxime, Cefdinir, Cefditoren, Cefetamet, > Cefixime, Cefmenoxime, Cefodizime, Cefoperazone, Cefotaxime, Cefpimizole, Cefpodoxime. Cefteram, Ceftibuten, Ceftiofur, Ceftiolene, Ceftizoxime, Ceftriaxone, f \ Ceftazidime, Cefpiramide, Cefsulodin. + i 4) Fourth Generation: eg. Cefelidine, Cefepime, Cefluprenam, Cefoselis, Cefozopran, f Cefpirome, Cefquinome. ‘et to be Classified: eg. Cefaclomezine, Cefaloram, Cefaparole, Cefcanel, Cefedrolor, Cefempidone Cefetrizole, Cefivitril, 160Chapter 10 Antibacterial Antibiotics Cefmatilen, Cefmepidium, Cefovecin, Cefoxazole Cefrotil, Cefsumide, Ceftioxide, Ceftobiprole, Cefuracetime, [1 Based on Route of Administration a) Oral Cephalosporins: eg. Cephalexin, Cephadrine, Cefadroxil, Cefachlor, Cefprozil, Loracarbef, Cefuroxine auetil, Cefpodoxime proxetil, Cefixime. b) Parenteral Cephalosporins: eg. Cephalothin, Caphapirin, Cefazolin, Cefamandole, Cefonicid ,Cefornaide, Cefuroxime, Cefotaxime, Ceftizoxime, Ceftrixone, Ceftazidime, Cefoperazone. Table 10.2 Structure of Cephalosporins Ry Generic Name R Rr First Generation Cephalexin < a -CH3 — ‘NH, Cephradine (ye a —CHs x, Cefadroxil -CHs 161Antibacterial Antibiotics Chapter 10 1 —cHoccn, Cephalothis g » ~ ephalothin _ ° — CH,OCCH, aan L \-s-on- to: _ ° C ) Cephaloridin { \ a ™ 2 1 =o, = Tw Cefazolin | ak vow » 7 AS / ‘ ¥ s ‘Second Generation Cefachlor (> r —/ ey, — 2 { \ o —cH =cicr, hn 5 | worl \, ~Cefamandole t 4 ‘OH ! cn, — -ol \ Cefonicid ie es / | oH CH,SOyH < 162Chapter 10 Antibacterial Antibiotics i cpa, | cu,coon La ° Cefuroxime es. ‘0 i] —cHocNnty NocH, Nt = CHROCONH: Cefoxitin l Vo s (COCHS at C; also) Cefotetan s \ ° Cefotaxime Hn c= V wo —cxyocers Nocit 163Chapter 10 Antibacterial Antibiotics ee Cefixime mk ‘ - —CH = CH, \ a ‘NOCH,CO,H = Ceftizoxime nh iG -H N’ I NOCH; A moO Gefviasine oe ees N’ I SS 5 NocHs —cHs N (\ we Cctacibine Si at \ toc Es wd \ gt on im he ES Cefoperazone t cee . I ‘CH; J T Cefivitril NC-CH=CH-S-CH, — 16410 Chapter Antibacterial Antibiotics x—N v0 o / Cefpiramide Ory. te \ $-CH, — Wy W0 ° Cefsulodin oun A NN, SoH ee Fourth Generation Generic Name R Cefepime O oe < cu, Z Cefpirome LK le, 2.1.2.2 Structure Activity Relationship ‘Acylamino group —_Dihydro thiazine —_— —— = Amino cephalospoanic acid : Fig.10.3 Essential Structural Features of Cephalosporins 165Antibacterial Antibiotics Chapter 10 Structure activity relationships of Cephalosporins are similar to penicillins. The cephalosporins are derivatives of 7-aminocephalosporanic acid and are closely related in structure to penicillin. 7-aminocephalosporinic acid (7-ACA) is used as precursor for many semisynthetic cephalosporins. The cis-stereochemistry at positions 6 and 7 are important. ‘They are relatively stable in dilute acid and are highly resistant to penicillinase. They have a B-lactam ring which is vital for activity. Bicyclic ring system, fused B- lactam and dihydrothiazine ring form a bicyclic system which is important for activity. The acylamino side chain may be altered. Other groups may be substituted for the acetoxy group which may or may not serve as good leaving group. The nature of the leaving group is important for activity. Better leaving group tend to give Cephalosporin C analogues with better activity. First-Generation Cephalosporins: Activity is comparably lower than penicillins but possess a broad spectrum of activity. Greater activity for Gram-positive organisms than Gram-negative organisms. The methyl group in Cephalexin is a poor leaving group (less active), but improves absorption. Cephalothin has an acetoxy as. a leaving group and 1-(Thiophen-2-yl) propan-2-one in its acylamino side chain. Despite being a good leaving group, the acetoxy moiety is susceptible to enzyme- catalysed hydrolysis. Cephaloglycin’s acylamino side chain is same as Ampicillin’s. Cephaloridine has pyridinium as a leaving group. Second-Generation Cephalosporins: Have increased activity against Gram- negative bacteria, but some have decreased Gram-positive activity. Many can cross the blood-brain barrier. Cefuroxime is also an example of an oximino cephalosporin. The presence of the imino methoxy group appears to increase stability against certain B-lactamases. Third-Generation Cephalosporins: Even better activity against Gram-negative bacteria. Some compounds have the same problem of decreased Gram-positive 166a Chapter 10 Antibacterial Antibiotics activity as the previous generation. Improved f-lactamase resistance and many can also cross blood-brain barrier. Fourth-Generation Cephalosporins: Better Gram-negative bacteria activity, B-lactamase resistance and many can also cross the blood-brain barrier. Gram: positive activity similar to the 1* generation. The fourth generation compound exists as zwitterion. They are also used against Pseudomonas aeruginosa. 2.1.2.3 Synthesis of 7- Amino cephalosporanic acid from Cephalosporin C ‘Noct woe == (ete na inweoon Ni, o J ‘cH,ococn, coon Cephalosporin C Move —ent~ (cxyye—w uv ma ° ne ° ‘cuococn, coon, s. Hove ~~ (crayon 1 on on ° cizococn, ‘coon | Cetintion s. 0 x = toc” aie ‘cu,ococn, ° bs coon ‘criocoon, 7- Amino cephalosporanic acid oa 167oe Chapter 10 Antibacterial Antibig, 2.1.2.4 Synthesis of Cephalexin cx.coon (Ct C0-Cot ce coon. | Tein ‘amine NH, a corey - Hel | (CH) CHOC Oe Eile, of ‘cH,ococt, Leo. Ce 7- Amino cephalosporanic acid Phenylglycine ° C\- = I la ce . Cephalexin COOH ‘cH,0CocH, 2.1.2.5 Synthesis of Cefazolin J tyChapter 10 Antibacterial Antibiotics , 4 w Mery Od. Cefazolin Fai 2.1.2.6 Synthesis of Cephalothin Cli oh t ad > fh 2 Thienyl acetylehloride LS Ht] Condensation 7 Amino cephalosporanicacid 2.1.2.7 Synthesis of Cefadroxil taereie "TIS a cH, Hel | (CH)OnCH,SICI ‘Amino ° acid cephalosporante acid (7- ADACA) i HCI NHCO.C(CH), ° He cH om 169ft = ™~ Chapter 10 Antibacterial Antibiotics . s. HO. a HN: TT NHCOCKH), t CH ° ° Butanol / H* Coosicityoct): ° 1 s. HO eaee —HN amie MLCO.C (CH, Jn, 4 ° Aen, ° coon | cr,coon ° 1 8. 0. | NH | ° Aon, Cefadroxil coon 2.1.2.8 Degradation of Cephalosporins i s. ' aN. i Rent : RONH _ to f re i (Ph ¥ o CHR Ce CHR of ‘cH,On aoaue coon coon — cet} ONCE Cephaosposin Desacey cephalosporin p-Lacamase 67 (00 HB -r0| © bam. 75 ° a 1 neontce: ‘ Ale me ET LOAN uh , o a cir / f° ey i i ZA FS — Rates accept Desacetyl -7- amino cephalosporanie acid lactone 170Chapter 10 Antibacterial Antibiotics 2.2 Aminoglycoside Antibiotics 2.2.1 Introduction Aminoglycoside antibiotics are from the Actinomycetes, particularly from the genus Streptomyces. Among the many antibiotics isolated from the genus, important one is Streptomycin. Several other closely related structures are Kanamycin, Neomycin, Gentamycin, Paromomycin, Tobramycin and Netilmycin. All the structures consist of amino sugars linked glycosidically. All have one amino hexose and some have a pentose. Additionally each aminoglycosides contains a highly substituted 1, 3 - diamino cyclohexane central ring. They are all basic, water-soluble compounds or mixtures of compounds. Mechanism of Action: They act directly on the bacterial ribosome to inhibit the initiation of protein synthesis and to interfere with translation of the genetic message. They bind to the 30S ribosomal subunit to form a complex that cannot initiate proper amino acid polymerization. Spectrum of Activity: Aminoglycoside antibiotics are broad-spectrum antibiotics, particularly effective against Gram-negative bacilli, Gram-negative and Gram-positive cocci except Staphylococci, tent to be less sensitive. Streptomycin is particularly active against Mycobacterium tuberculosis. Paromomycin is used in amoebic dysentery. A. Streptomycin on 4" H CHIN CHOW N=Methyl-L- glucosamine Streptidine - H Hot streptose onc H Streptomycin imi Chapter 10 Antibacterial Antibiotics Streptomycin is produced by a culture of Streptomyces griseus. It is official as Streptomycin sulfate salt, which is a white, odorless, hygroscopic powder, but stable towards light and air. It is freely soluble in water, slightly soluble in alcohol, but insoluble in most of the organic solvents. Hydrolysis ‘Acid hydrolysis Streptomycin > Steeptidine + _—_Streplobiosamine iscidc base) (Mono acidic Sisacharide) [ieee | : Hyerolysis ied N- Methyl - L- glucosamine (Diamine compound) Streptose cannot be isolated because of its instab Use: It is active against various Gram-positive and Gram-negative bacteria. It is effective against Mycobacterium tuberculosis. It is used in the treatment of TB along with other drugs PAS and Rifampicin. B, Neomyein B Neomycin B 172Chapter 10 Antibacterial Antibiotics Neomycin is a mixture of three substance isolated from Streptomyces fradiae. The three components A, B and C have been separated by counter-current distribution technique. Neomycin A is a disaccharide, which is a common degradation product of Neomycin B and C. Neomycin B and C consists of four carbohydrate units and differ only in the configuration of the amino-methyl group in the neosamine ring linked to the ribose unit. Neomycin as a sulfate salt is a white to slightly yellow crystalline hygroscopic and photosensitive powder, very soluble in water. Use: It is used for treatment of gastrointestinal infections, dermatological infections and acute bacterial peritonitis. C. Paromomycin Desoxy streptamine ‘Neosamine - B or C Paromomycin I : Ri= H; Ra = CHaNH2 Paromomycin If: Ri = CHsNH; Ra=H It was isolated from cultures of Streptomyces rimosus. It is more closely resembles Neomycin and Streptomycin in antibiotic, than it does Oxytetracycline, the antibiotic : ‘ 173