Eur JMed Chem (1994) 29,107-113 107

0 Elsevier, Paris

Conformational and structural features determining in d-o

antimalarial activity in some indolo[3,2-c]qwinolines,
anilinoquinolines and tetrahydroindolo[3,2-d]benzazepines

HL Kohl, ML Gal, Tl Ngiaml, JW Malt*

‘Department of Pharmacy, National University of Singapore, Kent Ridge Crescent, 0511 Singapore;
2Institute of Medical Research, Jalan Pahang, Kuala Lurnpur, Malaysia
(Received 7 June 1993; accepted 7 October 1993)

Summary - A series of indolo[3,2-clquinolines, anilinoquinolines and tetrahydroindolo[3,2-dlbenzazepines, which differ in the

conformational planarity of the key nuclei and the distance (N...N+) between the ring and side-chain nitrogen atoms, have been
synthesized and evaluated in vitro against a number of isolates of Plasmodium falciparum. The results show that differences in confor-
mational planarity of the indolo[3,2-clquinoline (rigid), anilinoquinoline (flexible) and tetrahydroindolo[3,2-d]benzazepine (semi-
rigid) nuclei have little effect on activity. However, the N.. .N+ distance is an important determinant of activity and analogues with a
distance comparable to the N.. .N+ distance of amodiaquine and chloroquine demonstrated antimalarial activity.

antimalarial activity / indolo[3Z2-c]quinoline / anilinoquinoline / tetrahydroindolo[3,2-d]benzazepine / SAR

Introduction ties of the conformationally rigid indolo(3,2-clquino-

Malaria is one of the most serious parasitic diseases
confronting developing nations of the world. The
WHO has estimated that some 100 million people are
clinically ill with malaria at any one time, and around
1 million (mostly young children in sub-Saharan
Africa) die from it annually [ 11. Until an effective
vaccine [2] becomes available, chemotherapy remains
the prime strategy in the fight against malaria. Even
with chemotherapy, there is an urgent need for novel
antimalarials to combat a parasite that is remarkably
adaptable to drug challenge. The appearance of drug-
induced resistant strains of plasmodia has rendered
several members of the fast-shrinking armamentarium
of antimalarial agents obsolete.
We have previously reported the antimalarial activi-
ty of some indolo[3,2-clquinolines 131. In that series,
activity appeared to be linked to the presence of a
basic side chain at position 9. The most active
compound, 3-chloro-%methoxy-9-(4-methyl-l-piper-
azinylmethyl)-llH-indolo[3,2-clquinoline 1, was
about lo”-fold more active than chloroquine when
tested in vitro against chloroquine-resistant plasmodia.
The antimalarial activity of indolo[3,2-clquinolines
is further explored in 2 directions in the present study.
Firstly, the importance of conformational planarity
was investigated by comparing the antimalarial activi-
line analogues 2a-d with those of the flexible 7-
chloro-4[4’-methoxy-3-substituted]anilinoquinoline
series 3a-d and the less rigid tetrahydroindolo[3,2-d]-
benzazepines 4a-d. Secondly, the importance of the
basic side chain in 2, 3 and 4 was considered by
synthesizing compounds with the following structural
variations: no basic side chain (2a, 3a, 4a); with a
monobasic diethylaminomethyl side chain (2b, 3b,
4b); with a dibasic 3’-dimethylaminopyrrolidin-l’-yl-
methyl side chain (2c, 3c, 4~); and with the dibasic
[N-(3-dimethylaminopropyl)-N-methyllaminomethyl
side chain (2d, 3d, 4d) (fig 1). Side chains c and d are
isosteric.
Chemistry
The syntheses of 2a [3], 2b [4], 3a [S], 4a [6] and 4b
[6] have previously been reported. O-Methylamo-
diaquine 3b was a gift from Parke-Davis/Warner-
Lambert. The synthesis of the remaining compounds
is outlined in scheme 1.
Chloromethylation [7] of 4nitroanisole 5 gave the
chloromethyl derivative 6, which on nucleophilic sub-
stitution with the appropriate amines gave 7c and 7d.
Catalytic reduction of the aromatic nitro function
yielded the corresponding 4-aminoanisoles SC and 8d
108

5 6 7 P
8 10

0 0
Cl
15 R = -N”-y--J-C”, Cl N Cl m N
0 Cl N 11 II
ml
12 3 tiJ\ 13

2 4
Scheme 1.

nucleus is a cyclized form of 7-chloro-4-anilinoquino-

line [4], from which important antimalarials, such as
amodiaquine 14 [9], have been obtained. Taking
amodiaquine as the lead compound and assuming that
the minimum energy conformation of amodiaquine is
its bioactive conformation, a knowledge of this is
essential for the rational design of the present series of
compounds.
At pH 7.4, the side-chain amino function of amodi-
aquine is protonated. Based on the pK, of 3a (7.59)
Fig 1. Structures of compounds lG4. [lo], it is probable that about 50% of the amodiaquine
molecules would have an unprotonated quinoline
nucleus at physiological pH. Cyclization of the
which were converted to the respective diazonium anilinoquinoline nucleus of 3a to give the indolo-
salts 9c and 9d with sodium nitrite under acidic condi- [3,2-clquinoline 2a results in a decrease in basicity of
tions and low temperature [4]. Subsequent reduction the quinolinyl nitrogen (pK, of 2a = 3.99) [lo]. As a
of the diazonium salts with stannous chloride [4] number of indolo[3,2-clquinolines have been shown
afforded the hydrazines 1Oc and 10d. No attempt was to possess antimalarial activity [3], it is reasonable to
made to isolate and characterize the intermediates 9c,
9d, 1Oc and 10d which were unstable.
The indolo[3,2-clquinolines 2c and 2d were obtained
by Fischer indolization of hydrazines 1Oc and 10d
with 4-keto-7-chloro- 1,2,3,4-tetrahydroquinoline 11
[8]. The anilinoquinolines 3c and 3d were prepared by
condensation of the respective arylamines SC and Sd
with commercially available 4,7-dichloroquinoline 12.
Similar indolization of hydrazines 1Oc and 10d with
8-chloro- 1,2,3,4-tetrahydro- 1-benzazepin-5-one 13
[6] gave the tetrahydroindolo[3,2-dbenzazepines 4c
and 4d, respectively.

Results and discussion

Drug design and proposed druglreceptor interaction

Cl ConfDrmer
s
The selection of the indolo[3,2-clquinoline nucleus as
the key structure in the present series of potential anti-
malarials is based in part on the recognition that the Fig 2. Conformers of amodiaquine.

assume that the quinolinyl nitrogen of these com-
pounds (as well as those of the anilinoquinolines,
including amodiaquine) is unprotonated in the bio-
active form at pH 7.4.
Molecular mechanical (MM) calculations in the
present work using an MMX force field [ll] have
shown that the proposed bioactive form of amodia-
quine has 2 stable conformers, A and B, which are
stabilized by intramolecular hydrogen bonding (fig 2).
Instead of an expected planar conformation, the
phenyl ring of both conformers is slightly twisted
from the plane of the quinoline ring, due to steric
interactions between the hydrogen atoms H,/H, and
H,/H,. Recent X-ray crystallographic studies [ 121
have favoured conformer B in the solid state, although
our data has shown B to be only slightly more stable
than A (AE = 0.33 kcal mol-1).
It is proposed that the 2 most important pharmaco-
phoric groups of amodiaquine are its positively
charged ammonium group at the side chain and the
unprotonated quinoline ring, which interact with
suitable binding sites on a hypothetical receptor via
electrostatic and hydrophobic interactions (fig 3). The
reactive moiety at the hypothetical receptor that inter-
acts with the protonated side chain of amodiaquine
must be anionic in nature, while that which interacts
with the quinoline ring via hydrophobic forces must
be flat in geometry and bear a hydrogen donor group
(such as OH) capable of hydrogen bonding with the
unprotonated quinolinyl nitrogen. The anionic and
hydrogen donor groups must necessarily be separated
by a distance comparable to the N...N+ distance
between the quinolinyl nitrogen and the side chain
nitrogen of amodiaquine.
It was noted at the onset that the N, .d .N3+ distances
of amodiaquine (conformer B, 8.30 A) and chloro-
quine (8.38 A) are very similar (table I), suggesting
that conformer B of amodiaquine is the bioactive
form. I,n contrast, the N...N+Odistance of mefloquine
(6.52 A) and quinine (6.55 A), although very close
to each other, are shorter, and that of another a$tive
anilinoquinoline antimalarial 15 [13] (14.24 A) is
much longer.
The different N.. .N+ distances of these potent quino-
line antimalarials suggest the following possibilities.
Firstly, there may be more than one anionic site at
the target macromolecule which could accomodate
molecules with varying N.. .N+ distances. Secondly,
the target macromolecule may be conformationally
labile, thus allowing a single anionic group to assume
different but quite definite distances from the hydro-
gen donor group.
In the design of the present series of compounds,
the importance of the N.. .N+ distance has been taken
into consideration. We began with compounds without
the side-chain amino function (2a, 3a, 4a), followed
by compounds with 1 or 2 side-chain nitrogens of
109
Planar
area
/ Hydrogen donor Anionic Additiona!
anionic
!T-ow site
site (?)
Fig 3. The pharmacophoric groups of amodiaquine.
varying N.. .N+ distances from the ring nitrogen. As
seen in table I, the N,. . .N,+ distance increases in the
sequence of the 3 (conformer B) -+ 2 + 4 series.
Dibasic side chains (c, d) were included in view of the
reported efficacy of 1 over analogues with only mono-
basic side chains [3].
The choice of 3 different ring systems (indolo-
[3,2-clquinolines 2, anilinoquinolines 3, and tetrahydro-
indolo[3,2-dlbenzazepines 4) allows the evaluation of
the effect of molecular planarity and conformational
lability on antimalarial activity. Table II gives the
angle of twist from planarity of the 3 ring systems
(represented by 2b, 3b, 4b). Compound 2b is seen to
be the most planar, followed by 4b and 3b. Con-
formationally, the anilinoquinolines 3 are labile, the
tetrahydroindolo[3,Zd]benzazepines 4 are semi-rigid
and the indolo[3,2-clquinolines 2 are the most rigid.
Structure-activity relationship
Table III gives the in vitro activity of the present
series of compounds against various isolates of
Plamodium falciparum. These isolates have been arbi-
tarily classified into Category 1 (isolates obtained
from venous blood of infected patients and tested
immediately) and Category 2 (isolates established in
the laboratory by continuous culture). Further differ-
entiation of Category 1 and 2 isolates was made on
the basis of their sensitivity to chloroquine and
mefloquine according to WHO guidelines. Category 1
isolates were generally sensitive to chloroquine and
mefloquine, unlike the Category 2 isolates.
Testing against a number of isolates instead of one
provided a more realistic assessment of antimalarial
efficacy. However, because the test compounds were
dispatched to the testing laboratory over a period of
1 yr, it was not always possible to test all the com-
pounds against the same plasmodial isolate at any one
time. Nonetheless, some useful generalizations could
be made from the available data.
Compounds without a basic side chain (2a, 3a, 4a)
generally have the lowest activity. This is in keeping
with our earlier observation on the indolo[3,2-c]
110

Table I. Interatomic distances (A) of basic nitrogen atoms in amodiaquine, 2bbd, 3b-d, 4b-d, 15, 1, chloroquine, mefloquine
and quinine at their minimum energy conformations computed from an MkIX force field with TCroutines.
Compound Conformationa Min energy (kcal mol-11 Nl.. .N3+b N,. . .N4+b NJ. . .N5+b
Amodiaquine A 17.85 10.09
B 17.52 8.30
2b - - 8.72
2c - - 8.80 12.05
2d - - 8.74 13.52
3b A 20.80 10.09
20.45 8.29
3c i 85.03 10.09 13.31
B 84.54 8.24 11.00
3d A 73.12 10.02 14.15
B 72.67 8.54 13.18
4b - - 9.54
4c - - 9.51 12.31
4d - - 9.58 13.23
15 - 14.24
1 8.71 11.14
Chloroquine 8.38
Mefloquine 6.52~
Quinine 6.55~

aAmodiaquine and the anilinoquinolines 3b-d have 2 stable conformations A and B. In conformer A, the basic ammo side
chain is orientated away from the quinoline ring. In conformer B, the side chain is orientated towards the quinoline ring (fig 2);
bsee figure 1 for numbering on N atoms; cN1.. .N,+ refers to the distance between the quinoline ring N and the protonated side-
chain ring N.

quinoline series [3] and gives support to our proposed
mode of interaction involving the ring nitrogen of
quinoline (or its isostere) and the side-chain nitrogen.
The tetrahydroindolo[3,2-dlbenzazepines (4) are
generally less active than the other 2 ring systems.
This may be attributed to the non-aromatic ring B of 4
in which being non-planar would weaken hydropho-
bic bonding involving this moiety and the flat area at
the proposed receptor site.
Table II. Degree of planarity for compounds 2b, 3b, and
4b.
Compound
2b -0.06”
3bb 41.31”
4b 22.36”
%4n appropriate torsion angle is chosen to best indicate the
angle of twist for each molecule; bconformer B.
The indolo[3,2-clquinolines 2 and anilinoquino-
lines 3 are found to be active against sensitive and
resistant isolates of P falciparum. It would appear that
the conformational characteristics and overall plana-
rity of these 2 ring systems are not critical to activity.
However, these factors may be important insofar as
they result in an optimum N.. .N+ distance for effec-
tive interaction with the receptor. As seen from table I,
the N,. . .N,+ distances of these compounds do not
deviate by more than 0.5 A from those of amodia-
quine and chloroquine. In contrast, the less active
tetrahydroindolo[3,2-d]benzazepines 4, which are
conformationally intermediate between the indolo-
[3,2-c]quinolinesO and anilinoquinolines, have N,. . .N,+
distances (9.5 A) which are longer than those of
amodiaquine and chloroquine.
It is interesting to note that although the indolo-
[3,2-clquinolines have N,. . .N,+ distances comparable
to that of amodiaquine (conformer B), they are in fact
structural analogues of conformer A of amodiaquine.
The nature of the side chain exerts a subtle effect
on antimalarial activity. The monobasic diethylamino-
methyl side chain, present in amodiaquine, appears to
 111

Table III. In vitro antimalarial activity of 2a-b, 3a-b, 4a-b, chloroquine (CQ) and mefloquine (MF) against different isolates
of P falciparum.
Concentration (picomoles) of drug in well (50 ~1) required to inhibit schizonts

Category 1 isolatesa CQ MT 2a 2b 2c 2d 3a 3b 3c 3d 4a 4b 4c 4d
Sensitive to CQ and MF
1 2 157 <28 <26 48 39 < 28 26 < 24 2000 476 320 300
2 4” 8 250 28 < 26 24 312 28 52 < 24 597 - - -
3 2 1000 <28 < 26 96 156 ~28 26 < 24 1200 - - -
4 : 0.5 1300 < 28 <26 48 78 28 207 < 24 -
2 02532 05 - - - - - - - - - 476
238 1300
84 2400
373

7 42--------- 952 804 1500

Category 2 isolate&
Resistant to CQ and MF
8 128 2000 < 28 415 192 500 28 < 26 200 1000 - - -
9 6”: 128 2000 < 28 26 192 312 28 170 48 1200 - - -
10 64 128 1000 <28 <26 96 623 < 28 < 26 24 120 - - -
11 z: 128 500 28 104 48
12 128 1000 114 52 96 628 226 827 2500 960 238 101 746

Resistant to CQ sensitive
to MF
13 16 0.5 - - - - - - - - - 59 50 187
14 i; 8 - - - - - - 119 101 <23
15 32 < 28 170 24 1 190 320 373
16 “6: 2 2000 28 - 192 312 <;S <26 -24 500 - - -
17 2 2000 < 28 - 96 623 ~28 < 26 24 120 - - -
acategory 1 isolates (l-7) were freshly obtained from venous blood of infected patients and tested immediately. Category 2
isolates (8-14) were established in the laboratory by continuous in vitro culture. bIf schizonts survived in 8 picomoles or more
of chloroquine per test well (50 pl), the isolate is said to be resistant to chloroquine. The same threshold value for mefloquine is
64 picomoles per well.

be optimum for both indolo[3,2-clquinolines
anilinoquinolines. Dibasic side chains such as c and d
did not give rise to improved efficacy as originally
intended.
An interesting compound to take note of is 3d,
which has a dibasic side-chain and is active against
several isolates. Compound 3d, in conformer A, is the
only compound in the present study which has a4basic
nitrogen (NJ whose N,. . .N,+ distance at 14.15 A is
close to the N,. . .N,+ distance (14.24 A) of 15. Com-
pound 15, an active antimalarial [ 131, has only one
side-chain nitrogen (N5) of sufficient basicity to be
protonated at physiological pH, unlike 3d which has 2
(N3 and N4). One wonders which of the 2 nitrogens of
3d is active, since N, (conformer B) is analogous to
the side-chain nitrogen of amodiaquine (conformer B)
whereas N, (conformer A) is analogous to N5 of 15.
The overall significance of this work is the obser-
vation that there are a number of compounds (for
example 2b, 2c, 3b, and 3c) which are more effective
than chloroquine or mefloquine when tested against
isolates resistant to these drugs. It indicates that struc-
and tural modification of existing antimalarials may still
be a viable approach to the design of new compounds
against drug-resistant plasmodia.
Conclusion
Using amodiaquine as a lead compound, a series of
indolo[3,2-clquinolines, anilinoquinolines and tetra-
hydroindolo[3,2-d]benzazepines have been designed
as potential antimalarial agents. These compounds
differ in the conformational planarity of the key nuclei
as well as the distance between the side-chain nitro-
gen and ring nitrogen functions. The latter (N...N+)
distance have been proposed to be an important deter-
minant of activity in amodiaquine. In vitro evaluation
of these compounds against various isolates of
P falciparum showed that conformational planarity
per se was not an important determinant of activity. It
was, however, important in determining the distance
between the 2 key nitrogen atoms of the side-chain
and the ring. Where the distance was greater than that
112

observed in amodiaquine, as observed in the tetra- General procedure for the preparation of hydrazines 10~ and 1Od
hydroindolo[3,2-dbenzazepines, antimalarial activity
was low. Hydrazines 1Oc and 10d were prepared from arylamines SC and
Sd according to the standard procedure [4]. They were unstable
in air and were always freshly prepared before use.
Experimental protocols
l-(5’-Hydrazino-2-methoxybenzyl)-3-dimethylaminopyrrolidine
Unless otherwise stated, materials were obtained from com- 1oc
mercial suppliers and used without further purification. Melting IR (neat): 3325, 3200, 1620 cm-r. NMR (CDCl,) 6: 1.02-2.83
points were determined with a Gallemkamp melting point (m, pyrrolidinyl H, N(CH,),), 3.18 (s, NH), 3.52 (s, Ar-CH3),
apparatus and were uncorrected. IR spectra were obtained in 3.67 (s, OCH,), 6.32-7.17 (m, aryl H).
pressed KBr discs on a Jasco IR-810 spectrophotometer. rH-
NMR spectra were recorded on a FT Jeol FX 90Q (90 MHz)
instrument and chemical shifts were reported as 6 relative to 5-Hydrazino-2-metho~-[N-(3’-dimethylaminopropyl)-N-methyl]-
tetramethylsilane when taken in deuterated organic solvents, or benzylamine 1 Od
sodium 3-(trimethylsilyl)propanesulfonate in the case of D,O IR (neat): 3350, 3225, 1635 cm-l. NMR (CDCl,) F 1.43-2.53
solutions. Mass spectra were determined on a Micromass 7035 (m, N-CH,, N-(CH,),-N), 3.37 (s, Ar-CH,), 3.65 (s, OCH,),
E double focussing mass spectrometer. Thin-layer chromato- 6.50-7.18 (m, aryl H).
graphy (TLC) was carried out on 5 x 20 cm Kieselgel G type
60 (Merck) plates. Elemental analyses (C, H, N, Cl) were 7-Chloro-4-[4f-methoxy-3~-(3”-dimethylaminopyrrolidin-I”-yl)-
performed on a Perkin-Elmer Auto-Analyser 240. methylJanilinoquinoline 3c
General method for the preparation of nitro amines 7c and 7d To a solution of SC trihydrochloride (1.26 g, 0.0035 mol) in
50 ml H,O was added 0.7 g (0.0035 mol) 4,7-dichloroquinoline
2-Chloromethyl-4-nitroanisole 6 [7] (5g, 0.026 mol) and an 12. The pH was adjusted to 3-4 using concentrated HCl and
excess (0.135 mol) of the appropriate amine (dimethylamino- the mixture heated on a water bath for 2 h, stirred at room
pyrrolidine for 7c and N,Wfl-trimethylpropanediamine for 7d) temperature overnight, and filtered. The precipitate was stirred
were retluxed for 18 h in methanol, after which the solvent was with excess concentrated ammonia, filtered and dried, yielding
removed by evaporation under reduced pressure. The residue 1.24 g of 3c (86% yield). Recrystallization from benzene gave
was dissolved in ether and washed with water until neutral. On 0.7 g of white solid (48% yield), mp 150-151.5”C. IR: 3400,
drying over anhydrous Na,S04, and removal of the solvent 1610 cm-l; NMR (CDC&) 1.28-2.92 (m, pyrrolidinyl H, N-
under reduced pressure, an oil was obtained. It was dissolved in CH,), 3.58 (s, Ar-CH,), 3.75 (s, OCH,), 6.50-8.23 (m, aryl H);
dry ether and acidified by dropwise addition of freshly pre- MS m/z 410.1869 (talc for C,,H&lN,O 410.1873). Anal talc
pared ethanolic HCl which precipitated the product as the HCl for C,3H,CIN40: C, 67.22, H, 6.62; N, 13.63; Found: C, 67.23;
salt. H, 6.58; N, 13.77.
2-[(3’-Dimethylaminopyrrolidin-l’-yl)methyl]-4-nitroanisole 7c
The HCl salt was obtained in 51% (4 g) yield, mp 192- 7-Chloro-4-14’-methoxy-3’-[N-(3”-dimethylaminopropyl)-N-
193.5”C, after recrystallization from ethanol/ether. IR: 2450, methylaminomethylJ}anilinoquinoline 3d
1650, 1590, 1510, 1335 cm-i; NMR (CD,OD) 6: 2.83 (s,
N(CH,),), 2.82 (s, Ar-CH,), 3.99 (s, OCH,), 2.08-3.01 (m, A mixture of Sd trihydrochloride (2.04 g, 0.0056 mol) and 12
pyrrolidinyl H, N-CH3), 7.12-8.27 (m, aryl H). MS m/z (1.12 g, 0.0056 mol)-was reacted together as described for the
279.1578 (talc for C,,H,,N303 279.1582). Anal talc for svnthesis of 3c: 2.12 e crude 3d was obtained (91% vield)
C4HZ1N303: C, 60.20; H, 7.58; N, 15.04. Found: C, 60.03; H, Which was recrystallizevd from benzene to give white feathery
7.61; N, 14.96. crystals mp 154.6-155°C. IR 3400, 1610 cm-l; NMR (CDCl,)
6: 1.42-3.42 (m, N(CH,), N-(CH,),-N), 3.45 (s, Ar-CH,), 3.77
2-[N-(3’-Dimethylaminopropyl)-N-methylamino]methy1-4-nitro- (s, OCH,), 6.50-8.33 (m, aryl H). MS m/z 412.2039 (talc for
anisole 7d C,,H,,ClN,O 412.2030). Anal talc for C2,H,,CIN,0: C, 66.90;
The HCl salt (3.4 g, 38.7%) was obtained after recrystallization H, 7.08; N, 13.57. Found: C, 66.62; H, 7.07; N, 13.42.
from ethanol/dry ether giving mp 219-220°C. IR: 2650, 2460,
1608, 1590, 1510, 1340 cm-i; NMR (CD,OD) 6: 2.85 (s, N- 3-Chloro-8-methoxy-9-[3 ‘-dimethylaminopyrrolidin-I ‘-yl]-
CH3), 2.92 (s, N(CH,),), 4.03 (s, OCH,), 4.47 (s, Ar-CH,),
7.15-8.37 (m, aryl H). MS m/z 281.1740 (talc for C1,HZsN30, methyl-IlH-indolo[3,2-clquinoline 2c
281.1740). Anal talc for C,,H2,N,03.2HCl: C, 47.46; H, 7.11;
N, 11.86. Found: C, 47.35; H, 7.10; N, 11.95. To a solution of 2.29 g (0.0126 mol) 4-keto-7-chloro-1,2,3,4-
tetrahydroquinoline 1118j and 3.71 g. (0.0141 mol) 1Oc in 13.4
General procedure for the preparation of arylamines SC and 8d ml absolute ethanol was added 2.9 ml concentrated HCl. The
mixture was refluxed for 18 h. On cooling, 2.31 g of crude 2c
Each nitro amine (7c, 7d) (0.0098 mol) was dissolved in etha- hydrochloride was obtained (34.4% yield). The HCl salt had
nol (100 ml) and hydrogenated at 50 psi, 29°C in a Parr hydro- mp 225-228°C after recrystallization from ethanol. The free
genator using PtO, (10% by weight) as a catalyst. When the base was recrvstallized from benzene giving mn 253°C. IR
uptake of hydrogen had ceased, the catalyst was filtered and the (base): 3425, j620 cm-i; NMR (HCl sait in D,Oj 6: 2.84 (m,
filtrate acidified with ethanolic HCl. Removal of solvent under N-CH2-CH,), 3.13 (s, N-CH& 4.0 (s, OCH,), 4.65 (s, AI-CHJ,
reduced pressure gave the arylamine as the HCl salt which 7.11-8.90 (m, aryl H). MS m/z. 408.1715 (talc for
was used without further purification for the next stage of C,,H2,C1N40 408.1717). Anal talc for C23H,,C1N40: C, 67.56;
reaction. H, 6.16. Found: C, 67.25; H, 6.20.

3-Chloro-8-methoxy-9-[N-(3~-dimethylaminopropyl)-N-methyl-
aminomethyl]-IIH-indolo[3,2x]quinoline 2d
A mixture of 11 (0.61 .g, 0.0034 mol) and 10d (1.01 g, 0.0038
mol) were reacted m-3.6 ml absolute ethanol and 0.8 ml
concentrated HCl as described for the svnthesis of 2c. Crude
2d trihydrochloride (0.73 g) was obtained in 41.3% yield, mp
256°C after recrystallization from ethanol. The free base was
recrystallized from benzene giving white feathery crystals, mp
258-261°C. IR (base): 3400, 1620 cm-l; NMR (HCl salt in
D,O) 6: 2.47 (m, CH,CH,CH& 2.99 (s, N-CH,), 3.13 (s, N+-
CH,), 3.44 (m, N-CH, and A&H*), 4.07 (s, OCH,), 7.33- 9.12
(m, aryl H). MS m/z 410.1871 (talc for C23H27C1N40
410.1873). Anal talc for CZ3H,,C1N,0: C, 67.22; H, 6.62; N,
13.63. Found: C, 67.11; H, 6.59; N, 13.46.
3-Chloro-9-metho~y-10-[(3’-dimethylaminopyrrolidin-l’-yl)-
methyl]tetrahydroindolo[3,2-dlbenzazepine 4c
A mixture of S-chloro-2,3,4,5-tetrahydro-1-benzazepin-5-one
13 [6] (0.89 g, 0.00455 mol), 1Oc (1.67g, 0.0063 mol), 20 ml
ethanol and 5 ml concentrated HCl was refluxed for 18 h. On
cooling, 1.19 g (49% yield) of crude 4c HCl was obtained as a
light-pink solid. It was treated with 25% NaOH to give 0.79 g
of the free base which was purified by column chromatography
(neutral alumina) with hexane/chloroform. The light-yellow
flakes (mp 86-88°C) obtained from the column were-converted
to the HCL salt (mn 223°C) bv fractional nrecinitation from
ether with ethanohcl HCl. Id (base): 3375, ‘1606, 1480 cm-t;
NMR (base in CDCl,) 6: 1.75 (m, CH,-CH,-CH,), 2.08 (s, N-
CH,), 2.27-2.80 ( m, pyrrolinidinyl CH,), 3.10 (m, N-CH,-
CH,), 3.33 (m, N-CH,), 3.58 (s, Ar-CH,), 3.78 (s, OCH,), 4.38
(s, NH), 6.58-7.45 (m, aryl H), 8.48 (s, NH). MS m/z 424.2030
(talc for C2,H&1N,0 424.2030). Anal talc for
C2,H,,C1N40.2HC1: C, 57.89; H, 6.28; N, 11.25. Found: C,
57.66; H, 6.32; N, 11.04.
3-Chloro-9-methony-lO-[N-(3’-dimethylaminopropyl)-N-
methylaminomethyl]tetrahydroindolo[3,2-d]benzazepine
A mixture of 0.77 g (0.0039 mol) 13, 1.50 g (0.0056 mol) lOd,
15.8 ml ethanol and 3.9 ml concentrated HCl was reacted as
described for the synthesis of 4c. The precipitated solid was
treated with 25% NaOH to give 1.0 g (48.8%) of 4d base (mp
88°C). Column chromatography of the base through neutral
alumina with hexane/chloroform gave a yellow solid which on
fractional precipitation from ether with ethanolic HCl gave 4d
HCl (mp 190°C). IR (base); 3375, 3250, 1600, 1500 cm-l.
NMR (base in CDClJDMSO) 6: 1.72-2.55 (m, N-CH,, CH,),
3.08 (m, N-CH,-CH,), 3.35 (m, N-CH2), 3.55 (s, Ar-CH,), 3.80
(s, OCH,), 4.78 (s, NH), 6.65-7.50 (m, aryl H), 9.32 (s, NH).
MS m/z 426.2191 (talc for &,H,,ClN,O 426.2186). Anal talc
for C&H,,ClN,O: C, 67.51; H, 7.32: Cl, 8.30. Found: C, 67.20;
H, 7.36; Cl, 8.24.
Conformational studies
The minimum energy conformation of each compound was
determined using an interactive molecular modelling program,
PC Model Version 4 [ 111, which incorporates MMX force field
113
for molecular mechanics calculations. As the compounds were
tested at physiological pH, their degree of protonation at pH
7.4 was considered. The pK, of the ring-nitrogens of he
indolol3.2-clauinoline and tetrahvdroindolo13.2-dbenzazevine
nuclei-in 2a-&rd 4a were found to be 3.99 ilb] and 2.62 1141
respectively. Therefore, they are expected to exist in the non-
protonated state at pH 7.4 in the series 2 and 4 compounds. As
explained earlier, the unprotonated quinoline nitrogen of the
series 3 compounds was considered as the bioactive form.
Intramolecular hydrogen bonding was taken into account
wherever possible.
Evaluation of in vitro antimalarial activity
The test compounds were evaluated against various P falcipa-
rum isolates using an in vitro microtechnique [ 151 with chloro-
quine and mefloquine as controls.
A stock solution (1280 log ml-t) of the hydrochloride salt of
the test compound or control was prepared in distilled water
(except for 2a, 3a and 4a which were prepared in 95% alco-
hol), sterilized by filtration and serially diluted (1280-0.5 up
ml-l) in 96-well sterile tissue culture plates using’ a solution’o7
RPM1 1640 and gentamycin (40 pg/ml). A quantity of 25 ~1 of
diluted malaria culture (erythrocytes suspended in growth
medium) was then added to each well, the plate was gently
shaken to mix the contents and incubated at 37°C in a candle
jar for 24 h. The presence of schizonts in each well was deter-
mined accordingly. Antimalarial activity was expressed as the
highest concentration of test compound in which schizonts
were observed.
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