SYNTHESIS AND ANTICONVULSANT PROPERTIES OF NEW I-PHENYL AND l-PHENYLAM INO-3-PHENYLPYRROLIDINE-2,5-DIONE DERIVATIVES
JO L A N T A O B N IS K A , A L F R E D Z E JC and A G N IE S Z K A Z A G O R S K A
D ep artm en t o f P h arm aceu tical C h em istry , C o lleg iu m M ed icu m o f the Jag iello n ian U n iv ersity , 9 M ed y czn a Str., 3 0 -6 8 8 K rakow , P oland
A b stra c t: A s erie s o f N - a r y l an d N -a m in o a ry l 3 - p h e n y l p y r r o lid in c - 2 ,5 - d io n e s w e re s y n th e s iz e d an d te sted fo r a n tic o n v u lsa n t a c tiv ity in th e m a x im u m e le c tro s h o c k s e iz u re (M E S ) a n d p en tetra z o l s e iz u re th re sh o ld (sc M et) te sts. S tru c tu re s o f th e n o v el co m p o u n d s w ere c o n firm e d by ele m e n ta l an d sp e c tra l a n a ly ses.
K e y w o r d s: a n tic o n v u lsa n t a c tiv ity , p y r r o ! id in e - 2 ,5 - d io n e , su c c in im id e s.
In search fo r a new c o m p o u n d s w ith p red ic
table a n tico n v u lsan t p ro p erties, o u r atten tio n w as d raw n to a gro u p o f 3 -p h e n y lp y rro lid in e -2 ,5 -d io - ne w ith d ifferen t su b stitu en ts at the n itrogen atom (1,2). P revious reports (3,4,5) show ed th e a n tic o n v u lsan t activity o f N -p y rid y l-3 - a ry lp y r ro lid in e - 2 ,5 - d io n e w h ich d is p la y e d p ro te c tio n a g a in s t M E S . In a d d itio n , it w as a ls o re p o rte d th a t a s ig n ific a n t e ffe c t on th e a c tiv ity o f th is c o m p o u n d s b e in g e x e rte d by th e p re s e n c e , p o sitio n an d c h a ra c te ris tic o f th e s u b s titu e n ts in p y rid in e m o ie ty . C ry s ta llo g ra p h ic in v e s tig a tio n s (6 ,7 ) s h o w e d th a t th e ir p h a rm a c o lo g ic a l p ro p e rtie s w e re c o n n e c te d w ith th e c o n fo r m a tio n . T h e c o n fo r
m a tio n o f p h a rm a c o lo g ic a lly in a c tiv e m o le c u le s d iffe rs fro m th a t o f m o le c u le s w ith a c o n firm e d a c tiv ity .
T h is rep o rt w ill p ro v id e fu rth er stru c tu re -a c - tivity studies on th e 3 -p h e n y lp y rro lid in e -2 ,5 -d io nes in w hich p y rid in e m oiety is rep laced by th e aryl ring w ith m ethyl o r ch lo ro su b stitu en ts [com p.
I - V I I ] o r by the p h en y lam in o o r d ip h en y lam in o m oiety [com p. V I I I - X I ] .
T h e req u irem en t fo r N H gro u p linking the p y rro lid in e -2,5 -d io n e system to the aro m atic ring w as also in vestigated. A cco rd in g ly , a series o f new N -a ry l su b stitu ted 3 -p h e n y lp y rro lid in e -2 ,5 -d io nes, w as sy n th esized and ev alu a ted fo r a n tico n v u l
sant activity.
A s the starting m aterials fo r the synthesis 3 -( 2 -c h lo r o p h e n y l)- , 3 -( 3 -c h lo r o p h e n y l)- , 3 - ( 4 - c h lo r o p h e n y l) -,3 -(3 - flu o r o p h e n y l)- , 3 -( 3 -b ro m o - p h e n y l) - and 3 -p h e n y lsu c c in ic acids sy n th esized by the m eth o d o f M iller et al (8), as m odified by L an g e et al. (9) w ere used. T h e acids w ere cyclized to I-p h e n y l or l-p h e n y la m in o -3 -p h e n y lp y rro lid i- n e -2 ,5 - d io n e d eriv ativ es [I—XI] by heating them at
ca. 190°-200°C fo r 1 .5 -2 h w ith a p p ro p riate aryla- m in e (S ch em e 1) o r w ith arylhydrazine.
E X P E R IM E N T A L
C h em istry
M eltin g p o in ts (°C) w ere un co rrected , 'H N M R sp ectra [I - X I ] w ere reco rd ed in C D C L solution by using T M S as an internal stan d ard (B ru k er V M 2 50 M H z N M R S pectrom eter). T he ch ro m ato g rap h y w as p erfo rm ed w ith M erck silica gel G F2 5 4 p reco ated T L C sheets using the follow ing d ev elo p in g system s: A - chloroform : aceton (9:1), B - chloroform : m ethanol: acetic acid (60: 10: 5).
S pots w ere d etec ted by th e ir ab sorption u n d er UV light and v isu alizatio n w ith 0.05 m ol L in 10%
HC1.
1-P h e n y l and l- p h e n y la m in o -3 - p h e n y lp y rr o - lid in e -2,5 -d io n e d eriv ativ es [I - X I].
G en eral procedure.
A n a p p ro p riate ary lam in e o r ary lh y d razin e (0.02 m ole) w ere d isso lv ed in 25 ml o f w ater and ap p ro p riate 2-s u b s titu te su ccin ic acid (0 . 0 2 m ole) w as g radually added. T h e m ixture w as h eate d in oil b ath w ith sim u ltan eo u s d istillatio n o f w ater. A fter co m p lete rem o v al o f w ater, the tem p eratu re o f the reactio n m ix tu re w as raised up to 1 9 0 -2 0 0 l’C and m ain tain ed fo r 1.5 h. T h e cru d e p ro d u ct w as recry stallized from ethanol.
P h y sico ch em ical data, 'H N M R data, yields and elem en tary an aly sis resu lts o f th e c o m p o u n d s sy n th esized w ere p resen ted in T ab le la , b.
P h a rm a co lo g y
C o m p o u n d s I - X I w ere p h arm aco lo g ically p re -e v a lu a te d w ithin the A n tiep ilep tic D rug D e v e lopm ent (A D D ) P rogram , E p ilep sy B ranch, N euro-
209
S c h e m e 1.
S c h e m e 2.
logical D iso rd ers P rogram N ational Institutes o f the N eu ro lo g ical and C o m m u n icativ e D iso rd ers and S tro k e (N IN C D S ), by using the testin g pro ced u res describ ed elsew h ere (10,11). P h ase I stu d ies in v o l
ved three tests: m axim um elec tro sh o ck seizure (M E S ), su b cu tan eo u s p en ty len etetrazo le (sc M et), and the n eu ro to x icity (T O X ). T h e M E S test is a m odel fo r g en eralized to n ic -c lo n ic seizures. In th is test, an electrical stim ulus o f 0 .2 s in d uration (50 m A in m ice at 60 H z) is d eliv ered via corneal electrodes. M ice are tested fo r 30 m in. an d 4 h w ith
th e fo llo w in g doses: 30, 100, 30 0 m g/kg o f test c o m p o u n d (12).
T he scM et is a m odel fo r co m p o u n d s that raise a seizure threshold. In scM et test a d o se o f m etrazol is 85m g/kg in m ice. T h is p roduces clo n ic seizures du rin g a p erio d o f at least 5 seconds in 97% o f tested anim als. T h e m etrazol is ad m in istrated sub- cu tan e o u sly . T he test c o m p o u n d s are ad m in istrated in trap erito n eally , su sp en d ed in 0 .5% m eth y lcellu - lose at th e d o ses o f 30, 100, 3 00 m g/kg (13).
N e u ro to x icity , in d u ced by a test co m p o u n d , is
T a b le la . E x p e rim e n ta l d a ta fo r c o m p o u n d s I - V I I
No. M olecular Form ula
W eight
Yield % Mp. [”C]
Analysis cal./found 'H N M R /C D C F/5 R,/
solvent
%C %H %N
I cI7h,a n , 265.3
73 120-122
77.05 76.93
5.75 5.66
5.28 5.38
2.53 (3H, s, CH-0, 2.7 5 -2 .9 0 (IH , d, imide), 3.35-3.42 (1H, q, imide), 4 .0 8 A 4 0 (1H, q, imide), 7 .1 0 -7 .6 0 (9FI, m, arom)
0.67A 0.82B
II C n H u O jN iC l.F 317.74
50 104-105
64.21 64.27
4.12 4.07
4.40 4.26
2.15 and 2.19 (3H, 2 s, C H ,), 2 .9 6 -3 .0 0 (1H, td, im ide), 3.40-3.51 (1H, dq, imide), 4 .2 0 - 4.32 (1H, q, imide), 6.99-7.40 (7H, m, arom)
0.80A 0.79B
I I I C .hH ^ A N .F 297.33
48 5 7 -6 0
72.79 72.30
5.43 5.29
4.72 4.48
2.32 (3H, s, CH j), 2.74-2.83 (1H, d, imide), 3 .14-3.24 (1H, d, imide), 3 .97-4.02 (1H, q, imide), 4.4 2 -4 .7 4 (2H, q, C H 2), 6.87-7.38 (8H, m, arom)
0.72A 0.84B
IV C n H A N . C l 299.75
65 148-150
68.06 68.05
4.70 4.63
4.67 4.51
2.18 and 2.21 (H , 2s, C H ,), 2 .9 7 -3 .0 4 (1H , td , im id e), 3 .3 8 -3 .5 0 (1H , dq, im i
d e), 4 .1 8 -4 .2 9 (1H , dq, im id e), 7 .0 8 -7 .4 5 (7H , m, arom )
0.78A 0.86B
V c,6h„o2n,c i2 320.70
58 132-134
59.87 60.02
3.45 3.43
4.36 4.12
2 .9 2 -3 .0 0 (1H, dd, im ide), 3 .33-3.42 (1H, q, imide), 4 .1 5 -4 .2 0 (1H, q, imide), 7.23-7.49 (8H, m, arom)
0.82A 0.79B
VI C n H i A N iB r 344.22
52 58-61
59.36 58.98
4.10 4.02
4.07 3.90
2 .1 5 -2 .1 8 (3H , d, CH.,), 2 .9 2 -2 .9 6 and 3 .0 1 -3 .0 6 (1H , dt, im id e), 3 .3 3 -3 .4 8 (1H , dq, imide), 4.1 4 -4 .2 4 (1H, q, imide), 7 .2 0 - 7.48 (8H, m, arom)
0.80A 0.85B
V II C ,7H , A N , B r 344.22
50 62-6 4
59.36 59.13
4.10 4.14
4.07 3.72
2.15-2.18 (3H, 2s, C H ,), 2.92-3.05 (IH , dq, imide), 3.23-3.49 (1H, dq, imide), 4.14—4.24 (1H, dq, imide), 7 .0 7 -7 .5 0 (8H, m, arom)
0.72A 0.83B
T a b le lb . E x p e rim e n ta l d a ta fo r c o m p o u n d s V 1 1 I-X I
No. M olecular Formula
W eight
Yield % Mp. m
Analysis cal./found 'H NMR/CDCl.,/8 Rr/
solvent
%C %H %N
V III C A ÏM A N , 342.40
60 112-116
77.06 76.85
5.31 4.92
8.34 8.19
2 .89-3.00 (1H, dd, C H um ide), 3.24-3.39 (1H, dd, C H 2imide), 4 .0 9 -4 .1 7 (1H, q, CH imide), 7.09-7.42 (15H, m, arom)
0.73A 0.84B
IX C |f, H |A N 2Cl 300.74
68 158-159
63.85 63.82
4.35 4.01
9.31 9.18
2 .91-2.94 (1H, dd, C H ,im ide), 3.27-3.41 (1H, dd, C H ńm ide), 4.37-4.45 (1H, q, CH imide), 6.30 (1H, s NH), 6.85-7.45 (9H, m, arom)
0.84A 0.94B
X C 22H |70 2N2C1 376.83
74 159-161
70.09 70.43
4.55 4.85
7.43 7.46
2 .9 1 -2 .9 4 (1H, dd, CH 2imide), 3.27-3.41 (1H, dd, C H jim ide), 4 .37-4.45 (1H, q, CH imide), 6.30 (1H, s NH), 6.85-7.45 (9H, m, arom)
0.79A 0.89B
XI C i A A N . C l 300.74
65 173-174
63.85 64.28
4.35 4.31
9.31 8.81
2.90-3.03 (IH , m, imide), 3.26-3.35 (1H, m, imide), 4.3 4 -4 .4 2 (IH , m, imide), 6 .69-6.80 (2H, m, arom ), 7.13-7.51 (7H, m, arom), 8.44 (IH , s, NH)
0.69A 0.87B
T a b le 2a. A n tic o n v u ls a n t S c re e n in g P ro je c t (A S P ) P h a se I T e st re su lts in m ic e (co m p . I - V I I )
Comp. Dose
Activity
Tox1” A SPdl
M ES"’ sc M et1’1
mg/kg 0.5 h 4 h 0.5 h 4 h 0.5 h 4 h class
30 0/1 0/1 0/1 0/1 0/4 0/2
I 100 1/3 0/3 0/1 0/1 0/8 0/4 1
300 1/1 0/1 4/5 0/1 0/4 0/2
30 0/1 0/1 0/1 0/1 0/4 0/2
II 100 0/3 0/3 0/1 0/1 0/8 0/4 3
300 0/1 0/1 0/1 0/1 0/4 0/2
30 0/1 0/1 0/1 0/1 0/4 0/2
III 100 0/3 0/3 0/1 0/1 0/8 0/4 2
300 1/1 0/1 1/1 0/1 0/4 0/2
30 0/1 0/1 0/1 0/1 0/4 0/2
IV 100 0/3 0/3 0/1 0/1 0/8 0/4 3
300 0/1 0/1 0/1 0/1 0/4 0/2
30 0/1 0/1 0/1 0/1 0/4 0/2
V 100 0/3 0/3 0/1 0/1 0/8 0/4 3
300 0/1 0/1 0/1 0/1 0/4 0/2
30 1/4 0/1 0/1 0/1 0/4 0/2
V I 100 3/3 0/3 0/1 0/1 0/8 0/4 1
300 1/1 0/1 1/1 0/1 0/4 0/2
30 0/1 0/1 0/1 0/1 0/4 0/2
V II 100 1/3 0/3 0/1 0/1 0/8 0/4 1
300 1/1 0/1 3/5 0/1 1/4 0/2
* T o n ic e x te n sio n
“'M a x im a l e le c tro s h o c k test (n u m b e r o f a n im a ls p ro te c te d /n u m b e r o f a n im a ls te sted ) h,S u b c u ta n e o u s p e n ty le n e te tra z o le test
clR o to ro d to x ic ity (n u m b e r o f a n im a ls e x h ib itin g to x ic ity /n u m b e r o f a n im a ls te sted ) c’T h e cla s sific a tio n is as fo llo w s:
1 -a n tic o n v u lsa n t ac tiv ity at d o s e s 100 m g /k g o r less 2 -a n lic o n v u ls a n t ac tiv ity at d o ses g re a te r th a n 100 m g /k g 3 - c o m p o u n d in a ctiv e a t 3 0 0 m g /k g
T a b le 2 b . A n tic o n v u ls a n t S c re e n in g P ro je c t (A S P ) P h ase 1 T e s t re su lts in m ice (c o m p . V I I I - X I )
Comp. Dose
Activity
T ox° A S P 1’
MES"’ sc M et1”
mg/kg 0.5 h 4 h 0.5 h 4 h 0.5 h 4 h class
30 0/1 0/1 0/1 0/1 0/4 0/2 *
V III 100 0/3 0/3 0/1 0/1 0/8 0/4 2
300 0/1 0/1 1/5 0/1 0/2 0/2
30 0/1 0/1 0/1* 0/1 0/4 0/2
IX 100 0/3 0/3 0/1 0/1 0/8 0/4 2
300 0/1 0/1 1/5 0/1 0/4 0/2
30 0/1 0/1 0/1* 0/1 0/4 0/2
X 100 0/3 0/3 0/1 0/1 0/8 0/4 3
300 0/1 0/1 0/1 0/1 1/4 0/2
30 0/1 0/1 0/1 0/1 0/4 0/2
X I 100 0/3 0/3 0/1 0/1 0/8 0/4 3
300 0/1 0/1 0/1 0/1 0/4 0/2
* T o n ic ex ten sio n
;l,M ax im a l ele c tro s h o c k te st (n u m b e r o f a n im a ls p ro te c te d /n u m b e r o f a n im a ls te ste d ) h,S u b c u ta n e o u s p e n ty le n e te tra z o le test
c>R o to ro d to x ic ity (n u m b e r o f a n im a ls ex h ib itin g to x ic ity /n u m b e r o f a n im a ls te sted ) cT h e cla s sific a tio n is as fo llo w s:
1 -a n tic o n v u lsa n t ac tiv ity at d o se s 100 m g /k g o r less 2 - a n tic o n v u ls a n t ac tiv ity at d o se s g re a te r th a n 100 m g /k g 3 - c o m p o u n d in a ctiv e a t 3 0 0 m g /k g
d etec ted in m ice using a ro to ro d test. U ntreated control m ice, w hen placed on a 6 r.p.m . rotation rod can m aintain th eir e q u ilib riu m fo r a pro lo n g ed tim e. N eu ro to x icity is in d icated by the inability o f a m ouse m ain tain in g e q u ilib riu m fo r o n e m inute in each o f the th ree su ccessiv e trials (14). R esu lts are given in T ab le 2a, b.
R E S U L T S
In a series o f l,3 - d ip h e n y lp y rro lid in e -2 ,5 - d io n es [I—V II] co m p o u n d s I, III, V I, and V II sh o w ed som e an tico n v u lsan t p ro p erties in phase I o f screen in g project. T h e l- (2 -m e th y lp h e - n y l) - 3 -p h e n y lp y rro lid in e -2 ,5 -d io n e I and three an alo g u es o f this c o m p o u n d III, V I, V II w ere found to be effectiv e in this screen in g procedure.
T h e m ost p o ten t in M E S test w as l- ( 2 - m e th y l- p h e n y l) -3 -( 3 -b ro m o p h e n y l)p y r ro lid in e - 2 ,5 -d io n e V I, w hich p ro tected all an im als after 0.5 h at 30, 100 and 3 00 m g/kg. T h e l- ( 4 - m e th y lp h e n y l) - 3 - (3 -flu o ro p h e n y l)p y rro lid in e -2 ,5 -d io n e III had an
tico n v u lsan t p ro p ertie s but only at a d ose o f 300 m g/kg. T h is c o m p o u n d s in the scM et test at a dose o f 3 00 m g/kg sh o w ed m arginal a n tico n v u lsan t properties. S om e activ ity w as reco rd ed in the scM et test also fo r co m p o u n d s I, V I, V II (active at 300 m g/kg at 4h).
T he presen ce and p o sitio n o f the m ethyl group in N -a ry l m oiety appears to be essen tial fo r the an tico n v u lsan t activity in this gro u p o f co m pounds.
T he m ost p o ten t in M E S test w ere c o m p o u n d s w ith the m ethyl g ro u p in p o sition 2 o r 4 o f the 1-p h en y l ring v/z., I, V I and V II. T h e ch an g e o f the brom atom fo r c h lo r su b stitu en ts on the 3 -p h e n y l ring in c o m p o u n d s IV and V m ade th em inactive. T his effect m ay b e is co n n ected w ith lo w er lipophilicity.
T he m eth y len e b rid g e in c o m p o u n d III d ecreased th e activity. In tro d u ctio n o f the second su b stitu en ts (-C1) in the 4 -p o s itio n o f the 1 -p h en y l ring led to an inactive co m p o u n d II.
In a series o f 1 -p h en y lam in o d eriv ativ es X , X I w ere d evoid o f an tico n v u lsan t activity. C o m p ounds V III, IX w ere active on ly in the scM et test at 0.5 h using 3 00 m g/kg. T h is resu lt su g g ests that th e N H group, linking I-p h e n y l ring w ith 3 -p h e n y - lp y rro lid in e -2 ,5 -d io n e m oiety, is p robably not ne
cessary fo r an tico n v u lsan t activity.
A ck n o w leg em en ts
T he authors w ish to th an k D r Jam es S tables fo r p ro v id in g p h arm aco lo g ical d ata th ro u g h the A n tiep ilep tic D ru g D ev elo p m e n t P rogram , E p ilep sy B ranch, N ational In stitu te o f N eurological D iso rders and S troke N atio n al In stitu te o f H ealth, B et- hesda, M ary lan d , U .S.A .
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Received: 7.12.2001
