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METABOLISME OBAT

 Salmah Orbayinah
TUJUAN UMUM
 Mahasiswa dapat menerapkan dan
mempertimbangkan faktor metabolisme obat
dalam pengobatan atau praktek kefarmasian,
dimana faktor metabolisme obat akan
berpengaruh pada perubahan jendela terapi dari
suatu obat. Pemahaman perubahan metabolisme
obat akibat interaksi obat sangat diperlukan
keberhasilan pengobatan dan pencegahan
kegagalan terapi.
Introductory Concepts

■ Secara biokimia  Metabolisme berarti katabolisme (memecah


senyawa besar menjadi senyawa kecil) dan anabolisme
(pembentukan senyawa kecil menjadi besar)
■ Tetapi metabolisme obat hanya berbicara katabolisme yaitu
pemecahan molekul obat
■ Pembentukan obat disebut  sintesis
■ Obat disintesis di laboratorium dan tidak pada sistem endogen
■ Obat larut lemak membutuhkan metabolisme lebih untuk menjadi
polar, mudah terionisasi dan mudah diekskresikan dengan
melalui mekanisme fase 1 dan 2 pada metabolisme obat.
What Roles are Played by Drug Metabolism?

■ Satu dari empat parameter farmakokinetik : absorption, distribution,


metabolism and excretion (ADME)
■ Elimination of Drugs: Metabolisme bersama dengan ekskresi disebut
sebagai eliminasi
■ Ekskresi secara fisik adalah mengeluarkan obat dari tubuh.
Organ ekskresi utama adalah ginjal. Ginjal akan bekerja baik pada senyawa-
senyawa metabolit polar dan senyawa yang terionisasi.
■ Secara umum, dengan metabolisme, obat menjadi lebih polar, mudah
terionisasi dan selanjutnya lebih mudah larut air dan meningkatkan
eliminasi
■ Dan dengan metabolisme, obat dapat mengalami deaktifasi, dan
kemudian detoksikasi atau detoksifikasi
■ Beberapa obat menjadi lebih aktif setelah metabolisme (Prodrugs)
■ Kadang-kadang obat menjadi lebih toksik dan karsinogenik
Transformation of Xenobiotics by Biological Systems
Classification of metabolites:

 Inactive metabolites
 Metabolites retain similar activity
 Metabolites with different activity
 Bioactivated metabolites (prodrug
technique)
Metabolite Examples and notes
activity
Inactive Routes that result in the formation of inactive metabolites are often referred to as detoxification.
(detoxification) OH O O
Phenol sulphokinase S
O OH
3'-Phosphoadenosine-5'-
Phenol phosphosulfate (PAPS) Phenyl hydrogen sulfate
Similar activity The metabolite may exhibit either a different potency or duration of action or both to the
to the drug original drug. CH3
O CH3 O
O H
N N N
Hydroxylation N-Demethylation
OH OH
Cl N Cl N Cl N

Ph Ph Ph
Diazepam Temazepam Oxazepam
(Sustained anxiolytic action) (Short duration) (short duration)

CH3
CONHNHCH CONHNH2
CH3
Different N-Dealkylation
activity
N N
Ipronazid Isoniazid
(Antidepressant) (Antituberculosis)
HO
NCOCH3 NHCOCH3 NH2

Other substances
responsible for Substances responsible
Toxic hepatotoxicity for methemoglobinamia
metabolites
OC2H5 OC2H5 OC2H5
N-Hydroxyphenacetin Phenacetin Phenetidine
(Hepatotoxic) (Analgesic)
1-Inactive metabolites: Some metabolites are
inactive,
i.e. their pharmacological active parent compound
become inactive.

Examples:
i) Hydrolysis of procaine to p-aminobenzoic acid and
diethylethanolamine results in loss of anesthetic
activity of procaine.

H2N-C6H5-COOH + Et2N-CH2CH2OH
Inactive metabolites

ii) Oxidation of 6-mercaptopurine to 6-mercapturic acid results in


loss of anticancer activity of this compound.
6-Mercaptopurine 6-Mercapturic acid (inactive)
2-Metabolites retain similar activity:
Some metabolite retain the pharmacological activity of
their parent compounds to a greater or lesser degree.
Examples:
i) Codeine is demethylated to the more active analgesic
morphine
ii) Phenacetin is metabolized to more active paracetamol
iii) Imipramine is demethylated to the equiactive
antidepressant desipramine.

3-Metabolites with different activity:


Some metabolites develop activity different from that of
their parent drugs.
Examples:
i) Iproniazid (antidepressant) is dealkylated to isoniazid
(antitubercular)
ii) Retinoic acid (vitamin A) is isomerized to isoretinoic
acid (anti-acne agent).
4-Bioactivated metabolites
(activation of inactive drugs):
Some inactive compounds are converted to active drugs
within the body.

These compounds are called prodrugs.


Prodrugs may have advantages over the active form
(active metabolite) as more stable, having better
bioavailability or less side effects and toxicity.
Examples:
i) Levodopa (antiparkinson disease) is decarboxylated
in the neuron to active dopamine
ii) The prodrug sulindac a new non steroidal
antiinflammatory drug (sulfoxide) is reduced to the
active sulfide
iii) Benorylate to aspirin and paracetamol
iv) The prodrug enalapril is hydrolysed to enalaprilat
Stereochemistry of Drug Metabolism
OH CH2COCH3 OH CH2COCH3

Ph H
H Ph

O O O O
R-(+)-Warfarin
S-(-)-Warfarin
OH Major route Minor route OH

CH3 OH H2 C H
OH CH2COCH3 OH H2C H CH3
HO H
Ph H Ph
H Ph
O O
O O O O
R,S-(+)-alcohol derivative R,R-(+)-alcohol derivative
S-6-Hydroxywarf arin

CH3 COOH
Metabolism
H H
COOH CH3
R-(-)-Ibuprofen
S-(+)-Ibuprofen
(inactive)
(active)
Sites of Drug Metabolism
 Liver: tempat utama metabolisme, karena banyak mengandung enzym yang
diperlukan,dibandingkan organ lain
The first-pass effect
Beberapa obat dimetabolisme melalui first-pass effect: : Isoproterenol, Lidocaine
Meperidine, Morphine, Pentazocine, Propoxyphene, Propranolol, Nitroglycerin,
Salicylamide
 Intestinal Mucosa: metabolism ekstra hepatik, mengandung CYP3A4 isozyme
 Levodopa, chlorpromazine and diethylstilbestrol dimetabolisme di GI tract
 Esterases and lipases ada di usus dan memegang peran penting hydrolysis
pada beberapa prodrugs ester
 Bacterial flora ada di intestin dan colon mengurangi beberapa azo and nitro
drugs (e.g., sulfasalazine)
 Intestinal b-glucuronidase bisa menghidrolisis konjugat glucuronide yg
dikeluarkan dari empedu, dan membebaskan obat bebas atau metabolitnya yang
memungkinkan mengalami reabsorption (enterohepatic circulation or recycling)
Enzymes Involved in Drug Metabolism
CYP450, Hepatic microsomal flavin containing monooxygenases (MFMO
or FMO) , Monoamine Oxidase (MAO) and Hydrolases

 Sitokrom P450 (Cytochrome P450, CYP)


Simplified apoprotein portion
merupakan keluarga besar enzym berjenis
hemoprotein yang berfungsi sebagai katalis
oksidator pada lintasan metabolisme
L
CH3 CH3
steroid,asam lemak,xenobiotik termasuk obat, HOOC
racun dan karsinogenik Berbagai reaksi kimiawi N N CH2
organik dipercepat oleh CYP, seperti reaksi Fe+3
N N
monooksigenasi,peroksidasi, reduksi,dealkilasi, CH3
HOOC
epoksidasi dan dehalogenasi reaksi tersebut CH3 CH2
secara spesifik ditujukan guna mengkonversi
O
senyawa substrat menjadi metabolit polar untuk H R
di ekskresi atau diproses oleh enzim lain pada Substrate binding site
metabolisme fase II menjadi senyawa
konjugasinya. Heme portion with
activated Oxygen
Enzymes Involved in Drug Metabolism

 Cytochrome P450 system: berlokasi di


Simplified apoprotein portion
endoplasmic reticulum.
 Cytochrome P450 suatu Pigment yang dapat
membentuk komplek dengan CO menjadi bentuk CH3
L
CH3
tereduksi dan di absorbsi pada panjang HOOC

maksimal 450nm N N CH2


Fe+3
 Cytochromes adalah hemoproteins (heme- N N
CH3
HOOC
thiolate) yang fungsinya memberikan electrons CH3 CH2
dengan merubah secara reversibly Fe dalam
O
heme antara bentuk 2+ dan 3+ dan berperan H R
sebagai elektron acceptor–donor Substrate binding site

 P450 bukan singular hemoprotein .lebih 1000


Heme portion with
telah teridentifikasi di alam dengan ~50 fungsi activated Oxygen
active pada manusia dengan substrat spesifik
Few Important CYP450 Isozymes

CYP Main functions


family
CYP1 Xenobiotic metabolism
CYP2 Xenobiotic metabolism, Arachidonic acid metabolism
CYP3 Xenobiotic and steroid metabolism
CYP7 Cholesterol 7α-hydroxylation
CYP11 Cholesterol side-chain cleavage, Steroid 11β –
hydroxylation, Aldosterone synthesis
CYP17 Steroid 17α-hydroxylation
CYP19 Androgen aromatization
CYP21 Steroid 21-hydroxylation
CYP24 Steroid 24-hydroxylation
CYP27 Steroid 27-hydroxylation
EC Recommended name Family/gene
1.3.3.9 * secologanin synthase CYP72A1
1.14.13.11 * trans-cinnamate 4-monooxygenase CYP73
1.14.13.12 * benzoate 4-monooxygenase CYP53
1.14.13.13 * calcidiol 1-monooxygenase CYP27
1.14.13.15 * cholestanetriol 26-monooxygenase CYP27
1.14.13.17 * -monooxygenase CYP7
1.14.13.21 * flavonoid 3'-monooxygenase CYP75
1.14.13.28 * 3,9-dihydroxypterocarpan 6a-monooxygenase CYP93A1
1.14.13.30 * leukotriene-B4 20-monooxygenase CYP4F
1.14.13.37 * methyltetrahydroprotoberberine 14-monooxygenase CYP93A1
1.14.13.41 * tyrosine N-monooxygenase CYP79
Cytochrome P450 (KM hal 67-68)

fp = NADPH cytochrome P450 reductase, or NADH cytochrome b5


reductase
Jalur Umum Metabolisme Obat
1. Reaksi Fasa I atau Reaksi fungsionalisasi, terdiri dari :
reaksi oksidasi, reduksi dan hidrolisis
1. Tujuannya adalah : memasukkan gugus fungsional
tertentu yang bersifat polar, seperti OH, COOH,NH2
dan SH ke struktur molekul senyawa
Hal ini bisa dicapai dengan :
a. secara langsung memasukkan gugus fungsional,
eg: hidroksilasi senyawa aromatik dan alifatik
b. Memodifikasi gugus fungsional yang ada dalam
struktur molekul
cont
 Contoh :
1). Reduksi gugus keton dan aldehid menjadi alkohol
2). Oksidasi alkohol menjadi asam karboksilat
3). Hidrolisis ester dan amida menjadi gugus COOH,OH dan
NH2
4). Reduksi senyawa azo dan nitro menjadi gugus NH2
5). Dealkilasi oksidatif dari asam N,O,S menghasilkan gugus
NH2, OH dan SH

2. Reaksi Fase II atau reaksi Konjugasi


General Metabolic Pathways (hal 66)
Oxidation
Hydrolytic Reactions  Aromatic moieties
 Esters and amides  Olefins
 Epoxides and arene oxides  Benzylic & allylic C atoms
by epoxide hydrase and a-C of C=O and C=N
 At aliphatic and alicyclic C
 C-Heteroatom system
C-N (N-dealkylation, N-oxide
Phase II - Phase I - formation, N-hydroxylation)
C-O (O-dealkylation)
Conjugation Functionalization C-S (S-dealkylation, S-oxidation,
desulfuration)
 Oxidation of alcohols and
Drug aldehydes
Metabolism
Reduction
 Glucuronic acid conjugation  Aldehydes and ketones
 Sulfate Conjugation  Nitro and azo
 Glycine and other AA  Miscellaneous
 Glutathion or mercapturic acid
 Acetylation
 Methylation
Metabolic reaction:

Phase I reaction Phase II reaction

Oxidation
Conjugation
Reduction

Hydrolysis
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 Oxidative reaction:
1) Oxidation of aromatic carbon atoms
2) Oxidation of olefins (C=C bonds)
3) Oxidation of Benzylic, Allylic carbon atoms &
carbon atoms alpha to carbonyl & imines
4) Oxidation of aliphatic carbon atoms
5) Oxidation of alicyclic carbon atoms

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6) Oxidation of carbon-heteroatom systems:
A. Carbon-Nitrogen system
 N- Dealkylation.
 Oxidative deamination
 N-Oxide formation
 N-Hydroxylation

B. Carbon-Sulfur system
 S- Dealkylation
 Desulfuration
 S-oxidation

C. Carbon-Oxygen systems(O- Dealkylation)

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7) Oxidation of Alcohol, Carbonyle and Acid functios.

8) Miscellaneous oxidative reactions.


 Reductive reactions:
1) Reduction of Carbonyl functions.(aldehydes/ketones)
2) Reduction of alcohols and C=C bonds
3) Reduction of N-compounds (nitro,azo & N-oxide)
4) Miscellaneous Reductive reactions.

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 Hydrolytic reactions
1) Hydrolysis of Esters and Ethers

2) Hydrolysis of Amides.

3) Hydrolytic cleavage of non aromatic heterocycles

4) Hydrolytic Dehalogination

5) Miscellaneous hydrolytic reactions.

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 Oxidation of aromatic carbon atoms
(aromatic hydroxylation):
R

OH

Arenol (major)

R R R OH
H2O
O
epoxide hydrase

OH
Arene oxide
Arene (highly reactive electrophile) Dihyrdrodiol
GSH
5-epoxide transferase

R
R OH OH

SG OH
Tissue toxicity in instances
when glutathione is depleted. Glutathione conjugate Catechol(min. Pro.)
(min.pro.)
 E.g. Epoxides of Bromobenzene and Benzopyrene.
27
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 Oxidation of olefins (C=C bonds):
 Oxidation of non-aromatic C=C bonds is
analogous to aromatic hydroxylation. i.e. it
proceeds via formation of epoxides to yield 1,2-
dihydrodiols.

O HO OH

H2O

N
N epoxide hydrase N
CONH2 CONH2 CONH2

Carbamazepine Carbamazepine-10,11 Trans-10,11


epoxide dihydroxy
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 Oxidation of Benzylic Carbon Atoms:
 Carbon atoms attached directly to the aromatic ring
are hydroxylated to corresponding Carbinols.
 If the product is a primary carbinol, it is further
oxidized to aldehydes and then to carboxylic acids,
 E.g.Tolbutamide
 A secondary Carbinol is converted to Ketone.

CH2OH CHO COOH


CH3
Alcohol
dehydrogenase

SO2NHCONHC4H9 SO2NHCONHC4H9
Corresponding Corresponding
aldehyde carboxylic acid.
Tolbutamide Prmary carbinol
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 Oxidation of Allylic carbon Atoms:
 Carbon atoms adjacent to Olefinic double bonds (are
allylic carbon atoms) also undergo hydroxylation in a
manner similar to Benzylic Carbons.
 E.g. Hydroxylation of Hexobarbital to 3`-hydroxy
Hexobarbital.
Allylic carbon atom
OH
3'

2'
H3C H3C
O O O O

HN N HN N

CH3 CH3

O O

Hexobarbital 3'-Hydroxy Hexobarbital


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 Oxidation of Carbon Atoms Alpha to
Carbonyls and Imines:

 Several Benzodiazepines contain a carbon atom (C-3)


alpha to both Carbonyl (C=0) and imino (C=N)
function which readily undergoes Hydroxylation.

 E.g. Diazepam
O
N N

3 OH

N N
IC IC

Diazepam 3-Hydroxy diazepam


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 Oxidation of Aliphatic Carbon Atoms
(Aliphatic Hydroxylation):
 Terminal hydroxylation of methyl group
yields primary alcohols which undergoes
further oxidation to aldehydes and then to
carboxylic acid.

H CH3 OH CH3

H3C C C C COOH H3C C C C COOH


H2 H H2 H
CH3 CH3

Tertiary alcohol metabolite


Ibuprofen

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 Oxidation of Alicyclic Carbon Atoms
(Alicyclic Hydroxylation):
 Cyclohexane (alicyclic) and piperidine (non-aromatic
heterocyclic) rings are commonly found in a number
of molecules.
 E.g. Acetohexamide and minoxidil respectively.
 Such rings are generally hydroxylated at C-3 or C-4
positions.
H2N H2N
N N

O N N 4' O N N OH

H2N
H2N

Minoxidil 4'-Hydroxy Minoxidil


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 Oxidation of Carbon-Heteroatom Systems:
 Biotransformation of C-N, C-0 and C-S system
proceed in one of the two way:

1. Hydroxylation of carbon atom attached to the


heteroatom and subsequent cleavage at carbon-
heteroatom bond.

E.g. N-, O- and S- dealkylation, oxidative


deamination and desulfuration.

2. Oxidation of the heteroatom itself.


 E.g. N- and S- oxidation.
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 Oxidation of Carbon-Nitrogen System:
 N-Dealkylation:
 Mechanism of N-dealkylation involve oxidation of α-
carbon to generate an intermediate carbinolamine
which rearranges by cleavage of C-N bond to yield
the N dealkylated product and the corresponding
carbonyl of the alkyl group.
H OH
O C
NH
N C N C +

H H
Carbonyl
N-Dealkylated
Carbinolamine metabolite
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 A tertiary nitrogen attached to different alkyl groups
undergoes dealkylation by removal of smaller alkyl
group first.

Example:

 Secondary aliphatic amine E.g. Methamphetamine.

 Tertiary aliphatic amine E.g. imipramine

 Tertiary alicyclic amine E.g. hexobarbital

 Amides E.g. Diazepam


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 N-Hydroxylation:-
Converse to basic compounds that form N-oxide, N-
hydroxy formation is usually displayed by non-basic
nitrogen atoms such as amide Nitrogen.

E.g. Lidocaine
CH3
CH3
O C2H5
O C2H5

N C C N
N C C N H2
H H2
OH C2H5
C2H5
CH3
CH3

N- Hydroxy Lidocaine
Lidocaine
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 Oxidation of Carbon-Sulfur Systems:
 S-Dealkylation:
The mechanism of S-Dealkylation of
thioethers is analogous to N-dealkylation .IT
proceed via α-carbon hydroxylation.
The C-S bond cleavage results in formation of
a thiol and a carbonyl product.
 E.g. 6-Methyl mercaptopurine.
SCH3 SCH2OH SH

N N N
N N N
+ HCHO
N N N
N N N
H H H

6-Methyl Hydroxylated
intermediate 6-Mercaptopuri9ne
Mercaptopurine
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 Desulfuration:
 This reaction also involves cleavage of carbon-sulfur bond
(C=S).

 The product is the one with C=0 bond. Such a desulfuration


reaction is commonly observed in thioamides such as
thiopental.

Thiopental
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Pentobarbital
 S-Oxidation:

 Apart from S-dealkylation, thioethers can also undergo S-


oxidation reaction to yield sulfoxides which may be further
oxidized to sulfones several phenothiazines.

 E.g. Chlorpromazine undergo S-oxidation.

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 Oxidation of Carbon-Oxygen Systems:
 O-Dealkylation:

 This reaction is also similar to N-Dealkylation and


proceeds by α-carbon hydroxylation to form an
unstable hemiacetal or hemiketal intermediate.

 Which spontaneously undergoes C-0 bond cleavage


to form alcohol and a carbonyl moiety.
OH H

R-O-CH2R' R-O-CH-R' R-OH + O=C-R'

Alcohol Aldehyde/ketone
Hemiacetal
Ether
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 Oxidation of Alcohol, Carbonyl and
Carboxylic Acid

 In case of ethanol, Oxidation to acetaldehyde is


reversible and further oxidation of the latter to
acetic acid is very rapid since Acetaldehyde is
highly toxic and should not accumulate in body.

Aldehyde
alcohol
CH3CHO CH3COOH
CH3CH2OH
dehydrogenase
dehydrogenase
Ethanol Acetaldehyde Aceticacid
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 Miscellaneous oxidative reactions:
 Oxidative aromatization /dehydrogenation:

 E.g. Metabolic aromatization of drugs is nifedipine.

COOCH3 COOH
NO2 NO2 CH2OH
CH3

NH N

CH3 CH3
COOCH3 COOCH3

Nitedipine Pyridine metabolite


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 Oxidative Dehalogenation:
 This reaction is common with halogen containing drugs
such as chloroform.

 Dehalogenation of this drug yields phosgene which


may results in electrophiles capable of covalent
binding to tissue.

Cl O
oxi. Covalent binding to tissues.
Cl Cl Cl Cl
-HCL
H
Chloroform Phosgene
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 Reductive reaction:
 Bioreductions are also capable of generating polar
functional group such as hydroxy and amino which can
undergo further biotransformation or conjugation.

 Reduction of carbonyls:
 Aliphatic aldehydes :

E.g. Chloral hydrate

Cl3C-CHOH2O Cl3C-CH2OH

Chloral hydrate Trichloroethanol


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Aliphatic ketones: E.g. Methadone.

O OH
C 2 H5 C 2 H5
H2 C H CH3 H2 C H CH3
C N C N
H3 C CH3 H3 C CH3

Methadone Methadol

Aromatic Ketone: E.g. Acetophenone.


O OH
CH3 C CH3
H

Acetophenone Methyl phenyl carbinol


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 Reduction of alcohols and C=C:
 These two reductions are considered together because
the groups are interconvertible by simple addition or
loss of a water molecule. Before an alcohol is reduced it
is dehydrated to C=C bond.

 Example – bencyclane . (antispasmodic)

O (CH2)3 N(CH3)2 OH

Bencyclane Bencyclanol Benzylidine cycloheptane

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 Reduction of N-compounds:
 Reduction of nitro groups proceeds via formation of
nitro so and hydroxyl amine intermediates to yield
amines.
RNO2 R-N=O R-NHOH RNH2
Nitro Nitroso Hydroxylamine Amine

 For E.g. Reduction of Nitrazepam.


H O
H O N
N

H2 N N
O2N N

Nitrazepam 7-Amino metabolite.


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 Reduction of azo compounds yield primary amines
via formation of hydrazo intermediate which
undergo cleavage at N-N bond.
R-N=N-R' R-NH-NH-R' RNH2 + NH2R'

Azo Hydrazo Amines


 E.g. Prontosil.
NH2 NH2

H2N N N SO2 NH2 H2N NH2 + H2N SO2 NH2

Prontosil 1,2,4-Triamino benzene sulfanilamide

 It is reduced to active Sulfanilamide.


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 Miscellaneous reductive reactions:
 Reductive Dehalogenation:
This reaction involves replacement of halogen attached
to the carbon with the H-atom. E.g. Halothane.
Br
CF3 H CF3-CH3 CF3-COOH
Cl
Halothane 1,1,1- Trifluroethane Trifluroacetic acid

 Reduction of sulfur containing functional groups:


C 2H5 S S C 2H5 C 2H5 S E.g. Disulfuram
N S S N N SH
C 2H5 C 2H5 C 2H5

Dsulfiram Diethyldithiocarbamic acid


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 Hydrolytic reactions:
1. The reaction does not involve change in the state of
oxidation of substrate.
2. The reaction results in a large chemical chain in the
substrate brought about by loss of relatively large
fragments of the molecule.
 Hydrolysis of esters and ethers:
Esters on hydrolyisis yield alcohol and carboxylic acid.
The reaction is catalyzed by esterases.

O O
R-C-OR'
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R-C-OH
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+ R'-OH
51
 Organic acid esters:
Esters with a large acidic group : E.g. Clofibrate
COOC2H5 COOH
Cl O CH3 Cl O CH3 C 2H5OH
CH3 CH3
+

Clofibrate Free acid metan

 Esters with a large acidic and alcoholic group:


E.g. Succinylcholine
H2C COOCH2CH2-N(CH3)2 Pseudo CH2COOH
+ 2
HOCH2CH2N(CH3)2
C COOCH2CH2-N(CH3)2 Choline- CH2COOH
H2 sterase
Suceinylcholine succinic acid choline
03-12-2010 KLECOP, Nipani 52
 Inorganic acid esters:
 Phosphates: E.g. Stilbestrol diphosphate

OH C2 H5 OH C2 H5
O P O O P O HO OH + 2 H3PO4
OH C2 H5 OH C 2 H5

Stilbestrol diphosphate stilbestrol

 Sulfates: E.g. Isopropyl methanesulfonate


CH3 O CH3 O
H3C O S CH3 H3C OH + HO S CH3
H O H O

Isopropyl methnesulfonate isopropanol methanesulfonic acid


03-12-2010 KLECOP, Nipani 53
 Hydrolysis of amides:
The reactions catalyzed by amides, involves C-N
cleavage to yield carboxylic acid and amine.
O O
R-C-NHR' R-C-OH
+ R'NH2

 primary amide with aliphatic substituent on N-atom:


E.g. Procainamide
C2H5 O C2H5
O
H2N N C C N H2N + H2N C C N
H2 H2 C2H5
H H2 H2 C 2H5
OH

Procanamide PABA
03-12-2010 KLECOP, Nipani 54
 Secondary amide with aromatic Substituent on N-atom:
E.g. Lidocaine
CH3 CH3
O C 2H5 C 2H5
N C N NH2 + HOOC C N
H H2 C 2H5 H2 C 2H5
CH3 CH3

Lidocaine 2,6 Xylidine N, N- Diethylglycine

 Tertiary amide: E.g. Carbamazepine

N N
CONH2 H

Carbamazepine Iminostilbene
03-12-2010 KLECOP, Nipani 55
 Hydrolytic cleavage of non
aromatic heterocyclics:
 Four – membered lactams: E.g. Penicillins
O O
H H
C N S CH3 C N
H2 S CH3
H2
N CH3 HO N CH3
O
COOH O
COOH

Penicillin G Penicinoic acid metabolite

 Five – member lactams: E.g. Succinimides

O
O N O COOH
CH3 NH2

Phensuximide Phenyl succinamic acid

03-12-2010 KLECOP, Nipani 56


 Hydrolytic dehalogenation:
Chlorine atoms attached to aliphatic carbons are
dehalogenated easily.
 E.g. Dichloro diphenyl trichloro ethane
H -HCL H
Cl Cl Cl Cl
CCl3 CCl2

DDT DDE

 Miscellaneous hydrolytic reactions:


Include hydration of epoxides and arene oxides,
hydrolysis of Sulfonylureas, Carbamates, Hydroxamates
and alpha Glucuronide and sulfate conjugates
03-12-2010 KLECOP, Nipani 57
Type of Oxidative Reactions by CYP 450 (hal 69)

Arenols Benzylic, allylic


Arene Oxides
OH aliphatic C
O Hydroxylation
Epoxides
O
C OH
C C
C H
C C
R N H R N OH

Miscellaneous "Activated Oxigen"


R N CH2R R NH + O CHR
Oxidations [FeO]3+

S C R O CH3 R N R N O
S P S CH3
O C R OH
O O-Dealkylation N-Hydroxylation
O P SH, S CH3 N-Dealkyaltion and
Desulfuration Oxidative Deamination
S-Dealkylation
and S-Oxidation N-Oxide Formation
■ Hydroxylation is the primary reaction mediated by CYP450

■ Hydroxylation can be followed by non-CYP450 reactions including


conjugation or oxidation to ketones or aldehydes, with aldehydes
getting further oxidized to acids

■ Hydroxylation of the carbon α to heteroatoms often lead to cleavage of


the carbon – heteroatom bond; seen especially with N, O and S,
results in N–, S– or O–dealkylation.

■ Must have an available hydrogen on atom that gets hydroxylated, this


is important!!!
Aromatic Hydroxylation (KM Hal 73)
R1 R1 R1

CYP450 Spontaneous

O
■ Mixed function oxidation of arenes to OH

arenols via an epoxide intermediate R1 R1

arene oxide Epoxide


Epoxide Hydrase
hydrolase Aromatase

■ Major route of metabolism for drugs with OH OH


OH OH
phenyl ring
■ Occurs primarily at para position R1

Glutathione
■ Substituents attached to aromatic ring
influence the hydroxylation OH
S
■ Activated rings (with electron-rich Glutathione

substituents) are more susceptible while R1

deactivated (with electron withdrawing Macromolecule


groups, e.g., Cl, N+R3, COOH,
OH
SO2NHR) are generally slow or Macromolecule
resistant to hydroxylation
H
H CYP2C19 H N
N N H
HO
O O CH3
O N O N
H H
Amphetamine
Phenytoin p-hydroxyphenytoin

O O
OH OH H
C CH O N CH3
ONa CH3
CH3
O
HO
Warfarin sodium
17-a-Ethinylestradiol Propranolol
HO O Ca+2
C
O
OH
O
CH3
F N
N CH3
H3 C N

HN C
O
O
Phenylbutazone
2
Atorvastatin
Cl O O Antihypertensive drug clonidine undergo little
H H3C N S
N N aromatic hydroxylation and the uricosuric
O OH agent probenecid has not been reported to
HN H3C undergo any aromatic hydroxylation (KM hal
Cl
Probenecid 71-72)
Clonidine

CH3 O
N

CH3
N N N Preferentially the more electron
Cl
S CH3
rich ring is hydroxylated (KM
hal 72)
Cl
Diazepam Chlorpromazine

NIH Shift: Novel Intramolecular Hydride shift named after National Institute of Health where
the process was discovered. This is most important detoxification reaction for arene oxides
R R R R
Spontaneous
Rearrangement NIH Shift
Arenol
+
H
H
O -
OH H
O OH
Arene Oxide
Oxidation of olefinic bonds (also called alkenes)
O OHOH
Epoxide hydrolase

Alkene Epoxide trans dihydrodiol derivative


■ The second step may not occur if the epoxide is stable, usually it is
more stable than arene oxide
■ May be spontaneous and result in alkylation of endogenous molecules
■ Susceptable to enzymatic hydration by epoxide hydrase to form trans-
1,2-dihydrodiols (also called 1,2-diols or 1,2-dihydroxy compounds)
■ Terminal alkenes may form alkylating agents following this pathway
O HO OH

CYP3A4 Epoxide hydrolase


N N N

O NH 2 O NH 2 O NH2

Carbamazepine Carbamazepine 10,11 epoxide Carbamazepine trans 10,11 diol


(Active) (Active & Toxic) (Inactive)

Q. Any similarities or dissimilarities with aromatic – NIH Shift, Conjugation


with macromolecules?
Benzylic Carbon Hydroxylation (hal 77-78)
■ Hydroxylate a carbon attached to a phenol group (aromatic ring)

R2 R2 ■ R1 and R2 can produce steric hindrance as they get larger and


R1 C H R1 C OH
more branched
■ So a methyl group is most likely to hydroxylate
■ Primary alcohol metabolites are often oxidized further to
aldehyde and carboxylic acids and secondary alcohols are
converted to ketones by soluble alcohol and aldehyde
dehydrogenase
O O O O O O
S CYP2C9 S
N N CH3 N N CH3
H H H H H
H3C C
HO
H
Tolbutamide Metabolism

ONa
N
CH3 Dicarboxylic acid is the
major metabolite
O

H3C
Tolmetin sodium
Oxidation at Allylic Carbon Atoms (78)
H H H H H OH
R1 C C C C R3 R1 C C C C R3
R2 H R4 R2 H R4

7
CH3 7CH2OH CH3 CH3
1 HO HO
6 2 OH OH OH OH
5 3
4
H3C + H3C + H3C
H3C
O C5H11 O C5H11 O C5H11 O C5H11
CH3 CH3 CH3 CH3

1-THC 7-Hydroxy-1-THC 6a-Hydroxy-1-THC 6b-Hydroxy-1-THC

H2C H2C
3
2 OH
H
HO HO
N N
1
H3CO H3CO

N N
Quinine 3-Hydroxyquinine
O-Glucuronide Cojugate

O O O
CH3 CH3 CH3

2' 3' O O
O O O O OH O
CH3 CH3 CH3

3'-Hydroxyhexabarbital 3'-Oxohexabarbital
Hexabarbital

Pentazocine
Hydroxylation at C a to C=O and C=N (hal 79)
CH 3 CH 3 H O
O O N
N N
O H O H OH
3 OH N-demethylation
Cl N
R C C R' R C C R' Cl N Cl N

H OH

(3S) N-Methyloxazepam Oxazepam


The benzodiazepines Diazepam
or 3-Hydroxydiazepam
are classic examples
(CH3 CH 2 )2 NCH2 CH 2 CH 3
with both functionalities N
O
N
O

3 3

Cl N O2 N N

Flurazepam Nimetazepam

CH2 CH 3 CH2 CH 3
HO
3
The sedative hypnotic 4 C6 H5 4 C6 H5
1
glutethimide possesses
O N O ON O
C a to carbonyl function H H
Glutethemide 4-Hydroxyglutethemide
Aliphatic hydroxylation (hal 80)
H H H
R1 C C C OH ■ Catalyzes hydroxylation of the ω and ω-1
H H H

H H H carbons in aliphatic chains
R1 C C C H
■ Generally need three or more unbranched
H H H H OH H
carbons
R1 C C C H
H H H

O H O H
N CYP450 N
Pentobarbital Metabolism O O
N N
O H OH O H

CH3
CH3 CH3
O
O O

CYP450 OH
Ibuprofen Metabolism OH OH +
H3C CH3
HOOC CH3
H 3C CH3
OH
Alicyclic (nonaromatic ring)
Hydroxylation (hal 80)

■ Cyclohexyl group is commonly present in many drug


molecules
■ The mixed function oxydase tend to hydroxylate at the 3 or
4 position of the ring
■ Due to steric factors if position 4 is substituted it is harder to
hydroxylate the molecules

OH
O O O O O O
S S
N N CYP450 N N
H H H H
H3C H3C

O O
Acetohexamide Metabolism
Oxidation Involving Carbon-
Heteroatom Systems (hal 80)
■ C-N, C-O and occasionally C-S
■ Two basic types of biotransformation processes:
1. Hydroxylation of a-C attached directly to the heteroatom (N,O,S).
The resulting intermediate is often unstable and decomposes with
the cleavage of the C-X bond:
H
H O O
R X Ca R X Ca R XH +

Usually Unstable
Oxidative N-, O-, and S-dealkylation as well as oxidative deamination
reaction fall under this category
2. Hydroxylation or oxidation of heteroatom (N, S only, e.g., N-
hydroxylation, N-oxide formation, sulfoxide and sulfone formation)
■ Metabolism of some N containing compounds are complicated by the
fact that C or N hydroxylated products may undergo secondary
reactions to form other, more complex metabolic products (e.g.,
oxime, nitrone, nitroso, imino)
C-N systems (hal 82)
■ Aliphatic (1o, 2o, 3o,) and alicyclic (2o and 3o) amines; Aromatic and heterocyclic
nitrogen compounds; Amides
■ Enzymes:
1. CYP mixed-function oxidases: a-C hydroxylation and N-oxidation
2. Amine oxidases or N-oxidases (non-CYP, NADPH dependent flavoprotein
and require O): N-oxidation

■ 3o Aliphatic and alicyclic H


amines are metabolized by H O O
oxidative N-dealkylation R1 N Ca R1 N Ca R1 NH +
(CYP)
R2 R2 R2
■ Aliphatic 1o, 2o amines are 3o or 2o amine Carbinolamine 2o or 1o amine
susceptible to oxidative
deamination, N-dealkylation H
and N-oxidation reactions H O
O
Ca Ca + NH3
■ Aromatic amines undergoes
NH2 NH2
similar group of reactions as
o
aliphatic amines, i.e., both N- 1 amine Carbinolamine Carbonyl Ammonia
dealkylation and N-oxidation
N-Dealkylation (Deamination)
H OH
CYP450 Spontaneous
R1 C N R3 R1 C N R3 R1 C O + HN R3
R2 R4 R2 R4 R2 R4
■ Deamination and N-dealkylation differ only in the point of reference; If the drug is R1 or R2
then it is a deamination reaction and If the drug is R3 or R4 then it is an N-dealkylation
■ In general, least sterically hindered carbon (a) will be hydroxylated first, then the next, etc.
Thus the more substituent on this C, the slower it proceeds; branching on the adjacent
carbon slows it down, i.e. R1, R2 = H is fastest.
■ Any group containing an a-H may be removed, e.g., allyl, benzyl. Quaternary carbon
cannot be removed as contain no a-H
■ The more substituents placed on the nitrogen the slower it proceeds (steric hindrance)
■ The larger the substituents are the slower it proceeds (e.g. methyl vs. ethyl). In general,
small alkyl groups like Me, Et and i–Pro are rapidly removed; branching on these
substituents slows it down even more

OH
CH3 CYP2C19 CH2 H
N N N N Spontaneous N N
CH3 CH3 CH3

Imipramine N-Dealkylation
Alicyclic Amines Often Generate Lactams (hal 83)

N OH N O
N
CH3 CH3
CH3 N N
N
Carbinolamine Cotinine
Nicotine

N N O
1 CH3 CH3
C6 H 5 O 2
C6 H 5 O C6 H 5 O Lactum metabolite
Cyproheptadine
3
H 3C N H 3C N OH H 3C N O
H H H
Phenmetrazine Carbinolamine 3-Oxophenmetrazine
intermediate

COOCH 3 COOH COOH


Hydrolysis

HN HN HN

O
Methylphenidate Ritalinic Acid 6-Oxoritalinic Acid
CH3
3oAmine drugs H3C CH3
N N
CH3
H CH3
CH3 CH3
N C
N CH3 NH2 N
H3C O
O O
CH3 CH3

Lidocaine Disopyramide Tamoxifen

CH3
N
O CH3
CH3 CH3 N CH3
N N
N
S CH3 N
CH3
CH3
Cl
Br
Diphenhydramine Chlorpromazine Benzphetamine Brompheniramine
CH3 CH3 CH3
Alicyclic Amine drugs N N N
H

O CH3
HO O OH O
O
CH3
Meperidine Morphine Dextromethorphan
2o & 1o Amines

O
CH3 CH2 NH3
CH3 CH3

HN NH 2 O
CH3
Methampetamine Ampetamine Phenylacetone

Cl Cl
NHCH 3 NH 2
O O

Ketamine Norketamine

Generally, dealkylation of secondary amines occurs before deamination. The rate of


deamination is easily influenced by steric factors both on the a-C and on the N; so it is
easier to deaminate a primary amine but much harder for a tertiary amine.
Exceptions: Some 2o and 3o amines can undergo deamination directly without dealkylation
(hal 94-95).

OH OH OH
O O O
O H H2 N O
Direct Oxidative
HN CH 3 Deamination CH 3
HN
CH 3 CH 3
Propranolol NH3
Aldehyde
Carbinolamine Metabolite

OH O OH
O Oxidative Deamination
O
H3C CH3 Through Primary Amine

HN CH 3 NH 2

O CH 3
H Primary Amine Metabolite
(Desisopropyl Propranolol)
N-Oxidation (86)
H H H OH
N N N O
Aromatic amines

1 aromatic amine Hydroxylamine Nitroso


H H H H
H H O
1° amines R C N R C N R C N O R C N
H OH O
H H H H
1 amine Hydroxylamine Nitroso Nitro

H H H
CH3 CH3 CH2
2° amines R C N R C N R C N
H OH O
H H H
2 amine Hydroxylamine Nitrone

H H
CH3 CH3
R C N R C N O
3° amines
CH3 CH3
H H
3 amine N-Oxide
■ The attack is on the unbonded electrons so 3o amines can be oxidized
■ Generally, only occurs if nothing else can happen, so it is a rare reaction
■ Performed by both amine oxidases and hepatic MFO’s
■ Good examples would include amines attached to quaternary carbons since
they cannot be deaminated

H H
H3C H3C
N CYP450 N
H OH
CH3 CH3
Cl Cl
Chlorphentermine N-Hydroxylation Hydroxylamine

H NH2 Nitroso
H3C
N
H
CH3
Nitro
Phentermine
Amantadine
Amides

C-N bond cleavage via a-C


hydroxylation (formation of
carbinolamide) and N-
hydroxylation reactions
Oxidation involving C-O System (O-Dealkylation)
H OH
CYP450 Spontaneous
R1 C O R3 R1 C O R3 R1 C O + HO R3
R2 R2 R2

■ Converts an ether to an alcohol plus a ketone or aldehyde


■ Steric hindrance discussion similar to N-dealkylation
OH
CH2
O
O N NH2
H3C
H 3C N
O

Sp
on
NH2

tan
50

eo
P4

us
CY

CH3
O OH
O N NH2 O N NH2
H 3C H 3C
H3C N H 3C N
O O
NH2 Trimethoprim O-Dealkylation NH2
CH3
O O
N O H3C
H OH
N CH3
N CH3

O
O O OH
O CH3
CH3 Cl
Codeine Phenacetin Indomethacin
O OH H
O N N
H3C O O N CH3
N
H3C N
O H3C CH3
O Metoprolol
NH2 Prazosin

■ One exception that appears to be a form of O-dealkylation is the


oxidation of ethanol by CYP2E1
■ In this case R3 is hydrogen instead of carbon to form the terminal
alcohol rather than an ether
■ The enzyme involved is CYP2E1 and has been historically referred to
as the Microsomal Ethanol Oxidizing System (MEOS)

H OH
CYP450 Spontaneous
H3C C OH H3C C OH H3C C O
H H H
Oxidation involving C-S System
H OH
CYP450 Spontaneous
■ S-Dealkylation R1 C S R3 R1 C S R3 R1 C O + HS R3
R2 R2 R2
Steric hindrance discussion similar to N-dealkylation
S O
■ Desulfuration R1 C R2 R1 C R2

O O
■ S-Oxidation R1 S R2 R1 S R2 R1 S R2
O
Sulfoxide Sulfone

O
CH3 CH2 OH CH2 SH
S S
N N N N
N N
N N N N
N N H
H H
6-(Methylthio)-purine 6-Mercaptopurine
COOH
O H O H O H
H3C S N S CH2C6H5 N N
S S O
N N N
O H CF3 O H O H
Methitural Thiopental Pentobarbital
2-Benzylthio-4-
trifluoromethyl benzoic acid

NO2 NO2
H3C O S H3C O O
P P
O O
H3C O H3C O
Parathione Paraoxone
N N N N
S CH3 O S CH3
O
O
CH3 CH3
S S
N N Ring Sulfoxide Ring Sulfone
S CH3

CH3
S
N N N N
Thioridazine
S CH3 S CH3

CH3
S CH3 S
O O O
Mesoridazine Sulforidazine
Oxidative Dehalogenation
H OH
CYP450 O Spontaneous O
R C Cl R C Cl R C R C
+H2O
Cl Cl Cl OH
+ +
■ Requires two halogens on carbon
H Cl H Cl
■ With three there is no hydrogen available to
replace
■ With one, the reaction generally won’t proceed
■ The intermediate acyl halide is very reactive
OH OH HCl OH

OH OH OH
NHCOCHCl2 NHCOCCl2 NHCOCCl
O2N O2N O2N
OH O
Chloramphenicol
Oxamyl Chloride
Derivative
OH
Tissue
Nucleophiles
OH
NHCOC OH Covalent Binding
O2N
O (Toxicity)
Q. What is Gray Baby Syndrome? Oxamic Acid
Derivative
Hepatic Microsomal Flavin Containing
Monooxygenases (MFMO or FMO)

■ Oxidize S and N functional groups


■ Mechanism is different but end products are similar to those
produced by S and N oxidation by CYP450
■ FMO’s do not work on primary amines
■ FMO’s will not oxidize substrates with more than a single charge
■ FMO’s will not oxidize polyvalent substrates

H3C H H H3C H H
N N MFMO N N
S CH3 S CH3
NH N N NH N O N
C C
N N
Cimetidine MFMO S-Oxidation

Q. What is the difference with MFO?


Non-Microsomal Oxidation Reactions

■ Monoamine oxidase (outer membrane of mitochondria, flavin containing enzyme )


■ Dehydrogenases (cytoplasm)
■ Purine oxidation (Xanthene oxidase)

H Monoamine oxidase
R1 C N H R1 C O + H N H
R2 R3 R2 R3

■ Two MAOs have been identified: MAO–A and MAO–B. Equal amounts are found in
the liver, but the brain contains primarily MAO–B; MAO–A is found in the adrenergic
nerve endings
■ MAO–A shows preference for serotonin, catecholamines, and other monoamines
with phenolic aromatic rings and MAO–B prefers non–phenolic amines
■ Metabolizes 1° and 2° amines; N must be attached to α-carbon; both C & N must
have at least one replaceable H atom. 2° amines are metabolized by MAO if the
substituent is a methyl group
■ b–Phenylisopropylamines such as amphetamine and ephedrine are not metabolized
by MAOs but are potent inhibitors of MAOs
Alcohol dehydrogenase Aldehyde dehydrogenase
R2
R2 R1 C O R1 C O
R1 C OH
R1 C O H OH
H

Metabolizes 1° and 2° alcohols and aldehydes containing at least one “H” attached to a-C; 1°
alcohols typically go to the aldehyde then acid; 2° alcohols are converted to ketone, which
cannot be further converted to the acid. The aldehyde is converted back to an alcohol by
alcohol (keto) reductases (reversible), however, it goes forward as the aldehyde is converted to
carboxylic acid; 3° alcohols and phenolic alcohols cannot be oxidized by this enzyme; No “H”
attached to adjacent carbon
Alcohol Aldehyde
OH
H2 Dehydrogenase H Dehydrogenase
Ethanol Metabolism C C C
H3C OH H3C O H3C O

Purine oxidation
O O O O
Xanthine Xanthine H
N oxidase N oxidase HN N N
HN HN HN
OH O
N N O N N O N N O N N
H H H H H H H
Hypoxanthine Xanthine Uric acid Uric acid
(hydroxy tautomer) (keto tautomer)
Molybdenum Containing
Reductive Reactions

■ Bioreduction of C=O (aldehyde and keton) generates alcohol


(aldehyde → 1o alcohol; ketone → 2o alcohol)
■ Nitro and azo reductions lead to amino derivatives
■ Reduction of N-oxides to their corresponding 3o amines and
reduction of sulfoxides to sulfides are less frequent
■ Reductive cleavage of disulfide (-S-S-) linkages and reduction
of C=C are minor pathways in drug metabolism
■ Reductive dehalogenation is a minor reaction primarily differ
from oxidative dehalogenation is that the adjacent carbon does
not have to have a replaceable hydrogen and generally
removes one halogen from a group of two or three
Reduction of Aldehydes & Ketones (hal 93)
H H
R C O R C OH R C O R1 C OH
H H R2 R2
Aldehyde 1 alcohol Ketone 2 alcohol

■ C=O moiety, esp. the ketone, is frequently encountered in drugs and


additionally, ketones and aldehydes arise from deamination
 Ketones tend to be converted to alcohols which can then be glucuronidated.
Aldehydes can also be converted to alcohols, but have the additional
pathway of oxidation to carboxylic acids
■ Reduction of ketones often leads to the creation of an asymmetric center
and thus two stereoisomeric alcohols are possible
■ Reduction of a, b –unsaturated ketones found in steroidal drugs results not
only in the reduction of the ketone but also of the C=C
■ Aldo–keto oxidoreductases carry out bioreductions of aldehydes and
ketones. Alcohol dehydrogenase is a NAD+ dependent oxidoreductase that
oxidizes alcohols but in the presence of NADH or NADPH, the same
enzyme can reduce carbonyl compounds to alcohols
O
+ O
H H
H
O H2N
H2N HO H
C C +
R1 R2 R1 R2 N+
N
R
R
Ketone Red Nicotinamide moiety Chiral Alcohol Ox Nicotinamide moiety
of NADPH or NADH + +
of NADP or NAD

O HO H H OH

OH H2C CH3 OH H 2C CH3 OH H2C CH3


C6H 5 C 6H5 C6H5
H H H
+
O O O O O O

R (+)-Warfarin R,S (+)-Warfarin R,R (+)-Warfarin

CH2 O CH3
O OH

N N
OH

OH O OH
O O OH
H3C
H 3C O
HO O HO O
H2 N
OH
O O
Naloxone Daunomycin Naltrexone
CH3
OH
CH3 H
OH C CH

C CH
H
O
HO

Norethindrone H 3b,5b-Tetrahydronorethindrone

H2 H2 H2
C CH3 C CH3 C CH3
CH C CH

NH2 O OH

Amphetamine Phenylacetone 1-Phenyl-2-propanol

OH OH OH
H H H
C H C CH3 C CH3
C CH3 C CH

NHCH3 O OH

(-)-Ephedrine 1-Hydroxy-1-phenyl- 1-Phenyl-1,2-propandiol


propane-2-one
Reduction of Nitro & Azo Compounds
(hal 95-96)

H H H H
O H H
R C N R C N O R C N R C N
O OH H
H H H H
Nitro Nitroso Hydroxylamine 1 amine

H H
R1 N N R2 R1 N N R2 R1 NH2 + H2N R2

Azo Hydrazo Two 1 amines

R N N NH R NH2 + N N
Azido Amine N2
 R1 and R2 are almost always aromatic
 Usually only seen when the NO2 functional group is attached directly to an
aromatic ring and are rare
 Nitro reduction is carried out by NADPH-dependent microsomal and soluble
nitroreductases (hepatic)
 NADPH dependent multicomponent hepatic microsomal reductase system
reduces the azo
 Bacterial reductases in intestine can reduce both nitro and azo

O O
H2N S O O
H2N
N S
H2 N
H2 +
N N NH2
H2N NH2
NH2
Prontosil Sulfanilamide 1,2,3-Triaminobenzene
H
O
N

O2N O
O O
O2N N S O
N N
N NNa
H N
Cl HO
N N
OH O
O

Sulfasalazine Dantrolene
Clonazepam
Reduction of Sulfur Containing Compounds (hal 96)

O O
Sulfoxide reduction (Cannot reduce a sulfone) R1 S R2 R1 S R2 X R1 S R2
Sulfoxide O
Sulfone
Disulfide reduction R1 S S R2 R1 SH + HS R2

H3 C H3C
CH3
S S
H3C N S H3C N
N CH3
S SH
S
N,N-Diethylthiocarbamic
Disulfiram
Acid
F O

OH

CH3

H3 C
S
O Sulindac
Hydrolytic Reactions

Hydrolyzes (adds water to) esters and amides and their isosteres; the OH from water
ends up on the carboxylic acid (or its isostere) and the H in the hydroxy or amine

■ Enzymes: Non-microsomal
hydrolases; however, amide hydrolysis
appears to be mediated by liver Table: Naming carbonyl - heteroatom groups
microsomal amidases, esterases, and R1 R2 Name Susceptibility
deacylases to Hydrolysis
■ Electrophilicity of the carbonyl carbon, C O Ester Highest
Nature of the heteroatom, substituents  C S Thioester
on the carbonyl carbon, and O

substituents on the heteroatom R1 C R2 O O Carbonate


+
influnce the rate of hydrolysis
C N Amide
■ In addition, Nucleophilicity of
O N Carbamate
attacking species, Electronic charge,
and Nature of nucleophile and its N N Ureide Lowest
steric factors also influence the rate of
hydrolysis
Amida hidrolisis

AMIDE HYDROLYSIS

RCONR'R" RCOOH+ HNR'R"

MICROSOMES AND CYTOSOL


Ester hidrolisis

ESTER HYDROLYSIS

RCOOR' RCOOH + R'OH

MICROSOMES AND CYTOSOL


The Reactions
O O
Ester hydrolysis R1 C O R2 R1 C OH HO R2

O O
Amide hydrolysis (slower) H
R1 C N R2 R1 C OH H2N R2
Carbonate hydrolysis
O O O
H
R1 O C O R2 R1 OH + HO C O R2 HO R2 + HO C OH O C O + O H
Carbonate Carbonic acid derivative Carbonic acid
Carbamate hydrolysis
O O R2 O
R2 R2 H
R1 O C N R1 OH + HO C N HN + HO C OH O C O + O H
R3 R3 R3
Carbonic acid
Carbamate Carbamic acid derivative

Urea hydrolysis
O O R2 O
R1 R3 R1 R3 H
N C N NH + HO C N HN + HO C OH O C O + O H
R2 R4 R2 R4 R3
Carbonic acid
Urea derivative Carbamic acid derivative
O O
H R2 R2
Hydrazide hydrolysis R1 C N N R1 C OH + H2N N
R3 R3
Hydrazide Hydrazine
Drug Examples
OH OH

O O O O OH
H3C
+
O H3C O
OH OH

CH3 H3 C O
N
Aspirin Salicylic Acid
O O O HO O O
CH3 CH3 O CH3
H3C H3C
N N N
Cl O +
O HO
Indomethacin
O O

H2N Cocaine Benzoylecgonine Methylecgonine


H O
N O N N
N CH3 H3C O
O N
CH3 Slow Hydrolysis H3C N
O
Procainamide NH2 Prazosin
H2N
CH3
H2 N OH H
N
N CH3
O Rapid Hydrolysis O
N CH3 O
O CH3 CH3
CH3
Procaine Lidocaine
Stereoselectivity of Hydrolysis

 Etomidate (Amidate, hypnotic): R-(+)-isomer is more rapidly hydrolyzed,


but S-(-)-isomer is more rapidly hydroxylated.
Phase II Conjugation Reactions
These reactions require both a high-energy molecule and an
enzyme.

The high-energy molecule consists of a coenzyme which is


bound to the endogenous substrate and the parent drug or the
drug’s metabolite resulted from phase I reaction.

The enzymes that catalyzed conjugation reactions are called


transferases, found mainly in the liver and to a lesser extent in
the intestines and other tissues.

Most conjugates are biologically inactive and nontoxic because


they are highly polar and unable to cross cell membrane.
Exceptions to this are acetylated and methylated
conjugates because these phase II reactions (methylation
and acetylation) does not generally increase water solubility but
serve mainly to terminate or reduce pharmacological activity (they
are usually pharmacologically inactive).
Conjugating molecules:
o 1- Glucuronic acid conjugation:
o It forms O-glucuronides with phenols Ar-OH,
alcohols R-OH, hydroxylamines H2N-OH,and
carboxylic acid RCOOH.
o It can form N-glucuronides with
sulfonamides, amines, amides, and S-
glucuronides with thiols.
o 2-Sulfate conjugation:
o It is less common.
o It is restricted to phenols, alcohols,
arylamines, and N-hydroxyl compounds.
o But primary alcohols and aromatic
hydroxylamines can form unstable sulfate
conjugates which can be toxic.
Conjugating molecules:
 3-Amino acid conjugation:
 By the formation of peptide link. With
glycine or glutamine.
 4- Glutathione conjugation:
 It reacts with epoxides, alkylhalides,
sulfonates, disulfides, radical species.
 These conjugates are converted to
mercapturic acid and mostly are excreted
in bile. It is important in detoxifying
potentially dangerous environmental
toxins.
Conjugating molecules:
5,6- Methylation and acetylation reactions:
 These decrease the polarity of the drugs except tertiary
amines which are converted to polar quaternary salts.
 The groups susceptible for these reactions are phenols,
amines, and thiols.
 O-methylation of meta-phenolic OH in catecholamines
 does not generally increase water solubility but serve
mainly to terminate or reduce pharmacological activity
(they are usually pharmacologically inactive).
7- Cholesterol conjugation:
 For carboxylic acids by ester link formation or for drug
with ester group by trans esterification.
8- Fatty acid conjugation:
 For drugs with alcohol functional groups by ester link.
There are six conjugation pathways:

1)-Glucuronidation: by glucuronyl transferase.


O O
HOOC HOOC
H H
+ R X
HO HO
HO OH O UDP HO OH X R

Glucuroinc acid UDP X = OH, NR 2, CO2H, SH, acidic carbon atom

Glucuronyl Transferease catalyses this conjugation reaction

2)-Sulfate conjugation
O O
HO S O P
Adenine
O OH O O
+ R X HO S X
O R
H2O3PO HO
X = OH, arylamine, NHOH
PAPS

Sulfotransferease catalyses this conjugation reaction


There are six conjugation pathways:

3)-Amino acid conjugation:


O Y O Y
H H
C CoA + C
R S H2N CO2H R N CO2H

Acyl coenzyme A H
Y = H or CH2CH2CO2H

N-acyltransferase catalyses the conjugation reaction

4)-Glutathione conjugation
O Y O Y
H H
C CoA + C
R S H2N CO2H R N CO2H

Acyl coenzyme A H
Y = H or CH2CH2CO2H

Glutathione S-transferase catalyses this conjugation


reaction
107
5)-Methylation
CH3
+
HO2C S
Adenine
O
NH2 + R X R X CH3

H2O3PO HO
X = OH, NH2, SH
SAM

Methyltransferase catalyses this conjugation reaction

6)-Acetylation
O O
C CoA + R X C R
H3C S R X

Aceyl CoA
Y =NH2, NHNH2, SO2NH2, CONH2

N-acyltransferase catalyses the conjugation reaction


Extrahepatic metabolism
 Refers to drug biotransformation that
takes place in tissues other than the
liver.

 The most common sites include the


plasma, GI mucosa, nasal passages,
lungs and kidneys. However,
metabolism can occur throughout the
body.
Factors influencing Drug
Metabolism

110
Factors influencing Drug Metabolism
 1-Chemical Structure :
 The chemical structure (the absence or presence of certain
functional groups) of the drug determines its metabolic
pathways.

 2-Species differences (Qualitative & Quantitative):


 Qualitative differences may result from a genetic deficiency of a
certain enzyme while quantitative difference may result from a
difference in the enzyme level.

 3-Physiological or disease state:


 1-For example, in congestive heart failure, there is decreased
hepatic blood flow due to reduced cardiac output and thus alters
the extent of drug metabolism.
 2-An alteration in albumin production can alter the fraction of
bound to unbound drug, i.e., a decrease in plasma albumin can
increase he fraction of unbound free drug and vice versa.
 3-pathological factors altering liver function can affect hepatic
clearance of the drug.
Factors influencing Drug
Metabolism
 4-Genetic variations:
 Isoniazid is known to be acetylated by N-
acetyltransferase into inactive metabolite.
 The rate of acetylation in asian people is higher or
faster than that in eurpoean or north american
people. Fast acetylators are more prone to
hepatoxicity than slow acetylator.
 5-Drug dosing:
 1- An increase in drug dosage would increase drug
concentration and may saturate certain metabolic
enzymes.
112
 2- when metabolic pathway becomes saturated, an
Factors influencing Drug
Metabolism
6-Nutritional status:
 1-Low protein diet decreases oxidative reactions or
conjugation reactions due to deficiency of certain
amino acids such as glycine.
 2-Vitamin deficiency of A,C,E, and B can result in a
decrease of oxidative pathway in case of vitamin C
deficiency , while vitamin E deficiency decreases
dealkylation and hydroxylation.
 3-Ca, Mg, Zn deficiencies decreases drug metabolism
capacity whereas Fe deficiency increases it.
 4-Essential fatty acid (esp. Linoleic acid) deficiency
reduce the metabolism of ethyl morphine and
hexobarbital by decreasing certain drug-metabolizing
Factors influencing Drug
Metabolism
7-Age:
1- Metabolizing enzymes (sp.glucuronide conjugation)are
not fully developed at birth, so infants and young
children need to take smaller dosesthan adults to
avoid toxic effects.
2-In elderly, metabolizing enzyme systems decline.

8-Gender (sex):
Metabolic differences between females and males have
been observed for certain compounds
Metabolism of Diazepam, caffiene, and paracetamol is
faster in females than in males while oxidative
metabolism of lidocaine, chordiazepoxide are faster in
Factors influencing Drug Metabolism
9-Drug administration route:

 1-Orally administered drugs are absorbed from


the GIT and transported to the liver before
entering the systemic circulation. Thus the drug
is subjected to hepatic metabolism (first pass
effect) before reaching the site of action.
 2-Sublingually and rectally administered drugs
take longer time to be metabolized than orally
taken drugs.Nitroglycerine is ineffective when
taken orally due to hepatic metabolism.
 3-IVadministration avoid first pass effect
because the drug is delivered directly to the
blood stream.
Factors influencing Drug Metabolism

10-Enzyme induction or inhibition


Several antibiotics are known to inhibit the
activity of cytochrome P450.
Phenobarbitone is known to be cytochrome
P450 enzyme inducer while cimetidine is
cyt. P450 inhibitor.
If warfarin is taken with phenobarbitone, it will
be less effective.
While if it is taken with cimetidine, it will be
less metabolized and thus serious side
effects may appear.
Strategies to manage drug
metabolism

117
Strategies to manage drug
metabolism
1)-Pharmaceutical strategies:
by using different dosage forms to either avoid or compensate for
rapid metabolism.
1-Sublingual tablets (through mucous mermbrane) by delivering
drugs directly to blood and bypassing first-pass effect as
nitroglycerine (antiangina drug).
2-Transdermal patches and ointments: provide continuous supply of
drug over extended period of time and are useful for rapidly
metabolizing drugs suchj as prophylactic nitroglycerine.
3- Intramuscular injections provide a continuous supply of drug over
extended period of time such as`lipid soluble esters of estradiol
and testosterone. Hydrolysis of these prodrugs produce a steady
supply of rapidly metabolized hormones.
4-Enteric coated formulation can protect acid-sensitive drugs as
erythromycin.
5-Nasal administration allows for the delivery of peptides such as
aerosols since they need only to penetrate the thin epithelial
layer to reach the abundant capillary beds
Strategies to manage drug
metabolism
2)-Pharmacological strategies
These involve the concurrent use of enzyme inhibitors to decrease drug
metabolism. Sometimes the concurrent use of an additional agent
doesn’t prevent metabolism but prevents the toxicity caused by
metabolites of the therapeutic agent.

1- Levodopa, the aminoacid precursor of dopamine, in the treatment o


Parkinson’s disease. Carbidopa , a dopa decarboxylase inhibitor

2- β-Lactam antibiotics activity is reduced by micoorganisms capable o


secreting β–lactamase enzymes. Clavulanic acid is a β–lactamase
inhibitor used in conjunction with penicillin antibiotics.
Strategies to manage drug
metabolism
3)-Chemical strategies
These are molecular modifications
involving the addition, deletion or
isosteric modification of functional
groups.
Examples are:
1-chlorpropamide was designed from
tolbutamide
2-Methyl testosterone was designed from
testosterone. 120
Prodrugs strategies
Prodrugs are used instead of active
form of the
drug to:
a) Enhance membrane permeability,
b) Reduce drug toxicity
c) Overcome /mask bad taste
d) Overcome acid sensitivity
e) Prolong (short) duration.

121
Advantages of Prodrugs
1- An increase in water solubility by using sodium succinate esters as
chloramphenicol succinate IV injection.

2- An increase in lipid solubility


a-Increase duration of action by using lipid soluble esters
b-Increase oral absorption as by using esters of the highly polar drugs or
N-methylation
C-Increase topical absorption of steroids by masking OH group as esters or
acetonides.
3-A decrease in water solubility to increase palatability as in
chloramphenicol palmitate

4- Decrease GI irritation (side effect) as in aspirin

5- Site specificity as in methyl dopa

6- Increased half-life and chemical stability as in cefamandole acetate a


stable prodrug, while the parent cefamandole is unstable solid dosage
form. Hetacillin is another prodrug (for ampicillin ).