Stereochemistry: A Source of Problems in

Medicinal Chemistry
Everardus J. Ariens
lnstitute of Pharmacology and Toxicology, University of Nijmegen, Geert Grooteplein Noord 21, 6525
EZ NlJMEGEN The Netherlands

I. Introduction . . . . . . . . . . . . . . . .
11. Isomeric Ballast .........
111. Actions of "Inactive"Isomers.. .................................... 453
IV. Drug Metabolism and Ste
V. Stereochemical Pitfalls in
VI. Conclusions ............................................

INTRODUCTION
Symmetry is a common aspect of nature at first view. On a molecular level,
however, asymmetry dominates, both in the building materials such as ami-
noacids and sugars, and in metabolic and regulatory processes in which en-
zymes and specific receptors are involved. As a consequence, the presence
of a chiral center in drugs and bioactive agents in general implies large dif-
ferences for the enantiomers both in the activity in the strict sense, and for
metabolic conversion and pharmacokinetics in general.
Structural requirements for the biological activity often imply the presence
of one or more chiral centers in the drug. Many of them are marketed as
racemates. The enantiomers must, particularly from the biological point of
view, be regarded as different substances. The neglect of stereochemistry in
the development and application of drugs and bioactive agents in general
leads to serious misconceptions and is a source of problems in pharmacoki-
netics.
At first view, symmetry is a very common phenomenon in nature. On the
molecular level, however, asymmetry dominates as illustrated by the chirality
of amino acids and sugars and by the stereospecificity of enzymatic reactions,
drug-receptor interaction, etc. This holds true for the whole field of biology.
It counts also for various types of messenger molecules, such as neurotrans-
mitters, hormones, allosteric modulators of enzyme activity, as well as for
xenobiotic, exogenous, messenger molecules such as drugs, insecticides and
weedkillers. These transfer specific information, chemically coded in suitable
molecular carriers, into biological objects.' Stereoselectivity in action is com-
mon. Chirality is not a requirement for bioactivity but in those cases, and
there are many, in which a chiral centre is present in the bioactive molecule,
usually great differences are found for the activities of the enantiomers. Un-
derstanding of the processes involved will be helpful in the development of
more active and selective agents and particularly in the proper use of ther-
apeutics and of bioactive agents in general.

Medicinal Research Reviews, Vol. 6, No. 4, 451466 (1986)

0 1986 by John Wiley & Sons, Inc. CCC 0198-6325B61040451-16$04.00
452 ARIENS

No doubt, the selectivity in action is based on a chemical complementarity

between the messenger molecule or pharmacon and its specific receptor-site,
the site of binding on the receptor molecule. In this complementarity, besides
the physicochemical characteristics of the groups in the molecule which par-
ticipate in the interaction, also their spatial arrangement, (their sterical con-
figuration), is essential. Stereospecificity in action can be counted for on the
basis of as few as three binding groups in the molecule, a three-point
interaction,1 2 3 A 5 . 6
The isomers do not differ in their chemical properties in solution. Once
bound to the specific, that is "stereoselective", site of action differences be-
tween the stereoisomers show up.
Separation techniques too are based largely on stereoselective "binding" of
one of the isomers to a proper substrate. This holds also true for stereoselective
analytical methods, such as radio-immuno- and radio-receptor assays7 and
chromatography via stereoselective adsorbents. In this respect also enantio-
selective determination based on derivatization with an enantiomer of a chiral,
possibly labeled or fluorescent, reagent opens perspective^.^,^ The two en-
antiomers in the racemic drug then form, usually easily separable and de-
tectable, diastereoisomers.
Another possibility is the use of pseudoracemates, 1:l mixtures of deuter-
ated (+) and non-deuterated ( - ) or non-deuterated (+) and deuterated ( - )
isomer^.^^,^^ Since there is plenty of evidence that deuterated and non-deu-
terated compounds often are metabolized in a different, even stereospecific,
way70,71,72 there are restrictions. This technique is only valid if the deuterium
is located in a stable position, free of metabolic attack.
If more than one center of asymmetry is located in one bioactive molecule
(the case of diastereoisomers) and in other cases of geometric isomers such
as: cis-trans-isomers, chair and boat configurations, epimers, isomerism on
basis of intramolecular sterical hinderance, etc. as a result of differences in
the intramolecular relationship between the various groups in the isomers,
these will differ physicochemically.Therefore, as a rule such isomers are more
easily separated than the enantiomers of compounds with a single center of
asymmetry. Separation of the latter is still a laborious task.
In organic synthesis, normally enantiomers are obtained in a proportion
1:l. For geometric isomers this proportion depends on the synthetic proce-
dure. Products of biological origin, such as hormones, antibiotics, etc. as a
rule are obtained in stereospecific form. Largely for economic reasons, syn-
thetic products such as drugs, insecticides, weed killers and in general in-

~ ~~~~~~

E.J. ARIENS was born on January 29, 1918, at Wijk bij Duurstede, The Netherlands. He studied
chemistry and medicine at the university of Utrecht. In 1950 he obtained the degree of doctor in sciences
(chemistry)(Ph.D.)and of doctor in medical sciences ( M .DJ. Dr. Ariens was promoted in 1954 to professor
and director of the Department of Pharmacology at the University of Nijrnegen. His many publications
concern chemical structure and biological activity, the mechanism of action of biologically active compounds,
and receptor theory. More recently he has studied drug design: the rational development and application
of drugs, the avoidance of toxicity. His major publications are: Book "Molecular Pharmacology" (two
volumes) and a series of 10 uolumes on "Drug Design." Textbook "Introduction to general toxicology"
appeared in Dutch (2973)' English, German, Italian, Spanish and lapanese. Indonesian in press. Recently
appeared "Stereochemisty and biological activity of drugs" (1983).
STEREOCHEMISTRY PROBLEMS 453

dustrial products usually are marketed as racemic mixtures. The users, such
as physicians, and in certain cases even the producers, too often unknowingly
or unaware apply mixtures of compounds (stereoisomers) in the supposition
that just one compound is involved.2

ISOMERIC BALLAST
In case of stereoselectivity in action only one of the components in the
mixture, the racemate, is truly active. The biologically more active isomer is
termed the eutomer, the less or inactive isomer the distomer. This is so
regardless of their -d- and -1- or -R- and -S- configuration but with regard to
a particular biological action. The degree of stereospecificity, that is the ratio
of the activities (affinities, potencies, etc.) of the enantiomers is termed the
eudismic r a t i ~ . ~The
, ~ ,distomer
~ in the mixture should be regarded as an
impurity, or "isomeric ballast", not contributing to the effect aimed at. It,
however, potentially contributes to the unwanted effects, the side-effects and
toxicity.
In medicine, there apparently is not too much concern about carrying along
with e.g. 50 mg of the agent with the therapeutic action, 50 mg of a second
agent with no contribution to the desired effect, although potentially harmful.
For certain types of therapeutics, such as P-adrenergic agents, P-adrenergic
blockers, anti-epileptics and oral anticoagulants, up to 90% of the products
on the market are in fact racemic mixtures. For antihistaminics, anticholin-
ergics, and local anesthetics, this holds true for about 50%,while on the whole
it concerns 20 to 25%of the therapeutics.'l In certain cases the differences in
activity of the enantiomers are well established.2~4~5,10,12,13J4~15
In other cases no information is available. The implications of the use of
mixtures of active and inactive isomers become clearer if one considers ap-
plication of, for instance, pesticides. Neglect of the "isomeric ballast", the
inactive isomer, constitutes a risk. Its presence implies chemical pollution-
be it of the milieu interne of man and animals-or of the environment in
general. Taken into account the growing apprehension on chemical pollution,
one has to be well aware of this situation.
The remarkable discrepancy between on one hand the high degree of purity
required for pharmaceuticals and on the other hand the acceptance of 50%
impurity, as long as isomeric ballast is involved should be a matter of serious
concern.
One at least might require that the presence of this impurity is harmless.
Time is ripe to consider marketing of really pure drugs.
Techniques for stereoselective synthesis and separation of enantiomers
by means of stereoselective anorganic- and biocatalyst are rapidly develop-
ing~16,17,18,19,20,21,22,22a,22b

ACTIONS OF "INACTIVE" ISOMERS

If only one of the enantiomers (the eutomer) is responsible for the desired
biological e.g. therapeutic action there is no reason why not the, in this sense
inactive one (the distomer) could be active in a different way. There is a whole
spectrum of possibilities in this respect, many of which have been confirmed
experimentally.
454 ARIENS

1. One isomer has the therapeutic action, the other one contributes to
the side-effects, or even is the main source thereof. d-Ketamine is
predominantly hypnotic and analgetic. The Z-isomer is the main source
of unwanted side-effect~.~~ The anorectic action of Fenfluramine, a
racemate, in use now for about 15 years is located in the S( +)-enan-
tiomer recently marketed as dextrofenfluramine (IsomerideB). This is
twice as active and has less disturbing side-effects (dizziness, drow-
siness, and sedation) reported for the racemate and to be ascribed to
the "inactive" R( -)-enanti~mer.~*
2. Both isomers contribute to the main effect such as the local anesthetic
action of the isomers of prilocaine while only one contributes to the
hemotoxicity.24
3. One of the isomers may have an additional advantageous action like
in the case of bupivacaine. Both isomers are local anesthetics but only
one, the ( -)-isomer shows a vasoconstrictive
4. The therapeutically non-active isomer counteracts a side-effect of the
therapeutically active isomer. In the diuretic indacrinone d is diuretic
and causes uric acid retention, 1 acts as an uricosuric. It antagonizes
the uric acid retention brought about by the diuretic isomer. This is
not as effectiveas it looks since the "natural" proportion 1:l between
the compounds, the isomers, in a mixture as a rule is far from optimal.
A study of various mixtures shows that a proportion of ld:81 is optimalz6
(Fig. 1).Comparable relations are found for the isomers of the diuretic
tienilic acid.27
5 . The isomers may have opposite effects. In some barbiturates the 1-
isomer is a depressant, the d-isomer a convulsant (Fig. 2).2s,29
Another example are the optical isomers of 1.4-dihydropyridine(BAY K8644).
+
The ( )-(4R)-enantiomer being a calcium entry-blocker while the ( - )-(45)-
enantiomer promotes the calcium In some cases isomers act as com-
petitive antagonists of each other. Mutual interaction between the enantiom-
ers occurs. Dependent on the affinities of the isomers to their common sites
of action the racemic mixture acts as a partial agonist.2In the narcotic analgetic
picenadol d acts as an agonist, f as an antagonist, the racemate, dl, as a partial
agoni~t.~' Similar relationships are reported for other agent^,',^^,^^ among
which pheromonesM and auxine-type plant growth substance^.^^
In those cases that the enantiomers have an affinity to common receptors
but differ in their intrinsic activity the racemate will behave as a "pseudo"
partial agonist. The characteristics thereof depend on the affinities and in-
trinsic activities of the individual enantiomers. In case of a racemic mixture
composed of an agonist and a competitive antagonist with equal affinities to
the receptors at saturation thereof with the racemate, only 50% will be acti-
vated. 35ar35b,35c
6. The isomers may have advantageous complementary action like in the
case of the a-p-adrenergic blocking "pseud~hybrid-drugs."~~
7. Stereoselectivity may be restricted to only one component in the bi-
ological action. The p-adrenergic blocking action of the p-blockers is
stereoselective the non-specific cardiodepressant and the local anes-
thetic action is not. This indicates a difference in the mechanisms of
STEREOCHEMISTRY PROBLEMS 455

A Uric acid (mgldl)

21
1-
10140 10180
0 -.
1010 10110 10120
-1 -
0A.M. ‘I
-3 P.M.
Figure 1. Change in plasma uric acid concentration (X 2 SD) of healthy humans (AM, n = 18,
PM, n = 17) at day 7 of daily treatment with the diuretic indacrinone. The dose is composed of
various (+)I( -) isomer ratios ranging from 10/0 to 10180 mg/mg (Tobert et al., Clin. Pharmacol.
Ther. 29,344 (1981)). 10/10 still causes uric acid retention. The ratio 10180results in a clear uricosuric
action.

action i n ~ o l v e d .Then
~ ~ , ~the
~ actions concerned can be separated by
suitable molecular manipulation. If the eudismic ratio for one, e.g. the
therapeutic, action clearly differs or even is inverse to that for other
components in the action, this too indicates different mechanisms of
action. In the last case the actions can be separated by separation of
the isomers.

r S( - ) depressant

R( + ) convulsant

RS convulsant

0‘ S( - ) antagonizes R( + )

barbituric acid DMBB

5-(1,3-dimethylbutyl)-5-ethyl
Figure 2. Stereospecificity of depressant and convulsive action of barbiturates (I. K. Ho, Ann.
Rev. Phamacol. Toxicol., 21,83, 1981). In general for barbiturates: S( - ) predominantly depressant
and R( + ) convulsive component in the action.
456 ARIENS

S-enantiomer R-enantiomer
Timolol L-714 465

P&-adrenergic
blockade strong very weak

reduction intra- strong slightly less

ocular pressure strong

Figure 2A. Richards et al., Br. 1. Clin. Pharmacot. 1985, 20,459.

The P-adrenergic blocking action must be considered as an undesired side-

effect in those cases in which other components in the action are exploited
therapeutically for example both the (-)- and the (+)-isomer of sotalol have
a class I11 anti-arrhythmic action while the P-adrenergic blocking action is
practically restricted to the (-)-isomer. Therefore ( + )-Sotalolis to be preferred
when the drug is to be used as a class I11 anti-arrhythmic agent.38aA com-
parable reasoning holds true for Timolol, the S-isomer which with its R-isomer
has in common a reduction of the intra-ocular pressure but is by far more
potent as a P-adrenergic blocker. Therefore, for the treatment of a glaucoma
the R-isomer of Timolol is preferable since it is practically free of the risky
adrenergic blocking action38b(fig. 2").
8. The most common situation is that in which one of the enantiomers
is clearly active while the other one is only poorly active. One has to
take into consideration that incomplete separation of the enantiomers
particularly counts for the distomer.
Presence of a small fraction of the eutomer in the distomeric compound
shows up as activity of the latter. This will be the larger, the higher the
eudismic ratio on the receptor system concerned is. The differences in activity,
if measured in vivo, as a matter of fact may also be due to differences in
distribution, particularly metabolic conversion and excretion. The implications
thereof again can differ for oral (stereoselective first-pass metabolism) and
parenteral administration which is the case for instance for the calcium-chan-
nel blocker Verapami1.38c,38dData obtained with isolated tissues concern
stereospecificity in action in the more strict sense.
The various examples given above make clear that the terms eutomer and
distomer and eudismic ratio can be used only in relation to a particular bio-
logical action. The enantiomer, that is the eutomer with the therapeutic action
may, like in the case of ketamine, be the distomer for unwanted actions.23
For indacrinone the eutomer for the diuretic action is the distomer for the
uricosuric action.26For certain barbiturates, the eutomer for the depressant
action is the distomer for the convulsant action and vice ~ e r s a *(Fig.
~ , ~2).
~
STEREOCHEMISTRY PROBLEMS 457

Stereoselectivityin biological action may, like in the cases mentioned before,

be related to the drug-receptor interaction, the pharmacodynamics of the
agent.
It may, however, be due as well to differences in pharmacokinetics, e.g.
in the rate of metabolic conversion or even in the pathways
and in differences in transport processes, including uptake and storage in
particular tissues. *1,15,42,43,44,45*46 Also on this level mutual interference be-
tween the isomers is possible.

DRUG METABOLISM AND STEREOCHEMISTRY

There are various possibilities for the enantioselective biochemical conver-
sion: 1) Substrate stereoselectivity. Only one isomer is converted by the en-
zyme. The second isomer may show an enzyme inhibitory action. 2) Product
stereoselectivity. The non-chiral substrate-with a pro-chiral center-is con-
verted preferentially to one of the possible enantiomers of the product. The
enantioselectivity in drug metabolism may be restricted to differences in the
rate of conversion. 3) Combination of 1 and 2 substrate-product stereoselec-
tivity resulting in formation of preferentially one of the possible diastereoiso-
mers too is possible. 4) Enzymatic inversion of only one of the isomers to
preferentially the other enantiomer. This can be considered as a particular
case substrate-product stereoselectivity.
The schemes Ia, b, and c exemplify the cases 1, 2 and 3. A wide variety of
various types of enantioselective metabolic conversion of drugs and xeno-
biotics in general has been reported in the literat~re.",'~,~~,~~,~~
Drug metabolism implies bioactivation and/or bioinactivation. Thus chirality
of bioactive agents not only directly has consequences for the type of action
and activity, but also indirectly because of stereoselectivity in metabolic con-
version.
Enantioselectivity in drug metabolism is observed in regular compounds
where a methyl group is introduced next to the site of enzymatic attack. Such
a group then interferes with the enzymatic action, maybe by sterical hind-
erance or otherwise. The introduction of such a protective group, an often
applied manipulation, usually prolongs action. It usually implies introduction
of a center of asymmetry. The isomers usually clearly differ in action and
metabolism. One isomer may serve as a substrate for, and the other as an
inhibitor of the enzyme. This is the case for e.g. acetyl-P-methylcholinewith
a ratio for the cholinergic action of the S( +)-/R( -)-isomers is 200. The rate
of hydrolysis by acetylcholinesterasefor the S( +)-isomer is about 50% of that
for acetylcholine. The R( -)-isomer is hardly hydrolysed and in fact acts as
an inhibitor of the enzyme .47*48 For acetyl-a-methylcholinethese ratios clearly
differ. The position of the center of asymmetry in the molecules is of impor-
tance (table l).48 Dexamphetamine, the S( +)-isomer of amphetamine, is a
more potent stimulant than levamphetamine, the R( - )-isomer, which is a
more potent blocker of norepinephrine re-uptake in the sympathetic nerve
endings. Amphetamine deamination is species dependent in its stereoselec-
tivity, R( - ) > S( + ) in the rabbit and S( + ) > R( - ) in guinea pig and man.47
Only the (-)-isomer of a-methyldopa has an antihypertensive action and
is less toxic than the racemate. Contrary to (+)a-methyldopa, which is in-
active, the ( -)-isomer is decarboxylated and hydroxylated in a stereoselective
458 ARIENS

Scheme l a .
PRODUCT-SELECTIVITY

prochiral center
* center of asymmetry Rl
product
one isomer
diazepam enzy matis
t oxydation S( - ) N-methyloxazepam
prochiral center CJ
.1 enzymatic
desmethyldiazepam S( - ) oxazepam
oxydation
dopamine dopamine, R( - ) norepinephrine
hydroxylase
rabbit ( - ) phenyl-n-propylcarbinol
propiophenone
+
Scheme l b .
SUBSTRATE-SELECTIVITY

R3 R2 R3 Rx
dl

-
d-isomer product same config.
* center of asymmetry as I-isomer; or
non-chiral e g . R, = R,

-
dl-acetyl-P-methylcholine Ach-esterase R( - ) .acetyl-P-methylchuoline
S( + ) P-methylcholine +
acetic acid
dl-prilocaine amidase S( + ) prilocaine
toluidine + l-methyl-l-aminopropyl-
acetic acid

way to ( - )-( aS:PR)-methylnorepinephrine, the active endproduct. The ( -)-

isomer of a-methyldopa serves as a prodrug, the (+)-isomer is not thera-
peutically active but contributes to the ~ide-effects.~~
The ( + )-isomer of deprenyl has only a weak MAO-b-inhibitory (antide-
pressant) action and is therefore therapeutically ineffective, it is converted to
+
S( +)-metamphetamine, which is converted to S( )-amphetamine both con-
tributing strongly to the undesired CNS stimulant side-effect. The RC - )-iso-
mer (Selegilineo) has a strong MAO-B inhibitory and thus antidepressant,
therapeutic, action. It is converted to R( -)-metamphetamine and R( -)-am-
phetamine respectively which have only a weak unwanted CNS-stimulant
~ide-effect~~ (fig. 3).
Both isomers of prilocaine have a local anesthetic action, only the R( -)-
isomer, however, is hydrolysed to toluidine, the cause of methemoglobin
STEREOCHEMISTRY PROBLEMS 459

Scheme lc.
SUBSTRATE- AND PRODUCT-SELECTIVITY

dl d-isomer one diastereomer

dl-a-methyldopamine dopamine, 2R( - ) a-methyldopamine

hydroxylase 1R2S(- ) a-methylnorepinephrine

Rl R

,%R2 R-+,R3 isomeras:

enzyme R-f-R2 R+R2
R3 R2 R3 R3
dl one isomer

dl-clidanac enzymatic S( + ) clidanac

chiral inversio?

* center of asymmetry
0 prochiral center

formation, an unwanted side-effect. The S( +)-isomer, which is not hydro-

~ ~ ,4~).~
lysed does not cause this s i d e - e f f e ~ t(fig.
The implications of introduction of a metabolically stabilizing methylgroup
in the critical 15-position of prostaglandines are illustrated in fig. 5. The methyl-
ester formation increases lipophilicity and thus facilitates absorption.
An interesting situation is that of the non-steroidal antiinflammatory agents
such as ibuprofen. The therapeutically inactive isomer is to a certain extent
converted metabolically in the body to the active one51,52,53 (fig. 6 ) . There still
are, however, good reasons to apply only the eutomer since the conversion
of the distomer to the eutomer takes place only partially and will depend on
the condition, e.g. liver function, of the patient.52Metabolic chiral inversion

Table I
Cholinergic (muscarinic) activities and rate of hydrolysis by acetycholinesterase of acetyl-p-
and acetvl-a-methvlcholine isomers
~~

"ileum ratio rates of hydr.

compound guinea pig ( + )4-) ACh = 100

acetylcholine 1.0 - 100

( + )acetyl-P-methylcholine 1.0 240 54
( - )acetyl-p-methylcholine 240 weak inhibition
( + )acetyl-a-methylcholine 28 8 97
( - )acetyl-a-methylcholine 230 78
"number of molecules equivalent to 1 molecule of acetylcholine. A. Beckett, Ann. N.Y. Acad.
Sciences, 144, 675, 1967.
460 ARIENS

STEREOSELECTIVE ACTION AND BIOTOXICATION

DEPRENYL

Selegiline (+I
(-)*
-
strong MAO-b inhibitor -
weak

oxydative dealkylation
methamphetamine methamphetamine

R(-) L weak
amphetamine
CNS stimulant
side-effect -
strong .1
S(+)
amphetamine

*antidepressant
antiparkinson

Figure 3. ( - ) Deprenyl (SelegilineR)is a strong Mao-b inhibitor and thus antidepressant, ( + )

deprenyl is only poorly active. This enantiomer, however, is metabolized to S( +) methamphe-
tamine and amphetamine with an undesired CNS stimulant action. The (-) enantiomer, Sele-
giline, is metabolized to the only poorly stimulant R( - ) amphetamines and therefore free of the
side effect (49). (G.P. Reynolds et al., Br. 1. Clin. Pharmacol., 6, 542, 1978.)

STEREOSELECTIVE BIOTOXICATION
LOCAL ANESTHETICS

prilocaine qua tacaine

R( - ) local anesthetic S(+) local anesthetic
hydrolyzed not hydrolyzed not hydrolyzed

no methemoglobinemia no methemoglobinemia

toluidine + oxidation -+ ultimate toxon -+ methemoglobinemia

Figure 4. Due to "packing" of the amide function by double methylgroup-substitution and the
consequential sterical hindrance quatacaine is resistant against hydrolysis and thus not biotox-
icated. The partial packing by one methyl group in prilocaine results in a center of asymmetry.
As often in similar situations only one of the isomers, here S( +), is sterically hindered. The
other isomer is hydrolyzed and thus biotoxicated (24). [T. Takada et al., Chem. Abstr., 67,72325
(1967)].
STEREOCHEMISTRY PROBLEMS 461

INTRODUCTION OF A STABILIZING MOIETY AND

A FACILITATING MOIETY IN PROSTAGLANDIN PGEz

bWH OH
/

H O H
PGE2
inhibitor gastric secretion
short action
t inactive if orally administered
prostaglandin 15-OH dehydrogenase
bioinactivation

f? - PG 15(S) 15-me-Ezmeester
~ C inhibitor gastric secretion
~ Q
prolonged action
active at oral administration

PG IS(R)15-me-E2me ester
inhibitor gastric secretion
spasmogen smooth muscle
active at oral administration

a stabilizing moiety
c] disposable facilitating moiety
Figure 5. Karim, S. M. M. et al. Brit. Med. J. 1, 143, 1973.

of antirheumatic 2-arylpropionic acids is further reported for benoxaprofen,

cicloprofen, elidanac, and n a p r ~ x e n . ~ ~ , ”
Stereoselectivity can differ for various closely related enzymes such as the
esterases and the mixed function oxidases and thus dependent on the dis-
tribution of these enzymes show species differences and even differ for var-
ious organs in one specie^.^,^^,^,^^ Monoamine oxidase A and B are closely
related but different enzymes which also differ in their distribution. R( -)
phenylethanolamine is metabolized preferentially by MAO-A whereas the
S( +) enantiomer is metabolized by MAO-B.

OOH

If 1-I bupr of eii’

lnversion of ibuprofen in humans
Administered, SIR-ratio Excreted SIR-ratio
95: 5 95: 5
6 : 94 80 : 20
50 : 50 70 : 30
Figure 6. Adapted from Kaiser et al. (51).
462 ARIENS

Table 11.
Stereochemical aspects, ( - )/( +) ratios, in pharmacokinetics after oral doses of pseudoracemic
propranolol in man and dog (55,69)

oral bioavailability 0.53 1.44

in urine
unchanged drug 0.52 1.50
glucuronides 3.50 1.76
side chain oxidation products 0.47 1.45
ring oxidation products 1.18 0.59

The ratios Mao-a/Mao-b inhibitory IC50values differ for the enantiomers.

For deprenyl they are 120 for ( + ), 75 for ( - ) and for N 425a compound related
to depreny18.8 for (+) and 0.5 for ( -).73 For S( +) amiflamine the inhibitory
ratio, KiMAO-A/MAO-Bis 500, for the R( -) enantiomer it is 27.58
Taken into account the differences in the various pharmacokinetic param-
eters of the enantiomers, the proportion thereof, initially 1:1, changes with
the time. This holds true for the concentration of ”the” drug as well as that
of the various metabolites. The enantiomers even may generate a different
spectrum of metabolites.55,59,60,61 After application of drugs as racemates, often
the drug and its metabolites excreted in the urine are optically active.
The stereoselective metabolic conversions and possibly distribution pro-
cesses resulted in a change in the original 1:l proportion of the enantiomers.
So for instance application of propranolol as a psuedoracemate to dogs and
man results in clear changes in the original 1:l ratio of the racemate in the
pharmacokinetic processing as summarized in table 2 (55,69). This also illus-
trates species differences for stereospecific metabolism. No doubt also in plasma
these proportions clearly differ from 1. In the time-plasmaconcentration curves,
representing the elimination of a racemate from plasma, one may expect a
steeper part related to the elimination of the metabolically more labile en-
antiomer followed by a more shallow part related predominantly to the more
stable enantiomer. If the role of stereoselectivity is neglected computerised
curve fitting will lead to ”good evidence” for an interesting multicompartment-
system with an intriguing set of pharmacokinetic constants. The investigator
not only fooled himself and his readers, but even the computer.

STEREOCHEMICAL PITFALLS IN PHARMOKINETICS AND

DRUG MONITORING
Taken into account the lack of difference in chemical properties of enan-
tiomers in solution, with the exception of immuno- and receptor-binding
assays and chiral derivatization8r9,the analytical methods used in the study
of pharmacokinetics as a rule do not differentiate between them. The con-
centrations measured in fact concern mixtures, not necessarily 1:1, of the
original compound and of the various metabolites.
Remarkable is the neglectance of stereochemical implications in clinical
STEREOCHEMISTRY PROBLEMS 463

pharmacology, where hardly any attention is paid to the inherent compli-

cations in pharmacokinetics.
It is more the rule than the exception that mixtures of two and even four
different compounds, the isomers, are treated as if just one compound was
Not withstanding the overwhelming abundance of
involved.62,63,64,65,66,67,68
evidence for the fact that different isomers usually differ in action, they often
are metabolized along different pathways and at different rates. Also the
transport, binding to proteins and storage in granula etc. are stereoselective.
The condition, for instance liver function, of the patient and genetic differ-
ences also count. The implications of stereochemistry for pharmacokinetics
have consequences too for therapeutic monitoring of patients treated with
racemic drugs. Conclusion: based on the relationship between the dose reg-
imen and the systemic concentration of what is erroneously regarded as “the
therapeutically active agent” and on the relation thereof with the therapeutic
response are misleading and possibly risky.
The flow of racemic drugs to the market is abundant. Of the chiral products
introduced in 1984 and 1985, if semisynthetic products such as penicillins,
cephalosporins and polypeptides are left out, about 50% are racemates. Note-
worthy is the frequency with which methyl groups located close to vulnerable
positions in the molecule, discussed before, are the basis for the asymmetry.
One wonders how much is known about these various products concerning
the differences in pharmacokinetics and pharmacodynamics, including toxi-
cology, of the enantiomers.
It is amazing how the authorities responsible for the acceptance, registra-
tion, of drugs in various countries accept impurities of 50% and more as long
as it concerns isomeric ballast. They never will accept 50% of a byproduct in
a drug even if it is considered (without proof) safe. The fact that it is not easy
to eliminate the impurity hardly will count. The authorities even go further.
They require pharmacokinetic date on the racemic drugs accepted by them.
A waste of time and money and a token of incompetence. In those cases that
the racemates are replaced in the future by the pure agents, the active en-
antiomers, requirement of complete repeat of preclinical and clinical inves-
tigation (including toxicology) thereof would underline the lack of insight in
the implications of stereochemistry in the field of drug development and
application.

CONCLUSION
The presumption that a mixture of two different chemicals such as two
enantiomers will behave as if only one agent is involved is not tenable. In a
straight forward empirical analysis of the relationship between the dose and
the therapeutic effect and in general the therapeutic efficacy, for practical
reasons stereoselectivity may be neglected. In pharmacokinetic studies, how-
ever, where concentrations of the drug applied and of its various metabolites
are measured, one has no right to talk about concentrations of “the” active
agent and ”its” various metabolites without differentiating between the en-
antiomers if a racemic mixture is involved.
In the study of bioactive agents it is preferable if not a requisite to use pure
464 ARIENS

agents, compounds with as little impurity as possible. Mixtures of compounds

may be an object of study, particularly if one wants to gain information on
the interaction between the compounds in the mixture. The use of mixtures
of compounds, without being aware of it, is reprehensible. In those cases
where racemates or mixtures of. isomers in general are used because of the
non-availability of the separate isomers, one at least should mention explicitly
that the data presented concern a mixture of compounds which possibly and
often definitely differ in their biological action including pharmacokinetics.
This may prevent the author of presenting fictitious pharmacokinetic con-
stants and the reader from accepting them.
In the journals covering pharmacokinetic research as well as studies on
therapeutic monitoring there is such an abundance of articles dealing with
racemic mixtures neglecting the implications of stereochemistry that many
pages would be required to summarize them. After being alerted the reader
and hopefully the referees easily will notice.
In the coming years many patents, also on inferior racemic drugs, are
expiring. This may be a good occasion to replace these by “new” ”DEXTRO”
-”or ”LAEVO-” products to be announced as: Twice as active! Free of bulk
impurity! No isomeric ballast”! Less side effects! Produced according to ad-
vanced standards of drug development! In some cases patent-protection may
be obtained on the rejuvenated drug.

Acknowledgement
The author is indebted to Mrs. A.R.H. Wigmans for her thorough and
patient assistance in editing this manuscript.

REFERENCES
1. E. Ariens, Molecular Pharmacology, Academic Press, New York London, 1, 3 (1964).
2. E. Ariens, Stereochemisty and biological activity of drugs, E. Ariens, W. Soudijn, and P. Tim-
mermans, Blackwell Scientific Publications, Oxford, p. 11 (1983).
3. L. Eason and E. Stedman, Biochem. J., 27, 1257 (1933).
4. F. Lehmann, J. Rodrigues de Miranda, and E. Ariens, Progress Research. E. Jucker, Vol. 20.,
Birkhauser Verlag, Base1 Stuttgart, 101 (1976).
5. P. Portoghese, Annual review of pharmacology, H. Elliot, W. Cutting, and R. Dreisbach, Vol.
lo., Annual Reviews, Inc., Palo Alto, California, 51 (1970).
6. F. Lehmann, Receptors and recognition. P. Cuatrecasas and M. Greaves, Vol. 5, Series A,
Chapman and Hall, London, 1 (1978).
7. S. Enna, Neurotransmitter binding, H. Yamamura, S . Enna, and M. Kuhar, Raven Press, 177
(1985).
8. H. Weber, H. Spahn, E. Mutschler, and W. Morke, Naunyn-Schmiedeberg’s Arch. Phurmacol.
Suppl., 325, R8 (1984).
9. J. Thomson, J. Holtzman, M. Tsuru, C. Lerman, and J. Holtzman, J. of Chromatography, 238,
460 (1982).
10. E. Ariens and A. Simonis, Ann. NY. Acad. Sci., 144, 842 (1967).
11. M. Simonyi, Med. Res. Rev., 4, 359 (1984).
12. P. Jenner and 8.Testa, Drug Metab. Rev., 2(2), 117 (1973).
13. W. Soudijn, Stereochemisty and biological activity of drugs, E. Ariens, W. Soudijn, and P.
Timmermans, Blackwell Scientific Publications, Oxford (1983), p. 89.
14. P. Timmermans, Stereochemisty and biological activity of drugs, E. Ariens, W. Soudijn, and
P. Timmermans, Blackwell Scientific Publications, Oxford (1983), p. 161.
15. P. Patil, D. Miller, and U. Trendelenburg, Pharmacol. Rev., 26, 323 (1975).
16. H. Mosher and J. Morrison, Science, 221, 1013 (1983).
STEREOCHEMISTRY PROBLEMS 465
17. W. Bartmann and E. Winterfeldt, Stereoselective synthesis of natural products. Excerpta Medica,
Amsterdam, 3 (1979).
18. H. Wijnberg, Chem. Eng. News., 58, 24 (1980).
19. S. Wilen, Enuntiomers, racemates and resolutions, Wiley, New York, (1981).
20. H. Kagan and J. Riaud, Topics in Stereochemistry, E. Eliel and N. Allinger, Wiley, New York,
10, 175 (1979).
21. J. Morrison, Asymmetric synthesis, Vol. 1,2,3, J. Morrison and J. Scott, Asymmetric Synthesis.,
Vol. 4., Academic Press, New York, (1983-1984).
22. A. Lakin, G. Tsao, and L. Wingard Jr. (1984) Ann. N Y . Acud. Sci., 434, 78 and 186 (1984).
22a. R. Porter and S. Clark, in Enzymes in organic synthesis, Ciba Foundation symposium 211, Pitman,
London (1985).
22b. J. Tramper, H. van der Plas, and P. Linko (eds.), in Biocatalysts in Organic Synthesis,
Proceedings of an International Symposium held at Noordwijkerhout, The Netherlands, 14-1 7 April
1985, Elsevier Science Publishers B.V., Amsterdam, 1 (1985).
23. P. White, J. Ham, W. Way, and A. Trevor, Anesthesiology, 52, 231 (1980).
24. T. Takada, M. Tada and A. Kiyomoto, Chem. Abstr., 67,72325 (1967).
25. C. Aps and F. Reynolds, Br. J. Clin. Pharmuc. 6, 63 (1978).
26. J. Tobert, V. Cirillo, G. Hitzenberger, I. James, J. Pryor, T. Cook, A. Buntinx, I. Holmes,
and P. Lutterbeck, Clin. Phurmacol. Ther., 29, 344 (1981).
27. W.Hoffman, 0. Woltersdorf, F. Novello, and E. Cragoe, J. Med. Chem., 24, 865 (1981).
28. H. Buch, F. Schneider-Affeld, and W. Rummel, Naunyn-Schmiedeberg's Arch. Phurmacol., 277,
191 (1973).
29. H. Downes, R. Perry, R. Ostlund, and R. Karler, J. Phurmacol. Exp. Ther., 175, 692 (1970).
29a. G. Franckowiak, M. Bechem, M. Schramm, and G. Thomas, Eur. J. Phurmac., 114, 223
(1985).
30. D. Zimmerman and P. Gesellchen, Ann Repts. Med. Chem., 17, 21 (1982).
31. K. Gotestam, Abstr. sut meeting "dextrofenfluramine and bodyweight control", First Int. Meeting
on bodyweight control, Montreux, (1985). Communication: Inst. Recherches Internationales
Servier, Neuilly s. Seine France.
32. V. Lotty and D. Taylor, Eur. J Pharmucol., 85, 211 (1982).
33. E. Ariens, Proceedings of the First International Phurmacological Meeting, Pergamon Press, Ox-
ford, 7, 247 (1963).
34. J. Vite, D. Klimetzek, G. Loskant, R. Hedden, and K. Mori, Natunuissenschuften, 63,582-583
(1976).
35. L. Luckwill and D. Woodcock, The chemistry and mode of action of plant growth substances, R.
Wain and F. Wightman, Buttenvorths Scientific Publications, London, 195 (1956).
35a. E. Ariens, in Drug Design, Academic Press, New York 1, 240, (1971).
35b. E. Ariens, in Molecular Phurmacology, E. Ariens, Ed., 1, 240, Academic Press, New York
(1964).
35c. E. J. Ariens, Arch. Znt. Pharmacodyn. Ther., in press (1986).
36. E. Ariens, Eur J. Clin. Pharmacol., 26, 663 (1984).
37. E. Ariens, Ann. NY. Acad. Sci., 139, 606 (1967).
38. E. Ariens, Drug Design, E. Ariens, Academic Press, New York, 1, 1-270 (1971).
38a. P. Taggart, P. Sutton, and R. Donaldson, Clinical Science, 69, 631 (1985).
38b. R. Richards and A. Tattersfield, Br. J. Clin. Phurmac., 20, 459 (1985).
38c. H. Echizen, B. Vogelgesang, and M. Eichelbaum, Clin. Pharmacol. Ther., 38, 71 (1985).
38d. M. Eichelbaum, G. Mikus, and B. Vogelgesang, Br. J Clin. Phurmac., 17, 453 (1984).
39. L. Low and N. Castagnoli, Annual Reports in Medicinal Chemistry, Academic Press, New
York, 13, 304 (1978).
40. P. Jenner, Concepts in drug metabolism, P. Jenner and B. Testa, Marcel Dekker Inc., New
York, 53 (1980).
41. N. Vermeulen and D. Breimer, Stereochemistry and biological activity of drugs, E. Ariens, W.
Soudijn and P. Timmermans, Blackwell Scientific Publications, Oxford, 33 (1983).
42. C. van Ginneken, J. Rodrigues de Miranda, and A. Beld, Stereochemistry and biological activity
of drugs, E. Ariens, W. Soudijn and P. Timmermans, Blackwell Scientific Publications,
Oxford, 55 (1983).
43. G. Pillai, J. Axelson, C. Kerr, and K. McErlane, Res. Commun. Chem. Pathol. Pharmacol., 43,
209 (1984).
44. E. Schillinger, L. Ehrenberg, and K. Lubke, Biochem. Pharmacol., 27, 651 (1978).
466 ARIENS

45. W. Schmidt and E. Janchen, J . Pharm. Pharmac., 29,266 (1977).

46. U. Kragh-Hansen, Pharmacol. Rev., 33, 17 (1981).
47. W. Bowman and M. Rand, Textbook ofi’harmacology, 2nd ed., BlackwellSdent. Pub]., Oxford,
1.1 and 43.51 (1980).
48. A. Beckett, Ann N Y . Acud. Sci., 144, 675 (1967).
49. G. Reynolds, Br. J. clin. Pharmuc., 6, 543 (1978).
50. B. Akerman and S. Ross, Acta pharmacol. toxicol., 28, 445 (1970).
51. D. Kaiser, G. van Geissen, R. Reisher, and W. Wechter, 1. Pharm. Sci., 65, 269 (1976).
52. A. Hutt and J. Caldwell, 1. Phrm. Pharmacol., 35, 693 (1983).
53. E. Lee, K. Williams, R. Day, G. Graham, and D. Champion, Br. 1. Clin. Pharmac., 19, 669
(1985).
54. P. Dayer, T. Leemann, J. Gut, Th. Kronbach, A. Kupfer, R. Francis, and U. Meyer, Biochem.
Pharmucul., 34, 399 (1985).
55. T. Walle, M. Wilson, U. WaIle, and S. Bai, Drug. Metab. Dispos., 11, 544 (1983).
56. G. Maksay, Z . Tegyey, and L. Otvos, 1. Pharm. Sci., 67, 1208 (1978).
57. G. Yost and B. Finley, Drug. Metab. Dispos., 13, 5 (1985).
58. M. Strolin Benedetti and P. Dosterd, TIPS., 6, 246 (1985).
59. L. Olanoff, T. Walle, U. Walle, T. Cowart, and T. Gaffney, Clin. Pharmacol. Ther., 35, 755
(1984).
60. 8. Silber, N. Holford, and S. Riegelaman, J . Pharm. Sc., 71, 699 (1982).
61. S. Caccia, M. Ballabio, and R. Ponte, Eur. 1. Drug metab. and pharmacokin., 3, 129 (1979).
62. J. McNeil, A. Anderson, and W. Louis, Br. 1. Clin. Pharmacol., 8, Suppl. 2, 157s (1979).
63. I. Wiedemann, H. Peil, H. Justes, S. Adamus, V. Brantl, and H. Lohmann, Drug Research,
35, 964 (1985).
64. J. Wagner, Clin. Pharmacol. and Therap., 37, 481 (1985).
65. H. Elliott, P. Meredith, D. Sumner, and J. Reid, Br. I . clin. Pharmac., 17, 573 (1984).
66. F. Keeley, D. Weiner, and R. Okerholm, Biopharm. Drug. Dispos., 4, 305 (1983).
67. Ch. Sum, A. Yacobi, R. Kartzinel, H. Stampfli, C. Davis, and C. Lai, Clin. Pharmacol. Ther.,
34, 427 (1983).
68. W. Ritschel, Drug Intell. Clin. Pharm., 14, 746 (1980).
69. T. Walle, Drug Metab. and Disp. 13, 279 (1985).
70. B. Foster, TIPS, 5, 524 (1984).
71. T. Baillie, Stable isotopes, McMillan Press Ltd., London, 1 (1978).
72. D. Kritchenski, Deuterium isotope effects in chemistry and biology, Ann. NY. Acad. Sci., 84, 1.
(1960).
73. H. Hazellhoff, J. de Vries, D. Dijkstra, W. de Jong, and A. Horn, Naunyn-Schmiedebergs
Arch. Pharmacol, 330, 50 (1985).
74. R. Allen, in Annual Reports in Medicinal Chemistry, D. Bailey, Ed., 20, 313, Academic Press,
New York (1985).
75. R. Allen, in Annual Reports in Medicinal Chemistry D. Bailey, Ed., 19, 315, Academic Press,
New York (1984).