Tetracyclines Aminoglycosides Macrolides Lincomycins Unclassified antibiotics Polypeptide antibiotics Fluoroquinolone antibacterial agents Sulfonamides & Sulfones

chemistry drugs mechanism of action and/or resistance of drugs Spectrum of activity Structure-activity relationships recognize drugs in this group
2

Tetracyclines
Broad-spectrum antibiotics from Streptomyces & synthesis

Chemistry

a hydrocarbon system that comprises four annelated six-membered rings It is from this tetracyclic system that the group name is derived
4

Pka3 R1 R2 R3 R4 H H NHMe 2 OH Cl
-

2.8-3.4

H3C

H N

CH3 O
-

Acid salt

OH

OH OH O

CNH2 O Pka1 O

CNH2 O

Pka2

pka: 2.8-3.4, 7.2-7.8, 9.1-9.7 amphoteric compounds forming salts with either acids (HCl) or bases (Na+, K+) In neutral solutions, these substances exist mainly as zwitterions The acid salts, which are formed through protonation of the enol group on C-2
5

7.2-7.8

Epitetracycline

Natural tetracycline

epimerization at C-4 in solutions of intermediate pH range. These isomers are called epitetracyclines Under acidic conditions, an equilibrium is established in about 1 day and consists of approximately equal amounts of the isomers epitetracyclines much less activity than the natural isomers, thus accounting for a decrease in therapeutic value of aged solutions
6

Epitetracycline

Anhydrotetracycline

Strong acids and strong bases attack OH on C-6, causing a loss in activity through modification of the C ring

H 3C

OH

NM e2 OH

5a
OH O OH OH O CN H 2 O

pH1 CH 3 NM e2 OH

11
OH O

12 11a

OH

OH O

CN H 2 O

CH 3

NM e2 OH

OH

OH OH O O

CN H 2 O

Anhydrotetracycline

Strong acids produce a dehydration through a reaction involving the 6-OH and the 5a-H double bond thus formed between positions 5a and 6 induces a shift in the position of the double bond between C-11a and C-12 to a position between C-11 and C-11a forming the more energetically favored resonant system of the naphthalene group found in the inactive anhydrotetracyclines 9

H3C

OH

NMe 2 OH pH3.5 OH OH O pH1 CNH2 O

H3C

OH

NMe 2 OH

OH

OH

OH OH O

CNH2 O

CH3

NMe 2 OH

CH3

NMe 2 OH

OH

OH OH O

CNH2 O OH O

OH OH O

CNH2 O

CH3

NMe 2 OH

CH3

NMe 2 OH

OH

OH OH O O

CNH2 O OH OH O

OH O

CNH2 O

Anhydrotetracycline

10 Epitetracycline Epianhydrotetracycline

H3C

OH

NMe 2 OH OH OH OH O CNH2 O CH3 O OH O OH OH O CNH2 O NMe 2 OH OH

-

CH3 O O-

NMe 2 OH

OH

OH O O

CNH2 O

Isotetracycline

Bases promote a reaction between the 6-OH and the ketone group at the 11-position causing the bond between the 11 and 11a to cleave and to form the lactone ring found in the inactive isotetracycline 11

O H3C OH O

NH2 O
-

O H3C OH

NH2 O
-

CH3 NH CH3 OH O

CH3 N CH3 O M O O

OH

O M O

OH

O M O

Stable complexes are formed by the tetracyclines with many metals including calcium, magnesium, and iron [Ca, Mg, Fe] chelates are very insoluble in water, accounting for the impairment in absorption of most (if not all) tetracyclines in the presence of milk calcium-, magnesium-, and aluminum-containing antacids [Ca, Mg, Al] 12 and iron salts [Fe]

Below pH 7

> pH 7

 Soluble alkalinizers, such as sodium bicarbonate

also decrease the gastrointestinal absorption of the tetracyclines Deprotonation of tetracyclines to more ionic species and their observed instability in alkaline solutions may account for this observation

13

 affinity of tetracyclines for calcium causes them to be laid down in newly formed bones and teeth as tetracycline-calcium orthophosphate complexes Deposits of these antibiotics in teeth cause a yellow discoloration that darkens (a photochemical reaction) over time Tetracyclines are distributed into the milk of lactating mothers and cross the placental barrier into the fetus The possible effects of these agents on the bones and teeth of the child should be taken into consideration before their use during pregnancy or in children under 8 years of age
14

Mechanism of action
Tetracyclines are specific inhibitors of bacterial protein synthesis bind to the 30s ribosomal subunit and prevent the binding of aminoacyl tRNA to the mRNA-ribosome complex Both the binding of aminoacyl tRNA and the binding of tetracyclines at the ribosomalbinding site require magnesium ions
15

Inhibition of bacterial protein synthesis by tetracycline

mRNA attached to 30S subunit of bacterial ribosomal RNA P (peptidyl) site of 50 S ribosomal RNA subunit contains nascent polypeptide chain Aminoacyl tRNA charged with the next amino acid (aa) to be added to the chain moves into the A (acceptor) site, with complementary base pairing between the anticodon sequence of tRNA and the codon sequence of mRNA Tetracycline: bind 30 S subunit, block tRNA binding to A site.
16

Ribosomal subunit
tRNA

Amino acid
17

2009

Steps in bacterial protein synthesis and targets of several antibiotics. Amino acids are shown as numbered circles. The 70S ribosomal mRNA complex is shown with its 50S and 30S subunits.
18

 In step 1, the charged tRNA unit carrying amino acid 8 binds to the acceptor site A on the 70S ribosome. The peptidyl tRNA at the donor site, with amino acids 1 through 7, then binds the growing amino acid chain to amino acid 8 (transpeptidation, step 2). The uncharged tRNA left at the donor site is released (step 3), and the new 8-amino acid chain with its tRNA shifts to the peptidyl site (translocation, step 4). The antibiotic binding sites are shown schematically as triangles. Chloramphenicol (C) and macrolides (M) bind to the 50 S subunit and block transpeptidation (step 2). The tetracyclines (T) bind to the 30S subunit and prevent binding of the incoming charged tRNA unit (step 1). 2009
19

Codon

4
translocation

2 3

20

Peptidyl transferase

21

Diagram of a polyribosome (polysome) Ribosomes move along the mRNA in the 5 to 3 direction. They function independent of each other
22

Inhibition of bacterial protein synthesis by tetracyclines.

Messenger RNA (mRNA) attaches to the 30S subunit of bacterial ribosomal RNA. The P (peptidyl) site of the 50S ribosomal RNA subunit contains the nascent polypeptide chain; normally, the aminoacyl tRNA charged with the next amino acid (aa) to be added to the chain moves into the A (acceptor) site, with complementary base pairing between the anticodon sequence of tRNA and the codon sequence of mRNA. Tetracyclines inhibit bacterial protein synthesis by binding to the 30S subunit and blocking tRNA binding to the A site.
23

 Tetracyclines inhibit bacterial protein synthesis by binding to the 30S bacterial ribosome and preventing access of aminoacyl tRNA to the acceptor (A) site on the mRNA-ribosome complex These drugs enter gram-negative bacteria by passive diffusion through the hydrophilic channels formed by the porin proteins of the outer cell membrane and by active transport via an energy-dependent system that pumps all tetracyclines across the cytoplasmic membrane. Entry of these drugs into gram-positive bacteria requires metabolic energy, but is not as well understood.
24

 Tetracyclines enter bacterial cells by two processes: passive diffusion and active transport active uptake of tetracyclines by bacterial cells is an energy-dependent process that requires adenosine triphosphate (ATP) and magnesium ions
25

Spectrum of activity
broadest spectrum of any known antibacterial agents bacteriostatic action

disadvantage in the treatment of lifethreatening infections aminoglycosides and/or cephalospolins usually are preferred for gram-negative and the penicillins for gram-positive infections
26

Spectrum of activity
Because of incomplete absorption and effectiveness against the natural bacterial flora of the intestine, tetracyclines may induce superinfections caused by the pathogenic yeast Candida albicans Parenteral tetracyclines may cause severe liver damage, especially when given in excessive dosage to pregnant women or to patients with impaired renal function.
27

 Acne. Tetracyclines have been used to treat acne. They may act by inhibiting propionibacteria, which reside in sebaceous follicles and metabolize lipids into irritating free fatty acids. The relatively low doses of tetracycline used for acne (e.g., 250 mg orally twice a day) apparently are associated with few side effects.
28

Resistance to Tetracyclines
Resistance is primarily plasmid-mediated and often is inducible. The three main resistance mechanisms are:

decreased accumulation of tetracycline as a result of either decreased antibiotic influx or acquisition of an energy-dependent efflux pathway production of a ribosomal protection protein that displaces tetracycline from its target, a "protection" that also may occur by mutation enzymatic inactivation of tetracyclines.
29

Resistance to Tetracyclines
Cross resistance, or lack thereof, among tetracyclines depends on which mechanism is operative. For example, S. aureus strains that are tetracyclineresistant on the basis of efflux mediated by tetK still may be susceptible to minocycline. Tetracycline resistance due to a ribosomal protection mechanism (tetM) produces crossresistance to doxycycline and minocycline because the target site protected is the same for all tetracyclines.

30

Ribosomal protection: involving elaboration of bacterial protein. These proteins associate with the ribosome, thus allowing protein biosynthesis to proceed even in the presence of bound tetracycline, although exactly how this works is not understood.

31

Structure-activity relationships
R1
7 8 9 10 6 11

O NH 2 O O

R2

H3C CH3 N R 3 R4 H H 3 OH
5a 11a 5 4a 12a 4 1 2

NH2 O

OH O OH O

OH

fewer than four rings are inactive or nearly inactive A-ring substituents 1. The enolized tricarbonylmethane system at C- 1 to C-3 must be intact for good activity 2. Replacement of the amide at C-2 with other functions, such as aldehyde or nitrile, reduces or abolishes activity

32

H3C

OH

NMe 2 H OH H2C N H O H HN

pyrrolidine

OH

OH OH O

CNH2 O

Tetracycline
H3C OH NMe 2 H OH

Rolitetracycline

OH O O Ring A 3. Monoalkylation of the amide nitrogen reduces activity proportionately to the size of the alkyl group 4. Amino-alkylation of the amide nitrogen, accomplished by the Mannich reaction, yields derivatives that are substantially more water-soluble than the parent tetracycline and are hydrolyzed to it in vivo (e.g., 33 rolitetracycline) OH OH O

CNHCH2 N

Ring A 5. The dimethylamino group at the 4-position must have the -orientation: 4-epitetracyclines are very much less active than the natural isomers 6. Removal of the 4-dimethylamino group reduces activity even further 7. Activity is largely retained in the primary and N-methyl secondary amines but rapidly diminishes in the higher alkylamines 8. A cis-A/B-ring fusion with -OH at C-12a is apparently 34 also essential

Ring A Ring B

9. Esters of the C-12a OH are inactive, with the exception of the formyl ester 10. Alkylation at C-11a also leads to inactive compounds, demonstrating the importance of an enolizable diketone functionality at C-11 and C-12 11. substantial loss in antibacterial potency resulting from epimerization at C-5a
35

12. Dehydrogenation to form a double bond between C-5a and C-11a markedly decreases activity, as does aromatization of ring C to form anhydrotetracyclines

36

3 1 2 3 4

13. 5-OH, as in oxytetracycline and doxycycline, may influence pharmacokinetic properties but does not change antimicrobial activity
37

R1
7 8 9

R2
6 11

H3C CH3 N R3 R 4 H H 3 OH
5a 11a 5 4a 12a 4 1 2

10 OH Ring C OH O OH O O 14. Acid & basic -stable 6-deoxy and 6-demethyltetracyclines retains activity 15.Reduction of the 6-OH causes change in the solubility properties of tetracyclines higher oil/water partition coefficients of the 6-deoxytetracyclines compared with the tetracyclines Hence, doxycycline and minocycline are absorbed more completely following oral 38 administration than oxytetracycline

NH2

3 1 2 3 4

7 8 9 10 6 11 5a 11a 5 4a 12a 4 1

3 2
2

- Polar substituents (i.e., OH) at C-5 and C-6 contribute decreased lipid versus water solubility to the tetracyclines - 6-position is considerably more sensitive than the 5-position to this effect - doxycycline (6-deoxy-5-oxytetracycline) has a much higher partition coefficient than either tetracycline or oxytetracycline
39

doxycycline much higher partition coefficient than either tetracycline or oxytetracycline

doxycycline (6-deoxy-5-oxytetracycline) mlogP -0.02

tetracycline
mlogP -1.25

oxytetracycline
mlogP -1.12

40

R1
7 8 9

R2
6 11

H3C CH3 N R3 R 4 H H 3 OH
5a 11a 5 4a 12a 4 1 2

NH2 O

Ring D

10

OH O OH O

OH

16.both strongly electron-withdrawing groups (e.g., chloro [chlortetracycline] and nitro) and strongly electron-donating groups (e.g., dimethylamino [minocycline]) enhance activity

17. introducing substituents at C-8 has not been

studied because this position cannot be substituted directly by classic electrophilic aromatic substitution reactions; thus, 8substituted derivatives are available only through total synthesis 41

Products
R1 Tetracycline Chlortetracyclin Oxytetracycline Demeclocycline Methacycline Doxycycline Minocycline H Cl H Cl H H NMe2 R2 CH3 CH3 CH3 H =CH2 H H R3 OH OH OH OH CH3 H R4 H H OH H OH OH H
42

3 3

3 3

Chlortetracycline

Tetracycline

tetracycline was obtained from fermentations of Streptornyces species, the commercial supply is still chiefly dependent on the hydrogenolysis of chlortetracycline
Chlortetracycline (prototype) no longer marketed in the 43 US.

Methacycline

Removal of the 6-OH increases the stability of ring C to both acids and bases preventing the formation of isotetracyclines by bases However, anhydrotetracyclines still can form by acid-catalyzed isomerization under strongly acidic conditions
44

Oxytetracycline
3 2 + 3

Demeclocycline

2 -

Meclocycline sulfosalicylate

45

methacycine

Doxycycline

produced by catalytic hydrogenation of methacycline 6-methyl epimer is more than three times as active as its -epimer absence of the 6-OH produces a compound that is very stable in acids and bases and that has a long biologic half-life In addition, it is absorbed very well from the GI
46

Minocycline most potent tetracycline obtained by reductive methylation of 7nitro-6-demethyl-6-deoxytetracycline lacks the 6-OH, it is stable in acids and base Minocycline is well absorbed orally
47

Lymecycline
It is approximately 5000 times more soluble than tetracycline base and is unique amongst tetracyclines in that it is absorbed by the "active transport" process across the intestinal wall, making use of the same fast and efficient mechanism by which carbohydrates are absorbed. (info: drug bank)
48

3 3

Lymecycline
H3C H3C OH H OH H N OH OH O OH O O N N CH3

Rolitetracycline

49

X CH3 H3C N O NH OH O

NMe 2 H OH

glycylcyclines

OH OH O

CNH2 O

X = N(CH3)2 X=H

9-(Dimethylglycylamino)minocycline (DMG-MINO) 9-(Dimethylglycylamino)-6-demethyl-6-deoxytetracycline (DMG-DMDOT)

a new class of second-generation tetracyclines highly active against bacterial strains that exhibit tetracycline resistance mediated by efflux or ribosomal protection determinants
50

 The glycylcyclines are synthetic analogues of the tetracyclines, with the most promising compound being the 9-tert-butyl-glyclyamido derivative of minocycline, tigecycline. The glycylcyclines exhibit antibacterial activities typical of earlier tetracyclines, and also display activity against tetracycline-resistant organisms containing genes responsible for efflux mechanisms or ribosomal protection. The glycyclcyclines also appear to be active against other resistant pathogens including methicillinresistant S. aureus and S. epidermidis, penicillinresistant S. pneumoniae, and vancomycin-resistant enterococci.
51

Tigecycline

(bio-loom)

52

H3C H3C OH

CH3 OH O OH H3C OH

H3C

CH3 OH NH O N NH HN H3C CH3 OH O NH2

OH OH O OH O HN N O OH O OH

OH O HN N

H3C

OH

OH OH O OH

OH O HN OH NH O OH OH OH

53

3 3

3 3 3 3

54

3 3

Etamocycline
(bioLoom)

3 3 3

55

Tetracycline

H3C H3C OH

CH3 OH O

OH OH O OH O HN HN Penimocycline O NH O N OH O

Penimocycline is an antibiotic obtained by Mannich reaction H3C between tetracycline and ampicillin.

H3C

Ampicillin
56

Aminoglycosides
term: mycin produced by Streptomyces micin produced by Micromonospora or other organism Others: semisynthetic or other Bacillus sp.
57

 1939: discovery of streptomycin, the first aminoglycosides antibiotic worldwide searches for antibiotics from the actinomycetes and, particularly, from the genus

Streptomyces

many antibiotics isolated from that genus: kanamycin neomycin paromomycin gentamicin tobramycin netilmicin Amikacin, a semisynthetic derivative of kanamycin 58A,

NH

Streptidine

NHCNH2 HO HO O O NH OH NHCNH2 O CH3 O OH NHCH3 CH O

OH

L-streptose

Properties

HOH2C HO

Streptobiosamine

N-methyl-L-glucosamine

absorbed very poorly (less than 1%) following oral administration kanamycin, neomycin, and paromomycin are administered orally for the treatment of GI infections potent broad-spectrum, also used for the treatment of systemic infections
59

Properties
undesirable side effects, particularly oto- and nephrotoxicity have led to restrictions in their systemic use for serious infections or infections caused by bacterial strains resistant to other agents systemic infection: must be given parenterally, usually by intramuscular injection

60

HO
3'

CH2NH2 O NH2 O HO NH2 O HO NH2 O

Tobramycin

CH2OH OH NH2

aminoglycosides: Chemistry named because structures consist of amino sugars linked glycosidically

All have at least one aminohexose and some have a pentose lacking an amino group (e.g., streptomycin, neomycin, and paromomycin)
61

HO
3'

CH2NH2 O NH2 O HO NH2 O HO NH2 O

NH

Tobramycin

Streptidine

NHCNH2 HO HO

Chemistry
NH OH O NHCNH2 O CH3 OH CH O

deoxystreptamine
HOH2C HO O OH

L-streptose

CH2OH OH NH2

NHCH3

Streptobiosamine

each aminoglycosides contains a highly substituted 1,3-diaminocyclohexane central ring: in kanamycin, neomycin, gentamicin, and tobramycin, it is deoxystreptamine in streptomycin it is streptidine 62

HO
3'

CH2NH2 O NH2 O HO NH2 O HO NH2 O

Tobramycin

Chemistry
strongly basic compounds

CH2OH OH NH2

exist as polycations at physiologic pH their inorganic acid salts are very soluble in water All are available as sulfates Solutions of the aminoglycoside salts are stable to autoclaving
63

Chemistry
high water solubility of the aminoglycosides contributes to their pharmacokinetic properties distribute well into most body fluids but not into the central nervous system, bone, or fatty or connective tissues concentrate in the kidneys and are excreted by glomerular filtration
64

Spectrum of activity
broad-spectrum antibiotics usefulness in the treatment of serious systemic infections caused by aerobic gram-negative bacilli Aerobic gram-negative and gram-positive cocci (with the exception of staphylococci) tend to be less sensitive the -lactam and other antibiotics tend to be preferred for the treatment of infections caused by these organisms Anaerobic bacteria are resistant to the aminoglycosides
65

 aminoglycoside and -lactam antibiotics exert a synergistic action in vivo against certain bacterial strains when the two are administered jointly carbenicillin and gentamicin are synergistic against gentamicin-sensitive strains of Ps. aeruginosa and several other species of gram-negative bacilli penicillin G and streptomycin (or gentamicin or kanamycin) tend to be more effective than either agent alone in the treatment of entero-coccal endocarditis The two antibiotic types should not be combined in the same solution because they are chemically incompatible Damage to the cell wall caused by the -lactam is believed to increase penetration of the aminoglycoside into the bacterial cell.
66

Mechanism of action
bind to the 30s ribosomal subunit 1. Block initiation of protein synthesis 2. Block further translation and elicits premature termination 3. Incorporation of incorrect amino acid (misreading)
67

 Misreading: resulting from failure of specific aminoacyl RNAs to recognize the proper codons on mRNA and the incorporation of improper amino acids into the peptide chain deoxystreptamine-containing aminoglycosides differ quantitatively from streptomycin in causing misreading at lower concentrations than are required to prevent initiation of protein synthesis, whereas streptomycin inhibits initiation and causes misreading equally effectively
68

Effects of aminoglycosides on protein synthesis.

69

 A. Aminoglycoside (represented by closed circles) binds to the 30S ribosomal subunit and interferes with initiation of protein synthesis by fixing the 30S-50S ribosomal complex at the start codon (AUG) of mRNA. As 30S-50S complexes downstream complete translation of mRNA and detach, the abnormal initiation complexes, so-called streptomycin monosomes, accumulate, blocking further translation of the message. Aminoglycoside binding to the 30S subunit also causes misreading of mRNA, leading to B. premature termination of translation with detachment of the ribosomal complex and incompletely synthesized protein or C. incorporation of incorrect amino acids (indicated by the X), resulting in the production of abnormal or nonfunctional proteins.
70

Mechanism of action
Spectinomycin, however, prevents
the initiation of protein synthesis but does not cause misreading

All aminoglycoside antibiotics except spectinomycin are bactericidal

71

Microbial resistance I
bacterial strains resistant to streptomycin, kanamycin, and gentamicin strains carrying R-factors for resistance to these antibiotics synthesize enzymes capable of
acetylating Phosphorylating or adenylylating key amino (NH2) or hydroxyl groups (OH) of the aminoglycosides
72

APH for aminoglycoside phosphotransferases, ANT for aminoglycoside adenyltransferases, AAC for acetyltransferases
AAC R NH 2 AcetyCoA APH R OH ATP R OH ATP ANT PPi ADP R O R CoASH O R O P O AMP
73

O C

H N

CH 3

OH

A. Sites of aminoglycoside phosphoryation by APHs

74

B. Sites of aminoglycoside adenylation by ANTS

75

C. Sites of aminoglycoside acetylation by AACs

76

-O-AMP
ANT (4') HO HO APH (3') CH2NH2 O R2 O HO

AAC (6')

O H N
AAC (3)

CH3

NH2 O NH2 O CH2OH OH ANT (4'')

77

Kanamycin B
ANT (2'')

HO

O O P O OH

APH (2'')

NH2

Resistance of aminoglycosides to specific inactivating enzymes

using chemical principles
First, target functional group is absent in a position of the structure normally attacked by an inactivating enzyme, then the antibiotic will be resistant to the enzyme Second, steric factors may confer resistance to attack at functionalities otherwise susceptible to enzymatic attack.
78

AAC (6')

HO
3'

CH2NH2 O NH2 O HO NH2 O HO NH2 O

ANT (4') HO HO APH (3')

CH2NH2 O R2 O HO NH2 O

AAC (3)

Tobramycin
Kanamycin B

NH2 O CH2OH OH ANT (4'')

ANT (2'') APH (2'')

HO

NH2

CH2OH OH NH2

tobramycin lack a 3-OH in ring I, are not inactivated by the phosphotransferase enzymes that phosphorylate that group in the kanamycins
79

R1 6' CHNHR2 O 3' NH2 O HO NH2 O HO O

Gentamicin C1 Gentamicin C2
NH2 Gentamicin C1a

R1 CH3 CH3 H

R2 CH3 H H

H3CHN

CH3 OH 4''

compare to Kanamycin B

lack a 3-OH in ring I, not inactivated by the phosphotransferase enzymes Gentamicin C1 (but not C1a or C2) is resistant to the acetylation the 6-NH2 in ring I
80

R1 6' CHNHR2 O 3' NH2 O HO NH2 O HO O

Gentamicin C1 Gentamicin C2
NH2 Gentamicin C1a

R1 CH3 CH3 H

R2 CH3 H H

H3CHN

CH3 OH 4''

compare to Kanamycin B

All gentamicins are resistant to the nucleotidyltransferase that adenylylates the secondary equatorial 4-OH because it is tertiary and has an axial orientation
81

CH2NH2 HO O HO OH O HO NH2

1N-L(-)-amino--hydroxybutyric acid (L-AHBA) derivative of kanamycin A

O NHCCHCH2CH2NH2 O OH O CH2OH OH

Amikacin

HO

St

ic r e

NH2

retains most of the intrinsic potency of kanamycin A resistant to all aminoglycoside-inactivating enzymes known except the aminoacetyltransferase that acetylates the 6NH2 and the nucleotidyltransferase that adenylylates the 4-OH of ring I 82

CH2NH2 HO O HO OH O HO NH2

1N-L(-)-amino--hydroxybutyric acid (L-AHBA) derivative of kanamycin A

O NHCCHCH2CH2NH2 O OH O CH2OH OH

Amikacin

HO

NH2

that introduction of the L-AHBA group into kanamycin A markedly decreases its affinity for the inactivating enzymes
83

Microbial resistance II
aminoglycosides requires uptake of the drug by an energy-dependent active process Uptake is initiated by the binding of the cationic aminoglycoside to anionic phospholipids of the cell membrane Electron transport-linked transfer of the aminoglycoside through the cell membrane then occurs Divalent cations, such as Ca2+ and Mg2+, antagonize the transport of aminoglycosides into bacterial cells by interfering with their binding to cell membrane phospholipids 84

Microbial resistance II
The resistance of anaerobic bacteria to the lethal action of the aminoglycosides is apparently due to the absence of the respirationdriven active-transport process for transporting the antibiotics.

85

1. Ring I is important for broad-spectrum antibacterial activity 2. Amino functions at 6 and 2 are particularly important as kanamycin B (6-NH2, 2-NH2) is more active than kanamycin A (6-NH2, 2-OH), which in turn is more active than kanamycin C (6-OH, 2-NH2)
HO HO CH 2 R 1 O R2 O HO N H2 O N H2

Structure-activity relationships

R1 K anamycin A K anamycin B K anamycin C NH 2 NH 2 OH

86

R2 OH NH 2 NH 2

Deoxystreptamine

R 1 6' CHN HR 2 O 3' N H2 O HO N H2 O

Gentamicin C 1 Gentamicin C 2
N H 2 Gentamicin C 1 a

R1 CH 3 CH 3 H

R2 CH 3 H H

3. Methylation at the 6-carbon of the 6-NH2 positions does not lower antibacterial activity and confers resistance to enzymatic acetylation of the 6-NH2 [Gentamicin C1 (but not C1a or C2) is resistant to the acetylation the 6-NH2 in ring I]
87

R 1 6' CHN HR 2 O 3' N H2 O HO N H2 O

Gentamicin C 1 Gentamicin C 2
N H 2 Gentamicin C 1 a

R1 CH 3 CH 3 H

R2 CH 3 H H

4. Removal of the 3-OH or the 4-OH or both in the kanamycins does not reduce antibacterial potency e.g. 3,4-dideoxykanamycin B or dibekacin gentamicins sisomicin netilmicin None of these derivatives is inactivated by phosphotransferase enzymes that 88 phosphorylate the 3-OH group

CH2NH2 HO O HO OH O HO NH2

1N-L(-)-amino--hydroxybutyric acid (L-AHBA) derivative of kanamycin A

O NHCCHCH2CH2NH2 O OH O CH2OH OH

Amikacin

HO

NH2

5. Few modifications of ring II (deoxystreptamine) functional groups are possible without appreciable loss of activity However, the 1-NH2 of kanamycin A can be acylated (e.g., amikacin), and activity is largely retained Netilmicin (1-N-ethylsisomicin) retains the antibacterial potency of sisomicin and is resistant to several additional bacteria-inactivating enzymes 89

R1 6' CHNHR2 O 3' NH2 O HO NH2 NH2 O O

R1 Gentamicin C1 Gentamicin C2 Gentamicin C1a CH3 CH3 H

R2 CH3 H H

2H CHN 3

HO

CH3 OH 4''

6. Ring III functional groups appear to be somewhat less sensitive to structural changes than those of either ring I or ring II Although the 2-deoxygentamicins are significantly less active than their 2-hydroxyl counterparts, the 2-NH2 derivatives (seldomycins) are highly active
90

R1 6' CHNHR2 O 3' NH2 O HO NH2 NH2 O HO O

R1 Gentamicin C1 Gentamicin C2 Gentamicin C1a CH3 CH3 H

R2 CH3 H H

H3CHN

CH3 OH 4''

3-amino group of gentamicins may be primary or secondary with high antibacterial potency 4-hydroxyI group may be axial or equatorial with little change in potency
91

NH

Streptidine

NHCNH2 HO HO O O OH NHCNH2 O CH3 O OH NHCH3 CH O NH

Streptomycin

OH

L-streptose

HOH2C HO

Streptobiosamine

N-methyl-L-glucosamine

Acid hydrolysis yields streptidine and streptobiosamine, the compound that is a combination of L-streptose and N-methyl-L-glucosamine Streptomycin acts as a triacidic base two strongly basic guanidino groups and the more weakly basic methylamino group 92

neosamine CNH2
HOH2C O O O

HO HO

R1

Deoxystreptamine
NH2 OH NH2

D-Ribose
O

R1 CH2NH2 CH2NH2 CH2OH CH2OH

R2 H CH2NH2 CH2NH2 H

R3 CH2NH2 H H CH2NH2

OH NH2 R2

Neomycin C Neomycin B
OH OH R3

Paromomycin I Paramomycin II

Neomycin: neamine is a combination of

deoxystreptamine and neosamine C

Paramomycin: D-glucosamine
93

HO HO

CH2R1 O R2 O HO NH2 O HO NH2

R1 Kanamycin A Kanamycin B Kanamycin C NH2 NH2 OH

R2 OH NH2 NH2

Deoxystreptamine

CH2OH OH NH2

Commercially available kanamycin is almost pure kanamycin A, the least toxic of the three forms kanamycins do not have the D-ribose molecule that is present in neomycins and paromomycins significant in the lower toxicity
94

CH2NH2 HO O HO OH O HO NH2

1N-L(-)-amino--hydroxybutyric acid (L-AHBA) derivative of kanamycin A

O NHCCHCH2CH2NH2 O OH O CH2OH OH

Amikacin

HO

NH2

semisynthetic aminoglycoside first prepared in Japan acylation of the 1-NH2 of the deoxystreptamine ring of kanamycin A with L-AHBA resists attack by most bacteria-inactivating retains about 50% of the original activity of kanamycin A against sensitive strains of gram-negative bacilli.
95

R1 6' CHNHR2 O 3' NH2 O HO NH2 NH2 O HO O

R1 Gentamicin C1 Gentamicin C2 Gentamicin C1a CH3 CH3 H

R2 CH3 H H

H3CHN

CH3 OH 4''

topical gentamicin be reserved for use in such infections and in the treatment of burns complicated by pseudomonemia

An injectable solution may be used for serious systemic and genitourinary tract infections chemically incompatible with carbenicillin, and the two should not he combined in the same intravenous 96 solution

HO

CH2NH2 O NH2 O HO O HO NH2 NH2

Tobramycin
O CH2OH OH NH2

active against most strains of Ps. aeruginosa, exceeding that of gentamicin by two- to fourfold closely resembles kanamycin B in structure: it is 3-deoxykanamycin B
97

CH2NH2 O
3'

R 1 6' CHNHR2 O NH2 O HO NH2 NH2 O O

Gentam Gentam Gentam

NH2 O HO

NH2 NHR O HO O

Sisomicin Netilmicin
CH3 OH

H CH2CH3

HO

H3CHN

CH3 OH 4'' R2

R1 Gentamicin C1 Gentamicin C2 Gentamicin C1a CH3 CH3 H

H3CHN

CH3 H H

1-N-Ethylsisomicin (Netilmycin) is a semisynthetic derivative prepared by reductive ethylation of sisomicin sisomicin and netilmicin resemble gentamicin C1a very few gentamicin-resistant bacterial strains are sensitive to sisomicin 98

 potency of netilmicin against certain gentamicinresistant bacteria is attributed to its resistance to inactivation by bacterial enzymes that adenylylate or phosphorylate genatmicin and sisomicin Netilmicin is inactivated by most of the bacterial enzymes that acetylate aminoglycosides, whereas amikacin is resistant to most of these enzymes Netilmicin and sisomicin exhibit pharmacokinetic and toxicologic properties similar to those of gentamicin
99

R CH2NH2 O NH2 O HO NH2 O HO O CH2OH OH NH2

HO HO

CH2R1 O R2 NH2 HO O HO NH2

Kanamycin Kanamycin Kanamycin

Dibekacin Arbekacin
NHR

H O CCHCH2CH2NH2 OH

Deoxystreptamine

CH2OH OH NH2

R1 Kanamycin A Kanamycin B Kanamycin C NH2 NH2 OH

R2 OH NH2 NH2

Dibekacin: 3,4-dideoxykanamycin B Arbekacin: semisynthetic derivative of dibekacin broad-spectrum active against some of gentamicin, kanamycin, tobramycin-resistant pathogens
100

OH NHCH3 H3CHN O HO OH OH O O OH CH3

spectinomycin

Solutions of spectinomycin, a hemiacetal, slowly hydrolyze on standing and should be prepared freshly and used within 24 hr It is administered by deep intramuscular injection

interferes with the binding of tRNA to the ribosomes and, thereby, with the initiation of protein synthesis not cause misreading of the messenger Bacteriostatic 101

The End

102