Asam lemak

sebagai
Oleh : agen antimalaria
Isti Istiqamah
anti jamur
Megawati
Andi Suryana anti bakteri
Adi
Edi Ilimu
Fatty acid as
antimalarial
agent
Malaria
 Malaria is one of the most important
tropical parasitic deseases.
 Estimated by WHO, there are about 300-
500 milion acute clinical malarial cases
every year and aroun 1 million deaths do
occure every year.
 Plasmodium falciparum is responsible for
the most severe and deadly form of the
desease and is to blame for 90% of
malaria-related deaths occuring in Africa.
Total population at risk of malaria
(WHO)

Global malaria mapper-2014 (WHO official website)

Malaria inpatient deaths (WHO)

Global malaria mapper-2014 (WHO official website)

The Life Cycle of the Malaria
Parasite: Plasmodium falciparum
The malaria parasite undergoes a significantly
intricate life cycle, requiring both human and
mosquito hosts to proliferate. Thus it is easier to
look at the life cycle in these three parts:
 sexual development (taking place mostly in the
mosquito-vector);
 Exo-erythrocytic development (taking place in
the human body)
 erythrocytic development (taking place in the
human body).
Antimalarial agent
H
H2C
H3C
N CH3
N
NH H
CH3 HO
H3CO

Cl N
N

Klorokuin Artemisinin Kuinin

FAS type
 Fatty acid biosynthesis in eukariot use a type I
fatty acid synthase (FASI) system.
 P. Falciparum occur in apicoplast use type II
FAS

Karena sistem biosintesis yang berbeda, ketika

kita mengganggu sistem biosisntesis asam
lemak pada P. Falciparum, sistem biosintesis
asam lemak pada manusia tak akan
terganggu.
Sistem biosintesis asam lemak pada P. Falciparum
merupakan suatu sistem yang sangat penting dan
berkaitan dalam beberapa proses seperti :
 Konstruksi membran sel
 Merupakan sumber energi yang sangat penting
 Memainkan peranan penting dalam transduksi
sinyal seperti pada proses asilasi protein
 Dibutuhkan dalam proses pertumbuhan
 Diferensiasi dan homeostasis dalam P. Falciparum

Hint : aktivitas biosintesis lipid akan meningkat pada

erythrocytic phases dari parasit.
Fatty acid biosynthesis in P.falciparum

Inhibited by isoniazid, triclosan

Inhibited by
thiolactomycin and derivatives
Kinetic mechanism of NADH-enoyl-ACP reductase from Brassica
napus
Tony Fawcett, Catherine L. Copse1, J. William Simon, Antoni R.
Slabas

 The cofactor, NADH, bound to ENR in a ratio

of 1 mol of enzyme to approximately 4 mol of
NADH.
 The native ENR enzyme is a tetramer with
each subunit having one NADH binding site,
capable of binding NADH in the absence of
the other substrate.
 substrate

enzyme

NADH

Therefore we conclude that cro-ACP (substrat) does

not bind to ENR in the absence of NADH and confirms
that the reaction mechanism must proceed via an
ordered ternary complex with NADH binding first.
 O

S ACP

A model has been proposed from recent crystallographic

studies that suggested the acyl chain of enoyl-ACP would
lie above the nicotinamide ring of the cofactor in the
substrate binding site, allowing the correct geometry for
hydride attack of the enoyl substrate
Mechanism of Triclosan Inhibition of Bacterial Fatty Acid
Synthesis
Richard J. Heath, J. Ronald Rubin, Debra R. Holland, Erli Zhang, Mark E.
Snow and Charles O. Rock
 Triclosan binds to the enoyl substrate site on
FabI, and tight binding of the drug requires
interactions between both the protein and
the NAD + cofactor (lock and key model)
 blocking enzyme activity through the
formation of noncovalent bisubstrate
complexes with NAD +.
 The binding studies suggested that the basis
for the potency of triclosan as a FabI
inhibitor was because of the formation of a
high affinity FabI-NAD+-triclosan ternary
complex.
 substrate
Product

enzyme

NADH

NAD+ + H

triclosan

No reaction
enzyme

NAD+
substrate

NADH
Structure of FabI-NAD+ -inhibitor ternary complex

The hydroxychlorophenyl ring stacks with the nicotinamide ring

of the NAD+ with an interplanar distance of 3.4 Å and contacts
Tyr-146 and Tyr-156 on the protein. The hydroxyl group of the
ligand forms hydrogen bonds with phenol of Tyr-156 and with
the 29-hydroxyl of the NAD1 ribose. The 2,4-dichlorophenyl ring
of triclosan sits in a hydrophobic pocket in contact with Met-
159. The 4-chloro substituent accepts a hydrogen bond from the
amide backbone amide nitrogen of Ala-95.
Mechanism
+
Nicotinamide ring of NAD

OH Tyr 146

O +
N

HO H
O
H O
H
O
Cl O
Cl

Tyr 156

Cl

O
O H
S
N OH
HO NH2
O
Ala 95 Met 159
Sintesis triklosan
Cl
O Cl
O
Cl OH
Cl Cl O Cl
Cl Cl Cl Cl
AlCl 3 basa/Cu katalis
O

RCO 3H

Cl Cl
+
ROH/H
R O + Cl O Cl Cl O Cl

O HO O
O
 Result
Among the many potential PfFabI inhibitors
tested so far triclosan remains the most potent
with an IC50 = 14 ng/ml (50 nM) but the
compound is not suitable for therapeutic use
because of human health and environmental
risks.

Therefore, it is evident that the plasmodial FabI

enzyme is a good target for treating malaria.
In vitro killing of intaerythrocytic forms of P.falciparum
Lakshmi M. Kumaratilake,* Brenton S. Robinson,* Antonio
Ferrante,* and Alf Poulos
This paper describes the in vitro killing of
intraerythrocytic forms of P. falciparum by
polyunsaturated fatty acids.

 The fatty acids were’nt toxic to either normal red

blood cell or parasitized red blood cells and didn’t
induce hemolylis.

 Microscopic examination, parasites exposed to

polyunsaturated fatty acids for 18 h underwent
degeneration and death.
found that immature and mature schizonts
were more susceptible to the fatty acid-
induced effects than were ring forms.
Effects of various fatty acids on growth inhibitions of P.
falciparum.

The degree of unsaturation was critical for the antiplasmodial effect of

the fatty acid towards the parasite
The oxidized forms of these fatty acids showed increased activity
against P.falciparum
Result

 Thefatty acid C22:6,n-3 caused > 90%

death of P. falciparum. The survived
parasites were in the ring form. In contrast,
C22.0- treated P. falciparum cultures were
very similar to the parasite grown in the
absence of fatty acids.
 Seleropyric acid

Good antiplasmodial activityagainst a KI multi drug resistance strain of

P. falciparum IC50 = 7.2 μg/ml
Suggest that lipid peroxidation was the most likely
mechanism responsible for the antiplasmodial activity
displayed by the polyunsaturated fatty acid. It’ll promote
oxidative damage to the infected membran cell of P.
Falciparum.
Δ5,9 very-long chain fatty acid
as farmakofor
Result
 This finding demonstrated, for the first
time, that the marine D5,9 fatty acids can
inhibit the FabI enzyme of P. falciparum,
which seems to be the potential
intracellular target of the fatty acids. This
fatty acid had almost no cytotoxicity on
mammal.
ASAM LEMAK SEBAGAI
ANTI JAMUR
 Pemanfaatan Asam lemak

Sumber www.eufic.org
 Asam Lemak Sebagai anti jamur

 Asam lemak dekanoat (10:0)/asam Kaprat mempu

menginhibisi 3 strain Candida albicans
 Asam (Z)-9-heptadecenoic (17:1) menghambat
pertumbuhan jamur phytophthora infestans dan
Idriella bolleyi. Mekanismenya : menggangu
membran sehingga terjadi pelepasan cairan
elektrolit seluler dan protein membran.
 Terbaru : asam 2-hexadecynoic dengan
mekanisme kerja menghambat proses asilasi asam
lemak khusunya sintesis triasil gliserol dan 6-
nonadecynoic dari akar Pentagonia gigantifolia ,
mekanisme kerja menghambat biosintesis
spingolipid Jamur. Aktif terhadap Cryptococcus
neoformans tp tdk aktif terhadap Candida albicans
Skema sintesis senyawa asetilenik

MEKANISMENYA???????
 Beberapa senyawa analog asam
asetilenik
Penyebab kenapa 2, 6 lebih aktif dari pada yg lain
Mekanisme reaksi obatnya???????
O
O H
O
2 - h e x a d e c y n o ic a c id
OH Aktif menghambat
C. Albicans dan C. neoformans
2 . 6 - h e x a d e c a d iy n o ic a c id O H
O
O
OH
5 - h e x a d e c y n o ic a c id

O H
O
2 . 9 - h e x a d e c a d iy n o i c a c id
9 - h e x a d e c y n o ic a c id
O
OH

O
O H
2 . 6 - n o n a d e c a d iy n o ic a c id
6 - n o n a d e c y n o ic a c id
 Asam tetradecanoic (asam miristat)
(14:0)

 Hasil uji in vitro mampu menginhibisi enzim N-

myristoyltransferase (NMT)
 Beberapa spesies jamur mensintesis N-myritoyl
protein menggunakan NMT(EC 2.3.1.97)
(Glycylpeptide N-tetradecanoyltransferase)
 Tetradecanoyl-CoA + glycylpeptide <=> CoA + N-
tetradecanoylglycylpeptide
Inhibitor (asam lemak)

Transfer asam miristic

 Asam lemak yang di isolasi
dari sponge
 α- methoxylated fatty acid seperti
(±)-2-methoxytetradecanoic acid (2-Ome-
14:0) dari spons Callyspongia fallax yang
berasal dari laut Karibia memiliki aktivitas
menghambat pertumbuhan jamur C.
Albicans dan Aspergilus niger.
 Satu mekanisme yang diajukan adalah
inhibisi NMT.
 Dengan Komputasi Biomolekul

 (±)-2-methoxytetradecanoic acid (2-Ome-

14:0) dilakukan uji efek inhibisi
menggunakan molekular modeling dengan
menggunakan enzim NMT dari Saccaromyces
cerevisine (Sc-NMT)
 Enantiomer S- 2-Ome-14:0 berikatan kuat
pada enzim NMT dari pada R- 2-Ome-14:0.
 Ditunjukkan dengan adanya ikatan hidrogen
yang terbentuk pada NH threonin 205 pada
sisi aktif NMT.
 O
Berikatan kuat dengan asam amino
O OH threonin pada Enzim NMT
(S)2-methoxytetradecanoic acid

O OH
(R)2-methoxytetradecanoic acid
 Hasil sintesis analog 2-Ome-
14:0
 2-methoxy-4-oxatetradecanoic disintesis
dan di uji aktivitasnya terhadap C.
Albicans, C. Neoformans dan A. Niger
 Bentuk S adalah inhibitor terbaik dr pada
R yg aktif terhadap C. Neoformans dan A.
Niger.
 Menariknya fungsi 4-oxa ternyata
menurunkan aktifitas induk 2-Ome-14:0
pada C. Albicans.
 O
O
O OH
2-methoxy-4-oxatetradecanoic acid

O
O OH
2-methoxytetradecanoic acid
 44

Contoh-contoh sterol
 45

KOLESTEROL DAN ERGESTEROL

 46

 KOLESTEROL terdapat di dalam jaringan dan

lipoprotein plasma yang secara khas
merupakan produk metabolisme hewan dan
terdapat dimakanan yang berasal dari
hewan seperti kuning telur, daging, hati, dan
otak
 ERGOSTEROL terdapat pada tumbuhan dan
ragi, jamur, prekursor vitamin D
 Keduanya merupakan steroid golongan
senyawa triterpenoid
 47

Biosintesis ergosterol
 Mevalonat yang merupakan senyawa 6 C, di
sintesis dari asetil koA
 Unit isopren dibentuk dari mevalonat dengan
menghilangkan CO2
 6 unit isopren mengadakan kondensasi untuk
membentuk intermediet, skualen
 Skualen mengalami siklisasi untuk menghilangkan
senyawa steroid induk yaitu lanesterol
 Ergosterol dibentuk dari lanosterolsetelah
melewati beberapa tahaplebih lanjut, termasuk
menghilangkan 3 gugus metil
 48

skualen
 49

Pembentukan streol dengan

asetat-malonat
 50

Sintesis lanosterol dari skualen

51
 52

jamur

Aspergilus
sp
c. albicans
 Definisi Fungal
Fungal Infections
Fungal infections are caused by microscopic organisms that
can invade the epithelial tissue. The fungal kingdom includes
yeasts, molds, rusts and mushrooms. Fungi, like animals, are
hetrotrophic, that is, they obtain nutrients from the
environment, not from endogenous sources (like plants with
photosynthesis). Most fungi are beneficial and are involved
in biodegradation, however, a few can cause opportunistic
infections if they are introduced into the skin through
wounds, or into the lungs and nasal passages if inhaled.
Diseases caused by fungi include superficial infections of
the skin by dermatophytes in the Microsporum, Trichophyton
or Epidermophyton genera. These 53
dermophytic infections
are named for the site of infection rather than the causative
organism.
54

Who am i??
 55

Akibat jamur
 56

Cell Membrane Disruption

Antifungal agents that disrupt the cell membrane
do so by targeting ergosterol, either by binding to
the sterol, forming pores and causing the
membrane to become leaky (as with polyene
antifungals), or inhibiting ergosterol biosynthesis (as
seen with azole antifungal agents).
Ergosterol is similar to mammalian cholesterol, thus
agents binding ergosterol may have a
cytotoxic effect in the host tissue. Ergosterol has two
conjugated double bonds that are lacking in
mammalian sterols.
57
58
 59

Jenis-jenis senyawa antifungal

 60

Amphotericin B / POLIENE
 AMPHOTERICIN B
 Class of Antifungal: Polyene
 Mechanism of Action: Binds to ergosterol
in fungal membrane causing membrane
to become leaky
 61

Azole antifungal
 Azole antifungal agents are the largest
class of synthetic antimycotics. About 20
agents on the market today. Some used
topically to treat superficial
dermatophytic and yeast infections.
Others used systemically to treat severe
fungal infections. Antifungal activity stems
from the presence of an aromatic five
member heterocyclic, either an imidazole
or a triazole.
 62

AZOLE MENGHAMBAT SINTESA

PEMBENTUKAN ERGOSTEROL
 These imidazoles and triazoles inhibit CYP P450 14 α-
demethylase in fungi. This enzyme is involved in the
conversion of lanosterol to ergosterol. Other P450s in
sterol biosynthesis may be affected. The basic
nitrogen of the azole ring forms a tight bond with the
heme iron of the fungal P450 preventing substrate
and oxygen binding. Inhibition of the C14α-
demethylase results in accumulation of sterols still
bearing a C14 methyl group changing the exact
shape and physical properties of the membrane
causing permeability changes and malfunction of
membrane imbedded proteins. They have a lower
affinity for mammalian P450's. The effect is fungistatic,
but may be fungicidal at higher concentrations.
 63

Enhibisi enzim
 64

Terbinafin
 Antifungal Agents Targeting Squalene Epoxidase: Ergosterol
Biosynthesis Inhibitors
 The allylamines have a more limited spectrum of activity
than the azoles and triazoles and are only effective against
dermatophytes. They are employed in the treatment of
fungal infections of the skin and nails.
 Mechanism of Action: These antifungal agents are
reversible, noncompetitive inhibitors of the first step in
ergosterol biosynthesis (see fig. 2), the conversion of
squalene to squalene-2,3- epoxide by squalene epoxidase.
The buildup of squalene in the cell membrane is toxic to the
cell, causing pH imbalances and malfunction of
membrane bound proteins.
 Ergosterol biosynthetic pathway.

Ghannoum M A , and Rice L B Clin. Microbiol. Rev.

1999;12:501-517
 Mechanisms by which microbial cells might develop resistance. 1, The target enzyme is
overproduced, so that the drug does not inhibit the biochemical reaction completely. 2, The
drug target is altered so that the drug cannot bind to the target. 3, The drug is pumped out by
an efflux pump. 4, The entry of the drug is prevented at the cell membrane/cell wall level. 5,
The cell has a bypass pathway that compensates for the loss-of-function inhibition due to the
drug activity. 6, Some fungal “enzymes” that convert an inactive drug to its active form are
inhibited. 7, The cell secretes some enzymes to the extracellular medium, which degrade the
drug.

Ghannoum M A , and Rice L B Clin. Microbiol. Rev.

1999;12:501-517
 Peta Konsep
67

Amphecirin b
flukanozole
azole terconazole
vorinacozole
ketonazole
Senyawa
antifungal 2,6
hexadecadiynoic
acid
terbinafin
jamur dapat di dirusak dengan dua
cara, yaitu menghambat enzim hingga
68

mencegah terbentuknya ergosterol dari

senyawa pembentuk yaitu skualen
2. senyawa 2,6 hexadecadiynoic acid
memiliki aktifitas antifungi yang lebih
baik dari obat jenis azole dengan
mekanisme sbg inhibitor enzim
3. ergosterol yang terbentuk dapat
dibocorkan dengan senyawa poliene
dimana gugus OH pada ergosterol
bereaksi dengan atom O karbonil pada
senyawa poliene/ amphicitirin B
 69

Lipid dan penggolongannya