CA2034224A1 - 1-amino-5-halogenouracils, process for their preparation, and central nervous system depressants containing same as active ingredient - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
Disclosed are a central nervous system depressant which comprises a 1-amino-5-halogenouracil represented by the formula [I]

wherein X represents a halogen atom, or a pharmaceutically acceptable salt thereof as an active ingredient;
a 1-amino-5-halogenouracil represented by the formula [II]

wherein X' represents chlorine, bromine or iodine, or a salt thereof; and a process for preparing a 1-amino-5-halogenouracil represented by the above shown formula [I] from a pyrimidine derivative represented by the formula [III]

wherein X represents a halogen atom and R represents a protective group.

Description

2~3~2~

l-AMINO-5~HALOGENOURACILS, PRO~ESS FOR THEIR PREPARATION, AND CENTRAL NERVOUS SYSTEM DEPRESSANTS CONTAINING SAME AS
ACTIVE INGREDIENT

TECHNICAL FIELD
The present invention relates to a central nervous system depressant containing a l-amino-5-halo~enouracil as an active ingredient, a novel l-amino-5-halogenouracil useful as a central nervous system depressant, and a novel process for preparing the 1-amino-5-halogenouracil.
BACKGROUND ART
Stress on human body tends to increase with the complication of social environment, and thus an increased number of patients complain of symptoms such as sleep duration disorder, insomnia and the like, which are considered to be induced by stress.
Although physiological phenomenon named sleep is very complicated and its mechanism has not been elucidated in detail, several nucleic acid-related compounds which affect sleep have been reported. Such compounds include the following:
(1) Uridine:
Sleep-promoting effect [see Biomed. Res., 4, 223 (1983); Neurosci. Res., 1, 243 (1984); Proc. Natl. Acad.
Sci. V.S.A., 81, 6240 (1984); Neuroscience Letters, 49, 207 (1984)];
(2) Uracil;
Hexobarbital induced sleep potentiating effect [see J. Am. Pharm. Assoc., 44, 56 (1955); ibid., 44, 550 (1955)];
~3) N3-Benzyluridine and its derivatives:
Hypnotic effect and/or pentobarbital induced sleep potentiating effect [see Chem. Pharm. Bull., 33, 4088 (1985); Japanese Patent Laid-Open Publication No.
207218/1987];
t4) N-Allyl or N-benzyl substituted derivati~es of uracil, thymine or 6-methyluracil:

2~3~24 Hypnotic effect and/or pentobarbital in~uced sleep potentiating effect [see Chem. Pharm. Bull., 35, 4982 (1987); Abstract of the Proceedings of the 108th Annual Meeting of Pharmaceutical Society of Japan, page 708 (1988)l.
It has been also reported that the N-allyl substituted derivative of uracil (Nl,N3-diallyl uracil) and uridine have anticonvulsive effect ~see Brain Res., 55, 291 (1973); Chem. Pharm. Bull., 35, 4928 tl987)]-However, these conventional compounds (i) have no hypnotic effect though they exhibit sleep-promoting effect (uridine and uracil), and (ii) must be administered in large doses ~320-752 mg/kg) to induce sleep by intraperitoneal injection, and thus they do not always have satisfactory effects.
An object of the present invention is to provide a central nervous system depressant which comprises as an active ingredient a compound exhibiting more potent central nervous system depressant effects such as 2~ hypnotic effect, sleep-promoting effect, anticonvulsive effect and the like than the conventional compounds.
DISCLOSURE OF THE INVENTION
We have conducted extensive research in order to find out a compound which has central nervous system sedative effects. As a resultl we have found that 1-amino-5-halogenouracil has excellent central nervous system sedative effects.
While l-amino-5-fluorouracil has hitherto been known as only one l-amino-5-halogenouracil, it has not been reported that the compound has central nervous system sedative effects [see Sci. Pharm., 52, 46 (1984)].
An object of the present invention is to provide a central nervous system aepressant which comprises a 1-amino-5-halogenouracil represented by the formula:

HN ~
~ ~ [I]
O N

wherein X represents a halogen atom, or a pharmaceutically acceptable salt thereof as an active ingredient.
Another object of the present invention is to provide a novel l-amino-5-halogenouracil represented by the formula:
1~ 0 Il X' ~N ~ ~ ~

1~ , lII]
N
O
NH~
wherein X' represents chlorine, bromine or iodine, or a salt thereof.
A further object of the present invention is to provide a process for preparing a l-amino-5-halogenouracil represented by the formula [I] shown abovewhich comprises reacting a pyrimidine derivative represented by the formula;
OR

3U N ~ [III]
1~ J
RO N
wherein X has the same meaning as defined above and R
represents a protective group, with an aminating agent to aminate the l-position of the pyrimidine derivative, and removing the protective groups.

4 ~34224 BEST MODE F~R CAR~YING OUT THE INVENTION
The present invention will now be explained in detail below.
The l-amino-5-halogenouracil which is an active ingredient of the central nervous system depressant of the present invention is the compound represented by the above shown formula ~I] (referred to hereinafter simply as "active ingredient of the present pharmaceutical composition").
In the formula [I], the halogen atoms represented by X include fluorine, iodine, bromine and chlorine. In this connection, the compounds represented by the formula [I] except for one in which X represents fluorine are novel compounds.
The active ingredient of the present pharmaceutical composition may be in the form of salts and includes acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid or phosphoric acid, or with organic acids such as citric acid, acetic acid, succinic acid, maleic acid, methanesulfonic acid or p-toluenesulfonic acid.
The active ingredient of the present pharmaceutical composition can be prepared, for example, by the method in which a pyrimidine derivative represented by the following formula [III] is used as a starting compound, which is reacted with an aminating agent (Z-NH2) to give an intermediate [IV] by the specific amination of the 1-position of the pyrimidine derivative (referred to hereinafter as "amination reaction"), and the protective groups represented by R are removed ~referred to hereinafter as "deprotection reaction"):

.

2034~24 OR X OR

1~ ~ Z-NH
RO N RO

[III] NH2 [IV]

HN

O N
NH2 ~I]
wherein X has the same meanings as defined above, R
represents a protective group and Z~NH2 represents an aminating agent.
The halogen atom represented by X in the starting compound may be selected appropriately so that it corresponds to X in the desired compound represented by the formula [I]~ The protective groups represented by R
include silyl groups such as trimethylsilyl, t r i e t h y 1 s i 1 y 1 , t - b u t y 1 d i m e t h y 1 s i 1 y 1 , methyldiisopropylsilyl and triisopropylsilyl, and alkyl groups such as methyl and ethyl. Particularly, the silyl groups are advantageously used.
The starting compound can be prepared by introducing 3~ the protective groups represented by R into a 5-halogenouracil according to a conventional method. For exampler when a silyl group is used as the protective group, protection can be accomplished by using 2- to 10-fold moles of the silylating agent in proportion to 1 mole of the 5-halogenouracil and reacting the mixture in a reaction solvent such as pyridine, picolinel diethylaniline, dimethylaminopyridine, dimethylformamide, 6 203~22~

acetonitrile, tributylamine, triethylamine or the like, which may be used alone or in admixture thereof, at a reaction temperature in the range of 0 to 50C for 1 to 30 hours.
After introducing the protective groups, the pyrimidine derivative represented by the above formula [III] is isolated and purified, if necessary, by a conventional isolation and purification means for nucleic acid bases such as distillation, adsorption chromatography with silica gel or the like, or recrystallization, and then subjected to amination reaction as the startin~ compound.
The aminating agent (Z-NH2) used for the amination reaction includes hydroxylamines. Specifically, there can be mentioned o-arylsulfonylhydroxylamines such as o-mesitylenesulfonylhydroxylamine, o-(2,4,6-triisopropylbenzenesulfonyl)hydroxylamine, and o-(2-nitrobenzenesulfonyl)hydroxylamine, o-nitrophenylhydroxylamines such as o-(2,4-dinitrophenyl)hydroxylamine and o-picrylhydroxylamine, o-mesitoylhydroxylamine and the like.
The amination reaction with these aminating agents can be accomplished by using 1- to 5-fold moles of an aminating agent, preferably 1- to 1.5-fold moles in proportion to 1 mole of the starting compound and reacting the mixture in a reaction solvent (a haloyenated hydrocarbon such as dichloromethane, dichloroethane, or chloroform, an ether type sol~ent such as tetrahydrofuran, or dioxane, or an aromatic hydrocarbon such as benzene, toluene, or xylene, which may be used alone or in admixture thereof) at a reaction temperature in the range of 0 to 50~C, preferably 0 to 30~C for 1 to 10 hours.
After the amination reaction, the intermediate [IV]
is subjected to deprotection reaction to obtain the 203~22~

active ingredient of the present pharmaceutical composition.
The deprotection reaction may be conducted according to any conventional method for the deprotection of the protective group used. For example, when a silyl group is used as the protective group, it can be removed by ammonium fluoride treatment or acidic or alkaline hydrolysis. Also, when an alkyl group such as methyl or ethyl is used, the alkyl group can be removed by acidic or alkaline hydrolysis.
The active ingredient of the present pharmaceutical composition thus obtained can be isolated and purified by an appropriate combination of conventional isolation and purification means, for example, chromatography such as adsorption chromatography or recrystallization which are applied to the isolation and purification of nucleic acid bases.
While the dose of the compound represented by the formula [IJ as the active ingredient of the present pharmaceutical composition depends on many factors such as the severity of patients or acceptabilities to the composition and finally should be determined by the judgement of doctors, it is generally in the range of 0.05 to 2 g per day for an adult patient, which is administered once or in portions. The route for administration of the composition may be any of appropriate routes such as oral or parenteral administration.
For oral administration, the compGsition may be in the form of a solid preparation such as powder, granules, capsules or tablets or a liquid preparation such as syrup or elixir. For parenteral administration, the composition may be in the form of injection, suppository, agent for external application or for inhalation. These preparations are prepared according to a conventional method with the addition of a pharmaceutically acceptable preparatory aid to the active ingredient of the present ~34~4 pharmaceutical composition. It is also possible to formulate the composition into a sustained release preparation by a well-known technique.
In the production of the solid preparation for oral administration, the active ingredient of the present pharmaceutical composition is mixed with an excipient such as lactose, starch, crystalline cellulose, calcium lactate, calcium monohydrogenphosphate, magnesium aluminometasilicate or anhydrous silicic acid to give a powder, or, if necessary, the powder is further mixed with a binding agent such as white sugar, hydroxypropylcellulose or polyvinylpyrrolidone, or a disintegrating agent such as carboxymethylcellulose or carboxymethylcellulose calcium for wet or dry granulation to give granules. In the production of tablets, these powders or granules, if necessary, mixed with a lubricant such as magnesium stearate or talc may be punched into tablets. Alternatively, these granules or tablets can be coated with an enteric bas e su ch a s hydroxypropylmethylcellulose phthalate or a methyl methacrylate copolymer to give enteric-coated preparations, or they can be coated with ethylcellulose, carnauba wax or a hydrogenated oil to give sustained release preparations. Further, in order to prepare capsules, powder or granules may be charged into hard capsules, or the active ingredient of the present pharmaceutical composition is first dissolved in glycerol, polyethylene ~lycol, sesame oil, olive oil or the like and next coated with a gelatin film to give soft capsules.
In order to prepare the liquid preparation for oral administrationl the active ingredient of the present pharmaceutical preparation and a sweetener such as white sugar, sorbitol or glycerol may be dissolved in water to give a clear syrup, or the syrup may be further mixed with an essential oil or ethanol to give an elixir or with gum arabic, tragacanth gum, polysorbate 80, or .

- . ~:

. ~ .

203422~

carboxymethylcellulose sodium to give an emulsion or a suspension. These liquid preparations may also contain flavoring agents, colorants, preservatives or the like, if desired.
In order to pxepare the preparation for injection, the active ingredient of the present pharmaceutical composition may be dissolved in distilled water for injection, if necessary, together with a pH adjusting agent such as sodium hydroxide, hydrochloric acid, lactic acid, sodium lactate, sodium monohydrogenphosphate or sodium dihydrogenphosphate, and an isotonizing agent such as sodium chloride or glucose, aseptically filtered and charged into ampoules, or these solutions may be mixed with mannitol, dextrin, cyclodextrin, or gelatin and lyophilized under vacuum to give injections which should be dissolved on use~ Furthermore, the active ingredient of the present pharmaceutical composition can be mixed w i t h l e c i t h i n , p o l y s o r b a t e 8 ~ , o r polyoxyethylenehydrogenated castor oil, and the mixture is emulsified in water to give an emulsion for injection.
In order to prepare the preparation for rectal administration, the active ingredient of the present pharmaceutical composition may be melted by heating together with a suppository base such as txi-, di- or mono-glycerides o~ cacao fatty acid or polyethylene glycol, poured into a mold and cooled, or the active ingredient of the present pharmaceutical composition may be dissolved into polyethylene glycol or soybean oil and coated with a gelatin film.
In order to prepare the preparation for external application, the active ingredient of the present pharmaceutical composition is added to white vaseline, beeswax, liquid parafin or polyethylene glycol and the mixture is kneaded, i necessary, under heat to give an ointment, or it is kneaded with an adhesive such as rosin or an alkyl acrylate polymer and then spread over 20~422~

~onwoven fabrics made of, for example, polyeth~lene to give a tape preparation.
The present invention will be illustrated below with reference to Synthesis Examples, Test Examples and Preparation Examples.
SYnthesis Example 1 l-Amino-5-fluorouracil 5-Fluorouracil was silylated as usual with hexamethyldisilazane, and then 8.8 g (32 mmole) of 2,4-ditrimethylsilyloxy-5-fluoropyrimidine thus obtained, distilled and purified was dissolved in dichloromethane (48 ml). To the solution was added 7.5 g (35 mmole) of mesitylenesulfonylhydroxylamine (MSH) under ice-cooling, and the mixture was reacted with stirring at room temperature for 4 hours.
After reaction, the reaction solution was concentrated under reduced pressure. Distilled water (200 ml) was added to the residue, and oily impurities were extracted with chloroform (50 ml) from the mixture.
2Q The aqueous phase was neutralized with a weakly basic resin and then concentrated to dryness under reduced pressure.
The crude crystal thus obtained was purified by sublimation under the conditions of 150C and 5 mmHg to give 2.9 g (yield, 62%) of 1-amino-5-fluorouracil.
The product was further recrystallized from 5Q%
ethanol to give colorless needle crystals.
M.P.: 196-199C (lit. 205-207C) Elementary analysis for C4H4N3O2F:
Calculated (%): C, 33.11; H, ~.78; ~, 28.96 Found (%): C, 33.15; H, 2.79; N, 28.78 Synthesis Example 2 l-Amino-5-bromouracil After the silylation of 5-bromouracil conducted in the same manner as in Synthesis Example 1, 8.8 9 t26 mmole) of 5-bromo-2,4-ditrimethylsilyloxypyrimidine distilled and purified was dissolved in 50 ml of 2~3~22~

dichloromethane. To the solution was added 6.7 g (31 mmole) of MSH under ice-cooling, and the mixture was reacted with stirring at room temperature for 4 hours.
After reaction, the reaction solution was concentrated under reduced pressure. To the residue were added distilled water (200 ml) and subsequently 2N-sodium hydroxide to adjust the pH to 6.0, and the solution was concentrated under reduced pressure.
The crude product thus obtained was collected by filtration and recrystallized from 50% ethanol to give 3 ~ (yield, 55~) of crystalline 1-amino~5-bromouracil.
M.P.: 214-215C
Elementary analysis for C4H4N3O2Br:
Calculated (~): C, 23.32; H, 1.96; N, 20.40 Found (%): C, 23.59; H, 1.97; N, 20.14 NMR spectrum (~, ppm, DMSO-d5):
5.54 (2H, s, NH2, disappeared by the addition of D20 ) 8.14 (lH, s, 6-H) 11.84 (lH, s, NH, disappeared by the addition of D20 ) Synthesis Example 3 l-Amino-5-chlorouracil Starting from 5-chlorouracil, 1-amino-5-chlorouracil was obtained in the same manner as in Synthesis E~ample ~.
M.P.: 224-225C (recrystallized from water) Elementary analysis for C4H4N3O2Cl:
Calculated (%): C, 29.74; H, 2.50; N, 26.01 30 Found (%): C, 29.83; H, 2.55; N, 25.93 NMR spectrum l~, ppm, DMSO-d6):
5.54 (2H, s, NH2, disappeared by the addition of D20) 8.09 (lH, s, 6-H) 11.86 (lH, s, disappeared by the addition of D2O) Synthesis Example 4 l-Amino-5-iodouracil 2~3~224 Starting from 5-iodouracil, 1-amino-5-iodouracil was obtained in the same manner as in Synthesis Example 2.
M.P.: 195-196C (recrystallized from water) Elementary analysis for C4H4N3O2I:
Calculated (%): C, 18.99; H, 1.59; N, 16.61 Found (%): C, 19.05; H, 1.57; N, 16.51 NMR spectrum (~, ppm, DMSO-d6):
5.51 (2H, s, ~H2, disappeared by the addition of D20 ) i3.08 (lH, s, 6-H) 11.69 (lH, s, di,sappeared by the addition of D2O) Test Example 1 Hypnotic effect The following compounds suspended in a physiological saline solution containing 0.5% carboxymethylcellulose were administered to ICR mice (male).
The time from the loss of righting reflex until the recovery of it (sleeping time) was measured. The results are shown in Table 1.
Table 1 Dose Route of No. of Mean Compound (mg/kg) Admini- Animals Sleep ng 1-Amino-5- 100 ip 2 53 fluorouracil l-Amino-5- 100 P 3 21 fluorouracil _ l-Amino-5- 112 ip 2 80 bromouracil l-Amino-5- 200 P 1 14 chlorouracil * ip = intraperitoneal; po = peroral Test Example 2 Anticonvulsive efect a) Thiosemicarbazide induced convulsion :
'~ .

13 203~22~

Thiosemicarbazide (20 mg/kg) was administered intraperitoneally to ICR mice (male), and 30 minutes thereafter the following compounds suspended in a physiological ,saline solution containing 0.5%
carboxymethylcellulose were administered subcutaneously.
The time required for initiating convulsion (initial convulsion time) was measured. Physiological saline containing no test compounds was used as a control. The results are shown in Table 2.
Table 2 : ~
Dose No. of Initial Convulsion Compound (mg/kg) Animals Time (min) l-Amino-5-fluorouracil 50 4 80.9 + 12.9*
1-Amino~5-bromouracil 50 8 118.3 + 6~9*~
Control 4 60.3 + 6.8 *: significant at a significant level of 5% or less **: significant at a significant level of 0.5% or less b) Picrotoxin induced convulsion The following compounds suspended in a physiological saline solution containing 0.5% carboxymethylcellulose were administered intraperitoneally to ICR mice (male).
Immediately thereafter, picrotoxin was also administered intraperitoneally in the same manner, and the number of convulsions per hour and lethality of the animals within 1 hour after the administration of picrotoxin were examined. Physiological saline containing no test compounds were used as a control. The results are shown in Table 3.

2~3~22~

Table 3 Compound Dose No. of ~No. of Lethality (mg/kg) Animals Convulsions (%) l-Amino-S- 10 1.3 ~ 1.0* 20 bromouracil l-Amino-5- 20 1.8 + 1.7* 0**
bromouracil Control _ 8 5.6 + 0.9 75 *: significant at a significant level of 0.1% or less **: significant at a significant level of 1% or less Test Example 3 Acute.toxicitY
l-Amino-5-bromouracil was suspended in a physiological saline solution containing 0.5%
carboxymethylcellulose. 500 mg/kg of the compound was administered intraperitoneally to 8 ICR mice, and lethality of the animals was observed for 1 week. As a result, all of the mice were survived with no lethality.
Preparation Example 1 Tablet l-Amino-5-bromouracil 10 g Corn starch 65 g Carboxymethylcellulose 20 g Polyvinylpyrrolidone 3 g Calcium stearate 2 q Total 100 9 Tablets each weighing 100 mg are prepared in a conventional manner. Each tablet contains 10 mg of 1-amino-5-bromouracil.
Preparation Example 2 Powder and Capsule 1-Amino-5-fluorouracil 20 g Crystalline cellulose 80 q Total 100 9 15 2034~24 Both the powders are mixed into a powder preparation. Separately, 100 mg of the preparation is charged into a No. 5 hard capsule to form a capsule preparation.
INDUSTRIAL APPLICABILITY
The active ingredient of the present pharmaceutical compositionr as apparent from the aforementioned Test ~xamples, has the following characteristics as compared with conventional compounds and is very useful as a central nervous system depressant.
(i) A little dose of the active ingredient of the present pharmaceutical composition induces central nervous system depressant effects such as hypno~ic effect or anticonvulsive effect.
In order to induce hypnotic effect by intraperitoneal administration, a dose of 320 to 752 mg is required for conventional compounds. For example, the doses of 433 mg, 320 mg and 752 mg per kg body weight were required for Nl,N3-diallyluracil, Nl-methoxymethyl-N3-benzyluracil and N3-benzyl-2',3',5'-tri-o-methyluridine, respectively.
On the other hand, l-amino-5-halogenouracil, the active ingredient of the present pharmaceutical composition, can induce sleep with a dose of about 1~0 mg per kg body weight.
(ii) The active ingredient of the present pharmaceutical composition can express its activity by oral administration. Hitherto, there have been reported no nucleic acid-related substances having hypnotic effect by oral administration.
Iiii) The active ingredient of the present pharmaceutical composition is a compound which has an extremely low toxicity. ~ The simultaneous production of both the l-amino derivative and the 1,3~diamino derivative which is a defec~ in the conventional method [Sci. Pharm., 52. 46 (1984)] is suppressed by the synthesis method of the present invention~ and the 1-16 2~3~224 position of pyrimidines can be specifically aminated togive l-amino-5-halogenouracil in a high yield.

. ~ .
.

Claims (3)

1. A central nervous system depressant which comprises a 1-amino-5-halogenouracil represented by the formula [I]

wherein X represents a halogen atom, or a pharmaceutically acceptable salt thereof as an active ingredient.

2. A 1-amino-5-halogenouracil represented by the formula [II]

wherein X' represents chlorine, bromine or iodine, or a salt thereof.

3. A process for preparing a 1-amino-5-halogenouracil represented by the formula [I]
wherein X represents a halogen atom, which comprises reacting a pyrimidine derivative represented by the formula [III]

wherein X represents a halogen atom and R represents a protective group, with an aminating agent to aminate the 1-position of the pyrimidine derivative, and removing the protective groups.