Bis(2-aminopyridine)s, preparation method therefor and use thereof for controlling parasitic infections

Substituted bis-2-aminopyridines of formula (1), wherein Q, R1,R2,R3,R4 are as defined in the specification. A method for preparing said compounds and the use thereof as a drug and in particular as active drugs for controlling parasiticc infections within red blood cells, e.g. malaria or babesiasis. ##STR1##

The present invention relates to substituted bis(2-aminopyridine)s, to a 
process for preparing them and also to the applications of 
bis(2-aminopyridine)s, substituted or otherwise, as a medicinal product, 
in particular as medicinal products which are active against 
intraerythrocytic parasitoses such as malaria or babesiosis (or 
piroplasmosis). 
Antimalarial and antibabesiosis activity is understood to mean the capacity 
to prevent the development of the parasite inside the erythrocyte and/or 
erythrocytic invasion, and to bring about the death of the parasites 
initially present. 
Malaria or paludism remains the most important parasitic disease rampant in 
the intertropical regions. The World Health Organization estimates that 
there are some 350 million cases of malaria worldwide, 90% of them in 
Africa, causing the death of 1.5 to 2.7 million people per annum. From 
1940 onwards, synthetic anti-folates such as Fansidar.RTM. 
(sulphonamide/pyrimethamine) and quinolines such as Nivaquine.RTM. 
(chloroquine) were employed to combat the disease. However, from the 1960s 
onwards, resistance appeared and necessitated the development of new 
active molecules, among them Lariam.RTM. (mefloquine) or Halfan.RTM. 
(halofantrine); nevertheless, these new molecules have also led to the 
emergence of new types of resistance, specific resistance but also 
cross-resistance. The emergence of chloroquine-resistant strains of 
Plasmodium falciparum in South-East Asia and in Latin America and the 
general expansion of these drug-resistance phenomena impose a limit on the 
effective treatment of malaria. In addition, the resistance to each of the 
new compounds, mefloquine and halofantrine, is also considerable and gives 
rise to a cross-resistance between these two substances. 
Other substances such as artemisinine and its derivatives (artemether, 
artether, artesunate and dihydroartemisinine) are also used in China and 
in South-East Asia. However, these substances have the drawback of 
possessing a short half-life and low water-solubility. 
Babesiosis, also known as piroplasmosis, is, like malaria, an 
intraerythrocytic parasitosis transmitted by tick bites, which affects 
many domestic and wild animal species and whose agent is a protozoan of 
the genus Babesia. There is thus a bovine babesiosis caused by Babesia 
bovis, Babesia bigemina, Babesia major and Babesia divergens which are 
responsible for considerable losses in cattle farming, in particular in 
developing countries. A canine babesiosis with Babesia canis and Babesia 
gibsoni, an equine babesiosis with Babesia equi and cabalii and even a 
rare but severe human babesiosis due to Babesia divergens and Babesia 
microti are also found. 
Few substances are active against Babesia, and they are generally toxic 
(imidocarb and amicarbalide). Other products prove active in vitro 
without, however, giving good results in vivo, such as tubercidin, 
tetracycline or menoctone. 
As early as 1984, McColm (Ann. Trop. Med. Parasitol., 78, (4), 345) 
stressed the increased resistance of Babesia towards classical 
antimicrobials such as tylosin, rifamycin and gramicidin D. 
Consequently, it was the Applicant's objective to provide new products 
which are active against intraerythrocytic parasites. 
The subject of the present invention is substituted bis(2-aminopyridine)s 
of general formula (I), 
##STR2## 
in which: Q represents: 
a C.sub.6 -C.sub.20 alkyl group, 
an optionally substituted aryl group, or 
a cycloalkyl group, and 
forms, between the 2 pyridine rings, a hydrocarbon linkage comprising in 
total from 6 to 34 carbon atoms, 
n is a number between 0 and 7, 
R.sub.1, R.sub.2, R.sub.3 and R.sub.4, which may be identical or different, 
represent: 
a hydrogen atom, 
an optionally substituted, linear or branched C.sub.1 -C.sub.6 alkyl group, 
an aryl group optionally substituted with one or more halogen atoms or 
C.sub.1 -C.sub.6 alkyl or C.sub.1 -C.sub.6 alkoxy radicals, 
a benzyl group, 
a thienyl group, preferably a 2-thienyl, 
a furyl group, preferably a 2-furyl, 
a halogen atom, 
an alkoxy or benzyloxy group, 
on condition that at least one of the radicals R.sub.1, R.sub.2, R.sub.3 
and R.sub.4 is not a hydrogen atom when Q represents a linkage 
(CH.sub.2).sub.m with m between 6 and 20, as well as their addition salts. 
Among the addition salts with acids, there may be mentioned those formed 
with inorganic acids such as hydrochloric, hydrobromic, nitric, sulphuric 
or phosphoric acids or organic acids such as acetic, propionic, maleic, 
benzoic, succinic, methanesulphonic, para-toluenesulphonic, fumaric and 
hydroxyethanesulphonic acids. 
The expression C.sub.6 -C.sub.20 alkyl denotes saturated or unsaturated, 
linear or branched alkyl groups. 
The expression aryl denotes, for example, a phenyl radical optionally 
substituted with one or more fluorine, chlorine or bromine atoms or alkyl 
or alkoxy radicals. 
The expression cycloalkyl can denote, for example, a cyclopropyl, 
cyclobutyl, cyclopentyl or cyclohexyl radical. 
The expression C.sub.1 -C.sub.6 alkyl denotes saturated or unsaturated, 
linear or branched alkyl groups such as, for example, methyl, ethyl, 
propyl, butyl, isopropyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, 
trifluoromethyl, vinyl and allyl groups. 
The expression alkoxy denotes C.sub.1 -C.sub.6 alkoxy groups such as 
methoxy, ethoxy, propoxy and butoxy groups. 
The expression halogen denotes a fluorine, chlorine or bromine atom. 
Although other bis(aminopyridine)s have been described, namely 
bis(4-aminopyridine)s (U.S. Pat. No. 4,206,215 and J. Med. Chem. 1984, 27, 
1457-1464) which display activity in the treatment of dental plaque, or 
bis(2-aminopyridine)s unsubstituted on the pyridine rings having 
iodination-catalyst activity (INOKUMA, Yakagaku, 1982, 31, (8), 515-19), 
the Applicant found, surprisingly, that both unsubstituted 
bis(2-aminopyridine)s and bis(2-aminopyridine)s bearing at least one 
substitution, as are defined above, possess antiparasitic activity and in 
particular an exceedingly advantageous and unexpected antimalarial and 
antibabesiosis activity; the substituted bis(2-aminopyridine)s display 
especially advantageous activity. 
In effect, the products of formula I as defined above, as well as the 
products of formula I in which R.sub.1, R.sub.2, R.sub.3 and R.sub.4 
represent hydrogen atoms, as well as their addition salts with acids, 
display advantageous pharmacological properties. They are endowed, in 
particular, with very good activity against intraerythrocytic parasitoses. 
Unexpectedly, these products block the erythrocytic proliferation of the 
parasite. In effect, these products interfere in the metabolism of the 
parasite by blocking the transport of choline, an essential precursor of 
the phosphatidylcholines which are constituents of the cell membranes and 
necessary for the proliferation of the parasites, which proliferation is 
accompanied by a considerable de novo synthesis of the phospholipids which 
are necessary for the biogenesis of the cell membranes. 
The compounds according to the invention display very strong activity in 
vitro against strains or isolates which are chemoresistant in a variety of 
ways. The activity is exerted mainly against the mature erythrocytic 
stages, the most intense phase of phospholipid synthesis. In particular, 
in murine systems infected with Plasmodium chabaudi or Plasmodium petteri, 
the compounds according to the invention eradicate the parasitaemia 
completely. 
The subject of the invention is, more especially, the products defined 
above, characterized in that, in the formula (I), R.sub.1, R.sub.2, 
R.sub.3 or R.sub.4 represents a methyl, chloro or phenyl group at 
positions 3, 4 and/or 5, and Q represents a C.sub.1 -C.sub.20 alkyl group, 
the hydrocarbon linkage between the two pyridine rings constituting a 
dodecane, a hexadecane, an octane or a decane. 
According to the invention, the products of formula I above and their salts 
may be prepared by a process which is characterized in that a derivative 
of formula (II) Hal--(CH.sub.2).sub.n --Q--(CH.sub.2).sub.n --Hal, in 
which Hal represents a halogen atom and n and Q have the general meaning 
stated above, is reacted in solution with a compound of formula (III): 
##STR3## 
in which R.sub.1, R.sub.2, R.sub.3 and R.sub.4 have the meaning given 
above, 
to obtain the product of general formula (I) in the state of the 
corresponding dihalide which, if desired, is alkalinized and converted to 
another salt of an acid. 
Under the preferred conditions of implementation of the invention, the 
process described above is carried out in the following manner: 
The reaction of a compound of formula II and a compound of formula III, 
present in a 1:2 ratio, the compound of formula II preferably being in 
slight excess, to bring about the total conversion of the aminopyridines 
(derivative of formula III), is carried out: 
in the presence of a solvent or mixture of solvents which is/are inert 
towards the reactants employed, the solvent being chosen from 
acetonitrile, aromatic hydrocarbons (toluene, xylenes), aliphatic ketones 
such as propanone, butanone and 3-methyl-2-butanone, alcohols (methanol, 
ethanol, propanol, butanol), dimethylformamide and common ethers, for 
example dioxane, monoglyme and diglyme, 
at room temperature or in the heated state (boiling point of the solvent, 
in particular), 
for a few hours to several days. 
The reaction may also be conducted without adding a solvent, provided one 
of the reactants is liquid at the reaction temperature and thus dissolves 
the other reactant. An advantage of the present process is that the 
dihydrohalide, in general hydrobromide or hydrochloride, of the desired 
bis(aminopyridine)s is obtained directly at the end of the reaction, and 
crystallizes directly in the pure state in the solvent. 
It is possible, however, to obtain the corresponding bases by displacing 
the hydrohalide ion, for example with an ion exchange resin or by 
precipitating a silver halide, and converting them to another salt of a 
pharmaceutically acceptable inorganic or organic acid. 
The collective products of formula (I) and their salts of acids which are 
acceptable from a pharmaceutical standpoint may be administered to man or 
animals as a medicinal product, in the form of pharmaceutical compositions 
comprising as active constituent an effective dose of at least one product 
of formula (I) or of an addition salt with an acid of such a compound, 
with, in addition, excipients and additives taken from those which are 
normally used in pharmacy. 
These compositions may be administered orally, parenterally or rectally, in 
particular in the form of tablets, dragees, capsules, solutions, syrups, 
emulsions or suspensions, pharmaceutical dosage forms capable of 
modulating the release of the active substance. 
Such compositions are generally administered at a dose of 0.5 to 50 mg/kg. 
Besides the foregoing arrangements, the invention also comprises other 
arrangements which will become apparent from the description which 
follows, which relates to examples of embodiment of the process which is 
the subject of the present invention. 
It should be clearly understood, however, that these examples are given 
only by way of illustration of the subject of the invention and in no way 
constitute a limitation of the latter. 
All the derivatives synthesized were subjected to an elemental analysis, 
with a maximum tolerance of 0.3% in the results obtained relative to the 
results calculated. 
Proton nuclear magnetic resonance spectra were recorded in solution in 
CDCl.sub.3 +TMS or in DMSO-d.sub.6 on a Brucker WP 200 instrument. The 
most characteristic chemical shifts are, where appropriate, shown in these 
examples. Except where specifically stated, all the products were isolated 
in the state of hydrobromides.

EXAMPLE 1 
1,1'-(1,12-dodecanediyl)bis(4-methyl-2(1H)-pyridinimine) dihydrobromide 
0.1 mol of commercial 1,12-dibromododecane is dissolved in 500 milliliters 
of methyl ethyl ketone, 0.15 mol of 4-methyl-2-aminopyridine dissolved in 
methyl ethyl ketone is introduced under argon and the mixture is left 
stirring at the refluxing temperature until completion of the reaction, 
which may be monitored by thin-layer chromatography and most generally 
takes 48 hours. After the precipitate has been filtered off and rinsed, 
the crude product is obtained in the form of its hydrobromide. The product 
may be recrystallized in a 9:1 isopropanol/methanol mixture. Beautiful 
white crystals, m.p. 206.degree. C., are thereby obtained in a yield on 
the pure product of 66%. 
As a variant, the product is prepared as follows: 
0.36 g (3.3 mmol) of 4-methyl-2-aminopyridine is dissolved in 5 ml of 
n-BuOH and refluxed under argon for 48 hours in the presence of 0.50 g 
(1.52 mmol) of 1,12-dibromododecane. After cooling of the reaction medium, 
the product crystallizes. 
It is recovered by filtration, rinsed with a little isopropanol and then 
ether and dried. 
0.30 g of salt is obtained (47% yield). 
Empirical formula: C.sub.24 H.sub.40 N.sub.4 Br.sub.2 ; molecular weight: 
436.28; .sup.1 H NMR (200 MHz, DMSO-d.sub.6): 8.27 (broad s, 4H, 
2.times.--NH.sub.2, exchangeable; 7.98-7.96 (part A of an AB system, 2H, 
12.times.H.sub.6); 6.81 (m, 4H, 2.times.(H.sub.3 +H.sub.5)); 4.08-4.05 (m, 
4H, 2.times.&gt;N--CH.sub.2 --); 2.31 (s, 6H, 2.times.--CH.sub.3); 1.23-1.63 
(m, 20H, 10.times.--CH.sub.2 --). 
EXAMPLE 2 
1,1'-(1,12-dodecanediyl)bis(3-methyl-2(1H)-pyridinimine) dihydrobromide 
0.1 mol of commercial 1,12-dibromododecane is dissolved in 500 milliliters 
of methyl ethyl ketone, 0.15 mol of 3-methyl-2-aminopyridine dissolved in 
methyl ethyl ketone is introduced under argon and the mixture is left 
stirring at the refluxing temperature until completion of the reaction, 
which may be monitored by thin-layer chromatography and most generally 
takes 48 hours. After the precipitate has been filtered off and rinsed, 
the crude product is obtained in the form of its hydrobromide. The product 
may be recrystallized in a 9:1 isopropanol/methanol mixture. Beautiful 
white crystals, m.p. 229.degree. C., are thereby obtained in a yield on 
the pure product of 38%. 
Empirical formula: C.sub.24 H.sub.40 N.sub.4 Br.sub.2 ; molecular weight: 
544.42; .sup.1 H NMR (200 MHz, DMSO-d.sub.6): 8.08 (broad s, 4H, 
2.times.--NH.sub.2 exchangeable); 8.00 (part A of an AB system, 2H, 
2.times.H.sub.6); 7.77 (m, 2H, 2.times.H.sub.5); 6.88-6.85 (part B of an 
AB system, 2H, 2.times.H.sub.4); 4.21 (m, 4H, 2.times.&gt;N--CH.sub.2 --); 
2.22 (s, 6H, 2.times.--CH.sub.3); 1.23-1.63 (m, 20H, 10.times.--CH.sub.2 
--). 
EXAMPLE 3 
1,1'-(1,12-dodecanediyl)bis(5-methyl-2(1H)-pyridinimine) dihydrobromide 
0.1 mol of commercial 1,12-dibromododecane is dissolved in 500 milliliters 
of methyl ethyl ketone, 0.15 mol of 5-methyl-2-aminopyridine dissolved in 
methyl ethyl ketone is introduced under argon and the mixture is left 
stirring at the refluxing temperature until completion of the reaction, 
which may be monitored by thin-layer chromatography. After the precipitate 
has been filtered off and rinsed, the crude product is obtained in the 
form of its hydrobromide. The product may be recrystallized in a 9:1 
isopropanol/methanol mixture. The derivative of the title is obtained in a 
40% yield, having an m.p. of 221.degree. C. 
Empirical formula: C.sub.24 H.sub.40 N.sub.4 Br.sub.2 ; molecular weight: 
544.42; .sup.1 H NMR (200 MHz, DMSO-d.sub.6): 8.08 (broad s, 4H, 
2.times.--NH.sub.2 exchangeable); 8.00 (m, 2H, 2.times.H.sub.6); 7.70 
(part A of an AB system, 2H, 2.times.H.sub.3); 6.80 (part B of an AB 
system, 2H, 2.times.H.sub.4); 4.20 (m, 4H, 2.times.&gt;N--CH.sub.2 --); 
1.2-1.63 (m, 20H, 10.times.--CH.sub.2 --). 
EXAMPLE 4 
1,1'-(1,12-dodecanediyl)bis(6-methyl-2(1H)-pyridinimine) dihydrobromide 
The reaction is performed under the conditions described above, starting 
from 6-methyl-2-aminopyridine. The product is purified by 
recrystallization in isopropanol. The derivative of the title, m.p. 
205.degree. C., is obtained in a 23% yield. 
EXAMPLE 5 
1,1'-(1,12-dodecanediyl)bis(5-phenyl-2(1H)-pyridinimine) dihydrobromide 
The procedure is as before, with dibromododecane and 
2-amino-5-phenylpyridine. The product obtained is purified by 
recrystallization in methanol. Its m.p.=185.degree. C. 
EXAMPLE 6 
1,1'-(1,12-dodecanediyl)bis(3-phenylmethoxy-2(1H)-pyridinimine) 
dihydrobromide 
The procedure is as above, with dibromododecane and 
2-amino-3-(phenylmethoxy)pyridine, which was synthesized by the method 
described by Rydzkowski R. in Tetrahedron Letters 26 (1985) 2571-1574. 
EXAMPLE 7 
1,1'-(1,12-dodecanediyl)bis(4-(4-methyl-phenyl)-2(1H)-pyridinimine) 
dihydrobromide 
The procedure is as above, with dibromododecane and 
2-amino-4-para-tolylpyridine, which is synthesized according to Chambron 
J.Cl. Strasbourg Tetrah. 1987 43 (5) 895/905. The product obtained is 
purified in the base state on a silica column with a benzene/ethyl acetate 
mixture. It is reconverted to the hydrobromide with a mixture of ether and 
an isopropanol solution of hydrogen bromide. It can then be 
recrystallized. 
EXAMPLE 8 
1,1'-(1,12-dodecanediyl)bis(3-n-butyl-2(1H)-pyridinimine) dihydrobromide 
2-Amino-3-n-butylpyridine is prepared according to Gassman J. Amer. Chem. 
Soc. 95 (1973).4453, and condensed as above with the corresponding 
dibromododecane. The desired product, m.p. 192.degree. C., is obtained. 
EXAMPLE 9 
1,1'-(1,12-dodecanediyl)bis(5-chloro-2(1H)-pyridinimine) dihydrobromide 
The reaction is performed under the conditions of Example 1, with 
2-amino-5-chloropyridine in methyl ethyl ketone. After recrystallizations 
in a 9:1 isopropanol/methanol mixture, the derivative of the title, m.p. 
239.degree. C., is obtained in an 18% yield. 
This product has the following empirical formula: C.sub.22 H.sub.34 N.sub.4 
Br.sub.2 Cl.sub.2. 
Molecular weight: 585.26. 
.sup.1 H NMR (200 MHz, DMSO-d.sub.6): 8.67 (broad s, 4H, 2.times.--NH.sub.2 
+exchangeable); 8.46 (m, 2H, 2.times.H.sub.6); 7.99 (part A of an AB 
system, 2H, 2.times.H.sub.3); 7.05 (part B of an AB system, 2H, 
2.times.H.sub.4); 4.09 (m, 4H, 2.times.&gt;N--CH.sub.2 --); 1.2-1.66 (m, 20H, 
10.times.--CH.sub.2 --). 
EXAMPLE 10 
1,1'-(1,12-dodecanediyl)bis(4-bromo-2(1H)-pyridinimine) dihydrobromide 
The procedure is as above, using 2-amino-4-bromopyridine (according to 
Larsen, Scott WO 93/25553) and the desired derivative, m.p. 230.degree. 
C., is obtained. 
EXAMPLE 11 
1,1'-(1,12-dodecanediyl)bis(3-chloro-5-trifluoromethyl-2(1H)-pyridinimine) 
dihydrobromide 
2-Amino-3-chloro-5-trifluoromethylpyridine, synthesized according to Haga 
Takahiro Heterocycles, 22 (1984) 1, 117-124, is used. By treatment 
according to the previous process, the derivative, m.p. 185.degree. C., is 
isolated. 
EXAMPLE 12 
1,1'-(1,8-octanediyl)bis(5-methyl-2(1H)-pyridinimine) dihydrobromide 
0.1 mol of commercial 1,8-dibromooctane is dissolved in 500 milliliters of 
methyl ethyl ketone, 0.15 mol of 5-methyl-2-aminopyridine dissolved in 
methyl ethyl ketone is introduced under argon and the mixture is left 
stirring at the refluxing temperature until completion of the reaction, 
which may be monitored by thin-layer chromatography and most generally 
takes 48 hours. After the precipitate has been filtered off and rinsed, 
the crude product is obtained in the form of its hydrobromide. The product 
may be recrystallized in a 9:1 isopropanol/methanol mixture. The 
derivative of the title is obtained in a 40% yield, having an m.p. of 
241.degree. C. 
EXAMPLE 13 
1,1'-(1,8-octanediyl)bis(4-methyl-2(1H)-pyridinimine 
0.1 mol of commercial 1,8-dibromooctane is dissolved in 500 milliliters of 
methyl ethyl ketone, 0.15 mol of 4-methyl-2-aminopyridine dissolved in 
methyl ethyl ketone is introduced under argon and the mixture is left 
stirring at the refluxing temperature until completion of the reaction, 
which may be monitored by TLC. After the precipitate has been filtered off 
and rinsed, the crude product is obtained in the form of its hydrobromide. 
The product may be recrystallized in a 9:1 isopropanol/methanol mixture. 
Beautiful white crystals, m.p. 248.degree. C., are thereby obtained in a 
yield on the pure product of 47%. 
As a variant, this product is prepared as follows: 
2 g of 2-amino-4-methylpyridine (18.5 mmol, 3 equivalents) and 6.16 mmol 
(1.7 ml) of 1,8-dibromooctane in 50 ml of butanone are introduced into a 
100-ml ground-necked Erlenmeyer equipped with a condenser denser and a 
magnetic stirrer. The mixture is heated to reflux for 36 hours and then 
left at 4.degree. C. overnight. The precipitate is filtered off and 
recrystallized in a methanol/isopropanol/ether mixture (48 h at 4.degree. 
C.). Beautiful white crystals, m.p. between 247.degree. and 250.degree. 
C., are thereby obtained in a yield on the pure product of 52%. 
.sup.1 H NMR in DMSO-D.sub.6 (.delta. in ppm): 1.25 (m, 8H); 1.65 (m, 4H); 
2.3 (s, 6H); 4.1 (t, 4H); 6.85 (m, 4H); 8.00 (d, 2H); 8.45 (s, 4H). 
EXAMPLE 14 
1,1'-(1,6-hexanediyl)bis(4-methyl-2(1H)-pyridinimine) dihydrobromide 
The reaction is performed under the conditions described above, with 
1,6-dibromohexane. After recrystallization in isopropanol, the derivative 
of the title, m.p. 253.degree. C., is obtained. 
EXAMPLE 15 
1,1'-(1,16-hexadecanediyl)bis(4-methyl-2(1H)-pyridinimine dihydrobromide 
The reaction is performed under the usual conditions, with 
1,16-dibromohexadecane, and, after recrystallization in an 
isopropanol/methanol mixture, the derivative of the title, m.p. 
167.degree.-170.degree. C., is obtained. 
As a variant, this product is prepared as follows: 
2.11 g of 2-amino-4-methylpyridine (3 equivalents) and 249 mg (0.65 mmol) 
of 1,16-dibromododecane in 5 ml of butanone are introduced into a 100-ml 
ground-necked Erlenmeyer equipped with a condenser and a magnetic stirrer. 
The mixture is heated to reflux for 36 hours; the solvent is evaporated 
off, and isopropanol is added, followed by ether, until the mixture 
becomes cloudy. The solution is left at 40.degree. for 2 days. 
The precipitate obtained is filtered off and dried in a desiccator. A 
product is thereby obtained in a 39% yield. 
.sup.1 H NMR in DMSO-D.sub.6 (.delta. in ppm): 1.2 (m, 24H); 1.65 (m, 4H) ; 
2.3 (s, 6H) ; 4.1 (t, 4H); 6.8 (m, 4H); 7.95 (d, 2H); 8.3 (s, 4H). 
EXAMPLE 16 
1,1'-(1,16-hexadecanediyl)bis(4-ethyl-2(1H)-pyridinimine) dihydrobromide 
Starting from 4-ethyl-2-aminopyridine described by Hansch and 
1,16-dibromohexadecane, the derivative of the title, m.p. 
105.degree./110.degree. C., is obtained. 
Using the method of Example 1 and starting from the various 
dimethyl-2-aminopyridines synthesized according to the literature (Dornow, 
Rohe, Chem. Ber., 93, 1960, 1093/1097; Wentrup, Mayov, J. Amer. Chem. 
Soc., 97, 1975, 7468/7477; Rao Venkateswarlu, J. Heterocycl. Chem., 12, 
1975, 731/732), the 1,1'-(1,12-dodecanediyl)bis(dimethyl-substituted 
2(1H)pyridinimine)s are obtained in the state of dihydrobromides, as 
follows: 
EXAMPLE 17 
1,1'-(1,12-dodecanediyl)bis(3,4-dimethyl-2(1H)-pyridinimine) 
EXAMPLE 18 
1,1'-(1,12-dodecanediyl)bis(3,5-dimethyl-2(1H)-pyridinimine) 
EXAMPLE 19 
1,1'-(1,12-dodecanediyl)bis(3,6-dimethyl-2(1H)-pyridinimine) 
EXAMPLE 20 
1,1'-(1,12-dodecanediyl)bis(4,5-dimethyl-2(1H)-pyridinimine) 
EXAMPLE 21 
1,1'-(1,12-dodecanediyl)bis(4,6-dimethyl-2(1H)-pyridinimine) 
EXAMPLE 22 
1,1'-(1,12-dodecanediyl)bis(5,6-dimethyl-2(1H)-pyridinimine) 
Using the same method and starting from the ethyl-2-aminopyridines 
synthesized according to the processes of the literature, such as Robison, 
J. Amer. Chem. Soc., 79, 1957, 2573/2576; Hansch, J. Org. Chem. 23, 1958, 
1924; Childress, J. Org. Chem., 23, 1958, 67; Yakhontov, Chem. Heterocycl. 
Compd., 3, 1967, 829/830, the 
1,1'-(1,12-dodecanediyl)bis(alkyl-substituted 2(1H)-pyridinimine)s are 
obtained. 
EXAMPLE 23 
1,1'-(1,12-dodecanediyl)bis(3-ethyl-2(1H)-pyridinimine) 
EXAMPLE 24 
1,1'-(1,12-dodecanediyl)bis(4-ethyl-2(1H)-pyridinimine) 
EXAMPLE 25 
1,1'-(1,12-dodecanediyl)bis(5-ethyl-2(1H)-pyridinimine) 
EXAMPLE 26 
1,1'-(1,12-dodecanediyl)bis(6-ethyl-2(1H)-pyridinimine) 
PHARMACOLOGICAL TESTS 
ANTIMALARIAL ACTIVITY IN VITRO TOWARDS PLASMODIUM FALCIUM 
Evaluation of the antimalarial activity in vitro is carried out according 
to the method of DESJARDINS, CANFIELD, HATNES and CHULAY (Antimicrob. 
Agents Chemother., 1979, 16, 710-718). A compound according to the 
invention is brought into contact for approximately 48 hours with human 
erythrocytes infected with Plasmodium falciparum. When a radioactive 
nucleic acid precursor, hypoxanthine, is added to the preparation, only 
erythrocytes infected with Plasmodium and not affected by the said 
compound incorporate this precursor. The ability or inability to 
incorporate the precursor hence reflects the viability of the 
malaria-infected cells. The results are presented in Table I below, in the 
form of an IC.sub.50 (or ED.sub.50) (concentration of compound capable of 
inhibiting in vitro the growth of the parasites by 50%). 
The derivatives according to the above examples are tested at three 
concentrations, 100, 33 and 10 nanomolar. 
Verification of the mechanism of action is carried out on the various 
derivatives according to the invention by studying the specific 
interference with phospholipid biosynthesis and measuring the biosynthesis 
of the different biomolecules, nucleic acids, proteins and phospholipids, 
on the basis of the incorporation of radiolabelled precursors, .sup.3 
H!hypoxanthine (see above), .sup.3 H!isoleucine and .sup.3 H!choline. 
The specificity in the precise realm of phospholipid metabolism is 
determined by comparison with the effect on the incorporation of .sup.3 
H!ethanolamine into phosphatidylethanolamine (Ancelin M. L., Vialettes F. 
and Vial H. J., 1991, Anal. Biochem., 199, 203-209). Under these 
conditions and among all of the derivatives described above, the compound 
of Example 1 is the one which proved to have the best specificity of 
action towards the biosynthesis of phosphatidylcholine, relative to the 
biosyntheses of the other macromolecules (nucleic acids, proteins, other 
phospholipids). 
ANTIMALARIAL ACTIVITY AND TOXICITY IN VIVO 
This activity is measured according to the test described by PETERS W. 
(Chemotherapy and Drug resistance in Malaria, 1970). A compound according 
to the invention is administered for 4 consecutive days to mice infected 
beforehand with malaria using Plasmodium vinckei, petteri or chabaudi. 
The said compound is dissolved in 0.9% aqueous NaCl solution or, where 
appropriate, in 10% aqueous gum arabic solution. The preparations are thus 
administered intraperitoneally or subcutaneously to male Swiss mice of 
average weight 30 to 40 g which have been infected beforehand 
intravenously with Plasmodium petterei or P. chabaudi (10.sup.6 infected 
cells). The compound is administered twice daily for 4 consecutive days, 
the first injection being performed 2 hours after infestation, and the 
second 10 hours after. The parasitaemia is determined by means of a smear 
on the day following the end of the treatment. 
The results are collated in Table II, in the form of an ED.sub.50 (dose of 
compound administered in vivo capable of a 50% inhibition of the growth of 
the parasites). This Table II also shows the toxicity measured according 
to the same method, the animals having received, under the conditions of 
administration used for the above tests, 2 injections daily for four days 
(semi-chronic toxicity); the results are expressed in the form of an 
LD.sub.50 (dose of compound bringing about the death of 50% of the 
animals). 
ANTIBABESIA ACTIVITY IN VITRO 
The screening method used for the antimalaria series and based on the 
incorporation of .sup.3 H!hypoxanthine by the parasite can be used to 
test the antimetabolite activity of such compounds on Babesia bovis. A 
close relationship is found between the degree of incorporation of .sup.3 
H!hypoxanthine in a standard measurement and the percentage of cells 
parasitized, determined by microscopic examination. The concentrations of 
the compounds causing a 50% inhibition of the incorporation of .sup.3 
H!hypoxanthine (ID.sub.50) are summarized in Table I below. 
TABLE I 
__________________________________________________________________________ 
Plasmodium 
Plasmodium 
Plasmodium 
Plasmodium 
Babesia 
IC.sub.50 in 
IC.sub.50 in 
IC.sub.50 in 
IC.sub.50 in 
bovis - 
vitro resp. 
vitro resp. 
vitro resp. 
vitro resp. 
activity 
Compound 
100 nM 
at 33 nM 
at 10 nM 
&lt;10 nM 
at 10 mM 
__________________________________________________________________________ 
Example 1 
+ + + + + 
Example 2 
+ + + + + 
Example 3 
+ + + + + 
Example 4 
+ + + - + 
Example 5 
+ + + - 
Example 6 
+ + + - + 
Example 7 
+ + + - - 
Example 8 
+ + + - + 
Example 9 
+ + + - - 
Example 10 
+ + + - + 
Example 11 
+ + + - + 
Example 12 
+ - - - 
Example 13 
+ + - - - 
Example 14 
+ - - - - 
Example 15 
+ + - - + 
Example 16 
+ - - - + 
Example 17 
+ + + - + 
Example 18 
+ + + - + 
Example 19 
+ + + + + 
Example 20 
+ + - - + 
Example 21 
+ + + + + 
Example 22 
+ + + - - 
Example 23 
+ - - - - 
Example 24 
- - - - 
Example 25 
+ + - - - 
Example 26 
+ + + - - 
__________________________________________________________________________ 
TABLE II 
______________________________________ 
Activity Activity 
in vitro in vivo Toxicity 
Toxicity 
IC.sub.50 ED.sub.50 mg/kg 
LD.sub.50 mg/kg 
LD.sub.50 mg/kg 
Compound 
nanomolar I.P. I.P. P.O. 
______________________________________ 
Example 1 
0.5 1.2 30 365 
Example 2 
1.8 3 20 &gt;500 
Example 3 
0.5 3 6.5 &gt;300 
Example 9 
1.4 4 10 &gt;200 
Example 23 
7.2 &gt;5 23 &gt;400 
Example 25 
3.8 &gt;5 7.5 &gt;200 
______________________________________ 
As is apparent from the foregoing, the invention is in no way limited to 
those of its embodiments and modes of implementation and application which 
have just been described more explicitly; it embraces, on the contrary, 
all the variants which may occur to the practitioner in the field, without 
departing from the scope or compass of the present invention.