DE2346334A1 - 3,5-dibenzyl-6-aminouracils - in vitro phosphodiesterase inhibitors, diuretics, bronchodilators etc. - Google Patents

Abstract

6-Aminouracil derivs. of formula (I) are new: (R1 = lower alkyl opt. substd. by one OH or alkoxy; R2 = halo, NO2 or lower alkyl; n = 0, 1 or 2). Alkyl and alkoxy are opt. unsatd. and pref. contain 1-4C. (I) have in vitro inhibitory activity against adenosine-3',5'-cyclophosphate phosphodiesterase (>100 times more active than theophylline) and also diuretic, platelet-aggregation, inhibiting and bronchodilating activities. Dose is 0.1-3 g/day.

Description

6-Aminouraci! Derivatives and process for their preparation

The invention relates to new and valuable 6-aminouracil derivatives the formula

(D

in which R is a lower alkyl radical that has a hydroxyl group or a lower alkoxy radical

2
can be, R represents a halogen atom, a nitro group or a lower alkyl radical and η represents O, 1 or 2. In-depth investigations of these compounds (I) have surprisingly shown that they have an excellent inhibitory effect against adenosine-3 ¹ , 5'-cyclophosphate phosphodiesterase and excellent pharmacological effects, for example a diuretic effect, an inhibitory effect on the agglomeration of platelets and in vitro Have bronchodilator effects.

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The invention accordingly relates to the new 6-aminouracil derivatives of the formula (I), pharmaceutical preparations which contain one or more of these compounds, and a method for producing the new 6-aminouracil derivatives (I).

The lower alkyl radical for which R in the formula (I) stands can be straight-chain or branched, saturated or be unsaturated and is advantageously an alkyl radical with up to 4 carbon atoms, e.g. a laethyl radical, ethyl radical, n-propyl radical, Isopropyl radical, allyl radical, n-butyl radical, isobutyl radical, tert-butyl radical and 2-methylallyl radical. These lower alkyl radicals can be substituted with hydroxyl groups and / or lower alkoxy radicals. Of the lower alkoxy radical is preferably an alkoxy radical with up to 4 carbon atoms, e.g. a methoxy radical, ethoxy radical, n-propoxy radical, allyloxy radical and n-butoxy radical. as Examples of suitable substituted lower alkyl radicals are ß-hydroxyethyl, γ-hydroxypropyl, ß-methoxyethyl, ß-Ethoxyethyl and ß-methoxypropyl should be mentioned.

2
The halogen atom for which R stands is chlorine, bromine,

Iodine and fluorine in question. The lower alkyl radical for which B ^ stands can be straight-chain or branched, saturated or unsaturated and has advantageous

up to 4 carbon atoms, e.g. methyl, ethyl, n-propyl, allyl, η-butyl, sec-butyl and 2-methylallyl.

2
In formula (I), R can be in any of o-, m- and p-

Position of the benzyl radical if η stands for 1,

and two substituents of R, which are the same or different, can be in any two of the o-, m- and p-positions of the benzyl radical if η is 2 stands.

The 6-aminouracil derivatives (I) can, for example, by reacting 6-aminouracil compounds of the formula

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(II)

Ir

in which R has the meaning given above, with a Compound of formula

^ 7-CH ₂ X (in)

ο
in which R and η have the meanings given above and X is a halogen atom, can be prepared in the presence of an inorganic base.

The halogen atom X in the formula (III) may be a chlorine atom, a bromine atom, an iodine atom or a fluorine atom.

The 6-aminouracil compounds of the formula (II) are known compounds and can easily be per se known processes, for example by the process described in "Journal of Organic Chemistry" 1879 ff., Is described.

The compounds of the formula (III) are also known and can be prepared by processes known per se e.g. according to the procedure described in "Journal of Organic Chemistry "1p_, (194-5) 228 ff., described will. These compounds (III) can generally be used in an amount of about 2 to 10 moles per mole of the 6-aminouracil compounds (II) can be used.

Examples of preferred inorganic bases are alkali hydroxides (e.g. sodium hydroxide, potassium hydroxide and lithium hydroxide) and alkali carbonates (e.g. potassium carbonate and sodium hydrogen carbonate). Advantageous v / ground them in aqueous solution. 509813/1065

It is also advantageous to use alkali metal alkoxides obtained by reacting monovalent alkali metals, for example sodium or potassium can be prepared with alcohols such as methanol or ethanol. In this case it can the alcohol can be used in excess, the alcohol also being effective as a solvent. in the In general, the inorganic bases are preferably in excess, in particular in an amount of about 2 to 10 molecular equivalents based on the 6-aminouracil compound (II) are used.

This reaction is generally rapid in a solvent. Aqueous alcohols, for example aqueous methanol and aqueous ethanol, and dimethylformamide can advantageously be used as solvents. The reaction is generally carried out at a ΤβπιρβΓ3ΐμΓ of about 50 to 120 ° C and to achieve better results at a temperature of about 70 ° to 90 ⁰ C.

The 6-aminouracil derivatives (I) formed in this way can easily be obtained by known methods from the Isolate and purify the reaction mixture, e.g. by extraction, recrystallization and chromatography.

The new 6-aminouracil derivatives (I) have in vitro a strong inhibitory effect on adenosine-3 ¹ , 5'-cyclophosphate phosphodiesterase (reference is made to the experiment described below) and excellent pharmacological effects, e.g. strong and long-lasting diuretic, the agglomeration of the blood platelets inhibitory and bronchodilator effects. They are therefore not only suitable as drugs, but also as reagents in biochemical tests.

The 6-aminouracil derivatives (I) can be used alone or in combination with pharmaceutically acceptable carriers administered. They are in the form of powders, tab-

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letten, solutions or emulsions orally or in the form of Injection fluids administered.

The pharmaceutical preparations containing one or more compounds according to the invention can by methods known per se which are customary for the production of powders, capsules, tablets, pills, injection liquids and the like. The choice of carrier depends on the mode of administration, the solubility of the compounds (I) and so on.

The dosage of the 6-aminouracil derivatives (I) depends, among other things, on the type and purpose of administration. A daily dose of about 0.1 to 3 g is advantageous for the Adults in the treatment of edema, thrombosis and bronchial asthma, for example.

In the following examples currently preferred embodiments of the invention described. In these Examples relate parts by weight to parts of volume as grams to cubic centimeters.

example 1

To a solution of 1.55 parts by weight of i-ethyl-6-aminouracil in 20 parts by volume of 95% ethanol there were 6 parts by volume of a 15% strength aqueous sodium hydroxide solution (15 g / 1O0 ml) and 2.4 parts by weight of benzyl chloride. * The mixture was refluxed with stirring for 3 hours and then under reduced pressure to Evaporated dry. The residue was shaken well with 50 parts by volume of chloroform and 50 parts by volume of water. The aqueous layer was then extracted with 50 parts by volume of chloroform. The chloroform layers were poured together and under reduced pressure for Evaporated dry. The residue was dissolved in 5 parts by volume of chloroform. The solution was to a with

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Abandoned column filled with 50 parts by weight of silica gel. The column was eluted with 500 parts by volume of chloroform. The fractions containing the desired compound were pooled and evaporated to dryness under reduced pressure. This gave 1.04 parts by weight of 1-ethyl-3,5-bisbenzyl-6-aminouracil in the form of a white powder.
UV absorption spectrum: ^ ^ k. ³¹¹⁰¹ 285 mu

Elemental analysis: Calculated for C ^^ H

Found:

71 C. 6th H 12th N NO: 71 , 62 6th , 31 12th , 53 2 , 69 , 43 , 35 example

To a solution of 3.4 parts by weight of i- (ß-hydroxyethyl) -6-aminouracil in 40 parts by volume of 95% ethanol, 10 parts by volume of a 16% strength (16 g / 100 ml) aqueous sodium hydroxide solution and 8.64 Parts by weight of p-nitrobenzyl bromide given. The mixture was refluxed with stirring for 15 minutes and then evaporated to dryness under reduced pressure. The ^e residue was shaken well with 50 parts by volume of chloroform and 50 parts by volume of water. The insoluble residue was washed well with boiling water and recrystallized twice from 300 parts by volume of acetone, 1.2 parts by weight of 1- (β-hydroxyethyl) -3,5-bis (p-nitrobenzyl) -6-aminouracil in the form from pale yellow needles melting at 225-227 ° C.

UV absorption spectrum : A f ^ ⁰¹¹⁰¹ 275 mu

Elemental analysis: Calculated for Cp ₀ J Found:

C. H N 54.40 4.33 15.84 54.34 4.30 16.52

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Example 5

In 100 parts by volume of 95% ethanol were 5.4-9 parts by weight i-ethyl-6-aminouracil dissolved. Became the solution 15 parts by volume of a 15% strength (15 g / 100 ml) aqueous sodium hydroxide solution and 6.48 parts by weight of p-nitrobenzyl bromide given. The mixture was refluxed with stirring for 15 minutes and then transferred to the in Example 1 described manner worked up. Here, 1.57 parts by weight of "1-Ethy 1-3,5-bis (p-nitrobenzyl) -6-aminouracil obtained in the form of pale yellow granules with a melting point of 215 to 217 ° C.

UV absorption spectrum : / [ f ^ i ^oroform 273.5 mn

Elemental analysis: H ₁₉ N ₅ O ₆ : 56 C. H 16 N Calculated for Cp ₀ 56 , 47 4.50 16 , 46 Found: example 4th , 62 4.71 , 31 •

7 parts by weight of i-ethyl-6-am.inouracil were dissolved in 80 parts by volume of 95% ethanol. 12 parts by volume of a 152 μl (15 g / 100 ml) aqueous sodium hydroxide solution and 3.64 parts by weight of p-chlorobenzyl chloride were added to the solution. The mixture was refluxed with stirring for 1 hour and then worked up in the manner described in Example 1. This gave 1-ethyl-3 _t 5-bis (p-chlorobenzyl) -6-aminouracil in the form of a syrup. This syrup was recrystallized from 5 cleared parts of ethyl ether, 1.1 parts by weight of white granules with a melting point of 157 ° C. being obtained.

UV absorption spectrum: ^ ^ ⁸¹¹⁰¹ 275 mu, 221 mu

Elemental analysis: (3 HN Eq

Calculated for C ₂₀ H ₁₉ N ₃ Cl ₂ O ₂ : 59.41 4.74 10.39 17.54 Found: 59.31 4.79 10.37 17.46

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Example 5

In 20 parts by volume of 95% ethanol were 1.55 parts by weight i-ethyl-6-aminouracil dissolved. 4 room parts were used as a solution a 15% strength (15 g / 100 ml) aqueous sodium hydroxide solution and then 2.81 parts by weight of p-methylbenzyl chloride given. The mixture was refluxed with stirring for one hour and then brought to the in Example 1 described procedure worked up. Here 1-ethyl-3,5-bis (p-methylbenzyl) -6-aminouracil in Obtained in the form of a yellow powder.

UV absorption spectrum: ^ ^A * HANOL ₂ 79.5 mu

Elemental analysis •
• 72 C. H 93 11 N Calculated for Cp ₂ H NO: 71 , 70 6, 96 11 , 56 Found:. 6th , 91 6, , 28 - example

6-aminouracil compounds of the formula (II) and compounds of formula (III) were in the presence of an inorganic base on the in the preceding examples Reaction described manner, the compounds shown in Table 1 were obtained, all of which are new and come under formula (I).

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Table 1

link

Elemental analysis

Gross Formula Calculated, Jo Found,

OO CaJ

1-isobutyl-3,5-bis-

(2,4-dichlorobenzyl) -

6-aminouracil

1- (ß-methoxyethyl) -3,5-bisb enzy1-6-aminouracil

1-allyl-3,5-bis (pm ethyIb en zyl) -6-aminouracil

1-methyl-3,5-bis (2,4-dimethylbenzyl) -6-aminouracil

UV melting

Sp ect rum point,

"\ Ethanol O _n Amax ^G ·

C 52.75 H 4.22 N 8.38 C128.32

C 69.10 H 6.34 N 11.48

C 73.65 H 6.72 N 11.18

C 73.25 H 7.22 N 11.12

C 52.49

H 4.40

N 8.21

Cl 27.96

C 69.23

H 6.12

N 11.11

C 73.62

H 6.54

N 10.95

C 73.04

H 7.11

N 10.85

283 mu

285

280 mu

282 mu

181JDis 183 C

CD CO CO

Example 7

The following are examples of the composition of practical formulations in which the compounds according to the invention for the treatment of edema, bronchial asthma, thrombosis and the like.

A. tablet

1) 1-Ethyl-3,5-bisbenzyl-6-aminouracil 20 mg

2) lactose "35 mg

3) corn starch 150 mg

4) microcrystalline cellulose 30 mg

5) magnesium stearate 5

240 mg per tablet

The components (1), (2), (3), 2/3 of the component (4) and half of the component (5) become good mixed. -The mixture is granulated. The rest Third of component (4) and half of component (5) are added to the granules. The crowd will pressed into tablets. The tablets so produced can be coated with a suitable enveloping agent, e.g. Sugar, to be coated.

B. Capsule

1) 1-ethyl-3,5-bis (p-chlorobenzyl) -6-aminouracil - 20 mg

2) lactose 102 mg

3) Microcrystalline Cellulose 70 mg

4) Magnesium stearate 8 mg

200 mg per capsule

The ingredients (1), (2), (3) and half of the ingredient (4) are mixed well. The mixture will granulated. The remaining half of the component (4)

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is added to the granules, and the mass is filled into gelatin capsules.

test

The inhibitory effect of representative 6-aminouracil derivatives (I) according to the invention on adenosine-3 ¹ , 5'-cyclophosphate phosphodiesterase was determined by the following method:

A mixture of 900 ml of fresh hind blood and 100 ml of a 3.8% strength (3 ^ 8 g / 100 ml) aqueous sodium citrate solution was centrifuged at 2000 rpm for 10 minutes
(Radius of the rotor 9 cm). The supernatant was separated. The lower layer was centrifuged again for 8 minutes at 3000 rpm, with another supernatant
was separated. The supernatants were pooled and centrifuged at 4000 rpm for 10 minutes, whereby the platelets were precipitated. The platelets thus obtained of the bovine blood were washed twice with 50 mL of brine and 25 of a 0.025-niolaren Tris-HCl buffer solution of p _H ml suspended 7.5. The suspension was turned off twice in a row
cycled rapid freezing in a dry ice-acetone bath and thawing at 37 ° C. The suspension thus treated was used as a bovine ^{platelet adenosine 3 1} , 5 ^l -cyclophosphate phosphodiesterase preparation.

0.1 ml of the prepared in the manner described
Preparation were mixed with 0.1 ml of a 0.02 molar aqueous solution of adenosine 3 ', 5'-cyclophosphate, 0.1 ml of a solution of 0.1 to 10 mmol of a solid compound in
Dimethyl sulfoxide, 0.2 ml of a 0.5 molar Tris-HCl buffer solution of p _H 7.5, o, 1 ml of a molar 0,02-
aqueous MgSO ^ solution, 0.1 ml snake venom (concentration 1 mg / ml) and water to make up the mixture

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mixed to 1 ml. The mixture was incubated at 37 ° C. for 15 minutes, after which 0.5 ml of an aqueous 10N sulfuric acid solution was added to the mixture. The amount of inorganic phosphate formed in the reaction mixture was then determined by the method described on page 8 ff. Of "Methods in Biochemical Analysis", Volume 3, Interscience Publishers, Inc., New York & London, 1956. The inhibition constant (Ki value) was calculated for the respective test compounds. The inhibition constants of the individual compounds were compared with the inhibition constants of theophylline, one of the most popular adenosine-3 ¹ »5 ¹ -cyclophosphate phosphodiesterase inhibitors, the determination being carried out under the above-mentioned conditions. The results obtained are shown in Table 2 below.

Table 2

Compound Inhibitory effect on obstructive blood

leaflet-adenosine-3 ¹ , 5'-cyclo-

- phosphate phosphodiesterase

1-ethyl-3,5-bisbenzyl- 175 times stronger than theophylline 6-aminouracil

1-ethyl-3,5-bis (p-108 times as potent as theophylline

chlorobenzyl) -6-aminouracil

1-Ethyl-3,5-ois (p-4-OO times as strong as theophylline

methylbenzyl) -6-aminouracil

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Claims (1)

Pat ent of sayings in which E is a lower one which is optionally substituted by a hydroxyl group or a lower alkoxy radical Alkyl radical, R is a halogen atom, a nitro group or a lower alkyl radical and η stands for 0, 1 or 2. 2. 6-aminouracil derivatives according to claim 1 with the formula mentioned there, in which R is a lower alkyl radical up to 4 carbon atoms, the one with a hydroxyl group or a lower alkoxy group with up to 4 carbon atoms can be substituted. 3. 6-aminouracil derivatives according to claim 1 and 2, wherein R an ethyl residue 1st. 4. 6-aminouracil derivatives according to claim 1 with the formula mentioned there, in which η stands for 0. 5. 6-aminouracil derivatives according to claim 1 with the formula mentioned therein, i
stands for 1 or 2. mentioned formula in which R stands for a halogen atom and η 6. 6-aminouracil derivatives according to claims 1 and 5 »wherein the Halogen atom is chlorine atom. 7. 6-aminouracil derivatives according to claim 1 with the there 2
, formula in which R is a lower alkyl radical with up to 4 carbon atoms and η is 1 or 2. 8. 6-aminouracil derivatives according to claims Λ and 7, wherein the S09813 / 1065 lower alkyl is methyl. 9. 6-aminouracil derivatives according to claim 1 with the there mentioned formula, in which R is a Mtrogruppe and η stands for 1. 10. 1-Ethyl-3,5-bisbenzyl-6-aminouracil 11. 1-Ethyl-3,5-bis (p -chlorobenzyl) -6-aminouracil 12. i-Ethyl-3,5-bis (p-methylbenzyl) -6-aminouracil 13. 1- (β-Hydroxyethyl) -3 »5- ^ iS (p-nitrobenzyl) -6-aminouracil . 1-Isobutyl-3,5-bis (2,4-dichlorobenzyl) -6-aminouracil 15 · Process for the preparation of 6-aminouracil derivatives according to claims 1 to 14, characterized in that one 6-aminouracil compounds of the formula ■ 0 R ¹ in d, he R has the abovementioned meaning with a compound of the formula. ρ
in which R and η have the abovementioned meanings and X is a halogen atom, is reacted in the presence of an inorganic base. 16. The method according to claim 15 »characterized in that an alkali hydroxide or alkali carbonate as the inorganic base is used. 509813/1065 17. Medicinal preparations containing at least one 6-aminouracil derivative according to claims 1 to 14 together with a pharmaceutically acceptable carrier. 509813/1065