Phase I – Phase II

• Phase I: Chemical modifications that introduce or uncover functional groups on a xenobiotic

that provide sites for Phase II metabolism

• Phase II: Synthetic reaction of a xenobiotic (or of a Phase I metabolite of a xenobiotic) with
an endogenous substance that results in introduction of polar, ionizable groups to enhance
water solubility and hence excretion

Phase II - conjugation Covalent "conjugation" to endogenous substances:

In subsequent phase II reactions, these activated xenobiotic metabolites are conjugated with
charged species such as glutathione (GSH), sulfate, glycine, or glucuronic acid. These reactions are
catalysed by a large group of broad-specificity transferases, which in combination can metabolise
almost any hydrophobic compound that contains nucleophilic or electrophilic groups.[1] One of the
most important of these groups are the glutathione S-transferases (GSTs). The addition of large
anionic groups (such as GSH) detoxifies reactive electrophiles and produces more polar metabolites
that cannot diffuse across membranes, and may, therefore, be actively transported.

Generalizations

1. Changes activity of xenobiotics

Quantitative (detox or activation)

Qualitative (disposition and effect)

2. Phase I products most often more polar and less lipophilic

3. Phase II reactions most often abolish biological activity and add to polarity

4. Biotransformation requires energy or energy equivalents

5. Enzyme systems are inducible

6. Enzyme systems are saturable, frequently by depletions of cofactors

7. Systems are age, gender, and species dependent and are affected by exgogenous environmental
factors

Biotransformation

• Water soluble xenobiotics are easier to eliminate ( t 1/2)

• Urine, feces but not exhalation

• If within barrier, no out

• Multiple enzymes (families)

• Constitutively expressed
 • Inducible

• Broad specificity

• Polymorphic (allelic variants)

• Stereo-isomer specificity: 6-OH in hormones:

Cytochrome P450 (CYP) Mixed Function Oxidases (MFO)

• Located in many tissues but highly in liver ER

• Human: 16 gene families

• CYP 1,2,3 perform drug metabolism

• 57 genes in the human genome

• Key forms: CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4

• Highly inducible

– Alcohol CYP2E1

– Dioxin/PCBs CYP1A

– Barbiturates CYP2B

• CYP genes have multiple alleles (2D6 has 53, and 2E1 has 13)

Oxidative Stress

1. Classic definition: The production of reactive oxygen in excess of antioxidant mechanisms

2. Modern definition: Altered homeostatic balance resulting from oxidant insult.

Definitions

• Toxicology is the quantitative and qualitative study of the adverse effects of toxicants on
biological organisms

• Toxicant is a chemical or physical agent that produces adverse effects on biological

organisms.

So Toxicology is the study of:

• How toxicants enter the organism

• How toxicants effect the organism

 • How toxicants are eliminated from (leave) the organism

All substances are toxic if taken in the wrong quantities

How toxicants enter organism

• Inhalation (mouth or nose to lungs) then into blood(+*)

• Ingestion (mouth to stomach) then into blood(+)

• Injection (cuts, punctures in skin) into blood

• Dermal absorption (through skin) into blood(+*)

+ Involve membrane transport

* Greatest threats in industry

KINETIC OF POISON

ABSORBTION : à DEPEND ON :

* FAT SOLUBLELITY

* PARTICLE SIZE

* BROAD OF SURFACE ABSORBTION

* CONTACT MANNER

* BLOOD CIRCULATION

PLACE OF ABSORBTION

* SKIN

* RESPIRATION

* G.I. TRACT

DISTRIBUTION :

DOSE

Vd = ------------------------

PLASMA CONCENTRATION

* RATE OF DETOXIFICATION

* CHARACTERISTIC PIOSON

* PROTEIN TISSUE / PLASMA BINDING

MECHANISM OF DETOXIFICATION

PHASE I :

* OXIDATIONà MAO, COMT

* REDUCTION à ENZIM MIKROSOMAL

- DECHLORINATION

- DEHALOGENATION

* HYDROLISIS

* HYDRATION

PHASE II :

* SULFATION à SULFOTRANSFERASE

* GLUCORONIDATION àGLUKOROSIL TRNSFERASE

* CONYUGATION à AS.GLUKORONAT

* ACETYLATION à

* METYLATION à

FACTORS TO INFLUENCE FOR ACTION OF POISON

1. DIFFERENCE OT THE SPESIES

CAT à SENSITIF PHENOL & ORGANOCHLORIN

à MORPHIN (STIMULATION)

2. AGE à DETOXIFIKASI AND EXCRETION

3. CHARACTERISTIC OF POISON à INSECTISIDE, LOGAM BERAT, GAS

4. DOSE OF POISON

5. CONTACT MANNER (LOKAL. SISTEMATIK)

6. CUMULATIVE EFFECT (DTT,GLIKOSIDA)

7. CONDITION (RESISTENSI) & SEX

FACTORS TO INFLUENCE FOR ACTION OF POISON...CONT

POISONING PROCESS

• ACUTE à 24 HOURS
 • SUBCHRONICà REPEATE UNTIL 90 DAYS

• CRONICà Up than 6 mounth

EFFECT AND ACTION OF POISON

5. TERATOGENIC EFFECT

6. MAIN DRUGS EFFECT TO BE MORE

7. WRONG IN DRUG APPLICATION

8. HYPERSENSITIF REACTION

9. ENZYM DAMAGE

- INSECTISIDE, METAL POISON