RU2036914C1 - N6-PHOSPHORYLATED DERIVATIVES OF 3-β- PHENYLISOPROPYLSYDNONEIMINE SHOWING ANTITUMOR ACTIVITY - Google Patents

Abstract

FIELD: organic chemistry. SUBSTANCE: products: N⁶-(methoxymethylphosphonyl)-3-β-phenylisopropylsydnoneimi- ne, empirical formula is C₁₃H₁₈N₃O₃P, yield is 79%, m. p. is 84-86 C; N⁶-(dimethoxyphosphoryl)-3-β-phenylisopropylsydnoneimine, empirical formula is C₁₃H₁₈N₃O₄P, yield is 67%, m. p. is 45.5-47.5 C; N⁶-(bis-dimethylamidophosphoryl)-3-β-phenylisopropylsydnon- eimine, empirical formula is C₁₅H₂₄N₅O₂P, yield is 83%, m. p. is 98-99 C; 3-β-phenylisopropylsydnoneimido-b-phenethylamidophosphate potassium, empirical formula is C₁₉H₂₂N₄O₃PK, yield is 70%, m. p. is 201 C (with decomposition); 3-β-phenylisopropylsydnoneimidomorpholinophosphate potassium, empirical formula is C₁₅H₂₀N₄O₄PK, yield is 60%, m. p. is 186-189 C; N⁶-(bis-ethyleneimidophosphoryl)-3-β-phenylisopropylsydnon- eimide, empirical formula is C₁₅H₂₀N₅O₂P, yield is 88%, m. p. is 90-92 C; 3-β-phenylisopropylsydnoneimidothiophosphoric acid disodium salt monohydrate, empirical formula is C₁₁H₁₂N₃O₃PSNa₂·H₂O, yield is 87%, m. p. is 238 C (with decomposition); 3-β-phenylisopropylsydnoneimido-b-oxyethylphosphate sodium monohydrate, empirical formula is C₁₃H₁₇N₃O₄PSNa·H₂O, yield is 56%, m. p. is 68-69 C; N⁶-(dimethoxythiophosphoryl)-3-β-phenylisopropylsydnoneimi- ne, empirical formula is C₁₃H₁₈N₃O₃PS, yield is 96%, m. p. is 53-55 C. EFFECT: improved method of synthesis. 6 tbl

Description

N

R₁·n·H₂O где, если Х есть атом кислорода и n= 0, то I. R₁= R₂= OCH₃ II. R₁=CH₃, R₂=OCH₃ III. R₁=R₂=N(CH₃)₂ IV. R₁= NHCH₂CH₂C₆H₅, R₂= 0^-К⁺ V. V. R₁= N

O R₂= O^-K⁺, R₂=0^-K⁺ VI. R₁=R₂= N (CH₂)₂ если Х есть атом серы, то VII. R₁=R₂=0^-Na⁺, n=1 VIII. R₁=OCH₂CH₂OH, R₂=0^-Na⁺, n=1 IХ. R₁=R₂=OCH₃, n=0 обладающие противоопухолевой активностью.The invention relates to the chemistry of 1,2,3-oxadiazoles and specifically relates to new derivatives of the series 3-β-phenylisopropylsidonimonimine of the general formula I:

N

R ₁ · n · H ₂ O where, if X is an oxygen atom and n = 0, then I. R ₁ = R ₂ = OCH ₃ II. R ₁ = CH ₃ , R ₂ = OCH ₃ III. R ₁ = R ₂ = N (CH ₃ ) ₂ IV. R ₁ = NHCH ₂ CH ₂ C ₆ H ₅ , R ₂ = 0 ^- K ⁺ VV R ₁ = N

OR ₂ = O ^- K ⁺ , R ₂ = 0 ^- K ⁺ VI. R ₁ = R ₂ = N (CH ₂ ) ₂ if X is a sulfur atom, then VII. R ₁ = R ₂ = 0 ^- Na ⁺ , n = 1 VIII. R ₁ = OCH ₂ CH ₂ OH, R ₂ = 0 ^- Na ⁺ , n = 1 IX. R ₁ = R ₂ = OCH ₃ , n = 0 with antitumor activity.

 The literature does not describe the biological activity of compounds of a number of sydnonimine containing both a phenyl isopropyl residue and a phosphoric acid residue.

проявляющий противоопухолевые свойства (1), но известный в медицинской практике как психотропный препарат (под международным названием сиднофен), обладающий психостимулирующим и антидепрессивным действием (2).In this regard, as the closest structural analogue was selected 3-β-phenylisopropylsidononimine hydrochloride of formula A:

exhibiting antitumor properties (1), but known in medical practice as a psychotropic drug (under the international name Sydnophen), which has a psychostimulating and antidepressant effect (2).

 Sidnofen has a relatively low antitumor activity due to the presence of both immunostimulating properties and the ability to increase the overall resistance of the body (inhibition of tumor growth does not exceed 26-40%).

 The purpose of the invention, new derivatives of the series 3-β-phenylisopropylsidnonimine with improved antitumor properties in the absence of psychotropic effects.

The goal is achieved by the chemical structure of the claimed compounds of General formula I, containing in the 3rd position of the oxodiazole ring a β-phenylisopropyl group and in the 5th position a group of the formula N-P (X) ^- , where X is an oxygen or sulfur atom with antitumor activity.

 The inventive compounds are atmospheric unpainted substances, soluble in water and organic solvents, insoluble in paraffin hydrocarbons, hygroscopic.

 The structure of the compounds is confirmed by elemental analysis and spectral characteristics (NMR, PMR, UV spectra).

 The antitumor activity of the claimed compounds was studied in experiments on 508 rats and 1223 mice using the indicated strains of tumors of mice and rats (in parentheses are the lines and sex of animals on which the strains were maintained and chemotherapeutic experiments were carried out, as well as the initial body weight of the animals): P388 lymphocytic leukemia (TWO 2 mice, VDF1, males, 20-22 g), L1210 lymphoid leukemia (TWO 2 mice, VDF1, males, 20-23 g), CM mouse sarcoma (non-inbred mice, females 18-21 g), Ca 755 adenocarcinoma mammary gland (mice C 57Bl6, females, 20-23 g), B 16 melanotic m B16 elanoma (mice VDF1, males, 20-23 g), CFL Lewis lung carcinoma (mice VDF1, males, 20-23 g), ACATOL colon adenocarcinoma (mouse BALB / s, males, 18-20 g), SI sarcoma Jensen (rats, non-inbred both sexes, 120-140 g), KSU of the Walker carcinosarcoma (rats, non-inbred, females, 120-140 g).

Leukemic tumor strains were transplanted intraperitoneally, dosing at (1.0-1.2) ^. 10 ⁶ C / mouse in 0.3 ml of 199 medium. Solid tumors were transplanted under the skin of the armpit or intramuscularly (CFL), injected with 0.3 ml of suspension of tumor tissue in 199 medium or isotonic sodium chloride solution. Treatment was started 72-96 hours after transplantation of solid tumors (except KSU) or 24 hours after inoculation of leukemia and KSU.

 The test compounds were dissolved in polyvinylpyrrolidone (IX) or in an isotonic sodium chloride solution (I-VIII) containing 10 mg or 1 mg in 1 ml, depending on the experimental conditions. Compounds were administered intraperitoneally 1 time per day for 5-9 days (or 3 times on the 1st, 5th and 9th days of the experiment at L 1210).

100 где М_к средняя масса опухоли в контроле;
М_о средняя масса опухоли в опыте.Animals with solid tumors were sacrificed 2 days after the treatment, and the antitumor effect was evaluated by calculating the inhibition index (V _m) by the formula
Y _t =

100 where M is _the average tumor mass in the control;
M _{about the} average mass of the tumor in the experiment.

100
и проценту излеченных животных, при этом излеченными считали животных, проживающих без рецидива в течение 2 мес.The therapeutic effect of leukemia was evaluated by the average life expectancy (LSS), determined by the formula:
SPG

one hundred
and the percentage of animals cured, while animals living without relapse for 2 months were considered cured.

 The chemotherapeutic index (CTI) was determined as the ratio of the maximum tolerated dose (causing the death of animals up to 10% or / and a smaller increase of no more than 15% compared to the control, which is observed constantly) to the minimum effective dose.

100, где МПД максимально переносимая доза.The cumulation index (IR) was calculated by the formula
IR

100, where MTD is the maximum tolerated dose.

The toxicity of compounds III and IX was assessed by determining the minimum dose that causes the death of the mouse after a single intraperitoneal injection in increasing doses (1 mouse per dose). The LD ₅₀ value for a single intraperitoneal administration to mice for the remaining compounds was determined by Litchfield and Walkoxon (comp. VI) or Kerber (compounds I, II, IV, V, VII, VIII) (3).

100, где А и А₁ средняя масса опухоли в начале опыта,
В и В₁ средняя масса животных в конце опыта,
С и С₁ средняя масса животных в начале опыта соответственно в подопытной и контрольной группах (4).Repeatability was assessed by the percentage of surviving animals in the group, as well as by calculating the coefficient of growth of body weight (K _p ) by the formula
K _p =

100, where A and A _{1 are the} average mass of the tumor at the beginning of the experiment,
B and B _{1 the} average mass of animals at the end of the experiment,
C and C _{1 the} average weight of animals at the beginning of the experiment, respectively, in the experimental and control groups (4).

When K _p with the sign "+" there was a more pronounced increase in body weight or less weight loss in experimental animals in the experiment compared with the control.

When K _p with the sign "-" the opposite relationship.

 The study of the antitumor activity of the claimed compounds was carried out in comparison with the well-known drug Sydnophene, exhibiting antitumor properties.

 The results of the study are presented in table. 1.

 Statistical processing of experimental data was carried out according to Belenky M. L. (3) with a probability level of 95% (TRULY-213 microcalculator) or using Student's criterion. The difference was considered significant at P ≅ 0.05 (the criterion for the reliability of the difference with control).

As can be seen from the table. 1, the claimed compounds I and II toxicity (LD ₅₀ are 90 mg / kg (I) and 113 mg / kg (II) and are comparable in activity to SI and KSU tumors with sydnophene (LD ₅₀ 123 mg / kg), but exhibit an effect on tumor Ca 755 (V _t 89% (I) and 80% (II)) and tumor SM / C _with 50% (II) and 48% (II) /, as well as leukemia P388 / SPM 163% ) (I) and 151% (II), significantly higher than Sydnophene / Y _t 31% (KSU) and 41% (SM), LNG 120% (P388) /.

 It should be emphasized that an increase in life expectancy (LSS) by 20% with leukemia P388 is below the limit of significance of the effect for this model, for a model for which an increase in LIV by 25% or more is considered significant (5).

Compound V is characterized by an 8-fold reduction in toxicity (LD ₅₀ 940 mg / kg) compared with sydnophen (LD ₅₀ 123 mg / kg), while it found activity comparable to sydnophen on SI, KSU and SM tumors and a significantly more pronounced activity on a Ca 755 tumor / _T 73% and for Sydnophene U _t 31% / and for P388 leukemia (SRF 144% and for Sydnophene SRF 120%).

 Thus, compounds III, IV, VIII, and IX, when antitumor activity is comparable to that of syndofen, are 3-5 times less toxic than sydnophen.

 Compounds I and II exhibit increased antitumor activity and new antileukemic properties with toxicity comparable to sydnophen.

 Compounds IV and V in the presence of increased antitumor activity and antileukemic properties are significantly less toxic than sydnophen.

The most active of a number of the claimed compounds was compound VI. When compared with sydnophene, it showed a significantly increased antitumor effect ( _T 83-100%) in all tumor models with reduced toxicity (LD ₅₀ 200 mg / kg, LD ₅₀ Sydnophen 123 mg / kg, and also found antileukemic properties (in Sydnophen they absent).

N

N

обладает высокой противоопухолевой и антилейкозной активностью благодаря выраженному цитостатическому действию на быстро пролиферирующие, в том числе опухолевые ткани.In addition, since this compound has the greatest resemblance to the antitumor drug thiophosphamide (2) that is widely used in world oncological practice, as this compound, as well as compound VI, which contains ethyleneimine groups in the molecule, which mainly determine antitumor activity, we considered it legitimate to additional comparison of compound VI and thiophosphamide. Thiophosphamide (N ^I , N ^II , N ^III -tri (ethylene) -thiophosphoric triamide) of formula B:

N

N

possesses high antitumor and antileukemic activity due to the pronounced cytostatic effect on rapidly proliferating, including tumor, tissues.

However, thiophosphamide is characterized by low selectivity of action on tumor cells due to high toxicity / with a single intraperitoneal injection of LD _{50 of} thiophosphamide into mice is 25 mg / kg (6) /.

 Thiophosphamide is characterized by a small therapeutic breadth of action and pronounced cumulation.

 The data obtained in the study of tolerance to the antitumor and antileukemic activity of compound VI, carried out in comparison with thiophosphamide (lyophilized thiophosphamide in ampoules of 0.01 each) are presented in table. 2-5.

Compound VI and thiophosphamide were administered in doses equally related to the LD _{50 of} each of them.

It was shown that compound VI with a single intraperitoneal administration to mice is 8 times less toxic than thiophosphamide (LD _{50 of} compound VI is 200 ± 22 mg / kg, thiophosphamide 25 mg / kg).

Of considerable interest are data from a comparative study of the tolerability of compound VI and thiophosphamide upon repeated use. Toxic effects (poor general condition, weight loss, death of some animals in the experiment) are observed with daily intraperitoneal injections of thiophosphamide in single doses of 0.125 LD ₅₀ for rats, 0.2 LD ₅₀ for mice with solid tumors (Table 2) and 0, 3 LD ₅₀ with leukemia (table. 3). A single dose of 0.25 LD ₅₀ for rats was fatal.

In contrast to thiophosphamide, animals tolerated compound VI well at doses that were toxic to thiophosphamide. The dose relative to LD ₅₀ , which is lethal for thiophosphamide (0.25 LD ₅₀ for rats and 0.4 LD ₅₀ for mice), is the maximum tolerated for compound VI.

 The life expectancy of mice with leukemia increased with increasing dose of compound VI, and in the case of increasing doses of thiophosphamide decreased.

 Evidence of less toxicity of compound VII is the results of determining the breadth of its therapeutic effect (CTI for compound VI is 20, for thiophosphamide -3).

 The cumulation index (IR) of compound VI (equal to 150) is significantly lower than that of thiophosphamide (IR = 900).

 With significantly better tolerance, compound VI exhibits antitumor activity equal to thiophosphamide with respect to Jensen's sarcoma and rat Walker's carcinosarcoma (Table 4). Both substances in optimal doses completely or almost completely inhibit the growth of these tumors. Compound VI causes a more inhibition than thiophosphamide inhibition of the development of B16 melanoma (up to 95% cf. with 73%), carcinoma 755 (up to 98% cf. 91%), Lewis lung cancer (up to 83% cf. 53%) and several less significant colon adenocarcinoma AKATOL (83% cf. 89%) tab. 5.

 Compound VI is significantly more active in mice with leukemia (Table 3). Life expectancy in mice with leukemia L 1210 treated with compound VI increases almost 3 times (with administration of thiophosphamide 1.6), in mice with leukemia P388 2.5 times (with administration of thiophosphamide 1.8 times). An increase in the dose of thiophosphamide leads to a decrease and then the complete disappearance of the antileukemic effect due to toxic manifestations.

 Thus, compound VI has a more pronounced or equal activity with thiophosphamide with significantly better tolerance.

 Since the studied compounds (I-IX) are phosphorylated derivatives of the original domestic psychotropic drug Sidnofen, it seemed necessary to evaluate their effect on the central nervous system (CNS) of animals in the experiment.

 The effect on the central nervous system was evaluated by indicators characterizing the central stimulating effect: the effect on the general condition and behavior of animals, the effect on spontaneous motor activity (8), behavioral depression caused by an unavoidable stressful situation (9), and the effect on the effects of reserpine (hypodynamia, hypothermia, blepharoptosis )

It is known (10, 11) that Sydnophene has a psychostimulating and antidepressant effect on the central nervous system. In an animal experiment, sydnofen at doses of 12.5 mg / kg intravenously and 25 mg / kg orally causes CNS excitation, which is expressed in increased audiogenic and tactile hyperreflexia, aggressive behavior, with increasing doses up to 50 mg / kg, these phenomena are amplified. The spontaneous locomotor activity of sidnofen in the indicated doses did not change significantly. At a dose of 25 mg / kg sidnofen to effectively antagonize central effects of reserpine, reducing ptosis score of 1.5, 1.5 hypothermia ^C., physical inactivity, and restores to a level corresponding to the spontaneous locomotor activity of control animals. In a model of behavioral depression in mice, Sydnophene at a dose of 12.5 and 25 mg / kg orally 2.5-3 times compared with the control group increases the number of active attempts to avoid in this situation.

The study of compounds IX at doses of 1/10 of their LD ₅₀ when administered orally showed that the compounds do not have a pharmacological activity spectrum that is characteristic of sydnophene. Elements of antidepressant activity are observed only in compounds I-VI in the test characterizing behavioral depression, and are not completely decisive for characterizing the psychotropic effect, since in other indicators (effect on locomotor activity and reserpine effects) they practically did not differ from the control level (tab. 6). Certain changes in the behavior of animals (hyperreflection, aggressiveness and lack of exercise), if they were noted, then only in doses exceeding 1/10 of the LD _{50 by} 2-2.5 times.

 Thus, the studies of the claimed compounds (phosphorylated derivatives of sydnophene) on the effect on the central nervous system show that compounds IX-IX do not have a stimulating effect on the central nervous system characteristic of sydnophene in animal experiments. The elements of antidepressant activity observed in compounds I-VI, although they indicate a certain activating effect, are manifested in doses exceeding sydnophen by 4-6 times.

 In conclusion, we can conclude that, as a result of studying the biological properties of the claimed compounds, it was found that for the first time in a series of sydnonimines, compounds were found that exhibit antileukemic properties along with antitumor properties, which was previously unusual for compounds of this series, while not possessing psychotropic properties characteristic of Sydnophene, a structural analogue of the claimed compounds.

 The inventive compounds are obtained in various ways.

N

Cl +
+ CH₃OH

H(CH₃)

I а (R₁ CI) I(R₁ R₂ OCH₃) IIа (R₁ CH₃) II(R₁ CH₃; R₂ OCH₃)
Исходные соединения Iа, Па новые, ранее неописанные в литературе, получают in situ взаимодействием N-нитрозо-2-( β-фенилизопропиламино)ацетонитрила с хлорокисью фосфора или дихлорфосфонатом в абс. ацетонитриле при 70^оС, т. е. в условиях, аналогичных описанным ранее для реакции N-нитрозо-2-(β-фенилизопропиламино)ацетонитрила с тиофосфорилхлоридом, приводящей в N⁶-(дихлортиофосфорил)-3-β -фенилизопропилмиднонимину (12).Compound I, II is prepared by reacting methanol with N ⁶ - (dichlorophosphoryl) -3-β-phenylisopropylimidonimonimine (1a) or previously described N ⁶ - (chloromethylphosphonyl) -3-β-phenylisopropylsidonimonimine (Pa) in benzene in the presence of triethylamine at room temperature :

N

Cl +
+ CH ₃ OH

H (CH ₃ )

I a (R ₁ CI) I (R ₁ R ₂ OCH ₃ ) IIa (R ₁ CH ₃ ) II (R ₁ CH ₃ ; R ₂ OCH ₃ )
The starting compounds Ia, Pa, previously not described in the literature, are prepared in situ by the reaction of N-nitroso-2- (β-phenylisopropylamino) acetonitrile with phosphorus oxychloride or dichlorophosphonate in abs. acetonitrile at ⁷⁰ ° C, ie, under conditions similar to those described previously for the reaction of N-nitroso-2- (β-fenilizopropilamino) acetonitrile tiofosforilhloridom resulting in ⁶ N -.. (dihlortiofosforil) -3-β -fenilizopropilmidnoniminu (12) .

 The inventive compound III is obtained by the interaction of Ia with an excess of dimethylamine in benzene at room temperature.

 The inventive compounds IV and V are prepared by reacting the corresponding primary and secondary amine with Ia in the presence of triethylamine in benzene at room temperature, followed by treating the resulting imidoamidochlorophosphate (IVa, Va) with potassium hydroxide in dioxane-water medium.

Intermediates IVa and Va are new, not described in the literature, and characterized by ³¹ P. NMR spectra.

 The inventive compound VI is prepared by reacting Ia with a slight excess of ethyleneimine in the presence of triethylamine (molar ratio of reactants 1: 2, 2: 2) in benzene at room temperature.

The inventive compound VII is obtained by the interaction of the previously described (12) N ⁶ - (dichlorothiophosphoryl) -3-β-lsidinonyminine (VII) with sodium hydroxide in a dioxane-water binary solvent (volume compound 1: 1.5) at room temperature.

N

+
+ NaOH

Заявляемое соединение IХ получают взаимодействием VIIа с метилом в присутствии избытка гидроксида натрия в бинарном растворителе метанол-диоксан (объемное соотношение 1,5:1) при комнатной температуре.The claimed compound VIII is prepared by reacting previously described (13) N ⁶ - (1,3,2-dioksofosfolan-2-yl) -3- β-fenilizopropilsidnonimina and sulfur in benzene at 60 ° ^C followed by treatment of the resulting thio derivative VIIIa (new, undescribed in the literature) sodium hydroxide according to the scheme:

N

+
+ NaOH

The inventive compound IX is obtained by the interaction of VIIa with methyl in the presence of an excess of sodium hydroxide in a binary solvent methanol-dioxane (volume ratio 1.5: 1) at room temperature.

PRI me R 1. Synthesis of N ⁶ - (methoxymethylphosphonyl) -3-β-phenylisopropylsidononimine (II).

 To 3.0 g (10 mmol) IIa and 1.7 ml (12 mmol) of triethylamine in 15 ml of abs. 0.46 ml (11 mmol) of abs. methanol in 4 ml of benzene. After 3 hours, the mixture was filtered, evaporated, the residue was reprecipitated from benzene twice with petroleum ether.

Obtain 2.33 g (79%) of II in the form of a colorless amorphous hygroscopic substance, soluble in water and organic solvents, so pl. 84-86 ^about S.

 Found, C, 52.59; H 6.33; P 10.16.

C ₁₃ H ₁₈ N ₃ O ₃ R.

 Calculated, C 52.88; H, 6.14; P 10.49.

³¹ P NMR Spectra (C ₆ H ₆ , δ ppm): 29.5.

PMR spectrum (CDSI, δ ppm): 1, 406, 1.415 (3Н, ∂.∂ RSN ₃ , ³ J (pH) 17.0 Hz); 1.56 (3H, ∂, CH ₃ ), 3.10 (2H, M, CH ₂ ), 3.504; 3,556 (3H, ∂.∂ OCN ₃ , ³ J (pH) 11.6 Hz), 4.65 (IH, m, CH), 7.0-7.3 (6H, m, C ₆ H ₅ , C ⁴ N). UV spectrum (H ₂ O): λ 317 nm (ε 6900).

Similarly, N ⁶ -dimethoxyphosphoryl) -3-β-phenylisopropylsidononimine (I) was obtained from Ia. Yield 67% T. pl. 45.5-47.5 ^about C (from a mixture of benzene-hexane). Colorless amorphous hygroscopic substance, soluble in water and organic solvents.

 Found, C 49.82; H 5.89; P 9.72.

C ₁₃ H ₁₈ N ₃ O ₄ R.

 Calculated, C 50.16; H 5.83; P 9.95.

Nuclear Magnetic Resonance Spectrum ³¹ P (C ₆ H ₆ , ∂ ppm): 6.9.

1 H-NMR spectrum (CDSl, δ ppm): 0.975, 0.980 (1: 2) (3H, ∂ ∂ CH ₃ ), 2.68 (2H, m, CH ₂ ), 3.685, 3.682, 3.712 and 3.717 (1 : 2: 1: 2) (6Н, ∂ ∂ ∂ ∂ ³ J (PH) 11.3 Hz), 4.60 (1Н, m, СН), 6.9-7.2 (5Н, m, С ₆ Н ₅ ), 7.96 (1H, s, C ⁴ H).

UV spectrum (H ₂ O): λ 310 nm (ε 9000).

PRI me R 2. Synthesis of N ⁶ - (bisdimethylamidophosphoryl) -3-β-phenylisopropylsidononimine (III).

 To a solution of 6.4 g (20 mmol) of Ia in 20 ml of abs. 4.0 g (88.7 mmol) of dimethylamine in 10 ml of benzene are added dropwise. After 5 hours, the mixture was evaporated, the residue was dissolved in 40 ml of benzene, the solution was filtered, the excess solvent was distilled off, the product was reprecipitated twice with benzene pentane.

Obtain 5.6 g (83%) of III in the form of an amorphous colorless substance, stable in air and in neutral aqueous solutions, soluble in organic solvents. T. pl. 98-99 ^about S.

 Found, C, 53.25; H 7.02; P 9.27.

C ₁₅ H ₂₄ N ₅ O ₂ R.

 Calculated, C 53.40; H 7.17; P 9.18.

Nuclear Magnetic Resonance Spectrum ³¹ P (C ₆ H ₆ , δ ppm): 20.7.

1 H-NMR spectrum (CDSl, δ n.a.): 1.53 (3H, δ CH ₃ ), 2.54, 3.53 (12H, ∂, ∂ CH ₃ ), ³ J (pH), 10.5 Hz ); 3.08 (2H, m, CH ₂ ), 4.57 (1H, m, CH), 7.0-7.2 (6H, m, C ₆ H ₆ , C ⁴ H).

UV spectrum (H ₂ O): λ 317 nm (ε 8100).

 PRI me R 3. Synthesis of 3-β-phenylisopropylsidonimonido-β-phenethylamidophosphate of potassium (IV).

 To a solution of 6.4 g (20 mmol) of Ia and 3.5 ml (25 mmol) of triethylamine in 15 ml of abs. 2.6 ml (21 mmol) of β-phenethylamine are added dropwise in benzene in 6 ml of benzene. After 5 hours, the solution is filtered, the mother liquor is evaporated to dryness. The residue was reprecipitated twice from benzene with pentane.

 6.5 g (80%) of IVa are obtained in the form of a viscous oil soluble in organic solvents.

Spectrum MR ³¹ P (C ₆ H ₆ , δ ppm): 18.0.

6.1 g (15 mmol IVa in 12 ml of dioxane are added dropwise with stirring to 1.68 g (30 mmol) of potassium hydroxide in 8 ml of water. After 3 hours, excess solvent is distilled off under reduced pressure. The aqueous solution is filtered and water is removed from the azeotropic mother liquor distillation with abs. benzene. The residue was washed with benzene, dissolved in 15 ml of chloroform, the solution is dried over calcium chloride, filtered and evaporated. The product was triturated with hexane, dried under Fischer gun over phosphorus pentoxide at 60 ° ^C.

5.5 g of IV are obtained (87% counting on IVa, or 70% counting on starting Ia). T. pl. 201 ^C. (dec.).

 Found, C 53.45; H 5.40; P, 6.90.

C ₁₉ H ₂₂ N ₄ O ₃ RK.

 Calculated, C 53.76; H 5.22; P, 7.30.

³¹ P NMR Spectrum (CHCl, δ ppm): 7.2.

1 H-NMR spectrum (CDL ₃ , δ ppm): 1.62 (3H, ∂ CH ₃ ), 2.7-3.0 (4H, m, CH ₂ CH ₂ ), 3.16 (2H, m, CH ₂ ), 4.71 (1H, m, CH), 7.0-7.25 (6H, m, C ₆ H ₅ , C ⁴ N).

UV spectrum (H ₂ O): λ 327 nm (ε 6600).

Similarly receive:
a) N ⁶ - (morpholidochlorophosphoryl) -3-β-phenylisopropylsidimino (Va).

Yield 75%
Nuclear Magnetic Resonance Spectrum ³¹ P (C ₆ H ₆ , δ ppm): 16.1.

 b) potassium (V) 3-β-phenylisopropyl sidonimonidido-morpholidophosphate.

Yield 60% counting on Ia. T. pl. 186-189 ^about S.

 Found, C, 45.93; H 5.38; P 7.53.

C ₁₅ H ₂₀ N ₄ O ₄ RK.

 Calculated, C 46.14; H 5.16; P 7.93.

³¹ P NMR Spectrum (CHCl ₃ , δ ppm): 7.5.

PMR spectrum (CDL ₃ , δ ppm): 1.54 (3H, ∂ CH ₃ ), 2.82 (4H, m, NCH ₂ ), 2.9-3.3 (2H, m, CH ₂ ), 3.53 (4H, m, OCH ₂ ), 4.68 (1H, m, CH), 7.0-7.3 (5H, m, C ₆ H ₅ ), 7.34 (1H, s , C ⁴ H).

UV spectrum (H ₂ 0): λ 326 nm (ε 8400).

PRI me R 4. Synthesis of N ⁶ - (bisethylenimidophosphoryl) -3- β-phenylisopropyl sidonimide (VI).

 To 4.5 g (14 mmol) of Ia and 4.2 ml (30 mmol) of triethylamine in 20 ml of abs. While stirring benzene, 1.8 ml (35 mmol) of ethyleneimine in 10 ml of benzene are slowly added dropwise. The mixture was incubated for 4 hours at room temperature, filtered and evaporated to dryness.

Obtain 4.1 g (88%) of VI in the form of a hygroscopic amorphous substance soluble in water and organic solvents, slightly soluble in ether and dioxane, unstable to acid reagents. Neutral aqueous solutions are stable for several weeks. T. pl. 90-92 ^about S.

 Found, C, 53.83; H 5.96; P 9.03.

C ₁₅ H ₂₀ N ₅ O ₂ R.

 Calculated, C 54.05; H 6.05; P 9.29.

NMR spectrum ³¹ P (C ₆ H ₆ , δ ppm): 28.3.

PMR spectrum (CDCl ₃ , δ ppm): 1.62 (3H, ∂ CH ₃ ), 2.0-2.2 (8H, m, NCH ₂ ), 3.16 (2H, m, CH ₂ ), 4.67 (1H, m, CH), 7.0-7.35 (6H, m, C ₆ H ₅ , ⁴ CH).

UV spectrum (H ₂ O): λ 312 nm (ε 8700).

 PRI me R 5. Synthesis of disodium salt of 3-β-phenylisopropylsidonimidothiophosphoric acid monohydrate (VII).

3.36 g (10 mmol) of VIIa in 8 ml of dioxane was added dropwise to 1.60 g (40 mmol) of sodium hydroxide in 12 ml of water, after 12 hours the solution was added 150 ml of acetone, the mixture was allowed to stand 48 hours at 0 ° ^C, filtered . 5 ml of methanol was added to the precipitate, the solution was filtered and evaporated under reduced pressure.

Obtain 3.14 g (87% based on the anhydrous salt) VII in the form of an amorphous hygroscopic substance, soluble in water and methanol. T, pl. 238 ^C. (dec.).

 Found, C 36.30; H 4.10; P 8.17.

C ₁₁ H ₁₂ N ₃ O ₃ PSNa ₂ ^. H ₂ O.

 Calculated, C 36.57; H 3.90; P 8.57.

Nuclear Magnetic Resonance Spectrum ³¹ P (H ₂ O, δ ppm): 35.7.

PMR spectrum (D ₂ O, δ ppm) 1.43 (3Н ∂CH ₃ ), 3.01 (2H, m, CH ₂ ), 4.65 (1H, m, CH), 6.9- 7.2 (5H, m, C ₆ H ₅ ).

UV spectrum (H ₂ O): λ 338 nm (ε 8100).

 PRI me R 6. Synthesis of 3-β-phenylisopropylsidonimonido-β-hydroxyethylphosphate sodium monohydrate (VIII).

Suspension 2.93 g (10 mmol) of N ⁶ - (1,3,2-dioxaphospholan-2-yl) -3-β-phenylisopropylsidononimine (VIIIa) in 15 ml of abs. benzene and 0.38 g (12 mmol) of sulfur are stirred for 60 min at 60 ^about C. After filtering and distilling off the solvent, the resulting St. yellow oil is wiped with abs. methanol.

Obtain 3.1 g (95%) N ⁶ - (2-thio-1,3,2-dioxophospholan-2-yl) -3-β-phenylisopropylsidononimine VIIIb in the form of a light yellow amorphous substance that dissolves in organic solvents. T. pl. 91.5 ^o C (from methanol).

³¹ P NMR spectrum (CHCl ₃ , δ ppm): 81.6.

PMR spectrum (CDSl ₃ , δ ppm): 1.58 (3H, ∂, CH ₃ ); 3.11 (2H, m, CH ₂ ), 4.29, 4.30 (4H, ∂. ∂ OCH ₂ CH ₂ O, ³ J (pH) 11.6 Hz), 4.71 (1H, m, CH), 7.0-7.2 (5H, m, C ₆ H ₅ ), 7.54 (1H, ∂ CH, ⁴ J (pH), 1.1 Hz).

IR spectrum (nujol, cm ^-1 ): 1630 (C = N), 3150 (C ⁴ H).

UV spectrum (CH ₃ CN): λ 323 nm (ε 8300).

To 3.0 g (9.2 mmol) VIIIb in 10 mL of dioxane at 60 ° ^C was added dropwise with stirring 0.40 g (10 mmol) of sodium hydroxide in 5 ml of water. The mixture is kept for 2 hours, the solvent is distilled off under reduced pressure. The residue was dissolved in 25 ml of chloroform. The solution was kept over freshly calcined sodium sulfate, filtered, and the solvent was distilled off. The product is dissolved in 5 ml of water, the solution is filtered through a dense layer of cotton wool, evaporated and the drying procedure is repeated.

Obtain 2.3 g of VIII (66), counting on VIIIb, or 56% counting on the original VIIIa, in the form of an amorphous hygroscopic substance, the tendency to dissolve of which in water decreases after dehydration, so pl. 68-69 ^about S.

 Found, C 40, 43; H, 5.30; P 7.64.

C ₁₃ H ₁₇ N ₃ O ₄ PSNa ^. H ₂ O.

 Calculated, C 40.73; H 5.00; P 8.08.

³¹ P NMR Spectrum (CDC ₃ , δ ppm): 52.8 and 53.1 (1: 1).

1 H-NMR spectrum (CDL ₃ , δ ppm): 1.38 and 1.39 (3H, ∂ CH ₃ ), 3.06 (2H, m, CH ₂ ), 3.6-4.0 (6H, m, POCH ₂ , CH ₂ O, H ₂ O), 5.40 (1H, s, OH), 7.0-7.2 (5H, m, C ₆ H ₅ ), 7.48 and 7.57 (1: 1) (1H, C ⁴ H).

IR spectrum (nujol, cm ^-1 ): 1640 (C = N).

UV spectrum (H ₂ O): λ 325 nm (ε 6500).

 PRI me R 7.

Synthesis of N ⁶ - (dimethoxythiophosphoryl) -3-β-phenylisopropylsidononimine (IX).

 To 6.7 g (0.12 mol) of potassium hydroxide in 30 ml of methanol, 10.0 g (29.7 mmol) of VIIa in 20 ml of dioxane are added with stirring. After 30 minutes, the solvent was distilled off in vacuo. The residue is dissolved in 50 ml of chloroform, the solution is washed with water, dried over calcium chloride. The oil formed after distillation of chloroform is rubbed with a small amount of triethylamine and washed with pentane.

9.3 g (96%) of IX are obtained in the form of an amorphous hygroscopic substance soluble in organic solvents. T. pl. 53-55 ^about S.

 Found, C, 47.90; H 5.49; P 9.61.

C ₁₃ H ₁₈ N ₃ O ₃ PS.

 Calculated, C 47.70; H 5.54; P 9.46.

Nuclear Magnetic Resonance Spectrum ³¹ P (CH ₃ OH), δ ppm 68.2.

PMR spectrum (СДСl ₃ , δ ppm): 1.62 (3Н, ∂СН ₃ ), 3.16 (2Н, m, СН ₂ ), 3.58 and 3.68 (3Н, ∂∂ OCН ₃ , ³ J (pH) 11.2 Hz), 4.77 (1H, m, CH), 7.05-7.25 (5H, m, C ₆ H ₅ ), 7.38 (1H, s, C ⁴ N).

UV spectrum (CH ₃ OH): λ 318 nm (ε 8600).

Claims (1)

N ⁶ Phosphorylated derivatives of 3-b-phenylisopropylsidonimonimine of the general formula I

where, if X is oxygen and n 0, then
I. R ₁ R ₂ OCH ₃ ;
II. R ₁ CH ₃ ; R ₂ OCH ₃ ;
III. R ₁ R ₂ N (CH ₃ ) ₂ ;
IV. R ₁ N HCH ₂ CH ₂ C ₆ H ₅ ; R ₂ - O ^- K ⁺ ;

VI. R ₁ R ₂ N (CH ₂ ) ₂ , if X is sulfur, then
VII. R ₁ R ₂ O ^- Na ⁺ , n 1;
Viii. R ₁ OCH ₂ CH ₂ OH; R ₂ O ^is Na ⁺ , n 1;
IX. R ₁ R ₂ OCH ₃ ; n 0,
possessing antitumor activity.

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