PERTEMUAN KE-2

MIKROBIOLOGI

ANTI
MICROBIAL
AGENTS
By: Rosario Trijuliamos Manalu, M.Si
FAKULTAS FARMASI
INSTITUT SAINS DAN TEKNOLOGI NASIONAL
Sejarah Penemuan Antibiotik

3 4
1 2
1920
Menemukan Lisozim
1888 Pada air mata manusia
Mengisolasi produk
1887 dari bakteri sebagai anti
Yang dapat melisis sel
1877 Penyakit Kolera = biotik: Pigmen Biru dari
Bakteri.
Staphyloccus yang di
Penyakit antraks dapat penyakit infeksi intes Bacillus pyocyaneus Tumbuhkan pada cawan
Dikurangi patogenisitas tinal dapat dicegah dapat menghambat Petri mengalami lisis di
nya, Setelah hewan uji dengan bakteri Bakteri. Sekitar koloni kapang
diinjeksi dengan bakteri Streptococcus Pyocyanase terbukti toksik Penicillium sp.
yang diisolasi dari tanah Sehingga tidak dapat
Dikembangkan menjadi
Obat yang efektif
PENDAHULUAN Secara klinis, agen antimikroba
mengeluarkan toksisitas selektif
terhadap parasit ke inang.
Umumnya, sifat toksisitas tersebut
menyerang struktur atau proses
mikroba yang berbeda dari sel
mamalia.
Sebagai contoh, beberapa agen menyerang
sintesis dinding sel bakteri, dan beberapa
lainnya pada fungsi ribosom bakteri 70S.
Beberapa antimikroba seperti penisilin, bersifat
nontoksik terhadap inang, kecuali inang
tersebut bersifat hipersensitif.
 ISTILAH
ANTIBIOTIK: antimikroba yang berasal dari mikroorganisme.
Antibiotik dihasilkan oleh kelompok bakteri dan fungi.
ANTIMIKROBIAL: berbagai senyawa yang memiliki aktivitas
antimikroba yang cukup sehingga dapat digunakan dalam
perawatan penyekit infeksi.
KEMOTERAPI: istilah yang biasa digunakan dalam terapi
kanker meliputi antibiotik, antimikroba atau obat yang
digunakan.
RESISTEN: organisme yang tidak terhambat
pertumbuhannya secara klinis meskipun konsentrasi
agen antimikroba sudah sesuai takarannya.
 ISTILAH
SENSITIF: mikroorganisme yang menunjukkan akan
terhambat pada konsentrasi antimikroba tertentu.

SUSCEPTIBLE: mikroorganisme yang menunjukkan mereka

akan terhambat pada konsentrasi tertentu secara klinis.
SPEKTRUM: kisaran dari mikroorganisme terhadap
antimikrobial yang aktif.
Narrow spectrum: agen yang hanya memiliki aktivitas
terhadap beberapa mikroorganisme
Broad spectrum: memiliki aktivitas terhadap
mikroorganisme yang lebih beragam
 Sumber Agen Antimikroba

1 2 3
Penisilin: dari Agen-agen antimikroba Manipulasi secara
Penicillium yang disintesis secara molekuler bertujuan
kimia ditemukan menemukan antibiotik
Cephalosporin: kapang diantara senyawa- atau kemoterapi untuk
Cephalosporium senyawa yang disintesis memperluas jangkauan
Streptomisin, untuk tujuan atau atau derajat aktivitas
tetrasiklin, pengujian efektivitas penyerangan terhadap
kloramfenikol, terapi terhadap hewan. mikroorganisme atau
eritromisin: dari untuk meningkatkan
Streptomyces sulfonamida ditemukan
karakter
akibat penapisan rutin
farmakologinya.
pewarna anilin.
What properties should
an antibiotic have?
 Selective toxicity is the most important
single attribute of an antibiotic

Antibiotics, like other chemotherapeutic agents,

need to be soluble in body fluids, in order to exert
their effect by penetrating the body tissues.
The compound must not be metabolised so quickly
that it is excreted from the body before having a
chance to act.

it should be sufficiently stable to have a good shelf

life, without special storage considerations.
if administered orally, it must not be inactivated by the
acid environment of the stomach, and must be capable
of being absorbed by the small intestine.
an antibiotic should not have any significant effect
on the resident microflora of the host.
How do antibiotics work?
1. Inhibition of cell wall synthesis
2. Disruption of cell membranes
3. Interference with protein synthesis
4. Interference with nucleic acid
synthesis
SELECTED ANTIBACTERIAL ANTIMICROBICS
I. Antimicrobics That Act on Cell Wall
Synthesis
 Mature peptidoglycan is held together by cross-
linking of short peptide side chains hanging off
the long glycan molecules.

 This cross-linking process is the target of two of

the most important groups of antimicrobics, the
β-lactams and the glycopeptides.
 β-Lactam Antimicrobics

 The β-lactam antimicrobics comprise the

penicillins, cephalosporins, carbapenems, and
monobactams.
 The β-lactam antimicrobics interfere with the
transpeptidation reactions that seal the peptide
crosslinks between glycan chains.

 They do so by interference with the action of

the transpeptidase enzymes which carry out this
cross-linking.
 Glycopeptide Antimicrobics
 Each of these antimicrobics inhibit assembly of
the linear peptidoglycan molecule by binding
directly to the terminal amino acids of the
peptide side chains.
 The effect is the same as with β-lactams,
prevention of peptidoglycan cross-linking.
 Two agents belong to this group: vancomycin
and teicoplanin. Both agents are bactericidal,
but are primarily active only against Gram-
positive bacteria.
II. Antimicrobics Acting on the Outer and
Cytoplasmic Membranes

 The polypeptide antimicrobics polymyxin B and

colistin have a cationic detergent-like effect.

 They bind to the cell membranes of susceptible

Gram-negative bacteria and alter their
permeability,
 Their spectrum is essentially Gram-negative; they
act against P. aeruginosa and other Gram-negative
rods.
III. Inhibitors of Protein Synthesis
Terbagi menjadi beberapa kelompok:
A. Aminoglycosides
B. Tetracyclines
C.Chloramphenicol
D. Macrolides
E. Clindamycin
F. Streptogramins
A. AMINOGLYCOSIDES

 are active against a wide range of bacteria,

but only those organisms that are able to
transport them into the cell by a mechanism
that involves oxidative phosphorylation.
 Thus, they have little or no activity against
strict anaerobes or facultative organisms that
metabolize only fermentatively (eg,
streptococci).
 Once inside bacterial cells, aminoglycosides inhibit
protein synthesis by binding to the bacterial
ribosomes either directly or by involving other
proteins.

 This binding destabilizes the ribosomes, blocks

initiation complexes, and thus prevents
elongation of polypeptide chains.

 The agents may also cause distortion of the site of

attachment of mRNA, mistranslation of codons,
and failure to produce the correct amino acid
sequence in proteins.
 Eukaryotic ribosomes are resistant to
aminoglycosides, and the antimicrobics are not
actively transported into eukaryotic cells.

 These properties account for their selective

toxicity and also explain their ineffectiveness
against intracellular bacteria such as Rickettsia
and Chlamydia.
 Gentamicin and Tobramycin are the major
aminoglycosides; they have an extended spectrum,
which includes staphylococci; Enterobacteriaceae;
and of particular importance, P. aeruginosa.

 Streptomycin and Amikacin are now primarily used

in combination with other antimicrobics in the
therapy of tuberculosis and other mycobacterial
diseases.

 Neomycin, the most toxic aminoglycoside, is used in

topical preparations and as an oral preparation
before certain types of intestinal surgery, because it
is poorly absorbed.
B. TETRACYCLINES
 The tetracyclines inhibit protein synthesis by
binding to the 30S ribosomal subunit at a point
that blocks attachment of aminoacyl-tRNA to
the acceptor site on the mRNA ribosome
complex.
 their effect is reversible

 they are bacteriostatic rather than bactericidal.

C. CHLORAMPHENICOL

It influences protein synthesis by binding to the

50S ribosomal subunit and blocking the action
of peptidyl transferase, which prevents
formation of the peptide bond essential for
extension of the peptide chain.

Its action is reversible in most susceptible

species; thus, it is bacteriostatic.
It has little effect on eukaryotic ribosomes,
which explains its selective toxicity.
D. MACROLIDES

The macrolides, erythromycin, azithromycin, and

clarithromycin

They affect protein synthesis at the ribosomal

level by binding to the 50S subunit and blocking
the translocation reaction.

Their effect is primarily bacteriostatic.

E. CLINDAMYCIN

Clindamycin is chemically unrelated to the

macrolides but has a similar mode of action
and spectrum with addition of anaerobes.
It has greater activity than the macrolides
against Gram-negative anaerobes, including the
important Bacteroides fragilis group.
F. STREPTOGRAMINS
They inhibit protein synthesis by binding to
different sites on the 50S bacterial ribosome.
 Quinupristin inhibits peptide chain
elongation

 Dalfopristin interferes with peptidyl

transferase
IV. Inhibitors of Nucleic Acid Synthesis
Terbagi menjadi beberapa kelompok:
A. Quinolones

B. Folate Inhibitor
C. Metronidazzole

D. Rifampin
A. QUINOLONES

The target is DNA topoisomerase (gyrase), the

enzyme responsible for nicking, supercoiling,
and sealing bacterial DNA during replication.

Bacterial topoisomerases have four subunits,

one or more of which are inhibited by the
particular quinolone.
Ex. ciprofloxacin, norfloxacin, and ofloxacin,
 The fluoroquinolones are highly active and
bactericidal against a wide range of aerobes and
facultative anaerobes.

 However, streptococci and Mycoplasma are only

marginally susceptible, and anaerobes are
generally resistant.

 Ofloxacin has significant activity against

Chlamydia; whereas ciprofloxacin is particularly
useful against P. aeruginosa.
B. FOLATE INHIBITORS

Bacteria must synthesize folate that humans

acquire in their diet

Folic acid is derived from para-aminobenzoic

acid (PABA), glutamate, and a pteridine unit.

The major inhibitors of the folate pathway

are the sulfonamides, trimethoprim, para-
aminosalicylic acid, and the sulfones.
1. Sulfonamides

 Competition with PABA disrupts nucleic acids.

 The effect is bacteriostatic.

 Their primary use is for uncomplicated urinary
tract infections caused by members of the
Enterobacteriaceae, particularly Escherichia
coli.
2. Trimethoprim-Sulfamethoxazole
 It competitively inhibits the activity of
bacterial dihydrofolate reductase, which
catalyzes the conversion of folate to its
reduced active coenzyme form.

 Activity against common bacteria and some

fungi
C. METRONIDAZOLE
 Action requires anaerobic or at least
microaerophilic growth conditions.

 The antibacterial action requires reduction

of the nitro group under anaerobic
conditions.
 The reduction products act on the cell at
multiple points; the most lethal of these
effects is induction of breaks in DNA strands.
D. RIFAMPIN

 Blocking of RNA synthesis occurs by binding to

polymerase

 This agent is active against most Gram-positive

bacteria and selected Gram-negative organisms,
including Neisseria and Haemophilus.

 The most clinically useful property of rifampin is

its antimycobacterial activity, which includes
Mycobacterium tuberculosis.
How does antibiotic
resistance work?
 Some bacteria are able to resist antibiotic
action by denying it entry to the cell.
 Others can pump the antibiotic back out of
the cell before it has had a chance to act, by
means of enzymes called translocases; this is
fairly non-specific, leading to multiple drug
resistance.
 Other bacteria are naturally resistant to a
particular antibiotic because they lack the
target for its action, for example,
Mycoplasma do not possess peptidoglycan,
the target for penicillin’s action.
 To avoid the action of an antibiotic, bacteria
may be able to use or develop alternative
biochemical pathways, so that its effect is
cancelled out.
 Many pathogens can secrete enzymes that
modify or degrade antibiotics, causing them to
lose their activity;

o we have already seen that penicillins can be

inactivated by enzymatic cleavage of their β-
lactam ring. Similarly, chloramphenicol can be
acetylated, while members of the
aminoglycoside family can be acetylated,
adenylated or phosphorylated, all leading to
loss of antimicrobial activity.
 Mutations may occur which modify bacterial proteins
in such a way that they are not affected by
antimicrobial agents.

 You will recall that streptomycin normally acts by

binding to part of the 30S subunit on the bacterial
ribosome; the actual binding site is a protein called
S12. Mutant forms of the S12 gene can lead to a
product which still functions in protein synthesis, but
loses its ability to bind to streptomycin.

 Similarly, mutations in transpeptidase genes in

staphylococci means they do not bind to penicillin
any more, so cross-linking of the cell wall is not
inhibited.
PENEMUAN SUMBER
ANTIBIOTIK BARU
 Penemuan sumber-sumber antibiotik
baru dilakukan dengan cara penapisan
atau skrining untuk menemukan
mikroorganisme penghasil antibiotik
 Proses penapisan melalui 2 tahap, yaitu
skrining primer dan sekunder
 Skrining Primer
1. Mencari sumber penghasil
2. Menumbuhkan mikroorganisme yang di
dapat
3. Mengisolasi dan mengoleksi
mikroorganisme
4. Uji kemamppuan isolat
Skrining Sekunder
1. Mendapatkan koloni mikroorganisme
terpilih
2. Mencari kondisi optimal untuk
pertumbuhan (suhu, pH, lama inkubasi,
media dll)
3. Identifikasi mikroorganisme (morfologi,
kimiawi, genetik)
4. Identifikasi substansi