RU2483066C1 - New antithrombotic and antiatherosclerotic agent and method for preparing it (versions) - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry and medicine, and concerns compounds that can be used to develop agents for cardiovascular disease. What is presented is the use of 2-[2-[5-(hydroxymethyl)-3-methyl-1,3-oxazolidin-2-ylidene]-2-cyanoethylidene]indolin-3-one of formula

as a biologically active compound exhibiting the properties of an exogenous nitric oxide donor, an activator of the enzyme soluble guanylate cyclase, a blood platelet aggregation inhibitor with antihypertensive activity for preparing cardiovascular diseases. The invention refers to a method for preparing 2-[2-[5-(hydroxymethyl)-3-methyl-1,3-oxazolidin-2-ylidene]-2-cyanoethylidene]indolin-3-one of formula I which is prepared by the reaction of α-cyano-β-(3-acetoxyindolyl-2)acrylic acid nitrile of formula

with N-methylaminopropane-2,3-diol (MeHN-CH₂CH(OH)CH₂OH (formula IIIa)) in a polar solvent on boiling. Also, the invention refers to a version of the method for preparing 2-[2-[5-(hydroxymethyl)-3-methyl-1,3-oxazolidin-2-ylidene]-2-cyanoethylidene]indolin-3-one of formula I characterised by the fact that N-acetylindoxyl and ethoxymethylene malondinitrile are reacted to form N-acetyl-3-hydroxy-2-dicyanovinylindole, then acetylated by acetic anhydride to N-acetyl-3-acetoxy-2-dicyanovinylindole, further reacted with an equimolecular amount of N-methylaminopropane-2,3-diol (IIIa) to form 1-acetyl-2-[2'-cyano-2'-(3"-methyl-5"-oxymethyloxazolidin-2"-ilidene)-ethylidene]indolinone-3 of formula

;

thereafter an acetyl group is removed in a methanol medium by the reaction of triton B representing 40% benzyl trimethylammonium hydroxide in methanol; the precipitation is washed, dried and purified by re-crystallisation from the mixture of dimethyl formamide and water to prepare the end product.

EFFECT: preparing the indolinone derivatives for preparing the agents for cardiovascular diseases.

3 cl, 4 tbl, 3 dwg, 11 ex

Description

The invention relates to the field of the pharmaceutical industry and medicine, and relates to compounds that can be used to create agents for treating cardiovascular diseases.

Cardiovascular disease remains the leading cause of death in humans, and one of the most seriously considered causes of this phenomenon is atherosclerosis and high levels of low density lipoproteins in the blood. Atherosclerosis is a chronic cardiovascular disease, expressed in the compaction of the arterial wall due to proliferation of connective tissue, the formation of atherosclerotic plaques, narrowing of the lumen of blood vessels, and deterioration of blood supply to tissues.

Dysregulation of cholesterol production leads to serious pathologies. When its total production (synthesized and supplied with food) exceeds the amount required to build and repair membranes, produce bile acids and steroids, the accumulation of cholesterol in the vessels (atheromatosis) can cause them to become clogged (atherothrombosis). Cardiovascular disease occurs due to high cholesterol and is especially caused by an increased level of low density lipoproteins (LDL).

A modern view of atherosclerosis is that it is a chronic inflammatory disorder. And anti-inflammatory drugs such as aspirin, which also has anti-aggregation properties, as well as angiotensin-converting enzyme inhibitors and anti-inflammatory cytokines, are useful for patients affected by atherosclerosis.

Hypertension is often associated with atherosclerotic lesions of the vascular wall and, therefore, studies aimed at studying the effect of antihypertensive agents on the development of atherosclerosis are important. According to some studies, calcium antagonists have proven anti-atherogenic properties, expressed in preventing the appearance of atherosclerotic lesions. Inhibitors of the angiotensin-converting enzyme also exhibit an anti-atherogenic effect in animal experiments. Calcium antagonists verapamil and amlodipine inhibit the atherogenic effect by inhibiting serum-induced cell proliferation, protein synthesis, and cholesterol accumulation in cells. Both drugs reduce the manifestations of atherosclerosis, showing antiatherosclerotic and antiatherogenic activity in cell culture. On the contrary, perindoprilat does not affect atherosclerotic parameters, and propranolol, moreover, even stimulates the accumulation of cholesterol, proliferative activity of cells and protein synthesis. According to the authors of [9], the data obtained on cell cultures can reflect the situation in vivo. A study of patients with coronary atherosclerosis in the treatment of calcium antagonists or propranolol showed that the latter in these conditions to some extent inhibits atherogenesis. Based on the studies, the authors constructed the following series of efficacy of the considered drugs: amlodipine> verapamil> perindopril> propranolol.

Among the drugs widely used for the treatment of atherosclerosis, statins, such as lovastatin, simvastatin, pravastatin, atorvastatin, fluvastatin, undoubtedly belong.

It is important to note that the mechanism of action of statins is associated with the inhibition of the rate-limiting stage of cholesterol biosynthesis - the conversion of β-hydroxy-β-methylglutaryl-CoA (HMG-CoA) to mevalonate, which is catalyzed by the enzyme - HMG-KoA reductase.

In a study [10], it was noted that despite the success of using statins in effectively lowering cholesterol and reducing mortality in cardiovascular diseases, about two thirds of patients treated with statins still continued to detect coronary artery damage.

In addition to, for example, increasing the level of high density lipids, chronic inhibition of the development of systemic and vascular inflammatory processes is considered as another promising area of treatment for atherosclerosis [8].

Thus, at present, a complex of drugs is used to treat atherosclerosis, since monotherapy is not possible. The main reason is the lack of a substance that has the most necessary spectrum of pharmacological activity: anti-inflammatory, antiaggregational, antihypertensive and hypocholesterolemic. The search for such a molecule is an urgent task of pharmacology.

In the past decade and a half in biology, events have taken place that have entailed fundamental changes in our ideas about the functioning of various biological systems. It was found that such a low molecular weight compound as nitric oxide (NO) is one of the universal and necessary regulators of the functions of cellular metabolism. Nitric oxide is involved in the regulation of blood vessel tone, inhibits platelet aggregation and adhesion on the walls of blood vessels, functions in the central and autonomic nervous system, regulating the activity of the respiratory system, gastrointestinal tract and genitourinary system. Nitric oxide plays an important role in neurotransmission, regulation of immunity, and protecting the body from bacterial damage.

Nitric oxide is known as a powerful inhibitor of platelet aggregation. Its formation helps maintain blood flow due to inhibition of thrombogenesis. Factors that regulate the occurrence and utilization of NO under endogenous conditions, ensure in many respects the normal functioning of the cardiovascular system.

In connection with the foregoing, the search for substances contributing to the emergence of NO as an agent that inhibits the spontaneous and induced ability of the platelets to adhere to blood is of great importance for modern pharmacology. Thus, the search for new effective compounds that may be in demand in medical practice was built in the present study on the synthesis and biological study of new nitric oxide generators.

In the indolinone-3 series, compounds are described that have antihypertensive activity [2, 4, 5, 6] and the ability to generate nitric oxide [1], but compounds that are capable of activating enzyme-soluble guanylate cyclase and exerting antiaggregatory action are not described.

The aim of the invention is the use of a derivative of indolinone-3 as a biologically active compound, which can be used to obtain funds for the treatment of cardiovascular diseases.

The goal is achieved by using a compound exhibiting the properties of an exogenous nitric oxide donor, an activator of the enzyme soluble guanylate cyclase, an inhibitor of platelet aggregation with antihypertensive activity.

The present invention relates to the chemistry of compounds of the indolinone-3 series and specifically relates to a derivative of this series - 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indolin- 3-one of the formula I:

Studies of anti-aggregation properties, effects on the activity of soluble guanylate cyclase and the generation of NO of compound I in vitro, as well as on the ability to exert hypotensive and antihypertensive effects in animals were carried out. The general toxicity and tolerability of the compounds of formula I in animals were investigated.

Studies have shown that it has pronounced anti-aggregation properties, releases NO, and activates highly soluble guanylate cyclase.

Example 1

The study of anti-aggregation properties was carried out by the effect on the platelet aggregation of rabbit plasma induced by ADP (adenosine diphosphate) and arachidonic acid. Platelet aggregation was evaluated by changing the optical density of the plasma. Studies were carried out both when administered intravenously in comparison with sodium nitroprusside, and when administered orally in comparison with acetylsalicylic acid. I was used in concentrations of 10 ^-3 -10 ^-7 (when administered parenterally) and in doses of 4-8 and 12 mg / kg when administered orally.

It was experimentally established that the compound of formula I significantly inhibits ADP-induced platelet aggregation.

To make a final conclusion about the ability of the studied compound to inhibit platelet aggregation, experiments were carried out in a living organism with its intravenous and oral administrations.

Compound I and sodium nitroprusside (as a reference drug) caused a unidirectional change in the platelet aggregation process - they suppressed it. However, there were significant quantitative and temporal differences in the effect. So, sodium nitroprusside actively (reliably) inhibited platelet aggregation 15 minutes after administration. This time point is the only one in which we were able to observe activity. After 1 h, the effect disappeared. Inhibition of platelet aggregation under the action of substance I was also observed 15 minutes after injection (as a tendency), reached its maximum values after 60 minutes and stopped by 3 hours of the experiment (Fig. 1).

Thus, these series of experiments showed the ability of a compound of formula I to inhibit ADP-induced adhesion of blood platelets after intravenous administration.

Significant inhibition of the aggregation process induced by ADP was detected by the first hour (30%), reached a maximum after 2 hours (40%) and, as a trend (15-25%), all 4 hours of observation continued (Fig. 2).

When using arachidonic acid (AA) as an aggregation inducer, experiments were also carried out under conditions of oral administration. We believed that such a formulation of experiments is fully justified by the possibility of prescribing the studied substance (in contrast to sodium nitroprusside) not only intravenously.

It turned out that the compound of formula I begins to act within 1 h after application, while the degree of initiation of aggregation of blood plates by arachidonic acid fell by a third. Even more significantly, the action of the inductor (AA) was suppressed by the second hour - more than twice. By the end of the observation period (4 hours), the effect of AA was comparable with the control (Fig. 3).

The effect of the compound of formula I persisted for more than 4 hours. By the end of the first hour, it was detected quantitatively as a trend (up to 10%), became statistically significant after 2 hours (32%) and continued to grow throughout the observation period (38% by 4 hours).

Thus, platelet aggregation induced by AA was effectively suppressed by the compound of formula I administered per os, and activity was manifested for more than two hours.

нейтрофилами человека, предварительно ослабленное развитием патологических процессов (гистеоцитоз, гипертоническая болезнь II ст.). Данная постановка опыта была выбрана в связи с тем, что для осуществления завершенного фагоцитоза, сопровождающегося полным уничтожением патогенных микроорганизмов, фагоциты, в том числе и нейтрофилы, образуют так называемые активные формы кислорода (АФК), к которым относятся радикалы кислорода и соединения, легко превращающиеся в такие радикалы. К АФК относятся супероксид анион радикал

, гидроксильный радикал (ОН^•) и гидроксиперекиси липидов (ROO^*). Все эти соединения имеют на внешней молекулярной орбитали один неспаренный электрон, что и определяет их высокую реакционную способность, приводящую к взаимодействию с молекулами клетки: белком, РНК, ДНК и др., сопровождающемуся потерей их биологической активности.We also investigated the effect of this compound on the formation of superoxide anion radicals induced by formyl-methionyl-leucyl-phenylalanine (FMLP)

human neutrophils, previously weakened by the development of pathological processes (hysteocytosis, hypertension II century). This experimental design was chosen due to the fact that for complete phagocytosis accompanied by the complete destruction of pathogenic microorganisms, phagocytes, including neutrophils, form the so-called active forms of oxygen (ROS), which include oxygen radicals and compounds that are easily converted into such radicals. ROS include superoxide anion radical

, hydroxyl radical (OH ^• ) and lipid hydroxyperoxides (ROO ^* ). All these compounds have one unpaired electron on the external molecular orbital, which determines their high reactivity, leading to interaction with cell molecules: protein, RNA, DNA, etc., accompanied by the loss of their biological activity.

It should be noted that along with protective functions (inactivation of bacterial toxins), oxygen radicals (with prolonged action) often have a detrimental effect on surrounding cells, up to their death. For this reason, the search for chemical compounds that briefly enhance the formation of ROS or suppress this process is an urgent task of modern pharmacology.

возникает высокореактивное соединение пероксинитрит (ONOO^•), который, по-видимому, эффективно влияет на липидные компоненты цитоплазматической мембраны клеток, в том числе остатки АА.After incubation of the compound of formula I with whole blood as a result of the interaction of NO with

a highly reactive compound peroxynitrite (ONOO ^• ) appears, which, apparently, effectively affects the lipid components of the cytoplasmic membrane of cells, including AA residues.

As a result of this, the release of AA from the phospholipid layer of plasma membranes may be impaired, and consequently, its subsequent metabolism with the formation of thromboxane A ₂ (TxA ₂ ) in platelets, which is known to be a powerful endogenous vasoconstrictor and proaggregant, which reduces the functional activity of blood platelets.

The data obtained suggest that one of the possible mechanisms of the anti-aggregation action of the compounds of formula I may be the activation of the formation of oxygen radicals in whole blood (“respiratory explosion”), which allows to reduce the level of thrombogenic factors.

A model of parenchymal bleeding characterizes hemostasis through stimulation of a thrombus-forming cascade of reactions. As a rule, it is rather difficult to obtain significant shifts in bleeding time on such models using resorptively acting compounds. Direct type anticoagulants (heparin, heparinoids) only at high doses promote prolongation of bleeding in such models. In this case, the effect is detected at different times (up to 3 hours after administration). Indirect anticoagulants (oxycoumarins, indandions, etc.) begin to increase bleeding, usually after a certain incubation period (from 12 to 48 hours after application). The latter is easily explained on the basis of the modern understanding of their mechanism of action: they stop the biosynthesis of plasma coagulation factors II, VII, IX, X, and the duration of the incubation period is determined by the number of these factors accumulated in the liver before administration of K-vitamin reductase inhibitors (oxycoumarins, indandions) .

In the control group of animals, the duration of parenchymal bleeding was 88.9 ± 1.2 s. The experimental groups of animals were taken into the experiment 60 minutes after the oral administration of the compound of formula I. At a dose of 8 mg / kg, the bleeding time increased by 71% compared to the initial values and after another 1 hour this effect intensified and amounted to 220.6 ± 3. 4 sec By the end of the experiment (240 min), this indicator did not differ from the control figures. At a dose of 16 mg / kg, the bleeding time doubled and was recorded only in the first 60 minutes of observation.

The discovered effect, apparently, can be explained by the previously established ability of the compound of formula I to inhibit the platelet link of hemostasis, which is realized through the generation of nitric oxide, regulation of the level of intracellular calcium in these blood cells, etc.

Another fact indicating the promise of this compound is the high LD ₅₀ value (8900 mg / kg), which corresponds to toxicity class V and allows it to be classified as practically non-toxic.

Summarizing all of the above (taking into account the experiments performed, which revealed the ability of NO donors to inhibit the induced platelet aggregation), it can be stated that the claimed new compound of formula I in the body of rabbits with parenteral (vein) application strongly inhibits the adhesion of blood platelets.

We can also consider the ability of a new NO donor (compound I) to be absorbed from the gastrointestinal tract and to show its anti-aggregation activity under enteral conditions. Of course, compared with intravasal administration, it appears a little later, but remains for a significantly longer period (the substance acted for more than 2-4 hours on AA-induced aggregation and about 3 hours on ADP-induced).

Test results

1. In vitro, the antiplatelet activity of the newly synthesized nitric oxide donor, a compound of formula I, is not inferior to that of acetylsalicylic acid.

2. The potential antiplatelet agent diendiamine of the indole series (compound of formula I) after intravenous administration to rabbits significantly suppresses the induced functional activity of platelets. The effect is equal to that of sodium nitroprusside, but exceeds its duration.

3. When administered orally to animals, the compound of formula I inhibits the action of ADP on platelets.

4. The compound of formula I neutralizes the effect of arachidonic acid on platelet activity.

5. A promising antiplatelet agent - a compound of formula I - for a short time enhances the reduced ability of human neutrophils to produce reactive oxygen species.

6. The compound of formula I statistically significantly inhibits hemostasis in rats on a model of parenchymal bleeding, while this effect is not dose-dependent.

7. A potential drug (compound of formula I) belongs to class V of practically non-toxic compounds.

Example 2

No less pronounced and specific is the effect of the compound of formula I on hemodynamic parameters and, in particular, on systolic blood pressure (SBP), as well as on the activity of soluble guanylate cyclase and on the generation of NO in in vitro experiments.

The experiments were performed on spontaneous hypertensive (SGC) and normotensive (NT) anesthetized male rats with the intravenous administration of the test compounds. The ability of the compound to lower blood pressure in SGC rats and to prevent spasmogenic reactions to norepinephrine (HA) and angiotensin I (AI) was evaluated.

The compound of formula I was used as a solution in 3% aqueous DMSO. Sodium nitroprusside and nitroglycerin were used as a comparison drug.

The introduction of the compounds of formula I in the dose range of 0.01-0.1-0.5 and 1.0 mg / kg caused a dose-dependent antihypertensive effect lasting from 30-40 seconds to 60 minutes or more. The minimum and short-term hypotensive effect was observed at a dose of 0.01 mg / kg (blood pressure decreased by 10-12 mm Hg for 30 s), and the maximum at a dose of 1.0 mg / kg (blood pressure decreased by 25-30 mmHg for 60 minutes).

Comparison of sodium nitroprusside caused a decrease in blood pressure similar to the maximum for compound I (decrease by 25 mm Hg) at a dose of 0.5 mg / kg for a duration of 20-30 minutes. The effect of nitroglycerin at a dose of 0.01 mg / kg was similar to the test compound (decrease by 10-15 mm Hg for 30-40 s), and at higher doses the decrease in blood pressure did not exceed 20-25 mm Hg. with a duration of 5-6 minutes (Table 1).

Table 1 The effect of the compounds of formula I in comparison with sodium nitroprusside and nitroglycerin on blood pressure in rat SGK Test substance Dose, mg / kg, iv The decrease in blood pressure, mm Hg (Δ) 10-20 s 5 minutes 30 minutes 60 min 120 min 0.01 -12 ± 5 * - - - - 0.1 -15 + 5 * -20 ± 7 * - - - Compound I 0.5 -10 ± 3 * -20 ± 8 * -25 ± 8 * -30 ± 7 * - 1,0 -12 ± 3 * -25 ± 7 * 30 ± 7 * -30 ± 8 * -30 ± 8 * Sodium 0.01 -5 -10 ± 3 * - - - nitroprusside 0.5 -10 ± 3 * -25 ± 5 * -30 ± 8 * -10 ± 3 * - 0.01 -5 -10 ± 3 * -3 - - Nitro 0.1 -7 ± 3 -20 ± 5 * -5 - - glycerol 0.5 -12 ± 3 * -25 ± 7 * -5 - - 1,0 -10 ± 3 * -25 ± 8 * -7 ± 3 - - * - the difference with the initial level is significant at P <0.05 (-) - no effect

In normotensive anesthetized rats, the effect of the compound of formula I on the pressor effects of norepinephrine (HA, 10-15 μg / kg), mesatone (M, 5 μg / kg) and angiotensin (A1, 5 μg / kg) was studied. The depressant effect of the compounds of formula I was evaluated relative to the pressor effects of the control introductions of these analyzers.

As studies have shown (Table 2), the compound of formula I in a dose range of 0.01-1.0 mg / kg in a dose-dependent manner antagonizes the vasoconstrictor effect of HA. So, with the introduction of the compound in doses of 0.01 and 0.1 mg / kg, the pressor effect of HA decreases by 25 and 50% and does not significantly change with a further increase in doses.

Compound I in the indicated doses does not significantly affect the pressor effect of M, however, at a dose of 0.1 mg / kg, it reduces the pressor effect of A1 by 25%.

Thus, the compound of formula I effectively and in a dose-dependent manner lowers the SBP in spontaneously hypertensive animals and surpasses the well-known drug nitroprusside in this regard. The compound also effectively antagonizes the effects of pressor amines on SBP, which indicates a specific effect on the regulation of blood pressure (Table 2).

table 2 The effect of the compounds of formula I upon intravenous administration on the pressor effects of HA and AI in normotensive, anesthetized rats Substance Dose mg / kg Inhibition of pressor reaction (in%) on Norepinephrine Angiotensin I The compound of formula I 0.01 25 0 0.05 35 10 0.1 fifty 25-30 0.5 60 thirty 1,0 55-60 30-40 Nitroglycerine 0.5 20-25 0 1,0 40-50 25-30 0 - no effect

In experiments on non-anesthetized SGC rats, the compound of formula I with the intragastric route of administration also caused a dose-dependent decrease in blood pressure, the duration of which increased in proportion to the dose increase (maximum duration of effect was 5 hours after a single administration at a dose of 50 mg / kg, Table 3).

The compound of formula I and arginine activate soluble guanylate cyclase from human platelets. Moreover, I is approaching L-arginine in activity. So, the average degree of enzyme activation when applying L-arginine at a concentration of 0.1 mM is 2.26 times, and when using I at the same concentration, 2.12 times (Table 4).

Compound I is characterized as low-toxic, well-tolerated, practically without causing changes in animals in general condition, behavior, without violating the life cycle, without causing them to reduce food intake, water and weight loss. The compound does not cause changes in the central nervous system, does not show neurotoxicity even in fairly high doses. Interaction with a number of pharmacological analyzers did not reveal a specific response in compound I, which, however, does not mean that it does not have a mediator mechanism of action.

Table 3 The effect of the compounds of formula I and nitroglycerin on blood pressure in non-narcotic SGC rats when administered orally (non-invasive measurement of blood pressure) Test substance Dose, mg / kg, by mouth BP reduction, mmHg one 2 3 four 5 6 Compound I one twenty* 10 0 0 0 0 5 17 * twenty* 32 * thirty* fifteen 0 10 23 * thirty* 37 35 17 * 0 25 26 * 32 * 45 * 45 * 25 * 0 fifty 25 * 42 * fifty* fifty* 25 * 0 Nitroglycerine 10 0 0 0 0 0 0 * - the difference with the initial level of blood pressure is significant at P <0.05 0 - no effect

Table 4 The effect of L-arginine and a compound of formula I on the specific activity of guanylate cyclase (HC, picamol cGMP / min / mg protein) of human platelets Sample number The initial specific activity of HZ Arginine (0.1 mM) Compound I (0.1 mmol) Specific activity Activation Specific activity Activation one 133 525 395 593 446 2 235 386 135 450 153 3 one hundred 147 146 146 146 four 140 444 317 265 189 5 211 286 136 256 122 Wed M ± m 174 ± 33 358 ± 38 226 ± 54 342 ± 86 212 ± 67

Acute toxicity of the compound in mice when administered orally is 8.9 ± 0.75 g / kg, cumulation when administered over 28 days is weak (less than 50% death).

Therapeutic doses for animals are in the range of 1/100 of the LD ₅₀ and are 4.0-8.0 and 12.0 mg / kg with intragastric administration.

The second object of the invention is a method for producing compounds of formula I.

The claimed compound contains as structural fragments: an indolinone cycle, a cyano group in the β-position of the side chain and a substituted cyclic fragment in which at one carbon atom there are two strong electron-donating groups - an ether oxygen atom and a substituted amino group. The inclusion of these groups in the cyclic system provides a high degree of conjugation of their lone electron pairs with double bonds of the side chain, which together with the cyclic indole nitrogen atom creates a system that is electronically similar to the guanidine system. The latter is of particular interest, since it is known that the guanidine fragment of arginine is the source of nitric oxide in a living organism.

The method of obtaining the compound, which is 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indolin-3-one of the formula I, consists in the interaction of nitrile α -cyano-β- (3-acetoxyindolyl-2) acrylic acid (II) with N-methylaminopropane-2,3-diol (IIIa) in a polar (preferably isopropyl alcohol) solvent under boiling according to the scheme:

The scheme of the proposed method

We recently described the first case of non-catalytic addition of amines to a cyano group [2, 4, 5], which consists in the interaction of compound II with primary and secondary amines, including primary alkanolamines, for example 1-aminopropane-2,3-diol (IVa) [2, 4] in an environment of a polar or non-polar solvent during boiling with the formation of 2- [2'-cyano-3'-diaminopropenylidene-2 '] indolinone-3 derivatives of the general formula VIII according to Scheme 1:

Scheme 1

As can be seen from Scheme 1, first deacetylation of starting compound II occurs (which consumes 1 mol of 1-aminopropane-2,3-diol (IVa)), accompanied by salt formation (with the formation of salt VIa) with the participation of intermediate V and amine IVa. When heated, salt VIa dissociates, and the released amine joins via the C≡N bond to form amidine VIIa, which is in tautomeric equilibrium with VIIIa [2, 4]. As our further studies showed, the interaction of compound II with β-ethanolamine (IVb) proceeds similarly (Example A).

The synthesis and physicochemical characteristics of 2- [2'-cyano-3'-amino-3 '- (2,3-dihydroxypropyl) aminopropenylidene-2'] indolinone-3 (VIIIa) are described in [2, 4]. ¹ H NMR spectrum.

Example A

2- [2'-Cyano-3'-amino-3 '- (β-hydroxyethyl) aminopropenylidene-2'] indolinone-3 (VIIIb).

A mixture of 2.5 g (10 mmol) of dicyanovinyl derivative II, 2.5 g (41 mmol) of β-ethanolamine IVb and 100 ml of isopropyl alcohol is boiled with stirring for 4 hours. The mixture is cooled, the precipitate formed is filtered off, washed with isopropyl alcohol and ether. 1.67 g (62%) of 2- [2'-cyano-3'-amino-3 '- (β-hydroxyethyl) aminopropenylidene-2'] indolinone-3 (VIIIb) are obtained. Mp 209-210 ° C (decomp. From a mixture of DMF-water, 4: 1).

IR spectrum, ν, cm ^-1 : 3350, 3240-3060, 2190, 1670, 1605.

Mass spectrum: M ⁺ 270.

Calculated,%: C 62.21, H 5.22, N 20.73.

C ₁₄ H ₁₄ N ₄ O ₂ .

Found,%: C 62.14, H 5.50, N 20.73.

¹ H NMR spectrum.

UV spectrum, λmax, nm (log ε): 217 (4.03), 231 (4.18), 257 (3.92), 286 (4.01), 310 pl (3.98), 379 (4.33), 490 pl (4.46), 509 ( 4.64).

Absolutely unexpected results were obtained in the interaction of compound II with N-substituted alkanolamines, such as N-methyl-β-ethanolamine (IIIb) (Example B) and N-methylglucamine (IIIc) (Example C). At first, apparently, the interaction of compounds II and IIIb with proceeds according to a scheme similar to scheme 1, however, the process does not stop at the stage of formation of compounds of the diendiamine structure, general formula VIII, but further oxazolidine cyclization occurs with the release of the ammonia molecule and the formation of 2- [2'-cyano-2 '- (3 ”-methyloxazolidin-2” -idene) ethylidene] -indolinone-3 IX and X. The indicated oxazolidine cyclization is unexpected and unpredictable, and the use of N-substituted alkanolamine. Theoretically, this factor remains unclear. Below is a diagram of the formation of compounds IX-X:

Scheme 2

Example B

2- [2'-Cyano-2 '- (3 ”-methyloxazolidin-2” -ylidene) ethylidene] indolinone-3 (IX).

A mixture of 2.5 g (10 mmol) of dicyanovinyl derivative II, 2.2 g (30 mmol) of N-methylaminoethanol IIIb and 100 ml of isopropyl alcohol is boiled with stirring for 3.5 hours. The mixture is cooled, the precipitate formed is filtered off, washed with isopropyl alcohol and ether. Obtain 1.2 g (44%) of 2- [2'-cyano- (3'-methyloxazolidinylidene-2 ') ethylidene] indolinone-3 (IX). Mp 222-226 ° C (decomp. From a mixture of DMF-methanol, 8: 1).

IR spectrum, ν, cm ^-1 : 3350, 2200, 1675, 1600.

Mass spectrum: M ⁺ 267.

Calculated,%: С 67.40, H 4.90, N 15.72 M. 267.

C ₁₅ H ₁₃ N ₃ O ₂ .

Found,%: C 67.61, H 4.89, N 15.96.

¹ H NMR spectrum.

UV spectrum, λmax, nm (log ε): 256 pl. (3.96), 282 (4.09), 310 pl. (4.03), 365 (4.36), 495 (4.41).

Example B

2- {2'-Cyano-2 '- [3 ”-methyl-5” - (1,2,3,4-tetrahydroxybutyl) oxazolidin-2 ”-ylidene] ethylidene} indolinone-3 (X).

A mixture of 2.5 g (10 mmol) of dicyanovinyl derivative II, 4 g (20 mmol) of N-methylglucamine VIIIc and 100 ml of isopropyl alcohol is refluxed for 3 hours with stirring. The mixture is cooled, the precipitate formed is filtered off, washed with isopropyl alcohol and ether. 1.8 g (45%) of 2- {2'-cyano-2 '- [3 ”methyl-5” - (1,2,3,4-tetrahydroxybutyl) oxazolidin-2 ”-ylidene] ethylidene} are obtained} -indolinone-3 (X). Mp 237 ° C (decomp. From a mixture of DMF-water, 4: 1).

IR spectrum, ν, cm ^-1 : 3430, 3320, 3180, 2190.1650, 1600.

Mass spectrum: M ⁺ not observed, 237 [M - OCH (CHOH) ₃ CH ₂ OH] ⁺ , 210 [M-CH ₃ NCH ₂ CH (CHOH) ₃ CH ₂ OH] ⁺ .

Calculated,%: C 58.91, H 5.46, N 10.85 M 387.

C ₁₉ H ₂₁ N ₃ O ₆ .

Found,%: C 59.02, H 5.47, N 10.65.

¹ H NMR spectrum.

UV spectrum, λmax, nm (log ε): 220 (3.85), 255 pl (3.65), 282 (3.81), 310 pl (3.73), 369 (4.07), 497 (4.13).

Thus, the originality, unusualness, and unpredictability of the preparation of compound I under the above conditions is that until now, cyclization of this type (with the elimination of ammonia) was not known in the series of conjugated diendiamines, and the possibility of such a closure is unexpectedly determined by the presence of a substituent at the nitrogen atom alkanolamine used.

The following example 3 illustrates the inventive method for producing a new compound I.

Example 3

2- [2- [5- (Hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indolin-3-one (I).

A mixture of 2.5 g (10 mmol) of dicyanovinyl derivative II, 2.1 g (20 mmol) of N-methylaminopropanediol IIIa and 100 ml of isopropyl alcohol is refluxed for 2 hours with stirring. The mixture is cooled, the precipitate formed is filtered off, washed with isopropyl alcohol and ether. 1.55 g (52%) of 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indoline-3-one (I) are obtained, T .pl. 236.5-237.5 ° C (decomp. From a mixture of DMF-water, 2: 1).

IR spectrum, ν, cm ^-1 : 3360, 3200, 2180, 1650.

Mass spectrum: M ⁺ 297.

Calculated,%: C 64.63, H 5.09, N 14.13 M. 297.

C ₁₆ H ₁₅ N ₃ O ₃ .

Found,%: C 64.93, H 5.11, N 14.26.

¹ H NMR spectrum.

¹ H NMR spectrum, (DMSO-d ₆ ), (δ, ppm, J / Hz): 3.94 (2H, t, J ₁ = J ₂ = 9.5, CH ₂ OH), 3.96 (2H, q, J ₁ = 7, J ₂ = 9.5, CH ₂ OH), 5.33. (1H, t, J ₁ = J ₂ = 6, CH ₂ OH ), 3.54-3.80 (2H, m, 4 ”-CH ₂ ), 4.91 (1H, oct, 5 ”-CH), 3.32 (3H, s, N-Me), 6.70 (1H, α-CH), 6.82, 7.22, 7.42, 7.54 (4H, m, H (4-7) ), 8.44 (1H, s, N (1) H).

In the ¹³ C NMR spectrum (DMSO-d ₆ ) of compound I, signals of aromatic carbon atoms C (4-7) are observed at 113.2, 118.5, 123.2 and 134.2, Quaternary aromatic C atoms at 121.1 and 151.4, C (1 ') - 113.2 , CN - 120.1, C (2 ”) - 55.5, C (2”) - 166.2, N (3 ”) Me - 34.3, C (4”) - 53.3, C (5 ”) - 80.0 (d, J = 156.3 Hz) and CH ₂ OH at 61.0 ppm.

UV spectrum, λmax, nm (log ε): 222 (4.09), 257 (3.95), 284 (4.07), 310 pl (4.00), 368 (4.34), 497 (4.40).

The synthesis of starting compound II is described in the literature [23, 24] (Scheme 3):

Scheme 3

When N-acetylindoxyl (XI) is reacted with dimethylformamide diethyl acetal, enaminoindolinone-3 XII is formed, which is subjected to acid hydrolysis to 1-acetyl-3-hydroxy-2-formylindole (XIII) without isolation (yield ~ 60 ÷ 70%). It should be noted that the C (1 ') - C (2) bond in the aldehyde is very easily cleaved and, with an increase in the processing time of intermediate XII with dilute HCl, N-acetylindoxyl XI is formed [1]. 2-Formylindole XIII is a very labile compound, does not withstand long-term storage, and to carry out the reactions of this compound in the formyl group with CH acids, it is first necessary to convert it to N-acetyl-3-acetoxy-2-formylindole (XIV) as quickly as possible. The reaction of compound XIV with malondinitrile in benzene in the presence of triethylamine gives dicyanovinylindole II. The following examples illustrate the synthesis method of starting compound II according to scheme 3 [1,3]:

1-Acetyl-2-formyl-3-hydroxyindole (XIII) [1]. To a suspension of 56 g (0.32 mol) of indoxyl XI in 640 ml of benzene, 198 ml (~ 1.24 mol) of diethyl acetal DMF were added and stirred for 1 h at 20 ° C. The resulting solution is boiled for 1 hour. Benzene is evaporated on a rotary evaporator at a temperature in the bath no higher than 60 ° C. After removal of benzene, the residue is dissolved in ~ 4 L of water and acidified (pH 2-3) 60 ml conc. HCl. After 5 min, ¹ * ( ¹ * After 1 h, the precipitate was filtered off, washed with water and isopropyl alcohol. N-acetylindoxyl was obtained in 51% yield.), The precipitate formed was filtered off, washed with water and isopropyl alcohol. The yield of compound XIII was 45.5 g (70%). Mp 122-123 ° C (from isopropyl alcohol).

IR spectrum, ν, cm ^-1 : 1700, 1620, 1580.

Mass spectrum: M ⁺ 203.

Calculated,%: C 65.02, H 4.47, N 6.89.

C ₁₁ H ₉ NO ₃ .

Found,%: C 64.73, H 4.50, N 6.90.

Spectrum ¹ H NMR (CDCl ₃ ): 10.24 (1H, br s, CHO); 2.79 (3H, s, NCOCH ₃ ); 7.34-7.91 ppm (4Н, m, arom. Protons).

UV spectrum, λmax (logε): 230 (4.22), 257 (4.03), 312 (3.95), 356 nm (3.89).

1-Acetyl-2-formyl-3-acetoxyindole (XIV) [1]. A solution of 45.5 g (0.23 mol) of aldehyde XIII in 130 ml of acetic anhydride is boiled for 10 minutes. It is cooled, the precipitate formed is filtered off, washed with acetic anhydride and ether. The yield of compound XIV was 31.8 g (58%). Mp 111-113 ° C (from ethyl acetate).

IR spectrum, ν, cm ^-1 : 1785, 1700.1670, 1610.

Mass spectrum: M ⁺ 245.

Calculated,%: C 63.67, H 4.52, N 5.71.

C ₁₃ H ₁₁ NO ₄ .

Found,%: C 63.66, H 4.52, N 5.64.

Spectrum ¹ H NMR (acetone-d ₆ ): 10.15 (1H, s, CH); 2.45 (3H, s, OCOCH ₃ ); 2.48 (3H, s, NCOCH ₃ ); 7.3 8-8.12 ppm (4H, m, arom. Protons).

Nitrile α-cyano-β- (3-acetoxyindolyl-2) acrylic acid (II) [3]. To a solution of 24.4 g (0.1 mol) of 2-formylindole XIV in 500 ml of benzene at 20 ° C (external cooling with water), 14.4 ml (0.1 mol) of triethylamine and 7.8 g (0) are added with stirring , 12 mol) malondinitrile. Stirred for 1.5-2 hours. The precipitate was filtered off, washed with benzene and methanol. 22.7 g (90%) of nitrile II are obtained, mp 210-212 ° C. (decomp., From acetone).

IR spectrum, ν, cm ^-1 : 3340, 2240, 2230, 1770, 1600.

Mass spectrum: M ⁺ 251.

Calculated,%: C 66.93, H 3.61, N 16.73.

C ₁₄ H ₉ N ₃ O ₂ .

Found,%: C 67.07, H 3.42, N 16.63.

Spectrum ¹ H NMR (acetone-d ₆ ): 2.47 (3H, s, OCOCH ₃ ); 8.17 (1H, s, CH); 10.39 (1H, s, NH); 7.19-7.68 (4H, m, arom. Protons).

¹³ C NMR Spectrum (DMSO-d ₆ + CD ₃ OD): 168.5 (q, J≈7 Hz, OCOCH ₃ ); 20.7 (q, J≈142 Hz, OSOSN ₃ ); 114.9 (d, J = 8.1 Hz, CN); 113.8 (d, J = 13.5 Hz, CN); 74.4 (d, J = 2 Hz, a- C), 144.3 (d, J = 166 Hz, β-CH); 138.8; 138.4 (m, C (2), C (7a)); 119.4 (m, C (3a)); 121.4 (d, J = 2 Hz, C (3)); 128.3, 121.4, 119.8, 113.9 (q, J ₁ = 160 ... 165 Hz, J≈8 Hz, C (4), C (5), C (6), C (7) )

The yield of 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indolin-3-one (I), counting on N-acetylindoxyl according to Scheme 3, is 13.5% (taking into account the recrystallization of the final product).

The bottleneck in the synthesis of compound II by this method is the synthesis of 2-formylindole XIII, namely the hydrolysis operation, which should be carried out in a strictly defined time frame. It is clear that when scaling up the process, it is possible to obtain contaminated 2-formylindole XIII with the product of its further hydrolysis - N-acetylindoxyl or there is a danger of isolation of pure N-acetylindoxyl. Therefore, in order to avoid technological difficulties associated with the isolation of intermediate 2-formylindole XIII when scaling the process, we modified the synthesis scheme of compound II (Scheme 4):

Scheme 4

This goal is achieved by the claimed method for producing compound II, which is a key intermediate for the synthesis of biologically active compound I.

The synthesis of starting compound II according to Scheme 4 begins, as in Scheme 3, with the interaction of N-acetylindoxyl with DMF acetal, but then, unlike Scheme 3, deaminylation of intermediate enaminoketone XII with triethylamine in methanol follows with the formation of enaminoketone XV with a yield of 55% [ one]. We described the acylation of enaminoketone XV [3] with various acyl halides. In contrast to the method [3], where acetylation of enaminoketone XV in acetyl chloride medium is described, we performed the acetylation process in DMF medium. As a result, the isolated chloride of 3-acetoxy-2-dimethyliminomethylindole (XVI) after dissolution in water was converted to 3-acetoxy-2-formylindole (XVII) in 63% yield against 25% by the method of [3]. The reaction of 3-acetoxy-2-formylindole (XXII) with malondinitrile in benzene in the presence of triethylamine gives dicyanovinylindole II in high yield. The following examples 4 and 5 illustrate the synthesis method of the starting compound II according to scheme 4.

2-Dimethylaminomethylene-indinolin-3 (XV) [1] is obtained from 7 g (40 mmol) of indoxyl XI and 24 ml (120 mmol) of acetal DMF in 80 ml of benzene under the conditions of synthesis of compound XIII. After removal of benzene, the residue is dissolved in 60 ml of methanol, 6 ml of triethylamine are added and boiled for 30 minutes. The methanol is evaporated. The residue was stirred with ethyl acetate, the precipitate was filtered off, washed with isopropyl alcohol and ether. The yield of enamine XV 4.1 g (55%). Mp 215 ° C (decomp., From isopropyl alcohol).

IR spectrum, ν, cm ^-1 : 3250-3100, 1675, 1615.

Mass spectrum: M ⁺ 188.

Found,%: C 70.33, H 6.44, N 14.94.

C ₁₁ H ₁₂ N ₂ O.

Calculated,%: C 70.19, H 6.43, N 14.88.

Spectrum ¹ H NMR (DMSO-d ₆ ), δ, ppm: 7.01 (1H, s, CH); 6.78-7.31 (4H, m, arom. Protons); 3.19, 3.35 (6H, two s, N (CH ₃ ) ₂ ); 8.85 (1H, s, NH).

UV spectrum, λmax (logε): 254 (4.05), 290 (4.03), 338 (4.32), 437 nm (4.12).

Example 4

3-Acetoxy-2-formylindole (XVII). 0.31 ml (4.4 mmol) of acetyl chloride are added dropwise to a suspension of 0.38 g (2 mmol) of enaminoketone XV [1] in 2.5 ml of DMF with cooling with water and stirring. The mixture was stirred at room temperature for 5 hours. After 2 hours, an additional 0.31 ml of acetyl chloride was added. They stand the night in the refrigerator (5-7 ° C). The precipitate formed is filtered off, washed with DMF and acetone. Obtain 0.4 g (75%) of 3-acetoxy-2-dimethyliminomethylindole chloride (XVI), which is dissolved in 25 ml of water and cooled with ice for 15 minutes. The precipitate formed is filtered off, washed with water and dried. 0.26 g (63%) of 3-acetoxy-2-formylindole (XVII) is obtained. Mp 145-147 ° C (from a mixture of water - isopropyl alcohol, 5: 1). The melting point of a mixed sample of a substance with a sample of the compound obtained by the method [3] does not show depression.

IR spectrum, ν, cm ^-1 : 3280, 1770, 1660, 1620.

Mass spectrum: M ⁺ 203.

Found,%: C 65.04, H 4.64, N 6.76.

C ₁₁ H ₉ NO ₃ .

Calculated,%: C 65.02, H 4.47, N 6.89.

Spectrum ¹ H NMR (CDCl ₃ ), δ, ppm: 9.90 (1H, s, CHO), 9.17 (1H, br.s, NH), 7.12-7.57 (4H, m, arom protons), 2.48 (3H, s, OCOCH ₃ ).

Example 5

Nitrile α-cyano-β- (3-acetoxyindolyl-2) acrylic acid (II). To a solution of 0.15 g (0.6 mmol) of 2-formylindole XVII in 5 ml of benzene at 20 ° C (external cooling with water), 0.01 ml (0.072 mmol) of triethylamine and 0.05 g (0.72) are added while stirring mmol) malondinitrile. Stirred for 0.5 hours. The precipitate was filtered off, washed with benzene and methanol. Obtain 0.15 g (99%) of nitrile II. Mp 210-212 ° C (decomp., From acetone). The melting point of a mixed sample of a substance with a sample of the compound obtained by the method [7] does not show depression.

The yield of 2- [2'-cyano-2 '- (3 ”-methyl-5” -hydroxymethyloxazolidin-2 ”-ylidene) ethylidene] indolinone-3 (I), counting on N-acetylindoxyl according to Scheme 4, is 12.5 % (including recrystallization of the final product).

We have developed methods for the synthesis of the claimed compounds - 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indoline-3-one (I) based on nitrile II according to schemes 3 and 4 they allow to obtain this compound with approximately the same outputs (13.5 ÷ 12.5%). Scheme 4 has some technological advantages over Scheme 3, however, both of these schemes involve the use of an expensive, inaccessible reagent - diethyl acetal dimethylformamide, which is used in large excess. In order to exclude the expensive reagent DMF diethyl acetal from the technological process, and also to increase the yield of the target product I, we were faced with the task of developing an alternative approach to the synthesis of compound I.

The goal is achieved by the claimed method of obtaining 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indoline-3-one (Scheme 5), which consists in the interaction N-acetylindoxyl (XI) with ethoxymethylene malondinitrile (XVIII) to form N-acetyl-3-hydroxy-2-dicyanovinylindole (XIX), which is then acetylated with acetic anhydride to N-acetyl-3-acetoxy-2-dicyanovinylindole (XX). The latter, when reacted with an equimolecular amount of N-methylaminopropane-2,3-diol (IIIa), turns into previously unknown 1-acetyl-2- [2'-cyano-2 '- (3 ”-methyl-5” -oxymethyloxazolidine-2 ” -ilidene) -ethylidene] indolinone-3 (XXI). The removal of the acetyl group is carried out by the action of Triton B (benzyltrimethylammonium hydroxide, 40% solution in methanol) in methanol with access to the claimed compound I:

Scheme 5

The following examples 6-8 and illustrate the synthesis method of the claimed compound I according to scheme 5.

Example 6

N-Acetyl-3-acetoxy-2-dicyanovinylindole (XX). To a suspension of 56 g (0.32 mol) of N-acetylindoxyl in 80 ml of pyridine, 39.04 g (0.32 mol) of ethoxymethylene malondinitrile (XVIII) is added. Then, with stirring and cooling with water, 44.8 ml (0.32 mol) of triethylamine are added, the temperature of the reaction mass is 25 ± 2 ° C. The mixture is stirred at this temperature for 1 h, N-acetyl-3-hydroxy-2-dicyanovinylindole (XIX) is planted with 160 ml of acetic anhydride under ice cooling. The temperature in the mass is not higher than 30 ° C. After 1 h at 20 ° C, the precipitate was filtered off, washed with acetic anhydride and acetone. It is dried in air for 1 hour. 79.6 N-acetyl-3-hydroxy-2-dicyanovinylindole (XIX) is obtained, to which 500 ml of acetic anhydride are added and boiled for 30 minutes with vigorous stirring. It is cooled, the precipitate is filtered off, washed with acetic anhydride and acetone. Air dried to constant weight. 65.34 (69.6%) g of N-acetyl-3-acetoxy-2-dicyanovinylindole (XX) are obtained. Mp 250-252 ° C (decomp. From acetic anhydride).

IR spectrum, ν, cm ^-1 : 2225, 1692, 1655, 1607.

Mass spectrum: M ⁺ 293.

Found,%: C 65.44, H 3.42, N 14.12.

C ₁₆ H ₁₁ N ₃ O ₃ .

Calculated,%: C 65.52, H 3.78, N 14.33.

¹ H NMR spectrum (DMSO-d ₆ ), δ, ppm: 2.32 (3H, s, OCOCH ₃ ), 2.85 (3H, s, NCOCH ₃ ), 7.48 (1H, t, H (5)), 7.68 1H, t, H (6)), 7.85 (1H, d, H (4)), 8.25 (1H, d, H (7)), (8.90 (1H, s, H (2 ')).

Example 7

1-Acetyl-2- [2'-cyano-2 '- (3 ”-methyl-5” -oxymethyloxazolidin-2 ”-ylidene) ethylidene] indolinone-3 (XXI). A mixture of 66.18 g (0.226 mol) of N-acetyl-3-acetoxy-2-dicyanovinylindole (XX), 47.5 g (0.452 mol) of N-methylaminopropane-2,3-diol (VIIIa) and 1400 ml of isopropyl alcohol are boiled with stirring for 1 h. Cool, the precipitate is filtered off, washed with isopropanol and dried at 80 ° C. Obtain 66.4 g (86.7%) of 1-acetyl-2- [2'-cyano-2 '- (3 ”-methyl-5” -oxymethyloxazolidin-2 ”-ilidene) ethylidene] indolinone-3 (XXI ) Mp 218-220 ° C (from methanol).

IR spectrum (in a tablet with KBr), ν, cm ^-1 : 2220, 1700, 1680, 1610.

Mass spectrum: M ⁺ 339.

Found,%: C 63.71, H 5.01, N 12.37. C ₁₈ H ₁₇ N ₃ O ₄ .

Calculated,%: C 63.71, H 5.05, N 12.38.

¹ H NMR Spectrum (DMSO-d ₆ ), (δ, ppm, J Hz): 2.42 (3H, s, NCOCH ₃ ), 3.39 (3H, s, N (3 ”) Me), 3.98 (2H , t, J ₁ = J ₂ = 9.5, CH ₂ OH), 3.80 (2Н, q, J ₁ = 7, J ₂ = 9.5, CH ₂ OH), 5.19 (1H, t, J ₁ = J ₂ = 6 , CH ₂ OH ), 3.55-3.90 (2H, m, 4'-CH ₂ ), 4.94 (1H, oct, 5'-CH), 7.20 (1H, H (1) '), 6.82, 7.23 (1H, d, H (4)), 7.59-7.67 (2H, m, H (5-6)), 8.07 ((1H, d, H (7)).

Example 8

2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indolin-3-one (I).

To the suspension 66.4 g (0.196 mol) of 1-acetyl-2- [2'-cyano-2 '- (3 ”-methyl-5” -oxymethyloxazolidin-2 ”-ilidene) ethylidene] indolinone-3 (XXI) 115.7 ml (0.254 mol) of Triton B are added to 800 ml of methanol and stirred at 20 ° C for 30 minutes. The precipitate is filtered off, washed with methanol, water and methanol. Dry at 80 ° C., 40.6 g (69.7%) of 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indoline- are obtained. 3-one (I). Recrystallized from 650 ml of a mixture of DMF-water 2: 1. Obtain 35.7 g of pure compound I. The yield of compound I on N-acetylindoxyl is 37%. The melting point of a mixed sample of the substance with samples of the compounds obtained according to schemes 3 and 4, does not show depression.

Spectra IR, mass - NMR, UV and polarograms of samples of compound I, obtained according to schemes 3, 4 and 5, are identical.

Conclusion: Effective original methods have been developed for the synthesis of a biologically active compound - 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indolin-3-one (I) .

Literature

1. Ryabova S.Yu., Trofimkin Yu.I., Alekseeva L.M., Bogdanova G.A., Sheinker Yu.N., Granik V.G. // Chemistry heterocycle. connection 1990. No. 11. S.1487-1494.

2. Ryabova S.Yu., Alekseeva L.M., Granik V.G., Faermark I.F., Schwartz G.Ya. Patent 2008308 of the Russian Federation "Derivatives of indolinone-3 with antihypertensive activity, and a method for their preparation."

3. Ryabova S.Yu., Trofimkin Yu.I., Alekseeva L.M., Khabarova L.S., Granik V.G. // Chemistry heterocycle. connection 1991. No3. S.343-348.

4. Ryabova S.Yu., Trofimkin Yu.I., Granik V.G., Faermark I.F., Schwartz G.Ya. Patent 2026287 of the Russian Federation “Derivatives of pyrrolin-2-one-4 or indolinone-3 with antihypertensive activity, and a method for their preparation”.

5. Ryabova S.Yu., Trofimkin Yu. I., Alekseeva L.M., Kerbnikova I.F., Schwartz G.Ya., Granik V.G. // Chem. - farm. journal 1995.Vol. 29 (9). S.22-29.

6. Khmelnitskaya E.Yu., Grigoryev NB, Ryabova S.Yu., Trofimkin Yu.I., Azimov V.A., Granik V.G. // Chemistry heterocycle. connection 2002. No. 11. S.1540-1546.

7. Feelish M. // J. Cardiovascular. Pharm. 1991. Vol. 17 (Suppl 3). P.25.

8. Hansson G.K. // Atheroscler. Thromb. Vasc. Biol. 2001. Vol.21 (12). P.1876.

9. Orekhov A.N., Tertov V.V., Pivovarova E.M. // Cardiology. 1998. Vol. 89. P.111.

10. Zadelaar S., Kleemann R., Verschuren L., et al. // Atheroscler. Thromb. Vasc. Biol. 2007. Vol. 27. P.1706.

Claims (3)

1. The use of 2- [2- [5- (Hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indolin-3-one of the formula I

as a biologically active compound exhibiting the properties of an exogenous nitric oxide donor, an activator of the enzyme, soluble guanylate cyclase, an inhibitor of platelet aggregation with antihypertensive activity to obtain an agent for the treatment of cardiovascular diseases. 2. The method of obtaining 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indoline-3-one of the formula I according to claim 1, characterized in that that nitrile α-cyano-β- (3-acetoxyindolyl-2) acrylic acid of the formula II:

subjected to interaction with N-methylaminopropane-2,3-diol of the formula IIIa:

in a polar solvent while boiling. 3. The method of obtaining 2- [2- [5- (hydroxymethyl) -3-methyl-1,3-oxazolidin-2-ylidene] -2-cyanoethylidene] indoline-3-one of formula I according to claim 1, characterized in that that N-acetylindoxyl and ethoxymethylene malondinitrile are reacted to form N-acetyl-3-hydroxy-2-dicyanovinylindole, which is then acetylated with acetic anhydride to N-acetyl-3-acetoxy-2-dicyanovinylindole, followed by its interaction with an equimolecular amount of N- methylaminopropane-2,3-diol (IIIa) to form 1-acetyl-2- [2'-cyano-2 '- (3 ”methyl-5” -oxymethyloxazolidin-2 "-Ilidene) -ethylidene] indolinone-3 of the formula XXI,

then the acetyl group is removed in methanol medium by exposure to Triton B, which is a 40% solution of benzyltrimethylammonium hydroxide in methanol, the precipitate is washed, dried and purified by recrystallization from dimethylformamide-water mixture to obtain the desired product.

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