Mechanisms of Toxicity

Biotransformation
Indri Garnasih, M.Si
Tazkiyah Izzati, S.Si
Randi, S.Si
Institut Teknologi Bandung

STEP I DELIVERY: FROM THE SITE OF

EXPOSURE TO THE TARGET
Ultimate chemical is: the chemical species that reacts with the

endogenous target molecule, ex: receptor, enzyme, DNA, protein,

lipid
The concentration of the ultimate toxicant at the target molecule

depends on the relative effectiveness of the process that increase

or decrease its concentration at the target site
The accumulation of the ultimate toxicant at its target is facilitated
by absorption, distribution to the site action, reabsorption and

toxication

Absorption versus Presystemic Elimination

Absorption: transfer chemical from the site of

exposure (external or internal body surface) into

the systemic circulation
The rate absorption related to:
Concentration of the chemical
Area of the exposed site
Psychochemical properties of the toxicant

 Presystemic Elimination
During transfer from the site exposure to the systemic
circulation, toxicant may be eliminated
The GI mucosa and the liver may eliminate a significant

fraction of a toxicant during its passage through the tissue,

decreasing systemic availability
Thus presystemic or first pass elimination reduce the toxic

effects of chemicals that reach their target site by way of

the systemic circulation

Distribution to and away from the

target
Toxicant exit the blood during the distribution phase, enter
the extracellular space, and may penetrate into cells

Chemical dissolved in plasma water may diffuse through

the capillary endothelium via aqueous intercellular spaces
and transcellular pores called fenestrae and/or across the
cell membrane

During distribution, toxicant reach their site or site of action, usually a

macromolecule on either the surface of the interior of a particular type of
cell

Chemicals also may be distributed to the site or site of toxication, usually

an intercellular enzyme, where the ultimate toxicant is formed

Some mechanism facilitate whereas others delay the distribution of

toxicants to their targets

Excretion versus Reabsorption

Excretion of the removal xenobiotics from the blood

and their return to the external environment

Excretion is a physical mechanism whereas
biotransformation is a chemical mechanism for
eliminating the toxicant
The route and speed of excretion depend largely on the
physicochemical properties of the toxicant

 The major excretory organs the kidney and the liver can efficiently

remove only highly hidrophilic, usually ionized chemicals such as

organic acids and bases
The reasons for this are as follow:
In the renal glomeruli only compounds dissolved in the plasma water
can be filtered
Transporters in hepatocytes and renal proximal tubular cells are
specialized for the secretion of highly hidrophilic organic acids and

bases
Only hydrophilic chemicals are freely soluble in the aqueous urine and
bile

Lipid soluble compounds are readily reabsorbed by trancellular

diffusion

 There are no efficient elimination mechanisms for

nonvolatile, highly lipohilic chemicals such as

polyhalogenated biphenyls and chlorinated hydrocarbon
insecticides

If they are resistant to biotransformation, such chemicals

are eliminated very slowly and tend to accumulate in the
body upon repeated exposure

 Three rather inefficient processes are available for the

elimination of such chemicals:
Excretion by the mammary gland after the chemicals is
dissolved in the milk lipids
Excretion in bile association with biliary micelles and/or

phospolipid vesicles
Intestinal excretion , an incompletely understood transport
from the blood into the intestinal lumen

Volatile, nonreactive toxicants such as gases and volatile

liquids diffuse from pulmonary capillaries into the alveoli

and are exhaled

 Reabsorption
Toxicants delivered into the renal tubules may
diffuse back across the tubular cells into the
peritubular capillaries
This process is facilitated by tubular fluid reabsorption,
which increases the intratubular concentration as well

as the residence time of the chemical by slowing urine

flow
Reabsorption by diffusion is dependent on the lipid
solubility of the chemical

 Toxicant delivered to the GI tract by biliary, gastric, and

intestinal excretion and secretion by salivary glands and
the exocrine pancreas maybe reabsorbed by diffusion
across the intestinal mucosa

Because compounds secreted into bile are usually organic

acids, their reabsorption is possible only If they are
sufficiently lipophilic or are converted to more lipid soluble
forms in the intestinal lumen

Toxication versus Detoxication

Biotransformation to harmful products is
called toxication or metabolic activation

Biotransformation that eliminate the ultimate

toxicant or prevents its formation are called

detoxication

Biotransformation of Xenobiotics
General Principle

 Xenobiotik: zat kimia asing/ berasal dari luar tubuh,

contoh: obat, pestidsida, polutan, alkaloid, metabolit
sekunder, dan racun tanaman
Lipophilicity: ciri fisik yang memungkinkan xenobiotik
diserap melalui kulit, pernafasan, atau perncernaan.
hidrophilicity: ciri fisik yang memungkinkan xenobiotik
dikeluarkan melalui organ2 ekskresi.

 Biotransformasi: perubahan xenobiotik dari zat larut dalam lemak

menjadi zat zarut dalam air

Ekskresi xenobiotik berdasarkan biotransformasinya, dikatalis oleh
enzim2 di hati dan jaringan lain
Xenobiotik memberi efek bagi system biologis (menguntungkan

atau merugikan)
Biotransformasi bergantung pada: jumlah xenobiotik yang diterima

Sifat dasar enzim biotransformasi

xenobiotik
Biotransformasi xenobiotik: mempertahankan
homeostasis, oleh enzim yang spesifik
Enzim: diinduksi oleh masuknya xenobiotik atau
diproduksi terus menerus
Contoh: sintesis dan konversi hormone steroid menjadi
larut dalam air yang dikatalis oleh cytochrome P450
Variasi enzim: variasi alel/allelozyme: perubahan asam
amino di sisi katalis
Pharmacogenetic: studi penyebab, proses, akibat
perbedaan heritate enzim biotransformasi xenobiotik

Metabolisme-biotransformasi
Metabolism: total proses yang dialami
xenobiotik, meliputi absorpsi, distribusi,
biotransformasi, dan ekskresi
Metabolit: produk dari biotransformasi
xenobiotik

Aspek stereokimia biotransformasi

xenobiotik
Banyak xenobiotik memiliki 1 atau lebih pusat
kiral sehingga memiliki 2 bentuk, disebut
enantiomer atau stereoisomer. 1 enantiomer
mengalami biotransformasi lebih cepat
daripada bentuk lainnya. Contoh: quinine
(inhibitor CYP2D6, enzim yang dihasilkan oleh
sitokrom P450)

Biotransformasi
Fase I dan Fase II
Fase I membuat toksikan lebih larut
dengan air co. Reaksi: hidrolisis, reduksi dan
oksidasi
Fase II membuat toksikan larut dengan air
sehingga dapat diekskresikan. Co. Reaksi :
glukuronidasi, sulfonasi, asetilasi, metilasi,
konjugasi dengan glutationin dan konjugasi
dengan asam amino.

Biotransformasi
Reaksi biotransformasi dapat terjadi tanpa melewati fase I atau
melewati fase I kemudian ke fase II, contohnya adalah morfin,
heroin dan codein semua dikonversi menjadi morphine-3glucuronide.
Pada morfin, reaksinya langsung menuju fase II, yaitu morfin
langsung dikonjugasikan dengan asam glucuronidase.
Pada heroin, terjadi reaksi hidrolisis (deasetilasi-FASE I) terlebih
dahulu yang kemudian diikuti dengan konjugasi produk fase I
dengan asam glucuronidase.
Pada codein, terjadi reaksi O-demetilasi (oksidasi oleh sitokrom
P450-FASE I) terlebih dahulu yang kemudian diikuti dengan
konjugasi produk fase I dengan asam glucuronidase.

Reaksi

Enzim

Lokasi

Fase I
Hidrolisis

Reduksi

Oksidasi

Esterase
Peptidase
Epoxide hydrolase
Azo- and nitro-reduction
Carbonyl reduction
Disulfide reduction
Sulfoxide reduction
Quinone reduction
Reductive dehalogenation
Alcohol dehydrogenase
Aldehyde dehydrogenase
Aldehyde Oxidase
Xanthine Oxidase
Monoamine Oxidase
Diamine Oxidase
Prostaglandin H synthase
Flavin-monooxygenase
Cytochrome P450

Mikrosom, sitosol,lisosom, darah

Darah, lisosom
Mikrosom, sitosol
Mikroflora, mikrosom, sitosol
Sitosol, darah, mikrosom
Sitosol
Sitosol
Sitosol, mikrosom
Mikrosom
Sitosol
Mitokondria, sitosol
Sitosol
Sitosol
Mitokondria
Sitosol
Mikrosom
Mikrosom
Mikrosom

Fase II
Glucuronide conjugation
Sulfate conjugation
Glutathione conjugation
Amino acid conjugation
Acylation
Methylation

Mikrosom
Sitosol
Sitosol, mikrosom
Miktokondria, mikrosom
Mitokondria, sitosol
Sitosol, mikrosom, darah

Distribution of Xenobiotic
Biotransforming enzyme
Lokasi enzim-enzim untuk biotransformasi :
Mostly in Liver (pada vertebrata), lainnya
adalah :
Kulit
Paru-paru
Nasal mukosa
Mata
Saluran gastrointestinal

Distribution of Xenobiotic
Biotransforming enzyme
Major routes of exposure to xenobiotics :
Selain lokasi2 enzim tersebut, rute lainnya
adalah ginjal, adrenal, pankreas, limpa,
jantung, otak, testis, ovarium, plasenta,
plasma, eritrosit, platelet, limfosit dan aorta.
Selain oleh beberapa organ dan jaringan,
biotransformasi juga dilakukan oleh mikroflora
yang berada di Usus.

Distribution of Xenobiotic
Biotransforming enzyme
Di HATI :
Reaksi biotransformasi umumnya terjadi pada
organel :
Retikulum endoplasma
Sitoplasma
Mitokondria
Nukleus
Lisosom

FIRST PASS ELIMINATION

FIRST PASS ELIMINATION di hati setelah oral
ingested xenobiotics
Tapi, first pass elimination juga bisa terjadi di usus
halus. Contohnya : oksidasi cyclosporine oleh
cytochrome P450 dan konjugasi morfin dengan
glucuronic acid di usus halus
First Pass Elimination di nasal mucosa (hanya untuk
inhaled xenobiotics).co. pada nasal epithelium banyak
terdapat enzim untuk biotransformasi seperti enzim
cytochrome P450, flavin monooxygenases,
glutathionine S-transferases dan carboxylesterases).
Level enzimnya sama dengan yang ada di hati.

Perbedaan Kapasitas Biotransformasi

Xenobiotics
Jaringan dan sel memiliki perbedaan dalam kapasitasnya untuk
biotransformasi xenobiotics berimbas pada toksisitasnya terhadap
xenobiotics. Contohnya adalah :
Asetaminofen dan karbon tetraklorida memiliki sifat hepatotoxic (merusak
hati) karena terjadinya aktivasi senyawa tersebut oleh enzim sitokrom
P450, menjadi metabolit yang reaktif di hati, menyebabkan nekrosis pada
centrilobular.
Perbedaan spesies juga menyebabkan perbedaan dalam biotransformasi
xenobiotik berimbas pada efek toxicologic dan pharmacologic.
Contohnya : Rate of hexoarbital biotransformation mencit > kelinci > tikus
> anjing.
Biotransformasi hexoarbital pada tikus dapat ditingkatkan dengan
diinduksi dengan cytochrome P450. Tetapi, dengan diinduksinya
cytochrome P450, perlakuan mencit dengan phenoarbital akan
meningkatkan efek hepatotoxic dari asetaminofen dan karbon
tetraklorida.