Note: Descriptions are shown in the official language in which they were submitted.
1 1 62187
PHARMACEUTICALL~ ACTIVE COMPOUNDS
This invention relates to a group of novel com-
pounds, their formulation as pharmaceutical compositions,
and their use in the therapy of disorders.
West German Offenlegungsschrift No: 2,748,260.6 .
describes compounds of the formula (A), and pharma-
ceutically acceptable salts thereof:
. R4
CO NH ~ ( 2)m ~ (CH )
( CH2 ) n /\~U
,~ Rl
R3 ~ (A)
wherein:
is a ~ 6 alkoxy group;
R2 and R3 are the same or different and are hydrogen,
halogen, CF3, hydroxy, Cl 6 alkoxy, C2 7 acyl, amino,
amino substituted by one or two Cl_6 alkyl groups 7 C
acyl amino, aminocarbonyl or aminosulphone optionally
substituted by one or two C1_6 alkyl groups, ~ 6
alkylsulphone or nitro groups;
i....
~.
l ~ 62187
X is either a nitrogen atom, in which case m+n is
3 to 5, m is 2 to 4 and n is 1 to 3; or X is CH in
which case m~n is 2 to 5, m is 1 to 5, and n is 0 to 4;
....... .....p is 0 to 3; and ~ -
R4 is hydrogen, Cl 6 alkyl, phenyl or phenyl-
Cl 6 alkyl, either of which phenyl moiety may be sub-
stituted by Cl 6 alkyl, Cl 6 alkoxy, CF3 or halogen,
and R5 is hydrogen; or R4 and R5 are attached to two
adjacent carbon atoms and form together with these
two carbon atoms a fused benzene ring, which benzene
ring may be substituted by Cl 6 alkyl, Cl 6 alkoxy,
C~3 or halogen; as dopamine antagonists having use
in the treatment of disorders of the gastro-intestinal
function and/or in the treatment of emesis.
A novel class of compounds has now been discovered
which is structurally distinct from the known compounds
of the formula (I) but also has useful pharmacological
activity, such as dopamine antagonist activity.
Accordingly the present invention provides a
compound of the formula (I):
~ /R4
CO-- ( CH2 ) r
( CH2 ) p_ -
1~ ~1 R 1
R3 ~
~2 (I)
7 ~ 6~187
and pharmaceutically acceptable salts and N-oxides
thereof, wherein:
Rl is a Cl_6 alkoxy group;
R2 and R3 are the same or different and are hydrogen,
halogen, CF3, C1_7 acyl, Cl_7 acylamino, Cl 6 alkyl-
S()n wherein n is 0, 1 or 2, nitro, Cl 6 alkoxy, hydroxy,
or amino, aminocarbonyl or aminosulphonyl optionally
substituted by one or two Cl 6 alkyl groups;
or Rl and R2 taken together are methylenedioxy
or ethylenedioxy in which case R3 is any one of the
groups given for Rl and R2 above;
R4 is hydrogen, Cl 4 alkyl or phenyl; or phenyl Cl-4 alkyl;
R5 is hydrogen or Cl 4 alkyl;
X is an oxygen or sulphur atom; or a sulphoxide
group >S~0;
p is l or 2; and
r is 1 or 2.
Within formula (I) there is a group of compounds
wherein Rl, R5, X, p and r as defined, R2 and R3 are the
same or different and are hydrogen, halogen, CF3, Cl 7
acyl, Cl 7 acylamino, Cl 6 alkyl-S(O)n wherein n is 0, 1
or 2, nitro, or amino, aminocarbonyl or aminosulphonyl
optionally substituted by one or two Cl 6 alkyl groups;
and R4 is hydrogen, Cl~ alkyl or phenyl and substitutes the carbon
atom in the ~ ring ad~acent the nitrogen.
Within the group of compounds defined in the pre-
ceding paragraph there is a further group of compounds
wherein the variables Rl, R4, R5, p and r are as defined,
R2 and R3 are as defined except that when alkyl-S()n,
n must be 2 (ie alkylsulphonyl), and X may only be an
oxygen or sulphur atom.
Suitable examples of the group Rl include methoxy,
ethoxy and n- and iso-propoxy. Preferably Rl is a
methoxy group.
Suitable examples of R2 and R3 include the following
atoms and groups: hydrogen; chlorine, bromine; CF3;
~ J 62187
-- 4
formyl, acetyl, propDnyl, n- and lso-butyryl; formylamino,
acetylamino, propionylamino, _- and iso-butyrylamino;
methyl, ethyl and n- and iso-propylsulphone, -sulphinyl or
-thia; nitro; methoxy, ethoxy and _- and iso-propoxy;
hydroxy; amino, aminocarbonyl and aminosulphonyl and amino,
aminocarbonyl, and aminosulphonyl substituted by one or
two methyl, ethyl, n- or iso-propyl groups.
When Rl and ~2 taken together are methylenedioxy or
ethylenedioxy, they are most suitably ethylenedioxy.
Particuarly suitable R2 and R3 groups include
hydrogen, halogen and amino; and as "intermediates", acyl-
amino and nitro, which can conveniently be converted to
the corresponding amino groups.
It is generally preferred that R2 i5 in the 4-position
relative to the carbonyl side chain, for greater activity
in the resultant compound of the formula (I). For the
same reason, it is generally preferred that R3 is in the
5-position relative to the carbonyl side chain.
Particularly preferred R2 groups include 4-amino and
4-acylamino. Most preferably R2 is 4-amino. Particularly
preferred R3 groups include 5-halo, such as 5~chloro.
In other useful compounds R2 is hydrogen, 4-halo (eg
chloro), or amino; and R3 is 5-Cl_6 alkyl S()n (such as
5-methylsulphony]./--s~phinyl or -thia) or 5-optionally
alkylated aminosulphonyl.
R4 may substitute any carbon in the X ring (apart
from the bridgehead carbon), but preferably substitutes the
carbon atom adjacent the nitrogen atom.
Suitable examples of R4 inc~ude hydrogen, methyl, ethyl, n- and
iso-propyl, and phenyl and benzyl. Phenyl groups in R4 may be substituted
by one or more Cl_4 alkoxy (such as methoxy), Cl_4 alkyl (such as methyl),
fluoro, chloro or CF3. Often R4 will be hydrogen, Cl_4 alkyl or phenyl.
~ost suitably R4 is hydrogen or methyL ~ -
Suitable examples of R5 include hydrogen, methyl, ethyl
and _- and iso-propyl. More suitably R5 is hydrogen or
methyl, preferably hydrogen.
Generally X is most suitably oxyaen.
(The X is >S~0 group has been referred to above as the
sulphoxide group. The skilled chemist will appreciate that
in naming a compound containing such a >S~0 group, the
I l 6~187
group can be referred to as "oxothia", and indeed this
terminolo~y has been used in Example 15).
Preferably p is 1.
Preferably r is 1.
It is generally pxeferred for higher activity that
the bond between the benzamide moiety and the cyclic
side chain (i.e. between the R5 substituted nitrogen
atom and the ring carbon atom to which this nitrogen
atom is joined) in the compounds of formula (I) ~s
axial (When used herein in relation to this bond axial
is'assigned with the bicyclic drawn with a trans rinb
fusion, and is also referred to as ~. Equatorial is,
similarly assigned, and is also referred to as ~.)
The pharmaceutically acceptable salts of the com- -
pound of the formula (I) include acid addition salts
with conventional acids such as hydrochloric, hydro-
bromic, phosphoric, sulphuric, citric, tartaric,
lactic and acetic acid and the like.
The pharmaceutically acceptable salts of the com-
pounds of the formula (I) also include quaternary
ammonium salts. Examples of such salts include salts
with compounds such as R6 ~ Y wherein R6 is Cl 6 alkyl,
phenyl - Cl 6 alkyl or C5 7 cycloalkyl, and Y is an
anion of an acid. Suitable examples of R6 include
methyl, ethyl and n- and iso-propyl; and benzyl and
phenyl ethyl. Suitable examples of Y include the
haLides such as chloride, bromine and iodide.
From the aforesaid it will be seen that suitably
the moiety of formula (II): --
C0 - NR5 -
'
R3
~ (II~
R2
l ~ ~2187
in a compound of the formula (I) will have the structure
(III):
.. . . .
CO ~
OCH3
Cl ~
I (III)
~ rom the aforesai.d it will also be appreciated
that preferably the moiety of formula (IV):
- r~7<
(IV)
i~ the compound of formula (I) is of the formula (V):
R~
~--~ 7 (V~
1 1 62~87.
.
- 7 -
Thus a preferred sub-group of compounds within
formula (I) is of formula (V~):
- ~ ~ - R `
/ N ~
CO - NH ~--C~
~ 3
Cl ~ (VI)
In formula (VI), suitably the bond indicated ~-
is axial.
Also in formula tVI) suitably R4 is hydrogen
or methyl~
Another preferred sub-group of compounds within
formula (I) is of formula (VII):
CO- NH ~7
X'
~ OCH3
Cl ~ ,JJ
(VII)
, NH2
wherein X' is S or S~O, preferably S.
In formula (VII), suitably the bond indicated __^
is axial.
I 1 62187
Also in foxmula (VII) suitably R4 is hydrogen
or methyl.
It will of course be realised that the compounds
of ~he formula (I) have chiral centres, -
and thus are capable of existing in a number ofsteroisomeric forms. The invention extends to
each of these stereoisomeric forms, and to mixtures
thereof (including racemates). The different
steroisomeric forms may be separated one from the other
by the usual methods, or any given isomer may be obtained
by stereospecific or asymmetric synthesis.
The invention also provides a process for the
preparation of a compound of the formula (I), which
process comprises reacting an acid of the formula
(VIII):
COOH
R3 ~ Rl
~ (VIII)
R2
or a reactive derivative thereof, with a compound of
the formula (IX):
HR5N ~ ~N ~ 4
SCH2)r,
(CH2)p _ X (IX)
l l 62187
. .
. g
wherein the variable groups are as defined in formula
(I); and thereafter if desired or necessary converting
a group R2, R3 or X in the thus formed compound-to another
group R2, R3 or X respectively.
"Reactive derivative" when used herein means a
derivative of the compound (VIII) which can be reacted
with the compound (IX) to form an amido linkage between
the acid group of the compound (VIII) and the amino
group of acid group of the compound (VIII) and the
amino group of the compound of the formula (IX).
Often this reactive derivative will be the acid
halide, such as the acid chloride, of the acid (VIII).
In such cases the reaction will normally be carried
out in an inert solvent, preferably in the presence of
an acid acceptor. The inert solvent can be any sol~ent
inert to both reactants, such as benzene, toluene,
diethyl ether or the like. The acid acceptor is
suitably an organic base such as a tertiary amine, e.g.
triethylamine, trimethylamine, pyridine or picoline, or
an inorganic acid acceptor, such as calcium carbonate,
sodium carbonate, potassium carbonate or the like.
It should be noted also that it is possible to use
certain acid acceptors as the inert solvent, for
example organic bases.
Another useful reactive derivative of the acid
(VIII) that may be used is an ester, such as a methyl,
ethyl, prop~l or butyl ester, in which case the ~action
is normally carried out by heating the reactants
together in an inert solvent such as ethylene glycol.
.
- 1 162187
-- 10 -- ,
The reaction may also be carried out by forming
an anhydride of ~he acid (VIII) in the usual manner,
and reacting tha* with the compound (IX); normally
a conventional mixed anhydride-will be used; or by
reacting the acid (VIII) and the compound (IX) in the
presence of a dehydrating catalyst such as a carbodi-
imide, for example dicyclohexylcarbodiimide.
It will be realised that in the compound of the
formula (I) the linkage between the -NR5- moiety and the
cyclic side chain may have an or ~ orientation
with respect to the ring of the bicyclic moiety to - ~ -
which it is attached. A mixture of a and ~ isomers
of the compound of the formula (I) may be synthesised
non-stereospecifically and the desired isomer separated
conventionally therefrom, e.g. by chromatography;
or alternatively the a or ~ isomer may if desired be
synthesised from the corresponding a or ~ form of the
compound of the formula (IX).
Alternatively, a mixture of the and ~ isomers
of the compound of the formula (IX) may be synthesised
non-stereospecifically and the desired isomer separated
conventionally therefrom~ e.g. by chromatography.
However, in this case it is generally more convenient
to react the mixture to give a mixture of a and ~
isomers of the compound of the formula (I) and to
separate these if desired as hereinbefore described.
l l 6218~
The intermediates of lormula tIX) may be prepared by
a process which depends on whether the a or ~ forms is
desired, or a mixture thereof.
To prepare a or axial compounds of formula (IX), a
compound of formula (X~`is reduced:
(CH2)~ ~ ~ (CH2)p
N3
(X) . . - .
.
This reduction is conveniently carried out with
lithium aluminium hydride under the usual conditions.
Compounds of formula (X) may themselves be pre-
pared by reaction of a compoùnd of the formula (XI):
X
I~"H
N ~
~` (XI)
OH
to effect conversion of hydroxy to azido, with inversion
of sterochemistry.
This reaction may be carried out in the general
manner described in A.K. Bose et. al., Tetrahedron
~etters 1977, 23, 1977). ~or example, the reaction
may be effected with triphenylphosphine, diethyl
l 1 6~187
- 12 --
azodicarboxylate and diphenylphosphoryl azide.
Compounds of the formula (XI) are important inter-
mediates, as from these compounds the ~ form of compounds
of the formula (IX), and ~/~ mixtures thereof, may be
prepared.
For example, reacti.on of a compound of formula
(XI) with a base and fluorenone yields a compound of
formula (X~I3: - -
X
( CH2 ) ~ ( CH2 ) p _
R ~ N ~
4 l ~ (XII)
~/
The compounds of formula ~XII) may be converted
to compounds of formula (XIII):
(CH2)r (CH2)p
R4
~ NOH (XIII)
with hydroxylamine. We have found that this reaction is
suitably carried out with pyridine and hydroxylamine
hydrochloride at reflux temperatures.
Reduction of a compound of formula (XIII).with
lithium aluminium hydride gives a mixture of axial
~.~) and equatorial (~) amines of formula (IX) where R5 is H.
1 1 6~187
_ 13 -
In constrast, a dissolving metal reduction of a com-
pound of formula (XIII) is substantially ~ereospecific and
yields mainly the ~ amines of formula (IX) (any minor prop-
ortions of the undesired isomer prepared in this process
can be separated in conventional manner). Such reductions
may suitably be carried out with sodium and amyl alcohol.
Returning now to compounds of the formula (XI) 9
these compounds may be prepared from a compound of for-
mula (XIV):
(CH2) ~ ( 2)p
O N ~
¦ ¦ H (XIV)
OCHO
by a process which depends on the nature of R4.
When R4 is hydrogen, the reaction may be carried
out by reduction with for example lithium aluminium
hydride.
When R4 is Cl 4 alkyl or phenyl, and substitutes the
carbon adjacent the nitro~en, the compound of formula
(XIV) is converted with first base, such as sodium hydro-
xide, and second a silylating agent, such as (CH3)3SiCl
or hexamethyldisilazane, to the silyl ether of formula
(XV):
X
( CH2 )( CH 2 ) p
r l~`H
O N ~ (XV)
H
" ~ i(CH3~3
1 1 6218~
_ 14 -
, which in turn is converted to the desired compound of
formula (XI) with a R4 metal complex, such as R4Li, and
subsequent reductionj for exampl.e with sodium borohydride.
The intermediates of formula (XIV) may themselves ~-
be prepared by treating a compound of formula (XVI):
X i.
(CH2)ri ( 2)p
O ~ N l OH
~ (XVI)
with formic acid.
Compounds of formula (XVI) may be prepared by
reduction, for example with sodium borohydride and
hydrogen chloride, of a compound of formula (XVII):
(CH2)r ~CH2)p
O ~ N ~ O
( ~ II)
, which compounds of formula (XVII) may themselves be
prepared by ~reaction of a compound of formula (XVIII):
X
(CH2)r - (C 2)p
H (XVIII)
with an alkylene halide, such as 4-brombut-l-ene,
using base.
1 162187
- 14a -
To obtain a compound of the formula (XI) wherein
R4 is Cl_4 alkyl or pheny~ or phenyl Cl 4 alkyl and substitutes the
X ring at the carbon other than the carbon adJacent the nitrogen,
a compound of formula (XIV)' is reduced, for example
with lithium aluminium hydride:
(CH2) ~ 2)p
4 ~ N ~
H (XIV)'
OCHO
(wherein R4 is Cl_4 alkyl or phenyl).
Such compounds of the fcrmula (XIV)' may be
prepared by the (XVIII) -~ (XVII) to (XVI)- (XIV) route
as described for compounds of formula (XIV), except
that the R4 substituent is present in each of the
compounds (XVIII), (XVII) and (XVI) to correspond to
the R4 in compounds (XIV)'. (It will of course be
appreciated that in such a reaction sequence the
(XVII) to (XVI) reduction will give a mixture of
isomers which can be separated by conventional means.)
1 J 621~7
_ 15 -
The aforesaid processes of course yield a compound
of the formula (IX) wherein R5 is hydrogen.
If a compound of formula (IX) wherein R5 is alkyl
is desired, it may simply be prepared from the-corres- ;
ponding R5 is hydrogen compound by alkylation. This
may be effected in any suitable manner, for example by
acylation with an anhydride followed by reduction with
lithium aluminium hydride.
It will be appreciated that any enantio~er, or
10 mixture of enantiomers, of the compounds of formulae . ~
(X), (XI), (XIV) and (XV) may be used provided it has
the depicted spatial relationship between carbons a and
b.
The intermediates of the formula (VIII) are known
compounds, or may be prepared in analogous manner.
The N-oxides of the compounds of formula (I) may be
prepared in conventional manner, for example by treatment
with a per-acid, such as m-chloroperbenzoic acid.
~ - ~ ~
I 1 ~21~
.
- - 16 - -
' The acid addition salts of compounds of the formula
(I) may be prepared in entirely conventional manner
, by reacting a comp~und of,the formula (I) in~bas,e form
with the chosen acid.
The quaternary ammonium s~lts of the'compounds
of the formula (I) may be prepared in conventional
manner for such salts, such as by reaction of the chosen
compound of the formula (I) with a compound R6Y as
defined. This reaction is suit~bly carried out in an
appropriate solvent such as acetone, methanol, ethanol 9 ' ' -
dimethylformamide and the like, at ambient or raised
temperature and pressure.
The interconversion of suitable groups R2 and R3
after formation of a compound of the formula (I)
or corresponding intermediate therefor may be carried
: out by conventional methods. By way of example nitro
groups may be reduced to amino groups in the normal
manner, and acylami~o groups may be converted to amino
groups also by conven~ional methods. Also a compound
of the formula (I) wherein R2 or R3 is halogen can be
prepared by a conventional halogenation of the cor-
responding compound of the fQrmula (I) wherein the
said R2 or R3 is hydrogen. Further, compounds wherein
R2 or R3 are Cl 6 alkylsulphonyl may be prepared by
oxidation,~of the corre~ ding Cl_6 alkyl sulphinyl ~ounds;
and su¢h Cl_6 alky1sulphinyl compounds may be
prepared by oxidation of~the corresponding Cl_~
alkylthia compound. In regard to X, ~ is S comp~unds
may be oxidised to X is S~0 compounds, suitably
1 1 62187
with sodium periodate. Accordingly it will be recalled that
compounds of the formula (I) containing a R2, R3 or X group
which is convertible to another R2, R3 or X group are useful
intermediates, and as such form an important aspect of the
invention .
As hereinbefore stated, the compounds of the formula
(I) are dopamine antagonists.
The co~pounds of the formula (I) may be used in the
treatment of disorders related to impaired gastro-intestinal
motility, such as retarded gastric emptying, dyspepsia, flatu-
lence, oesophagal reflux, peptic ulcer and emesis.
The invention therefore also provides a pharmaceutical
composition comprising a compound of the formula (I), or a
pharmaceutically acceptable salt thereof, together with a
pharmaceutically acceptable carrier.
Such compositions may be adapted for oral or parenteral
administration, and as such may be in the form of tablets,
capsules, oral liquid preparations, powders, granules, lozenges,
reconsitutable powders, injectable and infusable solutions or
suspensions and the like; the compositions may also be in the
form of suppositories and the like. Normally, orally admini-
strable compositions are preferred.
- 17 -
l 1 6~187
_18 _ -
,
.
Tablets and capsules for oral administration may
be in unit dose presentation form~ and may contain
conventional excipients such as binding agents, fillers,
tabletting lubricants, disintegrants, and acceptable
-wetting agents and the like. ~The tablets-may be coated
according to methods well known in normal pharmaceutical
practice. Oral liquid preparations may be in the form
of, for e~ample, aqueous or oily suspensions, solutions,
emulsions, syrups, or elixirs, or may be presented
in a dry product for reconstitution with water or
other suitable vehicle before use. Such liquid pre~
parations may contain conventional additives such as ~ -
suspending agents, emulsifying agents, non-aqueous
vehicles (which may include edible oils), preserva-
tives, and if desired conventional flavouring or
colouring agents, and the like.
~or parenteral administration, fluid unit dosage
\ forms are prepared utilizing the compound of the formula
(I)` and a sterile vehicle. The compound, depending
on the vehicle and concentration used, can be either
suspended or dissolved in the vehicle. In preparing
solutions the compound can be dissolved for injection
and filter sterilized before filling into a suitable
vial or ampoule and seaiing. ~dvantageously,
adjuvants such as a local anaesthetic, preservatives
and buffering agents can be dissolved in the vehicle.
Parenteral suspensions are prepared in substantially
the same manner except that the compound is suspended --
in the vehicle instead of being dissolved and sterilized
by exposure to ethylene oxide before suspending in
the sterile vehicle. Advantageously, a surfactant
or wetting agent is included in the composition to
'- . I J ~218.7
- - 19 -
,
facilitate uni~orm distribution of the compound.
As is common practice, the compositions will
usually be accompanied by written or printed directions
. fo~..use in *he medical treatme~nt concerned...~.;...`
It will of course be realised that the precise
dosage used in the treatment of any of the hereinbefore
described disorders will depend on the actual co~pound
of the ~ormula (I) used, and also on other factors
such as the seriousness of the disorder being treated.
The invention further provides a method of treat-
ment of maladies in humans comprising the administration - -
of an effective amount bf.a compound of the formula
(I) or a pharmaceutically acceptable salt thereof.
The "effective amount" will depend in the usual way on
a number of factors such as the nature and severity
of the malady to be treated, the weight of the sufferex,
and the actual compound.used.
However by way of illustration, unit doses will
suitably contain 0.1 to 20 mgs of the compound of
formula (I), for example O.S to 10 mgs.
~gain by way of illustration, such unit doses
will suitably be administered more than once a qay, for
example 2, 3, 4, 5 or 6 timeQ a day, in such a way
that the total daily dose is suitably in the range 0.01
2S to 10 mg/kg per day.
Compounds of the formula (I) have the ability to
potentiate the effect of conventional analgesics -
in migraine treatment when administered con~urrently
with the analgesic.
.
I ~ ~2187
. . .
, _ 20 _
, ' . - . ~ .
Thus the invention provides a pharmaceutical '
composition comprising a compound of the formula (I)
and an analgesic~
The compound of the formula (I) and the analgesic,
~suc~ as aspir~in or paracetamol~,-'~iIl'be presènt 'in the-
composition in amounts generally similar to their usual i'
effective dose.
The composition can be a combination product; for
' example a tablet or capsule containing both a compound
of the formula (I) and an analgesic for oral administration,
or a twin pack comprising the two active ingredients _
made up for separate administration.
The invention accordingly provides a method of
treatment of migraine comprising the adminîstration
to the sufferer of a compound of the formula (I) and
an analgesic.
The following Examples illustrate the pre~aration
of the compounds of formula (I) and the following
Descriptions illustrate the preparation of ~intermediates
thereto.
In the Examples, ('-) of course indicates that the
compound is a racemate. However, for the sake of con-
venience, the structural formula of only one enantiomer
has been shown.
. .
1 8 ~
- 21 -
.
- .
Description 1
(a) -(4-But-l-envl)morpholine-2~-dione (D.l~
O
I . i,
0~ ~N'~ O
¦ (D.l)
To a stirred solution of morpholine-2,6-dione (11.6 9)
in dry DMF (lOO ml) was added 80% sodium hydride (3.4 9).
On cessation of the gas evolution (ca. ~ hour), 4-bromobut-
l-ene (13.6 9) was added and the whole refluxed for 2
hours. On cooling, the reaction mixture was poured into
ice/water (300 n~ and extracted with ether (3 x 200 ml)
and ethyl acetate (20Q ml).
Fractional distillation ~afforded the 1-(4-but-1-
enyl)morpholine-2,~6-dione (D.l) (10.09, 60%).
b.p. 84-8/0.1 mm
n.m.r. (~, CDC13): 6.05-5.35 (m, lHI H2C=CH-CH2);
5.2-4.75 (m, 2H, H2C=CH-);
4.25 (S9 4H, 0(CH2CO)2N-);
3-73 (t, 2H, ~N-CH2-CH2_; J=7Hz);
2.5-2 (m, 2H, -CH2-CH2-CH=).
.
: . ^
1 ~162187
.
- 22 - -
.
.
(b) (-) l-aza 8~-For~ylox ~ 4-oxab - icvclor4?4 .Oldecane-
2-one (D.2)
- _ ~ ^ ~ N
HC0 ~
(D.2)
The 1-(4-but-1-enyl)morpholine-2,6-dione (D.l)
(5 9) was reduced with sodium borohydride (10.8 9)
in absolute ethanol (300 ml) at -15C for 5 hours, durin~
which time 3-5 drops of ethanolic hydrogen chloride
(4.5N) was added at 15 mimlte intervals.
The excess sodium borohydride was destroyed with
excess ethanolic hydrogen chloride (to pH 3) and the
ethanol removed by rotary evaporation. Addition of
saturated aqueous sodium bicarbonate solution and
extraction with methylene chioride gave the crude 1-
(4-but-1-enyl)-2-hydroxymorpholine-6-one (6.0 9).
The crude 1-(4-but-1-enyl)-2-hydroxymorpholine-
6-one was dissolved in 98% formic acid and the solution
stored overnight at ambient temperatures. The formic
acid was removed by rotary evaporation and the residue
taken up in methylene chloride, washed with aqueous
sodium bicarbonate solution and dried (MgS04). Removal
of the solvent gave the crude (-) l-aza-8-formyloxy~-H-4-
oxa bicyclo[4,4,0]decane-2-one (D.2) (5.6 g, ca. 100%).
n.m.r. (o. CDC13) 7.98 (s, lH, 02CH);
5.7-3.2 (m, 7H including 4.1;
s, 2H, -OCH2C-);
3.0-1.0 (m, 5H, remaining protons).
l 1 6~187
(c) (-? l-aza-8~-Hydr'oxy-6~-H-4-oxabicycloE4~4~o3decane
` (D '3') intermedia'te 'for'Compoun'd's l'and`2
/ N
HO ~ O
H
(D.3)
The crude (+) l-aza-8g-formyloxy-6~-H-4-oxabicyclo
[4,4,0] decane-2-one (D.2) (2.8 g) was reduced with lithium
aluminium hydride (0.8 g) in ether. Hydrolysis, filtration and
distillation afforded the (-) l-aza-8~-hydroxy-6~-H-4-oxabicyclo
[4,4,0] decane (D.3) (1.5 y, 65~ from D.l).
b.p. 85-96/0.1 mm.
n.m.r. (&, CDC13): 4.2-0.9 (m, 15 H, all protons).
(d) (-) l-aza-8a-Azido-6~-H-4-oxabicyclo [4,4, o3 decane (D.4)
intermediate for Compounds 1 and 2
/ ~~ N --~
~ ~ ~
N3 H
(D.4)
To a dry, stirred THF (30 ml) solution of triphenyl-
phosphine (2.75 g) and diethyl azodicarboxylate (1.9 g~ at 0C
was added (+) l-aza-8B-hydroxy-6~-H-4-oxabicyclo
- 23 -
._~
~ J ~2187
~4,4,0~ decane (D.3) (1.5 g). After stirring for 10 mins. di-
phenylphosphoryl azide (2.9 g) in dry THF (10 ml) was added
during 10 mins. and the mixture stirred at ambient temperature
for 3 days.
The THF was removed and replaced by methylene chloride.
The product was extracted into 5N hydrochloric acid (10 ml) and
washed with methylene chloride (3 x 50 ml). Basifieation and
saturation with potassium carbonate, re-extraetion with methylene
ehloride and removal of the solvent gave the crude ( ) l-aza-8~-
azido-6~-H-4-oxabieyclo[4,4,0~deeane (D.4) (1.6 g, 90~).
i.r. (fil ) (N3)
(e) (-) 8¢-Amino-l-aza-6a-H-4-oxabieyelo~4,4, oJ deeane (D.5)
intermediate_for Compounds l and 2
/ N
NH2 H
(D.5)
The crude (+) l-aza-8~-azido-6~-H-4-oxabicyclo[4,4,0]
deeane (D.4) (1.6 g) was redueed with lithi.um aluminium hydride
(0.3 g) in ether for 4 hours. Hydrolysis, extraetion with
methylene ehloride and solvent removal afforded the erude (-)
8~-amino-1-aza-6~-H-4-oxabieyelo [4,4,0~deeane (D.5) (1.5 g, ea.
100%).
- 24 -
l l 62187
Description 2
(a) (-) 'l-Az'a'-'6'~-H-'4-oxab'i'c'clo~4','4','03de'ca'n'è'-'8-one (D.6)
y
intermediate for Compounds 3 and 4
/--____ N--~
O ~--O
(D.6)
; To a stirred solution of (+~ l-aza-8~-hydroxy-6~-H-4-
oxabicyclo[4,4,0~decane (D.3) (2.8 g) and fluorenone (17 g) in
dry benzene (70 ml) was added potassium tert. butoxide (5.6 g)
and the solution stirred at room temperature for 30 mins. The
product was extracted into SN hydrochloric acid (25 ml), and
washed with methylene chloride (50 ml). The aqueous phase was
neutralised then saturated with potassium carbonate and extracted
with methylene chloride (3 x 100 ml). Removal of the solvent
gave the crude (+) l-aza-6~-H-4-oxabicyclot4,4,0~decane-8-one
(D.6) (3.0 g, ca 100~).
(b) (+) 1-Aza-6~-H-4-oxabicyclo L4,4, oJ decane-8-one oxime
-
tD.7) intermediate for Compounds 3 and 4
~ -N _-~
HON ~ 1/ ~
H
(D.7)
~1
- 25 -
l 1 6218~
An ethanolic (30 ml) solution of (+)1-aza-6~-H-4-oxa-
bicyclo[4,4,0]decane-8-one (D.6) (3.0 g) was treated with pyridine
(3 ml) and hydroxylamine hydrochloride (3 gm) and the mixture
heated under reflux for 30 mins. On cooling, the ethanol was
removed by rotary evaporation and the residue treated with dilute
potassium carbonate solution. The product was extracted with
methylene chloride and dried (K2CO3). Removal of the solvent
afforded the cru~e (-) l-aza-6~-H-4-oxabicyclo [4,4,0~decane-8-one
oxime (D.7) (3.0 g, ca 95%).
(c) (-) 8 -Amino-l-aza-6~-H-4-oxabicycloC4,4,0~ decane (D.8)
intermediate for Compounds 3 and 4
/ N
H2~
H
(D.8)
To a refluxing amyl alcohol (100 ml) solution of the
crude (-) l-aza-6~-H-4-oxabicyclo[4,4,0] decane-8-one oxime (D.7)
(3.0 g) was added sodium tca 3.0 g) portionwise over 1 hour. The
cooled reaction mixture was then treated with 5N hydrochloric
acid (ca 80 ml) and extracted with ethyl acetate. The acidic
aqueous layer was separated, neutralised and saturated with
potassium carbonate and re-extracted with methylene chloride
(4 x 150 ml). Distillation of the organic extracts gave the
(-) 8~-amino-1-aza-6~-H-4-oxabicyclo~4,4,0~decane (D.8) (1.5 g,
50~)-
b.p. 58-62/0.1mm.
~ - 26 -
~ J ~2187
Description 3
(a) (-) l-Aza-8~-hyaroxy-6~-H-4-oxabicyclo[4,4,03decane-
2-one (~.9) intermedi-ate for Compounds 5 and 6
~/~
HO ~ / O
H
(D.9)
To an ethanolic solution (50 ml) of (-) 1-aza-8~-formyl-
oxy-6~-H-4-oxabicyclo~4,4,0]decane-2-one (D.2) (7.0 g) was added
aqueous 2.5N sodium hydroxide solution (14 ml) and the mixture
stood for 10 mins. at room temperature. The ethanol was removed
by rotary evaporation and the residue extracted with methylene
chloride. Distillation of the organic extracts afforded the (-)
l-aza-8~-hydroxy-6~-H-4-oxabicyclo[4,4,0]decane-2-one (D.9) (4.1 g,
68%).
b.p. 80-90/O.lmm.
(b) (-) l-Aza-8~-hydroxy-2~-methyl-6~-H-4-oxabicyclo~4,4,0
decane (D.10) intermediate for Compounds 5 and 6
HO_ ~ N
~~0
H (D.10)
- 27 -
I J 62187
An immiscible mixture of (~ aza-8~-hydroxy-6~-H-4-
oxabicyclo~4,4,0]decane-2-one (D.9) (2.3 g) and hexamethyldi-
silazane (4 ml) was treated with 2 drops of conc. sulphuric acid
and subsequently heated to 80 until a single phase was obtained
(ca. ~ hour). On cooling, petroleum ether (40-60) (200 ml) was
added and the solution washed with aqueous sodium bicarbonate
(20 ml) and dried (MgSO4). Filtration and removal of the
petroleum ether gave the crude trimethyl silyl ether (3.0 g).
To a cooled (-30) stirred solution of the trimethyl
silyl ether in dry ether (50 ml) was added a 0.6M solution of
methyl lithium (40 ml) with subse~uent stirring at room tempera-
ture for 2 hours. Saturated sodium bicarbonate solution (20 ml)
was added, the aqueous layer separated and extracted with methyl-
ene chloride (3 x 50 ml). The combined organic extracts were
dried (K2CO3), filtered and concentrated.
Ice-cold ethanol (50 ml) was added followed by 4.5M
ethanolic hydrogen chloride (5 ml) and subsequently stirred at
0C with sodium borohydride (3.0 g) for 2 hours.
The ethanol was removed by rotary evaporation and the
residue extracted with methylene chloride (3 x 100 ml). Distil-
lation of the organic extracts gave the (+) l-aza-8~-hydroxy-2~-
methyl-6~-H-4-oxabicyclo~4,4,0~decane (D.10) (1.2 g, 55%).
b.p. 96/0.1 mm
n.m.r. (~, CC14): 4.2-0.87 (m, 17H, includes
0.92, d, 3H,> CHCH3 J-6Hz).
- 28 -
l 3 621~7
(c) (+) l-Aza-8~-az'ido'-2~-methyl-6q-H'-4-'oXabiCyclor4r4
decane '(D.~l'l)''in'te'r _diate' for Compounds 5~ahd 6
~ J~/
1~ o
N3 H
(D.ll)
Following the procedures outlined in Description 1, the
~ aza- ~-hydroxy-?.~-methyl-6~-H-4-oxabicyclo ~4,4,0~ decane
tD.10) (1.2 g) was converted to (-)-1-aza-8~-azido-2~-methyl-6~-
H-4-oxabicyclo~4,4,0~decane (D.ll). (1.05 g, 75~).
i-r. (film) ~(N ): 2090 cm 1.
(d) (+)8 -Amino-l-aza-2~-methyl-6~-H-4-oxabicyclo~4,4,0]
decane '(D.12) intermediate for Compounds 5 and 6
/ N CH3
NH2 H
(D.12)
Following the procedures outlined in Description 1, the
(-) l-aza-8~-azido-2~-methyl-6~-H-4-oxabicyclo E 4,0~decane
(D.ll) (1.05 g) was converted to (+) 8~-amino-1-aza-2~-methyl-6~-
H-4-oxabicyclo~4,4,0~ decane (D.12) (0.9 g, 100~).
29 -
,~i
18~
.
.
_ 30 -
.
, . ,. - ,
- Descri~tion 4
(a) 1-(4-But-l-envl~thiomorpholine-2.6-dione ~D.13) _
I I i
0 ~ N 0
- 1 '
~ (D.13)
Following the procedures outlined in Description
1, the thiomorpholine-2,6-dione (18.8 9) was converted
into l-(4-but-1-enyl)thiomorpholine-2,6-dione (D.13)
b.p. 106/0.4 mm (16 9, 60%).
_ 5 n.m.r. (~, CD~13): 6.1-5.35 (m, lH, H2C=CH-CH2-);
5.2-4.75 (m, 2H, H2C=CH-);
3.83 (t, 2H, J 7Hz, ~ NCH2-);
3.48 (s, 4H, S(C_2C0)2N-);
2.5-2 (m, 2H, -CH2-CH2CH=);
-6a-H
(b)` (+~ l-Aza-8~-formvlox ~ 4-thiabicyclor4,4,01decane-
2-one (D.14)
O - ~
Ho =
~ S
H (D.14)
.
l ~ 6~187
Following the procedures outlined in Description 1,
the 1-(4-but 1-enyl)thiomorpholine-2,6-dione (D.13) (7.2 g) was
converted into crude (+) l-aza-8~-formloxy-6~-H-4-thiabicyclo
[4,4,0]decane-2-one (D.14) (6.6 g, 79%).
n.m.r. (~ , CDC13): 7.98 (s, lH, OOCH);
5.7-1.2 (m, 12H including
3.3, s, 2H, -SCH2~-).
tc~ (+)1-Aza-8~-hydroxy-6~-H-4-thiabicyclor4,4,03decane
(D.15)
/ N
HO ~
(D.15)
Following the procedures outlined in Description 1, the
(-) l-aza-8~-formyloxy-6~-H-4-thiabicyclo[4,4,0~decane-2-one
(D.14) ~9 g) was converted into (+) 1-aza-8~-hydroxy-6~-H-4-
thiabicyclo ~4,4,03decane (D.15) (5.8 g, 80%).
b.p. 120/0.2 mm m.p. 130-3.
- 31 -
I J 62187
(d) (-? l-Az'a'-8~,~-'az'i'do-6~-H-4'-th'iabicyclo~4,'4, o3 decane
(D.16'~ intermed'iate for Compo'und's 7 and 8
-
/--~N
N ~ ~ ~
Following the procedures outlined in Description 1, the
(-) l-aza-8~-hydroxy-6~-H-4-thiabicyclo[4,4,0]decane (D.153
(2.5 g) was converted into a crude isomeric mixture of (+) l-aza-
8~-, and 1-aza-8~-azido-6~-H-4-thiabicyclo~4,4,0]decane (D.16)
(2.8 g, 95%).
i.r. (film) ~ (N3) 2090 cm 1
(e) (+) 8~, ~Amino-l-aza-6~-H-4-thiabicycloC4,4,0~decane
(D.17) intermediate for ComPounds 7 and 8
-
2 ~ \ ~
(D.17)
Following the procedures outlined in Description 1, the
(-) l-aza-8~, ~-azido-6~-H-4-thiabicyclo[4,4,03decane (D.16) was
converted into a crude isomeric mixture of (+) 8~, ~-amino-l-aza-
6~-H-4-thiabicyclo~4,4,0~decane (D~17) (2.4 g, ca 100%).
l l 621~7
Description 5
(a) (-) l-Aza-6Q~H-4-thiabicyclo[4,4,0~decane-~8-one (D.18)
ihtermediate for Compounds 9, 10, 19, 20 and 21
~/~~~ ~
O ~ S
(D.18)
Following the procedures outlined in Description 2, the
(+-) l-aza-8~-hydroxy-6~-H-4-thiabicyclo~4,4,0~decane (D.lS)
(prepared as in Description 15) (3.1 g) was converted into (-)
l-~za-6~-H-4-thiabicyclo[4,4,0]decane-8~one (D.18) (2.6 g, 80%).
(b) (+-) l-Aza-6~-H-4-thiabicyclo~4,4,03 decane-8-one oxime
(D.l9) intermediate for Compound 9 and 10
HON ~ ~ ~ ~
--S
(D.l9)
Following the procedures outlined in Description 2, the
(-) l-aza-6~-H-4-thiabicyclo[4,4,0~decane-8-one (D.18) (2.6 g)
was converted into the crude (-+) l-aza-6~-H-4-thiabicyclo ~,4,0]
decane-8-one oxime (D.l9) (2.8 g, ca 100~), used without purifi-
cation.
~ 33 -
. ~
l l 62187
(c) (+) 8~-Ami'no-l-az'a-'6~-H-4-thiab'icyclo'r4,4, o3 decane
(D.'2'0) in'termediat'e for Compounds 9 and 10
H2N-~N ~?
H
(D.20)
Following the procedures in Description 2, the (-)
1-aza-6~-H-4-thiabicyclo ~4,4,0]decane-8-one oxime (D.l9) (2.8 g)
was converted into the crude (~) 8~-amino-1-aza-6~-H-4-thia-
bicyclo~4,4,0~decane (D.20) (2.5 g, ca 100~).
? ~ - 34 -
9 1 62187
- 35 -~
Description 6
(a~ Aza-8~-HYdroxY-2~-phenyl-6a-H-4-oxabic~clo
[4,4,0]decane_(D.21) intexmediate for Com~ounds
11, 12, 13 and 14
Ph
H ~-
H (D.21)
To a cooled (0C) stirred solu~ion of bromobenzene
(6.2 g) in dry ether (50 ml) was added a 1.6M solution
of n-butyl lithium in hexane (25 ml) and the solution
was stirred at 0C for 30 minutes. On further cooling
to -30C, a solution of (~) l-aza-8~-trimethylsiloxy-
6a-H-4-oxabicyclo[4,4,0]decane-2-one (4.3 g crude)
(see Description 3(b)) in dry ether (25 ml) was added
over 5 minutes and the resu~ing solution stirred
to room temperature over 2 hours, during which time
a white precipitate formed. The stirred suspension
was then cooled to 0C and 25 ml of a saturated
aqueous potassium carbonate solution (25 ml), followed
by methylene chloride (100 ml) was added and the organic
upper layer separated, dried (K2C03), filtered and
concentrated in vacuo.
~16~87
- 36 -
Ice-cold ethanol ~100 ml) was then added followed
by 5N ethanolic hydrogen chloride (3.6 ml) and
subsequently stirred at 0 C with sodium borohydride
(4 g) for 2 hours.
The ethanol was removed by rotary evaporation
and the residue extracted with methylene chloride
(3 x 100 ml). Column chromatorgraphy (silica, ether
eluent) gave the (') l-aza-8~-hydroxy-2~-phenyl-6a-
H-4-oxabicyclo[4,4,0~ecane (D.21) as an oil (1.6 g,
24%).
(b) (') l-Aza-8a-azido-2~-phenyl-6a-H-4-oxabicyclo
[4,4,0]decane (D.22) intermediate for Compounds
11 and 12
-
N3
(D.22)
~ ollowing the procedures outlined in Description
1, the (i) 1-aza-8~-hydroxy-2~-phenyl-6a-H-oxabicyclo
[4,4p]decane (D.21) (1.6 g) was converted to (i) -1-
aza-8~-azido-2~-phenyl-6a-H-4-oxabicyclo[4,4,0]decane
(D. 22) (1.4 g, 75%).
1 8 7
- 37 -
I.r. tfilm) V(N3) : 2090 cm
N.m.r. (~, CC14) : 7.6-7.0 (m, 5H, aromatic
P C-H~
4.2-1.1 (m, 13H, remaining
C-H)-
(c) (') 8~-Amino-l-aza-2~-phenyl-6a-H-4-oxabicyclo
[4,4,0~decane (D.23) intermediate for ComPounds
11 and 1_
Ph
(D.23)
Following the procedures outlined in Description
1, the (~) 1-aza-8a-azido-2~-phenyl-6a-H-4-oxabicyclo
[4,4,0]decane (D.22) (1.4 g) was converted to (i) 8a-
amino-l-aza-2~-phenyl-6a-H-4-oxabicyclo[4,4,0]decane
(D~23) (1.3 g, 100%).
` ~ ~ 6~187
- 38 -
Description 7
(a) ~ Aza-2~-phenyl-6~-H-4-oxabicyclo[4,4,0]
decane-8-one (D. 24) intermediate for Compounds
13 and 14
Ph
N
~ ~ _O
H
(D.24)
Following the procedures outlined in Description
2(a), the (') 1-aza-8~-hydroxy-2~-phenyl 6a-H-oxabicyclo
[4,4,0]decane (D.21) (2.6 g) was converted to (~) 1-
aza-2~-phenyl-6a-H-4-oxabicyclo[4,4,0]decane-8-one
(D.24) (2.4 g, 92%).
(b) (~) l-Aza-2~-phenyl-6~-H-4-oxabicyclo[4,4,0]decane
-8-one oxime (D.25) intermediate for Compounds
13 and 14
Ph
HON
H
(D.25)
~ ~æl~7
- 39 -
Following the procedures outlined in Description
2(b), the (i) 1-aza-2~-phenyl-6~-H-4-oxabicyclo[4,4,0]
decane-8-one (D.24) (2.4 g) was converted to (~) l-aza-
2~-phenyl-6a-H-4-oxabicyclo[4,4,0]decane-8-one oxime
(D.25) purified by column chromatography (silica,
ether eluent) (2.3 g, 90%).
(c) (~) 8 ~Amino-l-aza-2~-phenyl-6~H~-oxabicyclo
[4,4,0]decane (D.26) intermediate for_Compounds
13 and 14
-
Ph
/ ~
H N
2 1 0
H (D.26)
Following the procedures outlined in Description
2(c), the (*) 1-aæa-2~-phenyl-6~-H-4-oxabicyclo
[4,4,0]decane-8-one oxime (D.25) (2.3 g) was converted
to (~) 8~-amino-1-aza-2~-phenyl-6~-H-4-oxabicyclo
[4,4,0]decane (D.26) (2.1 g, 95%).
1 ~ 62187
Example 1
(-~ 4-Acetamido-5-chloro-2-methoxy-N-('8~-1-aza-6~-H'-'4-
oxabieyelo G 4,01de'eyl)benzamide (1
~ -N
CONHl ~ ~ ~ O
~ OCH3
C1
NHCOCH3 (1)
To 4-acetamido-5-chloro-2-methoxybenzoyl chloride (3.4 g)
in toluene (150 ml) and triethylamine (3 ml) was added the crude
(-) 8G~amino-l-aza-6~-H-4-oxabicyelo L4,4,0~ decane (D.5) (1.5 g) in
toluene (20 ml). The reaetion mixture was stirred at room tempera-
ture for 4 hours, then treated with 2.SN aqueous sodium hydroxide
(10 ml). The toluene layer was separated, the aqueous layer
extracted with chloroform (3 x 100 ml) and the combined organie
extraets dried (K2CO3). The solvent was removed and ehromatography
of the produet (neutral alumina, Broekman II, ethylaeetate) gave
the (+) 4-aeetamido-5-ehloro-2-methoxy-N-(8~-1-aza-6~-H-4-oxa-
bicyelo [4,4,0~deeyl)benzamide (1) m.p. 189-92 (2.9 g, 80%).
n.m.r. (~ , CDC13): 8.27 (s, lH, aryl _);
8.12 (5 , lE~, aryl El);
8.2-7.2 (m, 2H, -CON_-CH~ ,
CH2CONH-Ar);
5.65-4.2 (m, lH, 8 -H);
4.].5-3 (m, 7H including 4.0 s, 3H,
-OCH3);
3-1.2 (m, 12H = CH2 ineluding
2.27, s, 3H, -COC_3).
- 40 -
'~
l J 62187
Example 2
( ) 4-Amino-5-chloro-2-methoxy-N-(8~-1-aza-6~-H-4-oxa bicyclo
L4,4,0~decyl)benzamide` (2)
~ N-
r~ O
CONH H
OCH3
J~ I
Cl ~
NH2
(2)
(1) (2.9 g) prepared as in Example 1) was refluxed with
an aqueous ethanol (water 5 ml, ethanol 50 ml) solution of
potassium hydroxide (1 g) for 2 hours. The mixture was then cooled
to room temperature and the ethanol removed by rotary evaporation.
Trituration with water gave a white solid which was collected and
recrystallised from ethylacetate yielding the (+) 4-amino-5-chloro-
2-methoxy-N-(8~-1-aza-6~-H-4-oxa-bicyclo[4,4,0]decyl)benzamide
(2) (1.35 g, 50%). m.p. = 190.
n.m.r. (~, CDC13): 8.3-7.8 (m, lH, ArCON~-C_=);
8.03 (s, lH, Acryl 6-_);
6.27 (s, lH, Acyl 3-_);
4.7-1.3 (m, 19H, includes
3.9, s, 3H, -OC_3).
- 41 -
I 1 62187
Example 3
(-) 4-Acetamido-5-chlo'ro-2-methoxy-N-(8~ aza-'6~-H-'4'-oxabicyclo
[4!4,0~decy1)benzami'de (3)
/ N
CONH ~ ~
OCH3
NHCOCH3
(3)
Following the procedures outlined in Example 1, the (-)
8~-amino-1-aza-6~-H-4-oxabicyclo[4,4,0] decane (D.8) (1.5 g) was
converted to (-) 4-acetamido-5~chloro-2-methoxy-N-(8~-1-aza-6~-H-
4-oxabicyclo t4,4,0]decyl)benzamide (3) (2.6 g, 70%) m.p. 188-9.
n.m.r. (S, CDC13): 8.18 (s, lH, aryl-_);
8.05 (s, lH, aryl-_);
7.9-7.4 (m, 2H, -CONHCH~ ,
CH3CONH-aryl);
4.2-0.8 (m, 20H, remaining H
including 3.87, s, 3H, OCH3 and
2.23, s, 3H, -COCH3).
~ - 42 -
, j
l 1 ~2187
Exam~e 4
(-) 4-Amino-~5-chloro-2-methoxy-N-(8 ~ aza-6~-H-4-oxabicyclo
C4,4,~ decyl)benzamide (4)
CONH~-- N\ /
~ OCH3 H
Cl J ~ J
NH2
(4)
(3) (2.6 g) (~repared as in Examplé 3) was refluxed with
an aqueous ethanol (water 5 ml, ethanol 50 ml) solution of
potassium hydroxide (1 g) for 2 hours. On cooling, the ethanol
was removed by rotary evaporation and the residue extracted with
chloroform (2 x 50 ml) and dried (K2CO3). Filtration and removal
of the solvent afforded the crude product, recrystallisation from
ethyl acetate-petrol gave (-) 4-amino-5-chloro-2-methoxy-N- ~ 1-
aza-6~-H-4-oxabicycl 4,4,0 decyl)benzamide (4) (1.3 g, 55%)
m.p. 195-7.
n.m~r. (S, CDC13): 8.00 (s, lH, aryl-6-_);
7.6-7.4 (m, lH, -CON_-CH=);
6.23 (s, lH, aryl 3-H);
4.6-1.0 (m, l9H, remaining proton
including 3.83, s, 3H, -OCH3).
- 43 -
I J 62187
Example 5
(-)4-Ace't'ami'd'o-'5-chlo'ro'-'2-methoxy-N-(8~-1-'aza-2~-methy1-6~-H-4-
oxabicyclo~4,4, o3 decyl')ben'z'amide (5)
CONH H
OCE~3
Cl/~
NHCOCH3
(5)
Following the procedures outlined in Example 1, the (-)
8~-amino-1-aza-2~-methyl-6~-H-4-oxabicycl~4,4,0~decane (D.12)
(0.9 g) was converted to 4-acetamido-5-chloro-2-methoxy-N-t8~-1-
aza-2~-methyl-6~-H-4-oxabicyclo[4,4,03decyl)benzamide (5) (1.0 g,
47%) m.p. 165-6.
n.m.r. (~, CDC13): 8.4-7.7 (m, 2H, -CON_-CH=,
CH3CONH);
8.28 (s, lH, aryl-H?;
8.14 (s, lH, aryl-_);
5.65-4.2 (m, lH, 8 -H);
4.2-3.0 (m, 8H including 4.0,
s, 3H, -OCH3);
2.9-0.8 (m, 13H, including 2.27
s, 3H, -COCH3) and 0.95, d, 3H,
=CH-CEI3, J=6Hz).
- 44 -
I 1 62187
Example 6
(+) 4 Amino-5-chloro-2-methoxy-~-(8~ aza-2~-methy1-6~-H-4-
_xabicyclor4,4,0~de'cy'1)b'en'z'amide`'(6)
/ ~ CH3
(~
CONH I ~ O
-OCH3
/J
C1/
NH2
(6)
Following the procedures outlined in Example 4, the (-)
4-acetamido-5-chloro-2-methoxy-N-(8~-1-aza-2~-methyl-6~-H-4-
oxabicyclo ~4,4,0~decyl)benæamide (5) (1.0 g) was converted to (-)
4-amino-5-chloro-2-methoxy-N-(8~-1-aza-2~-methyl-6~-H-4-oxabicyclo
~4,4,03decyl)benzamide (6) (0.7 g, 80~). m.p. 221-2.
n.m.r. (~, CDC13): 8.2-7.8 (m, lH, -CONH-CH=);
7.96 (s, lH, aryl 6-H);
6.24 (s, lH, aryl 3-H);
4.8-2.9 (m, llH includes 3.88,
s, 3H, -OCH3);
2.6-0.8 (m, 10H includes 0.96,
d, 3H, CH-C_3, J = 6Hz).
- 45 -
I l 62187
Example 7
(-) 4-Acetamido-5-chloro-2-methoxy-N-(8c~ aza-6~-H-4-
thiabicyclo ~,4,0~decyl)benzamide (7)
CONH
~OCH3 H
ll ,J
C~ ~
NHCOCH3
Following the procedures outlined in Example 1, the crude
(-) 8~,~-amino-1-aza-6~-H-4-thiabicyclo [4,4,0~decane (D.17) (2.4 g)
was converted to (+) 4-acetamido-5-chloro-2-methoxy-n-(8~r~-1-
aza-6~-H-4-thiabicyclo E,4,0~decyl)benzamide (7) (5.1 g, 90~).
m.p. 185-9 (E to Ac).
n.m.r. (~, CDC13): 8.3-7.4 (m, 4H, aryl-H, -CONHC~ ,
CH3CONH-Aryl);
5.6-1.2 (m, 20H, includes
4.0, s, 2H, -OCH3 [~-isomer~;
3.9, s, lH, -OCH3 ~-isomer~ ;
2.27, s, 3H, COCH3).
~ - ~6 -
~ J ~187
Example 8
(-) 4-Am'ino-5-chloro-2-methoxy-N-(8~ aza-6~-EI-4-thiabicyclo
~4,4, o3 decyl)benzamide (8)
~,/~ ~
CONEI H S
~ OCH3
Cl
NH2
(8)
Following the procedures outlined in Example 4, the (-)
4-acetamido-5-chloro-2-methoxy-N-(8~,~-1-aza-6~-H-4-thiabicyclo
[4,4,0]decyl)benzamide (7) (2.0 g) was converted to the crude (-)
4-amino-5-chloro-2-methoxy-N-(8~,~-1-aza-6~-H-4-thiabicyclo[4,4,0
decyl)benzamide. Fractional recrystallisation from ethyl acetate
afforded the pure (+) 4-amino-5-chloro-2-methoxy-N-(8~-1-aza-6~-H-
4-thiabicyclo [4,4,0~decyl)benzamide (8) (0.8 g, 45%). m.p. 183-6.
n.m.r. (~, CDC13): 8.2-7.8 (m, lH, ,CONH-CH=);
8.05 (s, lH, aryl 6-_);
6.27 (s, lH, aryl 3-_);
4.1-4.6 (m, 3H, aryl-NE12, 8~-_);
3.93 (s, 3H, -OCH3);
3.4-1.4 (m, 13H, remaining protons).
- 47 -
l ~ 62187
Example 9
(-) 4-Acetamido-5-'ch'loro-2-methoxy-N-(8~-1-aza-6~-H-4-thlabicyclo
[4,4,0~ decyl)ben'z-ami'de ~9)
CON ~ S
OCH3
Cl~ ~
NHCOCH3
(9)
Following the procedures outlined in Example 1, the crude
(-) 8~-amino-1-aza-6~-H-4-thiabicyclo[4,4,0~ decane) (D.20) (2.5 g)
was converted to (-) 4-acetamido-5-chloro-2-methoxy-N-(8~ aza-
6~-H-4-thiabicyclo[4,4,0]decyl benzamide (9) (2.2 g, 40%)
m.p. ~ 210-3.
n.m.r. ( , CDC13): 8.25 (s, lH, aryl EI);
8.17 (s, lH, aryl H);
8.0-7.5 (m, 2H, -CONH-CH~ ;
CH3CONH-aryl)
4.3-3.6 (m, lH, - 8~-H);
3.9 (s, 3H, -OCH3);
3.3-1.0 (m, 16H, remaining
protons including 2.27, s, 3EI, -C ~H3).
- 48 -
~ ~6`21~7
Example l_
(-) 4-Amino-5-chloro-2-methoxy-N-(8~ aza-6~-H-4-thiabicyclo
~4,4,03decyl)benzamide (10)
CONH ~ N
~ ~ OCH3 H
Cl~ ~
N 2
(10)
Following the procedures outlined in Example 4, the (+)
4-acetamido-5-chloro-2-methoxy-N-(8~-1-aza-6~-H-4-thiabicyclo
[4,4,03decyl)benzamide (9) (1~2 g) was converted to (-) 4-amino-
5-chloro-2-methoxy-N-(8~-1-aza-6~-H-4-thiabicyclo~4,4,0~decyl)
benzamide (10) (0.73 g, 65%). m.p. 206-8.
n.m.r. (~, CDC13): 8.03 (s, lH, aryl 6H);
7.7-7.3 (m, lH, ,CONHCH=);
6.23 (s, lH, aryl 3H);
4.6-3.7 (m, 6H, 8~-_, aryl
N-2 including 3.83, s, 3H, -OCH3);
3.5-1.0 (m, 13H, remaining protons).
- 49 -
.~
. --
- 50
Example 11
(~) 4-Acetamido-5-chloro-2-methoxy-N-(8a-1-aza 2~-
phenyl-6a-H-4-oxabicyclo[4l4~o]decyl)benzamide (11)
Ph
COWH ~ O
~ CH3
Cl ~ (11)
NHCOCH3
Following the procedures outlined in Example 1,
the (') 8a-amino-1-aza-2~-phenyl-6a-H-oxabicyclo
[4,4,0]decane (D.23) (1.3 g) was converted to (~) 4
acetamido-5-chloro-2-methoxy-N-(8a-1-aza-2~-phenyl-
6a-H-4-oxabicyclo[4,4,0]decyl)benzamide (11) (2.1 g,
80%) as an oil.
N.m.r. (~, CDC13) : 8.4-7.8 (m, 2H, CONH- and
aryl-H);
8.23 (s, lH, aryl-H );
0 8.08 (s, lH, aryl-H);
7.6-6.9 (m, 5H, aryl-H);
4.5-1.0 (m, l9H, including 4.0,
S, 3H-OCH3, 2.3, s, 3H,
-COCH3).
l 1 62187
- 51 -
Example 12
(i) 4-Amino-5-chloro-2-methoxy-N-(8a-1-aza-2~-phenyl-
6a-H-4-oxabicyclo[4,4,0]decyl)benzamide (12)
Ph
CONH H
ll I OCH3
Cl ~
NH2 (12)
Following the procedures outlined in Example 4,
the (i) 4-acetamido-5-chloro-2-methoxy-N-(8a-1-aza~
2~-phenyl-6a-H-4-oxabicyclo[4,4,0]decyl)benzamide
(11) (2.1 g) was converted to (~) 4-amino-5-chloro-
2-methoxy-N-(8a-1-aza-2~-phenyl-6a-H-4-oxabicycl~
[4,4,0~decyl)benzamide (12) (1.0 g, 50%), mp 145-7.
N.m.r. (~, CDC13) : 8.3-7.9 (m, lH, -CONH-CH= and
s, lH, aryl 6-H);
7.6-6.9 (m, 5H, aryl H);
6.3 (s, IH,aryl 3-H);
4.7-3.1 (m, llH, includes
3.93, s, 3H, -O CH3);
2.8-1.2 (m, 8H remaining C-H).
~ l 62187
. .
- 52 -
Example 13
(') 4-Acetamido-5-chloro-2-methoxy-N-(8~ aza-2~-
phenyl-6~-H-4-oxabicyclo[4,4,0]decyl)benzamide (13)
Ph
~ N
CONH ~-- ~--o
¦ H
~ OCH3
Cl ~
NHCOCH
3 (13)
Following the procedures outlined in Example 1,
the (~) 8~-amino-1-aza-2~-phenyl-6a-H-4-oxabicyclo
[4,4,0]decane (D.26) (2.1 g) was converted to (i) 4-
acetamido-5-chloro-2-methoxy-N-(8~-1-aza-2~-phenyl-
6~-H-4-oxabicyclo[4,4,0]decyl)benzamide (13) (3.2 g,
75%) as an oil.
N.m.r. (~, CDC13) : 8.4-7.5 (m, 2H, -CONH-CH=,
CH3CONH-);
8.25 (s, lH, aryl-H);
8.10 (s, lH, aryl-H);
7.4-7.2 (m, 5H, aryl-H);
4.7-1.0 (m, l9H, including s,
3H, 3.88, -OCH3; s, 3H, 2.22,
-COCH3).
2 1 87
- 53 -
Example 14
(+) 4-Amino-5-chloro-2-methoxy-N-(8~-1-aza-~-phenyl-
6a-H-4-oxabicyclo[4,4,0jdecyl)benzamide monohydrate (14)
Ph
CONH
~0
~ OCH3 H
C ~ (14)
NH2
Following the procedures outlined in Example 4,
the (~) 4-acetamido-5-chloro-2-methoxy-N-t8~-1-aza-
2~-phenyl-6a-H-4-oxabicyclo[4,4,0]decyl)benzamide
(13) (3.2 g) was converted to (i) 4-amino-5-chloro-2-
methoxy-N-(8~-1-aza-2~-phenyl-6~H-4-oxabicyclo[4,4,0]
decyl)benzamide monohydrate (14) (1.6 g, 50%) mp 138-45.
N.m.r. : 8.07 (s, lH, aryl 6-H);
7.7-7.2 (m, 6H, -CONH-CH= and
aryl -H);
6.28 (s, lH, aryl 3-H);
4.6-1.0 (m, 20H, includes 3.86,
s, 3H, -OCH3).
l ~ 62187
Example 15
(*J 4-Acetamido-5-chloro-2-methoxy-N-(8a,~-1-aza-
6a-H-4-oxothiabicyclo[4~4,0]decyl)benzamide (15)
CONH~
OCH3 H o
,J
Cl ~
NHCOCH3 (15)
To a stirred solution of (+) 4-acetamido-5-chloro-
2-methoxy-N-(8a,~-1-aZa-6~-H-4-thiabiCyCl0[4,4,0]deCyl)
benzamide (7) (2.0 g) in methanol (50 ml) was added
a solution of sodium periodate (1.5 g) dissolved in
the minimum quantity of water, and the resulting reaction
mixture stirred at room temperature for 3hours.
The solvent was removed in vacuo, water (10 ml)
added, followed by chloroform (100 ml). Separation
of the organic layer gave, on concentration and dryirig,
(*) 4-acetamido-5-chloro-2-methoxy-N-(8a,~-1-aza-
6~-H-4-oxothiabicyclo[4,4,0]decyl)benzamide (15)
(2.1 g, 100%) as a foam.
N.m.r. (~, CDC13) : 8.2-7.5 tm, 2H, -CONH-);
8.43 (br.s, lH, aryl-Hl;
8.27 (br.s, lH, aryl-H);
4.8-1.1 (m, 20H, including 3
singlets, 3H, -OCH3; 4.03,
4.00, 3.95, 2.3, s, 3H, -COCH3).
I 1 62~7
- 55 -
Example 16
(~) 4-Amino-5-chloro~2-methoxy-N-(8a,~-1-aza-6a-H-4-
oxothiabicyclo[4,4,0]decyl)benzamide (16)
CONH ~ ~ - ~ -
~ OCH3 H ~0
Cl ~ (16)
H2
Followtng the procedures outlined in Example 4,
the (l) 4-acetamido-5-chloro-2-methoxy-N-(8a,~-1-aza-
6a-H-4-oxothiabicyclo[4,4,0]decyl)benZamide (15)
(2.1 g) was converted into (~) 4-amino-5-chloro-2-methoxy-
N-(8a,~-1-aza-6a-H-4-oxothiabicyclo[4,4,0]decyl)benza
mide (1.0 g, 50%), mp 130-8.
N.m.r. (~, CDC13) : 8.3-7.4 (m, lH, -CONH-CH=);
8.06 (br.s, lH, aryl 6-H);
6.30 (br.s, lH, aryl 3-H);
4.7-1.1 (m, l9H, includes 3.94,
3.91, 3.87, 3 singlets, 3H.-OCH3)~
l l 62187
. . .
- 56 -
Examp1~17
(i) 4-Acetamido-5-chloro-2-methox~-N-(8~ aza-6a
-H-4-oxothiabicyclo[4,4,0]decyl)benzamide (17)
CONH - ~
OCH3 o
Il I .
Cl ~
NHCOCH3 (17)
Following the procedures outlined in Example 15,
the (i) 4-acetamido-5-chloro-2-methoxy-N-(8~-1-aza-
6a-H-4-thiabicyclo[4,4,0]decyl)benzamide (9) (1.1 g)
was converted to (i) 4-acetamido-5-chloro-2-methoxy-
N-(8~-1-aza-6~-H-4-oxothiabicyclo[4,4,0]decyl)benzamide
(17) (0.9 g, 80%). mp 245-6 (dec).
N.m.~ , CDC13) : 8.25 (s, lH, aryl-H);
8.15 (s, lH, aryl-H);
7.95-7.45 (m, 2H, -CONH-);
4.3-1.1 (m, 20H, including 3.95,
s, 3H, -OCH3; 2.3, s, 3H,
-COCH3).
l 1 62187
~ . .
- 57 -
Example 18
(+) 4-Amino-5-chloro-2-methoxy-N-(8~-1-aza-6a-H-4-
oxothiabicyclo[4,4,0]dec~1)benzamide(18)
CONH ~ S~
~ ~ CH3 H O
Cl ~
NH2 (18)
Following the procedures outlined in Example 4,
the (l) 4-acetamido-5-chloro-2-methoxy-N-(8~-1-aza-
6a-H-4-oxathiabicyclo[4,4,0]decyl)benzamide (17)
(0.9 g) was converted to (i) 4-amino-5-chloro-2-
methoxy-N-(8~-1-aza-6a-H-4-oxathiabicyclo[4,4,0]decyl)
benzamide (18) (0.4 g, 50%). mp 209-11
N.m.r. (~, CDC13) : 8.06(s, lH, aryl 6-H);
7.8-7.4 (m, lH, -CONH-CH=);
6.30 (s, lH , aryl 3-H);
4.7-1.1 (m, l9H, inlcudes 3.87,
s, 3H, -OCH3).
l ~ 62187
- 5B -
Example 19
(_)2-Methoxy-5-sulphonamido-N-(8~-1-aza-6~-H-4-thia-
bicyclo[4,4,0~decyl)benzamide (19)
~ N
CONH
OCH3 H
NH202S ~
(19)
To a solution of 2-methoxy-5-sulphonamido benzoic
acid (1.8 g) in dry DMF (30 ml) at 0C was added,
dropwise, a solution of oxalyl chloride (0.68 ml) in
chloroform (5 ml) over 20 minutes. On stirring for
a further 30 minutes, a solution of triethylamine (3 ml)
in chloroform (5 ml) followed by + 8~-amino-1-aza-
6~-H-4-thiabicyclo[4,4,0]decane (D 20) (1.2 g) in
chloroform ( 5 ml) was added and the whole stirred
for a further 2 hours at room temperature. Water
(10 ml) followed by saturated potassium carbonate
solution was added and the organic phase separated
and the aqueous phase extracted with chloroform
(2 x 100 ml), dried (K2C03) and the solvent removed
to yield an oil. Purification by column chromatography
(Alumina + 10% water; chloroform + 2% methanol) gave
the (+) 2-methoxy-5-sulphonamido-N-(88-1-aza-6~-H-
4-thiabicyclo[4,4,0]decyl)benzamide (19) (0.3 g,
11%), mp 216-8 (ethyl acetate).
116218~ ~
- 59 -
N.m.r. (~, CDC13) : 8.62 (d, lH, J=2.5 Hz, aryl 6-_);
7.98 (d,d, lH, J=2.5, 8.7 Hæ, aryl
4-H);
7.7-7.57 (m, lH, CON~
7.06 (d, lH , J=8.7Hz, aryl 3-_);
6.7-6.25 (m, 2H, -S02NH2);
4.5-3.7 (m, 4H, CONHCH< including
4.0, s, 3H, -OCH3);
3.4-l.o (m, 13H, remaining protons).
1 1 62187
- 60 -
Example 20
(~) 2,4-Dimethox~-N-(8~-l-aza-6~-H-4-thiabicyclo
[4,4,0]decyl)benzamide (20)
CON
OCH3 ~ S
CH3 (20)
To a stirred solution of 2,4-dimethoxybenzoic
acid (l.l g) in CH2Cl2 (lOO ml) was added oxalyl
chloride (0.48 ml) and DMF (ca 5 drops) and the whole
stirred until gas evolution ceased (ca 1 hour). The
reaction mixture was cooled to 0C and triethylamine
(2 ml) in C~2Cl2 (lO ml) followed by (+) 8~-amino-
l-aza-6~-H-4-thiabicyclo[4,4,0]decane(D 20) (0.85 g)
in CH2C12 (10 ml) added and the whole stirred at room
temperature for 2 hours. Aqueous sodium hydroxide
(2.5 N, 10 ml) was added and the o~ganic layer separated,
dried (K2C03) and evaporated to give an oil, which
was purified by chromatography (Alumina + lO~ water;
CH2Cl2) to give (+) 2,4-dimethoxy-N-(8~-l-aza-6~-
H-4-thiabicyclo[4,4,0]decyl)benzamide (20)(1.08 g,
65%), mp 130-2 (EtoAc/petrol).
`` 1162187
- 61 -
N.m.r. (~, CDC13) : 8.15 (d, lH, J = 8.6 Hz, aryl 6H);
7 . 75-7 . 45 (m, lH , CONH);
6.59 (d,d, lH, J = 8.6 Hz, 2.3 Hz,
aryl 5-H);
6.47 (d, lH, J = 2.3 Hz, aryl 3H);
4.8-3.7 (m, 7H, CON~CH= including
3.90, s, 3H, -OCH3 and 3.84, s,
3H, OCH3);
3.25-1.0 (m, 13~., remaining protons).
I 1 6~187
- 62 -
Example 21
(+) 4-Chloro-2-methoxy-N-(8~ aza-6~-E~ thia-bicyclo
~4,4,0]decyl)benz,amide (21)
~ N
CON
OCH3 H
Cl
(21)
Following the procedures detai~ed in Example 20,
the (+) 8~-amino-1-aza-6~-H-4-thiabicyclo[4,4,0]decane
(D 20) (0.85 g) was converted to (+) 4-chloro-2-methoxy-
N-(8~-1-aza-6~-H-4-thiabicyclo[4,4,0]decyl)benzamide
(21) (1.0 g, 60~, mp = 145-7 (E tOAc/petrol).
N.m.r. (~, CDC13) : 8.12 (d, lH, J = 8.3 HZ, aryl 6_);
7.75-7.4 (m, lH, -CONH);
7.12.6.93 (m, 2H, aryl 5_, 3H);
4.3-3.75 (m, 4H, CONHCH= including
3.93, S~ 3H, OCH3);
3.25-1.0 (m, 13H, remaining protons).
~ l l 62187
Bioloaical Data Section
~astric Activ tv
Increase in intraqastric pre~sure in the rat
- Intragastric pressure changes:were recorded from
previously starved conscious but retrained rats using
a saline filled catheter inserted into the lumen of the
stomach via a permanent gastric fistula. The catheter
was connected to a physiological pressure transducer
and pressure changes recorded on a hot wire pen
recorder. In each animal a pre-dose period of 40
minutes was allowed to obtain a measure of spontaneous
activity. An index of activity was obtained by measur-
ing the average height of pressure waves during 10minute periods. Values for 4 such periods were obtained
during assessment of spontaneous activity and for the
40 minute period after the subcutaneous administration
of the compounds. Students 't' test was applied to the
dîfference in average values obtained for spontaneous
and post-compound activity.
Table I shows the minimum dose for activity.
Table I
Compound of Dose ma/ka
Example No. Subcutaneouslv
2 0.5 0.5
4 1.0 n.t.
6 0.2 0.5
8 5.0 l~.t.
- 10 1.0 n.t.
n.t. = not tested
1 16~1g7
~ ~.
Table I (cont)
,
Compound of Dose mg/kg
Example No.
Subcutaneously Orally
14 1 n.t.
18 L n.t.
1 1 ~2~87
,
.
' Compounds were also tes*ed for their ability to
'inhibit apomorphine induced climbing in the mouse.
Inactivity in this test may be indicative of a low
propensity to-produce extra-pyramidal side-effects
S - in~Man. 'The results a're shown~n the-Table ~"~el-ow:
The'-me'thod used for this test is as set out below:
Table 2 - - ~- - -
_
Compound of-Example No Dose mg/kg ED50;~
4 30
6' 40 _
8 Ia at 25
12 30
14 Ia at 10 po
16 Ia at 10 po
. .. . .
Inhibition of apomorphine induced climbina in the mouse
.
The test is based on that described by Protais, P.,
Constantin, J. and Schwartz, J.C. (1976), Psychopharmacology,
50, 1-6.
10Apomorphine 1 mg/kg s.c. induces mice to climb the
wall of a wire'cage (inverted food hopper - 11 x 7.5 x
18 cm high). Mice acclimatised in their home cages in
groups of 5 are placed under the hoppers immediately
after the 'injection of apomorphine 1 mg/kg s.c. At
1510, 20 and 30 minutes after injection climbing behaviour
is scored. The mice are observed for 30 seconds and
scored according to the position in which they spend
'the majority of time, score 0 - four paws on floor of
cage; score''l - fore paws only on walls; score 2 - all
paws on wall of cage.
1 J ~187
The scores at all 3 times for each mouse are summed
for apomorphine only and for apomorphine plus compound
groups of mice.
Total scores are then calculated as the percentage
inhibition of climbing as follows:
rTotal scores of drug treated group 1 x 100
I i ition 100 LTotal scores of apomorphine only group
From graphical representation of the results ED50
vlaues (the dose that inhibits the apomorphine response
by 50%) are then obtained.
Toxicity
No toxic effects were observed in these tests.
l,b,
