Biotransformation of drugs

(Drug metabolism) (Drug metabolism)

PharmDr. Pavol Jeko
Center of Excellence Pharmacy
Department of Medicinal Chemistry
CADD Laboratory
Faculty of Pharmacy, Comenius University
Odbojarov 10, 832 32 Bratislava, Slovakia
Tel: + 421 250 117 221
e-mail: jezko@fpharm.uniba.sk; p.jezko@gmail.com
1
Drug metabolizms
it is a biochemical modification or degradation of
drugs
drugs are partly eliminated unchanged or partially
biotransformed by metabolic pathways and are biotransformed by metabolic pathways and are
excreted as metabolites
drug metabolism is also of great importance in
medicinal chemistry because it influences (in
qualitative, quantitative, and kinetic terms) the
deactivation, activation, detoxification, and
toxification of the vast majority of drugs
2
OBJECTIVE OBJECTIVE: :
conversion of lipophilic
drugs to more polar drugs to more polar
compounds that are easier
eliminated
3
XENOBIOTI XENOBIOTICS CS
substances that do not naturally occur in the body
(that enter the body ), are not necessary for the
physiological development and have no
nutritional value nutritional value
drugs (health)
agrochemicals (agriculture)
pesticides, herbicides, fertilizers
food additives
flavoring agents, coloring agents, preservatives, stabilizers
4
Places of drug biotransformation
liver: the majority of metabolic reactions
kidney: 2 phase
guts: 2 phase
blood (ester hydrolysis)
plasma (hydrolysis of esters) plasma (hydrolysis of esters)
within a cell:
endoplasmic reticulum (microsomes)
enzymes cytochrome P450 (CYP-450)
mitochondria
cell cytosol
5
CONSEQUENCES
of
BIOTRANSFORMATION
bioinactivation:
usually products are less active or inactive
metabolites metabolites
bioactivation:
in some cases, the metabolic process converts
non-active substance on its own active form
(prodrug)
6
TYPES OF BIOTRANSFORMATION
REACTIONS
drug metabolism takes place in two steps:
first phase - functionalization reactions
the molecule introduces a new functional group, usually polar (-COOH,-OH,-NH2)
the polar group can serve as a reaction point for the second reaction phase
second phase - conjugation reactions
includes the addition of endogenous molecules (glucuronic, sulphate) to the
metabolite of 1 phase
(prerequisite for conjugation reactions is the presence of a suitable chemical group
(-COOH,-OH,-NH2)
results of both phases are metabolites more soluble in water ,
and thus easier elimininated
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Type of reaction The reaction pathway
Oxidation aromatic or aliphatic hydroxylation
N- or S-oxidation
N-, O- or S-dealkylation
Reactions of 1 phase drug metabolism
Reduction Reduction NO
2
group to hydroxylamine and
amine
Reduction of the carbonyl to alcohol
Hydrolysis Ester on acid and alcohol
Amid on acid and amine
Hydrazide on acid and substituted hydrazine
Type of reaction Endogenous reagent or
substrate
Xenobiotic substrate
Glucuronidation Uridine-diphosphate of
glucuronic acid (UDPGT)
Carboxylic acid, alcohol,
phenol, amine
Sulfation 3-phosphoadenosine-5-
phosphosulfate (PAPS)
Alcohol, phenol, amine
Reaction of 2 phase drug metabolism
phosphosulfate (PAPS)
Acetylation Acetyl-CoA Amine
Glutathione
conjugation
glutathione Epoxides, compounds
with chlorine atom
Metylation S-adenozylmethionine Phenols, amines, thiols
Aminoacid Glycine,glutamine Carboxylic acid
1. OXIDATION 1. OXIDATION
incorporation of polar groups in the molecules of the drug or substitution
of one polar group by another for increase of the polarization
according to the character of reaction, metabolic oxidation is divided
into several types:
A. Oxidation of alkyl-compounds
B. Oxidation of alcohols, aldehydes, acids B. Oxidation of alcohols, aldehydes, acids
C. Oxidative hydroxylations of aromatic compounds
D. Oxidative N and O dealkylation
E. Oxidation to Noxids
F. Oxidative deamination
G. Oxidation of sulfids to sulfoxids and sulfones
H. Oxidative desulfuration
I. Oxidative dehalogenation
J. Oxidative opening of the ring
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1. a) Oxidation of alkyl 1. a) Oxidation of alkyl- -compounds compounds
it is the most common type of oxidation on the side-alkyl chain (if
a compound has aliphatic, aromatic or heterocyclic character)
drugs containing aliphatic unbranched alkyls are
easily oxidized; oxidizing of alkyls decreases with ramifications of the easily oxidized; oxidizing of alkyls decreases with ramifications of the
alkyl chain
oxidation usually begins on the last carbon through
the primary alcoholic group and proceed finally to acid
can take place on the penultimate carbon (beta-
oxidation), which are metabolised type of fatty acid compounds in the
body
11
oxidation of pentobarbital: metabolites with prim.
aliphatic hydroxyl or carboxyl group at the terminal methyl group
are produced; or the sec. alcoholic group on the penultimate carbon
atom of the alkyl is metabolised
N
H
O CH
CH
3
O
N
H
O CH
CH
3
O
N
H
O
CH
3
CH
3
O
N
H
O
CH
3
CH
3
O
OH
N
H
N
H
O
CH
3
C
H
2
CH
3
O
O
OH
N
H
N
H
O
CH
3
CH
3
O
O
OH
O
1. 1. b ) b ) Oxidation alkohols, aldehydes, acids Oxidation alkohols, aldehydes, acids
aliphatic primary alkohols aliphatic primary alkohols
light oxidation of alcohols (prim. drug) to aldehydes (COH, sec. metab.)
and to the acid (-COOH, final metab.)
ahk oxidcia cez aldehydy (COH) a na kyseliny (COOH)
tertiary alcohols are not so much metabolized
mainly are creating glucuronide conjugates
13
alcohols
1. 1. c) c) Oxidat Oxidative ive hydroxyl hydroxylation ation of of aromatic aromatic
compounds compounds ( (rings rings) )
phenolic substances are produced, the final metabolite is eliminated as
a conjugate with glucuronic acid or sulfuric acid, which is well soluble in water
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phenolic substances are produced
1. 1. d) d) Oxidative Oxidative N N and and OO dealkylation dealkylation
common reaction on ethers and alkylamino-drugs
etheric bond is quite strong, so simple aliphatic ethers
are often excreted unchanged, the complex ethers are metabolised partially only
phenolic ethers are cleaved to the phenolic part and aldehyde.
15
alcohols
alcohols
drugs containing a tertiary or secondary amine (NH
2
) are
partially metabolized to secondary and primary amines, and
aldehyde
it is assumed that N-metyl-derivates are dealkylated
through the N-oxides and N-hydroxymetyl-derivates by the
scheme:
16
1. 1. e) e) Oxidation Oxidation to to N Noxids (N oxids (Noxidation) oxidation)
drugs with tertiary amino-group
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alcohols
1. f) Oxidative deamination
drugs, which contains the primary aliphatic amines are metabolised by monoamine
oxidase (MAO). These reactions vary according to the nature of the amine.
phenylethylamine-type drugs are oxidized to aldehyde and acid and ammonia:
substances like aryl-isopropylamine are oxidized to ketone
18
alcohols
1. 1. g) g) Oxidation Oxidation of of sulfides to sulfoxids sulfides to sulfoxids
and and sulfones sulfones
thioethers type drugs - this type of biotransformation is
common in the group of phenothiazines
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alcohols
1. 1. h) h) Oxidative desulfurization Oxidative desulfurization
some sulphur-compounds are oxidized by oxidative desulfurization,
the sulfur atom will be refund for oxygen atom in drug molecule (SO)
(some thiobarbiturates are oxidized to barbiturates)
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alcohols
1. i) Oxidative dehalogenation
halogenated compounds are converted to acid
halothane is partially metabolised by the scheme
DDT
21
alcohols
alcohols
1. j) Oxidative opening of the ring
complete destruction of the drug molecule
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1.k) Oxidation of double bonds
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alcohols
alcohols
2. REDUCTION 2. REDUCTION
uncommon metabolic pathway
reduction takes place in liver microsomes, in the presence of
reductase
function groups: nitro and azo-groups, aldehydes, ketones,
arsenic compounds
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2. a) Aromatic nitro 2. a) Aromatic nitro- -compounds compounds
are reduced (in the presence of nitro-reductase) to aromatic amines
(as intermediates can be nitroso- a hydroxylamines derivates)
amino-compounds are excreted as acetyl-derivates
25
2. b) 2. b) Azo Azo- -compounds compounds
are reduced (in the presence of azo-reductase) to aromatic amines
(probably in the first step is produced hydrazo-benzene)
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alcohols
alcohols
alcohols
2. c) Aldehydes and ketones 2. c) Aldehydes and ketones
aldehydes are reduced to primary alcohols
alcohols
ketones are partly eliminated unchanged
in some cases are reduced to secondary alcohols, and are excreted from the
body in the form of glucuronic acid conjugate
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alcohols
cyclic ketones
alcohols
compounds which contains arsenic
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alcohols
3. 3. HYDROLY HYDROLYS SI IS S REAC REACTTI IONS ONS
reactions, in which substances with a functional group is
broken down into simpler products
A. hydrolysis of esters
hydrolysis of amides B. hydrolysis of amides
C. hydrolysis of anilides
D. hydrolysis of nitriles
E. hydrolysis of carbamates
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3. a) Hydrolysis of esters
all esters are splited by esterase on the acid and alcohol part
hydrolysis of esters is realised in blood or in liver (depends on drug
character)
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alcohols
3. b) Hydrolysis of anilides
first is N-dealkylation, next step is hydrolysis of anilide bond
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alcohols
alcohols alcohols
3. c) Hydrolysis of amides
by amidase (in the liver).
32
alcohols
alcohols
3. d) Hydrolysis of nitriles
on the acid (only with aromatic derivates)
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alcohols
4. C 4. CONJUGA ONJUGATION TION REAC REACTIONS TIONS
condensation reaction
metabolic reactions in which the drug, respectively its metabolite with hydroxyl
(OH), carboxyl (COOH) or amino (NH2) group in the body is conjugated with
hydrophilic - the body's own compound : glucuronic acid, sulphate, glycocoll,
acetic acid, etc. acetic acid, etc.
according to the nature body chemical component, we can divide conjugate
condensation reactions to conjugation:
A. glucuronic
B. sulfate
C. glycocollic
D. acetylation
E. methylation
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4. a) 4. a) Glucuronic Glucuronic conjugation conjugation
glucuronic acid reacts with drugs containing hydroxyl (alcohols, phenols),
carboxylic, sulphydryl (SH) and partially amino group
glucuronic conjugation mechanism - a reaction occurs between the metabolite
and glucuronic acid in the active form
glucuronic acid reacts in the activated formas
uridine diphosphate glucuronic acid (UDP), uridine diphosphate glucuronic acid (UDP),
- binds to the drugs with the active semiacetyl hydroxyl group, it means
with hydroxyl group bound to the phosphoric acid
N
N
O O
OH
H
OH
OH
H
OH
H
H
O
OH O
P O P O
O
OH
O
OH
O H OH
O
OH
Kyselina uridn-difosfoglukurnov (UDP-glukurnov)
uridine diphosphate glucuronic acid
UDP reacts with the alcohol (with long aliphatic chains) or phenol and forms
ether-glucuronides
O
OH
H
OH
OH
H
OH
H
H
O
OH
O
R OH
O
OH
H
OH
OH
H
OH
H
H
O
OH
O
UDP
+
R
terglukuronid
examples:
menthol, borneol, camphor, paracetamol, morphine, estriol
ether-glucuronide
UDP UDP reacts reacts with with the aliphatic or aromatic carboxyl-groups and forms
ester-glucuronide
O
OH
H
OH
OH
H
OH
H
H
O
OH O
R COOH
O
OH
H
OH
OH
H
OH
H
H
O
OH O
O
R
Esterglukuronid
UDP
+
examples:
ester-glucuronide
benzoic acid, salicylic acid, acetylsalicylic acid
UDP forms with aromatic amines N-glucuronides
UDP forms with sulfhydryl groups S-glucuronides
UDP forms conjugates also with the body's own substances such as steroid
hormones
4.b) Sulfate conjugation
sulfate conjugation is conneted with
aliphatic and aromatic hydroxyl-group-
drugs (alcohols and phenols) and
aromatic amine compounds, which
provide sulfamates
own process occurs in the liver
3-phosphoadenosine-
5-phosphosulfate
own process occurs in the liver
it is few-steps process and sulphonation,
and agent in the final phase works
3 -phosphoadenosine-5-phosphosulfate
sulfate conjugation of the drug are
eliminated mainly phenolic nature such
as phenols, cresols, naphthols,
morphine, etc.
38
by sulfate conjugation are
eliminated some of the body's own
substances such as hormones
(estrone, Androsteron)
4.c) 4.c) Glycocollic Glycocollic condenzation condenzation
by this way are predominantly metabolised aromatic acid as conjugation ingredients except
glycocoll (glycine) also apply to amino acids (cysteine, ornithine, glutamine)
reaction: acid reacts with glycocoll or amino acids in the active form, it means binding to
coenzyme A (CoA):
R
OH
O
R
S
O
N H
2
O
OH
R N
H
O
OH
O
CoA-SH
N-acyl-glykokol
CoA
+
+
CoA-SH
4.d) Acetylation
closely related to the glycocollic conjugation, as acetylation agent is
acetylcoenzyme A, which reacts with the amino group of the drug
reaction occurs in the liver and kidneys
acetylation is important conjugation reaction, especially for drugs with
a prim. amino group (histamine, p-aminobenzoic acid, sulfonamides)
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4.e) Methylation
this conjugation reaction is unique in humans (common reaction in animals)
common reaction with primary and secondary amines, unique at drugs with hydroxyl or
sulphydryl group
donor of the methyl group is activated S-adenosyl methionine:
most common methylation is reaction with most common methylation is reaction with
catecholamines (adrenaline, noradrenaline),
histamine, nicotinic acid )
N-methylation pyridin derivates morphine, codeine, barbiturates
S-methylation mercaptopurine, dimercaptopropanol
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PRODRUG PRODRUG
inactive inactive prodrug prodrug form form is is metabolised metabolised in in organism organism
on on the the active active drug drug form form
42
Strategies Strategies for for development development
of of prodrug prodrug form form
improvement improvement of of solubility solubility or or better better preparation preparation
of of drug drug form form
improvement improvement of of per os per os absorption absorption and and distribution distribution
high high specifity specifity and and lower lower toxicity toxicity high high specifity specifity and and lower lower toxicity toxicity
stability or stability or prolongation prolongation of of drug drug release release
better tolerance by patients
protectiong group must be stable against stomach acid
and enzymes, but after absorption should be sufficiently
labile, for release of aktive drug form (metabolite)
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Metabolizms of selected drugs
OCOCH
3
COOH
OH
COOH
eliminated without changes (2 22%)
OH
CONHCH
2
COOH
conjugation
with glycine
Acetyl-
salicyl
acid
Salicyl acid
45-91%
OH
COOH
O H
Gentisic acid 2-4%
esters and ether
glucuronids
NHCOCH
3
OH
OH
N O H COCH
3
NHCOCH
3
H
Paracetamol
Acetaminophen
ox.
paracetamol cysteine
intermediate
glutathion cleavage
O-glucuronide 60%
O-sulphate 30%
p-aminophenol
(nephrotoxic)
NHCOCH
3
O
OH
NHCOCH
3
S peptidy
OH
S C
H
2
C
H
COOH
NH
2
OH
NHCOCH
3
S C
H
2
C
H
COOH
NHCOCH
3
N-acetyl-benzoquinone-
imine
hepatic peptides
hepatotoxic metabolite
paracetamol
mercapturate
O
OH
OH
OH
C H C
H
2
OH
NH
2
OMe
OH
OH
OH
C H
OH
OH
CH
2
OH
OH
OH
OH
OMe
COMT
COMT
MAO
in the periphery
in CNS
C H C
H
2
OH
NH
2
OH
C H C
H
2
OH
NHCH
3
OH
OH
C H C
H
2
OH
NHCH
3
OMe
C H
OH
CHO
C H
OH
COOH
C H
OH
COOH
Noradrenaline
(Norepinephrine)
COMT
Adrenaline
(Epinephrine)
The The end end
no no drug drug is is metabolised metabolised only only by by one one pathway pathway,, but but there there are are
more more of of this this pathways pathways at at the the same same time time
so, so, for for the the drug drug metabolims metabolims, , there there are are no no general general rules rules
e.g. sulphonamides are metabolised side by side with oxidative hydroxylation and next
step is sulphate conjugation, next reaction is acetylation, the mutual relationship of step is sulphate conjugation, next reaction is acetylation, the mutual relationship of
metabolites may vary according to the total amount of drug metabolized
e.g. small amount of phenole-derivate is eliminated by conjugation with sulphate, but
when there is an excess of phenole-derivates in the body, there is the higher part of its
conjugate with glucuronic acid
47
48