CS270067B1 - N- [2- (2-Phosphonylethoxy) ethyl3 derivatives of heterocyclic bases and methods for their preparation - Google Patents

Abstract

The subject of the solution are N-C2-(2-phosphonylethoxy)ethyl derivatives of purine and pyrimidine heterocyclic bases and methods of their chemical synthesis. These substances belong to the field of biochemicals and substances with useful biological effects. This new group of substances from the group of acyclic nucleotide analogues is prepared by reacting the sodium salt of the respective heterocyclic base with dialkyl esters of 2-(2-chloroethoxy)ethylphosphonic acid. The intermediately formed diethyl esters are converted to the resulting products either directly by reaction with halogenotrimethylsilane or are first partially hydrolyzed with alkaline hydroxides to monoethyl esters, providing the subject substances also by reaction with halogenotrimethylsilane.

Description

The present invention provides N- [2- (2-phenylethoxy) ethyl] derivatives of heterocyclic bases and methods for their preparation.

Among the group of N-phosphonylalkoxyalkyl derivatives of heterocyclic bases are numerous important substances with possibilities of practical use. For example, N- (2-phosphonylmethoxyethyl) derivatives of purine bases (adenine, 2,6-diaminopurine and guanine) are important virostatics (Czech copyright certificates

Nos. 263951, 263952 and 264222) with specific activity against DNA viruses. O Fosfonylmethylethe- ^Ry 9- (2,3-dihydroxypropyl) adenine chemosteriiizační also have an effect on certain species of insects (Author's Certificate no. 23o341). All the substances mentioned so far contain a characteristic phosphonylmethoxy functional group (—CH 2 P (θ) (θ · Ό 2) ·

The present invention relates to novel compounds, N- [2- (2-phosphonylethoxy) -ethyl] derivatives of heterocyclic bases of the general formula I b-ch ₂ ch ₂ och ₂ ch ₂ p (o) (oh) ₂ (i), where B is the purine residue -9-yl or purin-7-yl, substituted in position 6 by hydroxy, amino, methylthio or chlorine and in position 2 by hydrogen or amino, or a residue uracil-1-yl or cytosin-1-yl,.

The invention furthermore relates to a process for the preparation of the compounds of the formula I in which the sodium salt prepared by reacting a heterocyclic base is alone or protected by acylation, preferably benzylation, amino groups or O-alkylation, preferably methylation, hydroxy groups, with equimolar amounts of sodium hydride. an aprotic solvent, preferably dimethylformamide, is reacted in the same solvent at 2 ° C to 14 ° C with one to three molar equivalents of 2- (2-chloroethoxy) ethylphosphonic acid diethyl ester of formula II.

C1CH ₂ CH ₂ OCH ₂ CH ₂ P (O) (OC ₂ H ₅ ) ₂ (li), the resulting intermediate of formula III b-ch ₂ ch ₂ och ₂ ch ₂ p (o) (oc ₂ h ₅ ) ₂ ( ΠΙ), where B has the same meaning as in formula I, is isolated by crystallization or preferably by chromatography on silica gel, reacted with bromotrimethyl silane in an aprotic solvent, preferably dimethylformamide, acetonitrile or chlorinated hydrocarbons, at a temperature of 20 ° C to 40 DEG C., after evaporation of the solvent, the mixture is decomposed with water and the product of formula I is preferably isolated by chromatography on an ion exchanger or hydrophobized silica gel.

The invention furthermore relates to a process for the preparation of compounds of the formula I in which the compounds of the formula III are reacted with a 5 to 2 M solution of alkaline hydroxides in water or mixtures thereof with methanol, ethanol or dioxane containing at least 50% by volume. of water, at temperatures of 20 ° C to 100 ° C, the mixture is neutralized with a mineral or organic acid, or preferably with a cation exchange resin in acid form and monoesters of general formula IV '

O

B-CH ₂ CH ₂ ^OC H2 ^C H2P- ^OC 2 ^H ₅ OH (IV).

where B has the same meaning as in formula I,

CS 27oo67 B1 isolated by ion exchange chromatography are reacted with bromotrimethylsilane under the above conditions and the compounds of formula I are isolated as described above.

A key compound for the process for the preparation of the compounds of formula 1 according to the invention is the diethyl phosphonic acid ester of formula II. This substance is preferably prepared by reacting triethyl sulphite with excess bis (2-chloroethyl) ether at the boiling point of the mixture and distilling off the ethyl chloride formed. This substance has also been described in the literature (L. Marier, M. M. Crutchfield, Phosphorus Sulfur 5f 45 (1978)). After completion of the reaction, diester II is isolated by fractional distillation in vacuo. It is sufficiently stable and can be stored at room temperature.

The heterocyclic base is used for the reaction in the form of the sodium salt, which can be prepared preferably directly from a suspension of the base in an appropriate solvent (preferably dimethylformamide) by adding one molar equivalent of sodium hydride (a commercial 50% to 60% dispersion of sodium hydride in paraffin can be used directly). In most cases, the base is used without modification; in special cases (eg to increase solubility or when several possible N-isomers may be formed in the reaction) it is recommended to use heterocyclic bases suitably substituted with protecting groups or bases otherwise modified, which may be further chemically reacted after the reaction. of the resulting intermediate or the corresponding compound of formula I to provide the final product of the desired structure. Thus, for example, adenine derivatives or 6-methylthiopurine give almost exclusively the N (9) -isomers of the compounds of the formula I in dimethylformamide under these conditions, and the corresponding bases can be used directly; however, hypoxanthine preferably gives the N (7) -isomer, so that its N (9) -substituted derivative of formula I is most readily prepared by deamination of the adenine phosphonate of formula I with nitrous acid or its ester. Although cytosine and its 5-substituted derivatives can be used directly for the reaction with the compound of formula II, for example, the previous benzoyl of the 4-amino group appears to be more preferred. In the case of uracil and thymine or other 4-oxopyrimidine bases, a mixture of the N (1) - and N (3) -isomers of substance I is formed under the above-mentioned conditions. with compound II of the sodium salts of the corresponding 4-methoxy-2-pyrimidones, which are readily available and, after isolation of the corresponding product of formula I, the 4-O-methyl protecting group can be cleaved by acid hydrolysis. Also for guanine, the use of the base alone or its N-acetyl derivative is unsuitable not only because the yield of the compound of formula III (B-guanin-9-yl) is generally low, but mainly because the reaction produces a mixture of N (9) - and N The (7) -isomer of the compounds of formula III. It is much more convenient to use the sodium salt of 2-amino-6-chloropurine instead of guanine; in this case the alkylation with the compound of formula II preferably takes place in the N (9) position and the resulting diester of formula III (B-2-amino-6-chloropurin-9-yl) can be easily hydrolyzed with dilute acid (eg hydrochloric acid) and subsequent reaction with converted with bromotrimethylsilane to the guanine derivative of formula 1 (β-guanin-9-yl).

After condensation and evaporation of dimethylformamide in vacuo, the crude product of formula III can be easily extracted with e.g. chloroform and in some cases obtained in pure state by direct crystallization, e.g. from methanol. In all cases, silica gel chromatography can be used. The intermediate of formula III is usually crystalline, well characterized by both UV spectra, NMR spectra and melting points. Its yields are around 50%, in some cases they are higher. An increase in yield can sometimes be achieved by using an excess of reagent II, especially in those cases where the reaction proceeds slowly and may be accompanied by partial decomposition of the compound of formula II (elimination of hydrogen chloride).

When using a dimethyl ester similar to the compound of formula II in the process for the preparation of the compounds of formula I in the above-mentioned manner, N-methyl derivatives of the corresponding heterocyclic base can be formed. Therefore, it is always preferable to use a diethyl ester of formula II for this reaction. However, the reaction is not limited to this substance; likewise, other dialkyl or aralkyl esters of the formula III can be prepared from the corresponding synthons of the formula II.

the cleavage of the ethyl ester bonds in the compounds of formula III is most preferably carried out by reaction with bromotrimethylsilane (A. Holy, I. Rosenberg, Collect. Czech. Chern. Commun. 47, 3447 (1982)). This reaction can be carried out in chloroform, dichloromethane or other chlorinated hydrocarbons or in dimethylformamide, but as practically the most suitable due to its chemical nature.

CS 27oo67 B1 has dissolution properties and a boiling point of acetonitrile. A slight excess of reagent (ca. 20%) is used for the reaction and is performed at room temperature. After evaporation of the solvent in vacuo and removal of excess reagent by co-distillation with acetonitrile or another inert solvent, the residue is quenched with water (preferably in the presence of ammonia or a volatile buffer, e.g. triethylammonium hydrogen carbonate). An bromine-contaminated salt of the compound of formula I is obtained. Desalting and purification of the compound of formula I is most preferably performed by ion exchange chromatography either directly on the anion exchange resin or by sequential chromatography on the cation exchange resin and the anion exchange resin (the procedure is determined by the properties of the heterocyclic base in the compound of formula 1). If the final chromatography is carried out, for example on an anion exchanger in acetate form, and the compound of the formula I is eluted with acetic acid, the product is obtained in the form of an acid after evaporation of the eluate, which can in many cases be purified by additional crystallization from water. In other cases where the compound of formula I is obtained after chromatography in the form of a salt (eg ammonium or triethylammonium), it is suitable to convert it to an alkaline (sodium, lithium) salt, preferably by cation exchange chromatography in the appropriate cycle. These salts are obtained by precipitating concentrated aqueous solutions with acetone or ethanol and are completely stable on storage. The compounds of the formula I are also sufficiently stable in the form of free crystalline acids (usually forming hydrates), provided that chemical changes do not result in chemical changes on the heterocyclic basis of the compounds of the formula I.

For the same purpose, iodotrimethylsilane can be used instead of bromotrimethylsilane under completely analogous reaction and work-up conditions. However, for practical reasons (price and availability of the iodine derivative), this option is not advantageous.

The hydrolysis of one of the ester bonds can also be carried out in an alkaline medium. However, under these conditions, i.e. by reacting the compounds of formula III in aqueous alkaline hydroxide or mixtures thereof with ethanol, methanol, dioxane and the like at room temperature or elevated temperature, only one of the two ester functions is cleaved so that the reaction product is the compound of formula IV. ‘

This route is justified in those cases where the compound of formula III contains acyl (e.g. benzoyl) protecting groups which are simultaneously cleaved by this reaction. However, the compounds of formula IV can no longer be well isolated by chromatography on silica gel and their purification requires the use of ion exchange chromatography under similar conditions as in the purification of the compounds of formula I. To cleave the remaining ester-linked group, . Prior to the reaction, the compound of formula IV must be dried well, in which case an excess (20 to 25%) of reagent must be used to achieve complete conversion.

In contrast to N- (2-phosphonylmethoxyethyl) derivatives, many of which have antiviral activity (Czech Patent Nos. 263951 and 264222), the compounds of the formula I do not show any significant activity against viruses. However, they can be used, for example, as chemosterilants of insects. Oral administration of dilute aqueous solutions of compounds of formula I ad libitum, e.g. to some species of bugs (Pyrrhocoris apterus L.), both last instar nymphs and adults, will significantly reduce the number of eggs laid, depending on the concentration of the substance. , to the complete cessation of laying and at the same time to a significant reduction to the complete suppression of their hatchability. This effect depends on the structure of the compounds of formula I (character of the heterocyclic base) and is distinguishable from simple toxicity. An overview of this effect is given in Table 1. Chemosterilization effect In this insect species, substances formally similar to the compounds of formula I, i.e. phosphonylmethyl ethers of glycerol and 9- (2,3-dihydroxypropyl) adenine, have also been found (Czech author's certificate No. 233626 ). However, the effect of the compounds of formula I is in no way predictable, since the compounds of formula 1 ee differ fundamentally from the above group both in the presence of a characteristic phosphonylethyl ether function and in particular in that they do not contain an asymmetric carbon atom. effects in phosphonylmethyla rivals.

An important property of the compounds of formula I is their chemical and biological stability. The phosphonic acid bond is non-cleavable by common enzymes degrading eg phosphoric acid monoesters and is stable to dilute organic and mineral acids, as well as to ammonia, amines and dilute aqueous alkalis.

CS 27oo67 Bl

In the following, the invention is illustrated by way of non-limiting examples of processes for the preparation and use of the compounds of the formula I.

Example 1

A mixture of bis (2-chloroethyl) ether (60 ml) and triethyl phosphite (18 ml) was heated for 1 h at 16 ° C in a 25 ml flask, dissolved in 10 ml of benzene and applied to a column of 500 g of silica gel. The column was eluted with benzene-ethanol (99: 1) and the individual fractions (30 ml each) were tested by chromatography on silica gel plates in benzene. The product-containing fractions (detection by spraying p-nitrobenzylpyridinium bromide followed by exposure to ammonia vapor) are combined, evaporated in vacuo and dried at 13 Pa over phosphorus pentoxide. 52% of 2- (2-chloroethoxy) ethylphosphonic acid diethyl ester are obtained as a colorless oil. For ΟθΗ ₁₈ Ο1Ο ₄ Ρ (244.7) calculated 14.49% Cl, 12.66% P; found 14.3o% Cl, 12, βο% P. Mass spectrum 245 (M * + 1),

A suspension of 13.5 g of adenine and 3.4 g of sodium hydride in 70 ml of dimethylformamide was stirred at 80 DEG C. for 1 hour with exclusion of moisture, then a solution of 22.5 g of 2- (2-chloroethoxy) ethylphosphonic acid diethyl ester was added dropwise over 2 hours. in 200 ml of dimethylformamide and the mixture was stirred for 12 h at 80 ° C with exclusion of moisture. The mixture is then evaporated at 50 DEG C./2 kPa and the residue is evaporated twice under the same conditions in 200 ml of toluene. The residue was extracted with boiling chloroform (1 L), filtered through a pad of celite, washed with 200 mL of chloroform, and the filtrate was concentrated in vacuo at 30 ° C to a volume of about 200 mL. This solution was applied to a 500 g silica gel column (30 to 40 °) and the column was washed with chloroform (1 L). Elution with a mixture of chloroform and methanol (9 and 1), evaporation of the combined product fractions (by chromatography on silica gel plates in chloroform-methanol, 4: 1; Rf 0.55) and crystallization of the residue from ether (petroleum ether to turbidity) gave 9- [2- (2-diethoxyphosphonylethoxy) ethyl] adenine (III, β-adenin-9-yl) in a yield of 19.5 g (62%). Mp 60 ° C. For ^{C 13 H} 22 ^ 5 ° 4 ^P ( ³⁴³ · ⁴ ) calculated 45.46% C, 6.46% Η, 20.0% N, 9.9% P; found 45.0%, 6.5%, 0.20% N, 8.93% P.

This product was dissolved in 600 ml of acetonitrile, 38 ml of bromotrimethylsilane was added, and the mixture was allowed to stand in a sealed flask for 24 hours at room temperature. It is evaporated at 40 DEG C./2 kPa, 200 ml of acetonitrile are added and the mixture is evaporated again under the same conditions. 500 ml of a 0.4 M aqueous solution of triethylammonium hydrogen carbonate (pH 7.5) are added to the residue and the pH of the solution is kept constant by adding triethylamine between 8 and 9 (about 1 h). The mixture is then evaporated at 40 DEG C./2 kPa and the residue is evaporated under water under the same conditions with water (three times 10 ml). This residue is dissolved in 300 ml of water and applied to a column of 500 ml of cation exchange resin (e.g. Dowex 50 X 8) (n ⁺ - form). The column is washed with water until the acid reaction of the eluate decreases and the UV absorption is lost at 26 nm.

It is then eluted with 2.5% aqueous ammonia and the eluate absorbed at 26 nm is collected. After evaporation in vacuo the residue dissolved in EE loo ml of water, applied onto a column 4oo ml of medium basic anion exchange resin (e.g. Sephadex A-25 (HCO @ ^- form)). The column is eluted with water until the UV absorption of the eluate decreases at 26 nm and then with a gradient of 0 to 4 M triethylammonium hydrogen carbonate (pH 7.5) (2 l each). The elution process is monitored by continuous measurement of UV absorption at 26 nm. The product fractions are evaporated in vacuo, the residue is taken up in water (three times 10 ml) and 100 ml of cation exchange resin (e.g. Dowex 50 x 8) (H ⁺ -form) are applied to a column of 10 ml of water. The column is eluted with water, the eluate absorbing at 26 nm is evaporated in vacuo and 200 ml of acetone and 200 ml of ether are added to the residue dissolved in 20 ml of water. The precipitated product is filtered off with suction, washed with ether (200 ml) and dried at 13 Pa over phosphorus pentoxide. 17 g (84%) of the sodium salt of 9- (2- (2-phosphonylethoxy) ethyl) adenine-3-phosphate (1, β-adenin-9-yl) are obtained. Electrophoretic mobility E

Up 0.75 (based on uridine- (pH 7.5). For C _g HN ₅ ° ₄ PNa ₂ (313.4) calculated 21.14% N, 9.37% P;

20.66% N, 9.12% P. UV spectrum (pH 2): max 257.7 nm (e _mnx - 114oo). R _f 0.17 (2-propano-aqueous ammonia-water, 7: 1: 2).

Example 2 A

To a suspension of 4.3 g of N-benzoylcytosine in 80 ml of dimethylformamide was added 46 mg of sodium hydride, and the mixture was stirred at 20 ° C for 1 h. A solution of 6 g of 2- (2-chloroethoxy) ethylphosphonic acid diethyl ester (see Example 1) in 20 ml of dimethylformamide was added and the mixture was stirred at 0 ° C for 20 h.

CS 27oo67 Bl 5 under a chlorocalcium cap. The resulting solution was evaporated to dryness at 40 DEG C./13 Pa and the residue was extracted with 500 ml of boiling chloroform. This ee extract was evaporated to dryness in vacuo and dissolved in 200 ml of ο, sodného sodium methoxide in methanol. The solution is allowed to stand overnight at 20 DEG C., neutralized by adding cation exchange resin (e.g. Dowex 50 θ) in the H @ ⁺ form, the suspension is basified with triethylamine, filtered off with suction and the ion exchange is washed with 300 ml of methanol. The filtrate is evaporated in vacuo, the residue is dissolved in 200 ml of water and extracted with 3 x 50 ml of ether, the aqueous layer is concentrated to half volume in vacuo and the solution is applied to a 300 ml column of cation exchange resin (eg Dowex 50 x 8) (H ⁺ - form). The column is eluted with water (500 ml) and methanol-water (3: 7) until the UV absorption at 26 nm has decreased to the original value. It is then eluted with 2.5% aqueous ammonia and the eluate absorbed at 26 nm is collected. This was evaporated in vacuo, the residue was taken up in ethanol (2 x 50 ml) and finally dried at 20 ° C / 13 Pa over phosphorus pentoxide for 24 h to give 4.2 g (66%) of the monoester of formula IV (β-cytosine-1 -yl). Electrophoretic mobility (see Example 1) o, lo, Rp, - o, 34 (TLC, chloroform-methanol, 4: 1).

To this residue were added 10 ml of acetonitrile and 10 ml of bromotrimethylsilane, the mixture was allowed to stand at 20 DEG C. for 48 hours and evaporated in vacuo. The residue is re-evaporated with 10 ml of acetonitrile. dissolved in 200 ml of 0.4M triethylammonium hydrogen carbonate and after 2 h evaporated in vacuo. The residue was evaporated with methanol (3 x 10 ml) and dissolved in 10 ml of water. This solution is applied to a column of 200 ml of cation exchange resin (e.g. Dowex 50 X 8) (H ^T -form), eluted with water until the acidic reaction of the eluate decreases and then with 2.5% aqueous ammonia. The ammoniacal eluate is evaporated to dryness in vacuo, dissolved in 50 ml of water, basified with ammonia and applied to a column of 200 ml of a strongly basic anion exchange resin (e.g. Dowex X 2) (acetate). The column was eluted with water (500 ml) and then 1M acetic acid. The UV-absorbing acidic eluate was evaporated to dryness, the residue was co-distilled with water in vacuo (4 x 20 ml), dissolved in a small amount of hot water, 5 volumes of ethanol and ether were added until cloudy. The product which crystallizes out in the refrigerator is filtered off with suction, washed with ether and dried at 13 Pa over phosphorus pentoxide. 2.1 g of 1- [2- (2-phosphonylethoxy) ethyl] cytosine (1, β-cytosin-1-yl) are obtained. R_ - 0.17 (2-propanol-conc. Ammonia-water, 7: 1: 2) E _IT -0.85. Mp 18 ° C. For C_H, .N_O_P j? up o (263.3) calculated 36.49% C ,. 5.36% H, 15.96% N, 11.79% Pj found 34.46% C, 6.36% H, 15.97% N, 10.5% P. UV spectrum (pH 2): Λ __- 28o nm (£ __- loooo).

m & x m & x

Example 3

A suspension of 3, 6 g of 2,6-diaminopurine and. in 20 ml of dimethylformamide. The mixture was stirred at 10 ° C for 18 h under a chlorocalcium cap and evaporated at 60 ° C / 13 Pa. The residue was co-distilled twice with 50 ml of toluene under the same conditions, and the residue was extracted with 200 ml of boiling chloroform. The suspension was filtered through a pad of celite and the filtrate was evaporated in vacuo. The product III (B-2,6-diaminopurin-9-yl), which was eluted with chloroform-methanol (92.5: 7.5), was obtained by evaporation in vacuo and drying as an amorphous foam in a yield of 4.6 g (64 %). Rf-0.4 (TLC on silica gel, chloroform-methanol, 4: 1). i * ·

This substance was dissolved in 10 ml of acetonitrile, 10 ml of bromotrimethylsilane was added, and the mixture was allowed to stand at room temperature for 48 hours. It is evaporated at 40 DEG C./2 kPa, co-distilled with 50 ml of acetonitrile under the same conditions and the residue is dissolved in 200 ml of 0.4M triethylammonium bicarbonate, pH 7.5. After 30 min, evaporate to dryness in vacuo and the residue was evaporated in vacuo with methanol (3 x 10 ml). The residue is applied in 500 ml of water to a column of 200 ml of cation exchange resin (e.g. Dowex 50 Xa) (H ⁺ form) and the column is washed with water until the acidic reaction of the eluate and UV absorption are lost at 26 nm. The column was then eluted with 2.5% aqueous ammonia and the UV-absorbing eluate was collected. After evaporation in vacuo, the crude product in 25 ml of water alkalized with ammonia to PpH 8.5 is applied to a column of 200 ml of a strongly basic anion exchange resin (e.g. Dowex 1X2) (acetate) and the column is eluted with water until the UV absorption decreases at 26 nm. original value. Ionex is suspended in 2M formic acid and the suspension is brought to the boil with stirring. It is filtered while hot and ionex

CS 27oo67 B1 is washed with 1 l of boiling 2M formic acid. The combined filtrates were evaporated in vacuo and the residue was co-distilled with water (5 x 100 ml) in vacuo. The residue is dissolved in boiling water, 6 volumes of ethanol and ether are added until turbid. The product crystallizes out in a refrigerator, which is filtered off with suction, washed with ether and dried in vacuo. Yield 3.0 g of 9-C2- (2-phosphonylethoxy) ethyl] -2-aminoadenine, m.p. For C _g H _g N ₆ PO ₄ (3o2,3) calculated 35.75% C, 5 oo% H, 27.8% N, 10.27% P; found 35.46% C, 4.8o% H, 27.75% N, 10.34% P. EyP (see Example 1) o, 66, Rp o, o7 (PC, 2-propanol-conc. aqueous ammonia -water, 7: 1: 2). UV spectrum (pH 2):

Λ 256 nm, 287.6 nmjA, 267.6 nm.

max mln

Example 4

To a solution of 3.32 g of 6-methylthlopurine in 80 ml of dimethylformamide was added 0.48 g of sodium hydride, and the mixture was stirred at 50 ° C for 1 h. 6.4 g of 2- (2-chloroethoxy) ethylphosphonic acid diethyl ester (see Example 1) are added and the mixture is heated at 110 DEG C. for 18 h, with the exception of moisture. It is evaporated at 40 DEG C./13 Pa, extracted with 400 ml of hot chloroform and the extract is evaporated in vacuo. The residue was applied to a column of 400 ml of silica gel in chloroform, eluting the column first with chloroform (1 L) and then with chloroform-methanol (95: 5). The product fractions (according to TLC in chloroform-methanol, 9: 1, Rf -0.68) are combined, evaporated in vacuo and dried at 13 Pa over phosphorus pentoxide. The yield of the amorphous foam of III (B-6-methylthiopurin-9-yl) is 4.0 g (53.5%). This product was dissolved in 80 ml of acetonitrile, and after adding bromotrimethylsilane (8 ml), the mixture was allowed to stand at room temperature for 48 h. It is evaporated at 40 DEG C./2 kPa, codistilled under the same conditions with 50 ml of acetonitrile and the residue is dissolved in 10 ml of 0.4M triethylammonium hydrogen carbonate (pH 7.5). After standing for 30 minutes, it is evaporated in vacuo, co-distilled with methanol (4 x 50 ml) and the residue in 50 ml of water is applied to a column of 200 ml of cation exchange resin (e.g. Dowex 50 x 8) (H ⁺ -form). The column is eluted with water until the acidic reaction decreases and the eluate is UV-absorbed. The column is then washed with 2.5% aqueous ammonia and the W-absorbing fraction is collected at 26 nm. The residue of this eluate is dissolved in 20 ml of water and applied to a column of 10 ml of cation exchange resin (e.g. Dowex 50 X 8) (Na ⁺ form). The column is washed with water, the UV-absorbing eluate is evaporated in vacuo to a volume of about 10 ml, 200 ml of ethanol and 200 ml of ether are added, the precipitated product is filtered off with suction, washed with ether and dried at 13 Pa over phosphorus pentoxide. 2.8 g of sodium salt of 9-C2- (2-phosphonylethoxy) ethyl] -6-methylthiopurine (1, β-6-methylthiopurin-9-yl) are obtained. Melts to 26 ° C. ^ up Example 1) - o, 73.

For C, H.A'PO.SNa. (362.4) calculated 33.14% C, 3.61% H, 15.47% N, 8.57% P, 8.85% S; 1U 1 «3 4 4 &

found 33.34% C, 3.75% H, 15.44% N, 8.93% P, 9.9%; % S.

Example 5

A mixture of 2.52 g of 4-methoxy-2-pyrimidone and .48 g of sodium hydride in 80 ml of dimethylformamide was stirred at 20 DEG C. for 1 hour and a solution of 6.2 g of 2- (2-chloroethoxy) ethylphosphonic acid diethyl ester was added. in 20 ml of dimethylformamide. The mixture was stirred under a chlorocalcium cap for 20 h at 10 ° C and then evaporated to dryness at 50 ° C / 13 Pa. The residue was co-distilled with toluene (2 x 50 ml) under the same conditions and extracted with boiling chloroform (500 ml). The extract was evaporated in vacuo, and the residue was chromatographed on a column of 300 g of silica gel with chloroform and then with chloroform-methanol (98: 2). Evaporation of the product fractions under vacuum and dried at 13 Pa over phosphorus pentoxide to yield 2.8 g of compound III (B-4-methoxy-2-pyrimidone-1-yl) as a colorless oil, R _p - o, 7o (TLC chloroform-methanol, 9: 1). This product was dissolved in 60 ml of acetonitrile, 6 ml of bromotrimethylsilane was added, and the mixture was allowed to stand at room temperature for 24 hours. It is evaporated at 40 DEG C./2 kPa, the residue is codistilled with 50 ml of acetonitrile and dissolved in 10 ml of 0.4M triethylammonium hydrogen carbonate pH 7.5. After 1 h, the solution was evaporated in vacuo and co-distilled with methanol (4 x 50 mL). The residue is refluxed in 100 ml of 80% aqueous acetic acid for 8 hours, evaporated at 40 DEG C./2 kPa and co-distilled with water (5.times.5 ml) under the same conditions. The residue is applied in 20 ml of water to a column of 15 ml of cation exchange resin (e.g. Dowex 50 X β) (1β ^- form) and the column is eluted with water. The eluate fraction absorbing at 26 nm is collected, evaporated in vacuo and the residue is stirred in 80 ml of ethanol. 80 ml of acetone are added and the mixture is stirred for 1 hour. The product is filtered off with suction, washed with ether and dried at 13 Pa over phosphorus pentoxide. 1.6 g of the lithium salt of 1- [2- (2-phosphonylethoxy) ethyl] uracil are obtained

CS 27oo67 B1 (i, B-uracil-1-yl), R _p - o, 21 (PC, 2-propanol-conc. Aqueous ammonia-water, 7: 1: 2), E o, 96 (see Example 1 ). UV spectrum (pH 2)! A _max 26o nm (t _max “loooo). For

Up ^ 8 ^ 11 ^ 2 ^ 6 ^^ 2 (276 L) 'VPO ^ teno 34.8% C, 4.0% H, 10.15% N, 11.24% P; found 34.52% C, 4.21% Η, ’1ο, ο6% N, 11.2o% P.

Example 6

A mixture of 3.4 g of 2-amino-6-chloropurine, 5.6 g of potassium carbonate, 6 g of 2- (2-chloroethoxy) ethylphosphonic acid diethyl ester and 100 ml of dimethylformamide was stirred at 100 DEG C. for 30 hours and filtered while hot. The precipitate is washed with 50 ml of dimethylformamide and the filtrate is evaporated at 50 DEG C./2 kPa. The residue is extracted with boiling chloroform (3.times.20 ml), the filtrate is evaporated at 40 DEG C./2 kPa and the residue in 50 ml of chloroform is applied to a 200 ml column. silica gel in chloroform. The column was washed with chloroform and the elution was monitored by thin layer chromatography on silica gel plates in chloroform-methanol (9: 1). A fraction of 9- [2- (2-diethoxyphosphonylethoxy) ethyl] -2-amino-6-chloropurine (R0.0.0) is obtained, which is evaporated and dried at 13 Pa over phosphorus pentoxide. Yield 3.2 g (42%) of a colorless foam. Further elution of the column with chloroform gave similarly 1.0 g (13%) of 7-C2- (2-diethoxyphosphonylethoxy) ethyl-2-amino-6-chloropurine (Rp 0.28).

The residue of the 9-isomer (3.2 g) was dissolved in 30 ml of dioxane, 30 ml of 1M sodium hydroxide was added, and the mixture was allowed to stand at room temperature for 24 hours. It is neutralized by adding a cation exchange resin (e.g. Dowex 50 X 8) (H ⁺ -form), basified with triethylamine, filtered, washed with 50 ml of water and evaporated to dryness in vacuo. The residue was heated at reflux with 80 ml of 1M hydrochloric acid, neutralized with aqueous ammonia and evaporated to dryness in vacuo. This residue in 20 ml of water is applied to a column of 200 ml of cation exchange resin (e.g. Dowex 50 X 8) (H ⁺ form) and eluted with water until the acidic reaction disappears and the eluate is absorbed at 26 nm. The column was then eluted with 2.5% aqueous ammonia and the eluate was collected at 26 nm. After evaporation in vacuo, the residue is dried at 13 Pa over phosphorus pentoxide, suspended in 60 ml of acetonitrile, 6 ml of bromotrimethylsilane are added and the mixture is left to stand for 24 hours at room temperature in a closed flask. The mixture was evaporated in vacuo, the residue dissolved in 10 ml of 0.4M triethylammonium hydrogen carbonate, evaporated in vacuo and co-distilled with methanol (4 x 50 ml). This residue is applied in 25 ml of water to a column of 200 ml of weakly basic anion exchanger (e.g. Sephadex A-25) (HCO-form) and eluted with water until the UV absorption of the eluate decreases at 26 nm. The column is then eluted with a linear gradient of triethylammonium hydrogen carbonate. , pH 7.5 (0 to 0.3 M in 1 L), and collect the major fraction of the eluate absorbing at 26 nm (eluting at 0.15 to 0.25 M). After evaporation in vacuo, the residue was codified with methanol (5 x 50 ml), dissolved in water and applied to a column of 50 ml cation exchange resin (e.g. Dowex 50 X 8) (Na ⁺ form). Elute with water, collect the eluate absorbed at 26 nm and evaporate in vacuo. The residue was codified with ethanol (2 x 50 ml) and precipitated from methanol (10 ml) with ether (200 ml). It is filtered off with suction, washed with ether and dried in vacuo. 1.4 g of sodium salt of 9- [2- (2-phosphonylethoxy) ethyl] guanine (I, B-guanin-9-yl), -0.68, Rp - o, o9 are obtained -conc.

ammonia — water, 7: 1: 2 $. For C "H N O PNa" (347.3) calculated 31.12% C, 3.48% H, y i »o o λ.

20.17% N, 8.94% P {found 31.3% C, 3.76% H, 19.82% N, 8.79% P.

In the same manner, the 1,10-7-isomer (see above) was prepared as the sodium salt of 7- [2- (2-phosphonylethoxy) ethyl] guanine (1, B-guanin-7-yl). Yield 0.36 g, and Rp values are the same as for the 9-isomer. For C ₉ H _{12 N 5} O ₅ Na ₂ (347.3) calculated 31.12% C, 3.48% 14.20, 17% N, 8.94% P; found 3o, 85% C, 3.61% Η, 2o, o6% N, 8.82% P.

Example 7

To a solution of 1.0 g of 9-2- (2-phosphonylethoxy) ethyl-adenine sodium salt in 50 ml of 80% aqueous acetic acid was added 4 ml of 3-methylbutyl nitrite, and the mixture was stirred overnight in a sealed flask at room temperature. It was evaporated in vacuo and the residue was co-distilled with water (5 x 25 ml). The residue is applied to a column of 150 ml of cation exchange resin (e.g. Dowex 50 X 8) (H-form) in 20 ml of water and the column is eluted with water. The product fraction absorbing at 26 nm is collected, evaporated in vacuo and dissolved in the required amount of water while hot. Ten times the volume of ethanol and ether (same volume as ethanol) are added. The product crystallizes out in a refrigerator, which is filtered off with suction, washed with a mixture of ethanol and o

CS 27oo67 B1 ether (1: 1), ether λ dried in vacuo. 0.56 g of 9-2- (2-phosphonylethoxy) ethyl hypoxanthine are obtained, nol-conc.

o, 96 (see Example 1), R r

- 0.16 (paper chromatography in 2-propa-ammonia-water, 7: 1: 2). For (288.3) calculated 37.5% C, 4.54% H,

19.44% N, 10.77% P; found 37.87% C, 4.85% H, 19.41% N, 11.7% P.

Example 8

Determination of chemosterilization effect of substances.

Pyrrhocoris apterus L. was reared under constant conditions at a temperature of 26 1 ° C in a photoperiod of 18 h light + 6 h dark and at a relative humidity of 75 + 10%. Linden seeds and drinking water were used as food. Basic breeding in o, 5 1 jars. Last instar nymphs aged up to 6 h were taken for testing. Solutions of test compounds of formula I (sodium salts) were prepared in drinking water and administered during the test instead of drinking water. .

The test itself was performed on Petri dishes with a diameter of 12 to 15 cm with five pairs of animals or on dishes with a diameter of 6 to 10 cm with individual pairs. The bottoms of the dishes were covered with filter paper, the dishes contained a drinker with a test solution of the compounds of formula I and linden seeds. The experiment was evaluated for toxicity, the length of the last instar, the course of the imaginary ecdysis, the onset of oviposition and the number of eggs laid in each clutch. The laid eggs were collected and placed in petri dishes for incubation under the above conditions. Hatchability was monitored at the time of hatching in control experiments, the second reading was taken 5 days later (to rule out possible prolongation of embryogenesis).

Under these conditions, the last instar length of the control subjects averaged 8 days; this length was not affected by any of the monitored substances. The course of undressing into an adult was not affected. Reduction of fertility and hatchability for individual substances of general formula I It is shown in Table 1. '

Table 1

Effect of compounds of formula I (sodium salt) on the fertility of Pyrrhocoris apterus L. after continuous oral administration to last instar nymphs

C. B (concentration,%) Average number of eggs laid per female Average number of hatched eggs per female Hatchability (%) Control 166.1 + 14.4 143.9 + 16.1 86.6 1 Adenin-9-yl lo ^-5 O O O lo ⁶ 92.3 ± 10.8 42.7 ± 7.7 46.3 2 2,6-Diaminopurin-9-yl lo ⁸ O O O 3 Guanin-9-yl lo ' ⁶ - O O O I ⁷ . 139.3 ± 17.8 121.1 ± 15.4 86.9 4 6-Methylthiopurin-9-yl lo “ ⁷ O O O lo “ ⁸ 112.8 ± 16.4 86.1 + 12.1 76.3 5 Uracil-1-yl lo ⁴ O O O lo ' ⁵ 93, C + 15.3 31.5 4- lo, 5 33.7

CS 27oo67 Bl

Cytosin-1-yl lo ^-4 lo ^-5

O

112.1 ± 11.8

O

79.7 + 14.8

About 71.1

Claims (3)

OBJECT OF THE INVENTION. N- [2- (2-Phosphonylethoxy) ethyl] derivatives of heterocyclic bases of general formula I b-ch ₂ ch ₂ och ₂ ch ₂ p (o) (oh) ₂ (I), wherein B is the residue purin-9- yl or purin-7-yl, substituted in position 6 by hydroxy, amino, methylthio or chlorine and in position 2 by hydrogen or amino, or a residue uracil-1-yl or cytosin-1-yl. 2. A process for the preparation of compounds of formula I according to claim 1, characterized in that the sodium salt prepared by reacting a heterocyclic base either alone or protected by acylation, preferably benzoylation, amino groups or O-alkylation, preferably methylation, hydroxy groups, with an equimolar amount of sodium hydride in an aprotic solvent, preferably dimethylformamide, in darkness indicating that solvent at temperatures of 2O ° C to 14o C to react with from one to three molar equivalents of diethyl 2- (2-chloroethoxy) ethyl phosphonate of the formula II is passing ₂ CH ₂ OCH ₂ ch ₂ p (o) (oc ₂ h ₅ ) ₂ (II) formed intermediate of general formula III. . B-CH ₂ CH ₂ OCH ₂ CH ₂ P (O) (OC ₂ H _g ) _{2 (m χ} where B has the same meaning as in formula I, is isolated by crystallization or preferably by chromatography on silica gel, given at a temperature of 2o to 4o ° C to the reaction with bromotrimethylsilane in an aprotic solvent, preferably dimethylformamide, α-cetonitrile or chlorinated hydrocarbons, after evaporation of the solvent the mixture is quenched with water and the compound of formula I is isolated, preferably by chromatography on ion exchanger or hydrophobized silica gel. 3. A process for the preparation of compounds of formula I according to item 1, characterized in that the compounds of formula III according to item 2 are reacted with a 0.5 to 2.0 mM solution of alkali hydroxides in water or mixtures thereof with methanol, ethanol or dioxane containing at least 50% by volume of water, at temperatures of 20 ° C to 100 ° C, the mixture is neutralized with a mineral or organic acid, or preferably with a cation exchange resin in acid form, and monoesters of general formula IV $ 'B-CH, CH, OCH CH P-OCH-H (IV) A & 4 ě | 4L O. . OH wherein Li has the same meaning as in formula I, isolated by ion exchange chromatography, are reacted with bromotrimethylsilane, and the compounds of general formula I are further isolated according to the method of point 2.