RU2712036C2 - Indolinone derivatives having melatonin receptor ligand properties and use thereof - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention relates to indolinone derivatives having the properties of melatonin receptor subtypes MT1, MT2 and enzyme-quinone reductase 2 (MT3).EFFECT: compounds can be used in treating ophthalmic diseases, such as glaucoma.15 cl, 1 tbl, 34 ex

Description

The invention relates to indolinone derivatives having the properties of melatonin receptor ligands of the MT1, MT2 subtypes and the enzyme –quinone reductase 2 (MT3). The compounds may find use in the treatment of ophthalmic diseases such as glaucoma.

Currently, in many countries, melatonin preparations are produced, which are either drugs or are related to dietary supplements. So, for example, the well-known dosage form of the biogenic amine melatonin (N-acetyl-5-methoxytryptamine) - melaxen is used in sleep disorders in patients with cerebrovascular accident and is used in complex antitumor therapy, in the treatment of arterial hypertension and neurodegenerative diseases. The wider introduction of the use of melatonin is its rapid metabolism in the body, especially during daylight hours, as well as the non-selective effect associated with the presence of numerous melatonin binding sites in the body. So in animals 3 subtypes of melatonin receptors are distinguished: transmembrane GPCR receptors MT1 and MT2 and low affinity MT3 receptor, which, according to some studies, is an enzyme, quinone reductase 2. In addition, melatonin is able to bind to nuclear ROR / RZR receptors. Despite numerous studies, the question of exactly what physiological functions melatonin regulates, which through each of these receptors, still remains open.

The local use of melatonin in the form of instillations is known to have an effective effect on inflammatory diseases of the choroid of the eye of various origins, often affecting other structures of the eye, including the retina, and leading to blindness. The use of corticosteroids in rather high doses for treatment is harmful to the health of patients (see N. B. Chesnokova et al. Effectiveness of topical administration of melatonin in the form of instillations in case of experimental uveitis in rabbits, VIII Russian National Ophthalmological Forum, Collection of scientific papers of a scientific and practical conference with international participation, edited by V.V. Neroev, Moscow, September 22-24, 2015, vol. 2, p. 929-933).

Derivatives of melatonin acylated to the first position of indole are known, their analogues and their use as medicines and cosmetics for depression, stress, irritation of schizophrenia, epilepsy and other diseases of the central nervous system, tumor diseases, psoriasis, acne, mycosis and glaucoma (see WO 9706140 / A61K31 / 00; publ. 02.20.1997 /).

 Da Settimo, Federico et al. (Novel, Highly Potent Aldose Reductase Inhibitors: Cyano (2-oxo-2,3-dihydroindol-3-yl) acetic Acid Derivatives, Journal of Medicinal Chemistry, 2003, 46 (8), 1419- 1428 (English) describes Z-1-carbethoxy-2-hydroxy-3-indolinylidene-cyanoacetate and its 2-oxo-3-yl acetate tautomeric form, which may be substituted with fluorine, bromine, nitro and methoxy atoms, as aldol reductase inhibitors Inhibitors may find application in the treatment of cataracts in the form of eye drops.

In EP2261206A1 / A61K31 / 4045, publ. August 17, 2011 / describes indolinone derivatives of the general formula (I)

where the group —̶̶R1 represents a group = CHC (O) O- (C ₁ -C ₆ alkyl), = COOH, CH ₂ CN, X-R2- represents H, C ₁ -C ₆ alkyl, benzyl, R3-hydrogen , halogen, C ₁ -C ₆ alkyl, OS ₁ -C ₆ alkyl, NO ₂ , CN, in the form of E and Z isomers or mixtures thereof, as active ingredients for pharmaceutical compositions for the treatment of gastrointestinal disorders.

SN102659662A / C07D209 / 38, publ. September 12, 2012 / describes the stereoselective synthesis of 2,3-dihydro-3-hydroxy-2-oxo-1H-indole-3-acetonitriles, possibly mono- or disubstituted in the indole ring with halogen atoms, an alkyl or alkoxy group. The method consists in the condensation of isatin with α-substituted acetic acid under the conditions of stereochemical basic catalysis.

M.S. Volkova et al. (Synthesis of novel MT3 receptor ligands via an unusual Knoevenagel condensation, Bioorganic & Medicinal Chemistry Letters, 2012, v. 22, 7578-7581) describes the preparation of N- [3-hydroxy-3- (cyanomethyl) -2-oxo-2, 3-dihydro-1H-indol-5-yl] acetamide (compound 7) and N- [3- (cyanomethyl) -2-oxo-2,3-dihydro-1H-indol-5-yl] acetamide (compound 4) reduction of cyano (5-nitro-2-oxo-2,3-dihydro-1H-indol-3-ylidene) acetic acid (compound 3) or cyano (5-nitro-3-hydroxy-2-oxo-2,3- dihydro-1H-indol-3-ylidene) acetic acid (compound 6).

When compounds 3 and 6 are reduced with zinc in acetic acid and acetic anhydride for 1 hour, the nitro group, double bond and acetylation of the formed amino group are simultaneously reduced. The synthesis is carried out similarly to obtain N- [1-methyl-3- (cyanomethyl) -2- oxo-2,3-dihydro-1H-indol-5-yl] acetamide (compound 12). Compounds 4 and 12 possess the properties of MT3 ligands of the melatonin receptor subtype.

WO2007122219 / A61K31 / 404, publ. 11/01/2007 / describes derivatives of 3- (aminomethylidene) -2-indolinone substituted at the 5th position with a sulfamide group as inhibitors of cell proliferation

WO2004 / 037247A1 / C07D209 / 96, publ., May 6, 2004 / describes the preparation and use of substituted methyl or ethyl (1,2-dihydro-2-oxo-3H-indole-3-ylidene) acetate as intermediates for the preparation of spirocyclopropanindoles possessing antiviral activity. The starting acetates can be substituted in the phenyl part of the indole with bromine, chlorine, nitro, trifluoromethoxy, methyl.

Lozinskaya NA et al. (Syntheses of spiroindole melatonin analog via 2- (indolin-3-ylidene) acetonitrile cycloadditions, Mendeleev Communications, 2014.24 (5)) describe 2- (2-oxo-1,2-dihydro-3H-indole-3-ylidene ) acetonitriles, which can be substituted at the 5th position of the indole with a methoxy group, as intermediates for the preparation of the corresponding spiroindole analogs of melatonin according to the following scheme.

R –H, OMe, R1 –H.

Given the great potential of analogues of melatonin receptor ligands, the search for new melatonin ligands of various subtypes and their synthesis, as well as the study of activity, is an urgent task.

The present invention relates to new indolinone derivatives of the general formula (I)

or their stereoisomers and pharmaceutically acceptable salts,

,where A means

,

R1 is H, benzyl, allyl, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkylcarbonyl, C6-aryloxycarbonyl, benzoyl, C ₁ -C ₆ alkyloxycarbonyl, C ₁ -C ₆ alkylcarbonyl,

m means 1-3,

 R2 is H, C1-C5 alkyl, C1-C5 alkoxy, halogen in combination with nitro, CN, NHC (O) C1-C5 alkyl, NHC (O) C3-C6 cycloalkyl, NHC (O) OC1-C5 alkyl, NHC (O) OC3- C6 cycloalkyl, NHC (O) C1-C5 haloalkyl, NHC (O) OC1-C5 haloalkyl, COR6, where R6 is OH, OC1-C5 alkyl, NH2, NH (C1-C5 alkyl) or N (C1-C5 alkyl) 2,

or R2 is C1-C5 alkyl, C1-C5 alkoxy, halogen, as the only substituent, wherein R3 is OH, OC1-C5 alkoxy, C1-C5 alkyl (CO) O-, C1-C5 alkyl O (CO) O-, C1-C5 haloalkyl (CO) O-,

R3 means H, OH, OC₁-FROM₅alkoxy, C₁-FROM₅alkyl (CO) O-, C₁-FROM₅alkyl O (CO) O-, C₁-FROM₅haloalkyl (CO) O-,

R4 means CN; COR6, where R6 is OH, OC₁-FROM₅alkyl; or C (O) NH₂; C (O) N (C₁-FROM₅alkyl)₂,

The new compounds of formula (I) can be used as MT ₁ , MT ₂ and MT ₃ ligands (quinone reductase 2 enzyme) of the melatonin receptor subtype and have inhibitory activity against MT _{3 of} the melatonin receptor subtype (quinone reductase 2 enzyme). the new compounds of formula (I) can be used to obtain drugs for the treatment and prevention of ophthalmic diseases, in particular with glaucoma.

 The object of the present invention is also the use of compounds of General formula (I)

or their stereoisomers and pharmaceutically acceptable salts,

или

,where A means

or

,

R1 is H, benzyl, allyl, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkylcarbonyl, C6-aryloxycarbonyl, benzoyl, C ₁ -C ₆ alkyloxycarbonyl, C ₁ -C ₆ alkylcarbonyl,

m means 1-3,

R2 means H, C₁-FROM₅alkyl, C₁-FROM₅alkoxy, nitro, CN, halogen, NHC (O) C₁-FROM₅alkyl, NHC (O) C₃-FROM₆cycloalkyl, NHС (О) ОС₁-FROM₅alkyl, NHC (O) OC₃-FROM₆cycloalkyl, NHC (O) C₁-FROM₅haloalkyl, NHС (О) ОС₁-FROM₅haloalkyl, COR6, where R6 is OH, OC₁-FROM₅alkyl, NH₂NH (C₁-FROM₅alkyl) or N (C₁-FROM₅alkyl)₂,

R3 means H; CN; COR6, where R6 is OH, OC₁-FROM₅alkyl; or C (O) NH₂; C (O) N (C₁-FROM₅alkyl)₂,

R4 means CN; COR6, where R6 is OH, OC₁-FROM₅alkyl; or C (O) NH₂; C (O) N (C₁-FROM₅alkyl)₂,

R5 means H, OH, C₁-FROM₅alkoxy, C₁-FROM₅alkyl (CO) O-, C₁-FROM₅alkyl O (CO) O-, C₁-FROM₅haloalkyl (CO) O-

as ligands MT ₁ , MT ₂ and MT ₃ (enzyme quinone reductase 2) subtype of melatonin receptors

The use of compounds of the general formula (I) for the manufacture of a medicament for the treatment and prevention of ophthalmic diseases, in particular for glaucoma, is proposed.

The use of a compound of formula I as an active ingredient for the preparation of a pharmaceutical composition suitable for the treatment and prevention of an ophthalmic disease, in particular glaucoma, is proposed.

The invention also relates to a pharmaceutical composition comprising an effective amount of a compound of formula (I) and excipients conventionally used to treat ophthalmic diseases, such as a solvent or carrier, preservatives.

The composition may be in the form of drops. The solvent medium may be water.

The composition may also contain, for example, a physiologically tolerant buffer, polyethylene glycol.

The preparation of compounds of formula (I) was carried out by reducing the compounds of formula (II) in several ways according to the scheme:

where R ’means H, C₁-FROM₅alkyl, C₁-FROM₅alkoxy, nitro, COOH, CN, halogen, COR6, where R6 is OH, OC₁-FROM₅alkyl, NH₂or N (R₇)₂where R₇means C₁-FROM₅alkyl

или

,A 'means

or

,

R3 ’ means H; CN; COR6, where R6 is OH, OC₁-FROM₅alkyl; or C (O) NH₂; C (O) N (R7)₂where R7 means C₁-FROM₅alkyl

R4 ’ means CN; COOR6, where R6 is OH, OC₁-FROM₅alkyl; or C (O) NH₂; C (O) N (R7)₂where R7 means C₁-FROM₅alkyl

R5 ’means H, OH

R9 is C ₁ -C ₅ alkyl, C ₃ -C ₆ cycloalkyl, OC ₁ -C ₅ alkyl, OC ₃ -C ₆ cycloalkyl, C ₁ -C ₅ haloalkyl, OC ₁ -C ₅ haloalkyl,

R1, R2, A, m have the above meanings.

For condensation products not containing nitro groups, reduction was carried out with zinc dust under the influence of hydrochloric acid in a two-phase system of an aqueous solution of hydrochloric acid / ethyl acetate. At the end of the reaction was judged using thin layer chromatography. After termination of the reaction, the organic layer was separated, the aqueous layer was extracted with ethyl acetate. The organic extracts were combined, washed with cold water, dried over anhydrous sodium sulfate, and the solvent was removed in a vacuum of a rotary evaporator.

The recovery of compounds containing one or more nitro groups was carried out in three ways. According to the first method, zinc dust was quickly added to a solution of the condensation product of isatin with CH-acid compounds in a carboxylic acid R9COOH containing the corresponding acid anhydride (R9C (O)) ₂ O at room temperature. At the same time, a significant heating of the reaction mixture was observed, then the solution was boiled for 2 hours. After completion of the reaction, the mixture was cooled to room temperature, the solvent was removed in a vacuum of a rotary evaporator. The product was purified by column chromatography on silica gel.

In the second method, a solution of hydrogen chloride in 1,4-dioxane was slowly added dropwise to a solution of the condensation product of isatin with CH-acid compounds in 1,4-dioxane containing a calculated amount of zinc dust at room temperature. In this case, a significant heating of the mixture was observed. After complete instillation, the reaction mixture was stirred for 3 hours. Then, the solution was purified from the precipitate by filtration, and the solvent was removed in a vacuum of a rotary evaporator. The obtained solid residue was dissolved in tetrahydrofuran, a base (for example, piperidine or triethylamine) and an acylating agent R9C (O) Hal, or (R9C (O)) ₂ O were added. The reaction mixture was boiled for 1.5 hours, then cooled to room temperature , the solvent was removed in a vacuum of a rotary evaporator, the obtained solid precipitate was washed with water, extracted with ethyl acetate. The organic extracts were combined, dried over anhydrous sodium sulfate, the excess solvent was removed in a vacuum rotary evaporator.

The reduction of compounds containing groups easily hydrolyzed was carried out catalytically in a hydrogen atmosphere in a suitable solvent. After complete hydrogenation, the solvent was removed in a vacuum of a rotary evaporator.

Compounds of formula (II), in turn, were obtained by condensation of isatins of formula (III) with CH-acid compounds of formula CH ₂ (COOH) R8 or CH ₂ (R8) ₂ in the presence of catalytic amounts of a base (e.g. triethylamine or piperidine) in pyridine medium, ethanol (by boiling) or 1,4-dioxane (at room temperature), according to the scheme:

,

,

where R8 is CN; COOR6, where R6 is H, alkali or alkaline earth metal, C₁-FROM₅alkyl; or C (O) NH₂; C (O) N (R7)₂where R7 means C₁-FROM₅alkyl

R ’, R1, A’, m have the above meanings.

If the reaction was carried out at room temperature, the mixture was stirred for 4 hours, then the calculated amount of concentrated hydrochloric acid was added. After that, if it is necessary to obtain a product containing a double C = C bond at position 3 of indolinone, the reaction mixture was left for 4 days. The precipitate was filtered off, washed with cold water to a neutral pH of the washings, and dried in air. If it is necessary to obtain a disubstituted product at position 3, after the addition of acid, the mixture was condensed in a vacuum of a rotary evaporator, dissolved in a small amount of ethyl acetate and poured into cold water. The precipitate was filtered off, washed with cold water, and dried in air.

If the reaction was carried out in pyridine or ethanol while boiling, then to obtain a product containing a double C = C bond at position 3 of indolinone, boiling was carried out for 30-60 hours, to obtain a disubstituted at position 3 of the product, boiling was carried out for 5-30 hours. Control over the completeness of the reaction was carried out using thin-layer chromatography. After the reaction, the mixture was cooled to room temperature and the product was isolated by various methods. Then, if necessary, hydrolysis and decarboxylation of the obtained intermediate product were carried out. For this, after the completion of the reaction, the reaction mixture was poured into water and left at room temperature for 4 days. The precipitate was filtered off, washed with cold water, and dried in air. Decarboxylation was carried out in 2-ethoxyethanol at the boil for 4 hours.

Compounds of formula (III) were prepared from anilines of formula (IV) according to the scheme:

,

,

where R means H, C₁-FROM₅alkyl, C₁-FROM₅alkoxy, COOH, CN, halogen, COOR6, where R6 is H, alkali or alkaline earth metal, C₁-FROM₅alkyl, NH₂or N (R₇)₂where R₇means C₁-FROM₅alkyl

R ’, R1, m have the above meanings.

To do this, first aniline (IV) was reacted with chloral hydrate and hydroxyamine hydrochloride in the presence of sodium sulfate in a dilute hydrochloric acid solution. For this, all components were mixed in a round-bottom flask and mixed at a temperature of 60 ° C for a day. Then the precipitate was filtered off, washed with cold water and dried in air. Thus obtained 2- (hydroxyimino) -N- (R-phenyl) acetamide was dissolved in small portions in concentrated sulfuric acid while keeping the temperature of the mixture in the range of 55-70 ° C, then the mixture was heated to 80 ° C and kept at this temperature for 10-15 minutes. Depending on the product to be obtained, the resulting mixture was cooled to room temperature or to 0 ° C in an ice bath. After cooling to room temperature, the mixture was poured into ice. When cooled to 0 ° C, potassium nitrate was added to the mixture in small portions, keeping the temperature of the reaction mixture in the range of 0-15 ° C. After complete addition, the ice bath was removed, the mixture was stirred for another 0.5 hours. After this, the mixture was also poured into ice. In both cases, the precipitate was filtered off, washed with a small amount of cold water to a neutral pH of the washings, and dried in air. To obtain other nitro derivatives, the isatins obtained by the previously described method were nitrated in two different ways.

In the first method, the obtained isatins were again dissolved in concentrated sulfuric acid, adding isatin to the acid in small portions, then the mixture was cooled in an ice bath to 0 ° C. After small portions, potassium nitrate was added, keeping the temperature in the range of 0-15 ° C. After complete addition, the mixture was stirred for another 0.5 hours at room temperature, then poured into ice. The precipitate was filtered off, washed with a small amount of cold water to a neutral pH of the washings, and dried in air.

In a second method, the resulting isatins were dissolved in acetic / propionic anhydride. A solution of acetyl / propionyl nitrate in acetic / propionic anhydride was added dropwise to such a solution using a dropping funnel, which was prepared as follows: a solution of concentrated nitric acid was slowly added to acetic / propionic anhydride to prevent overheating. After the acetyl / propionyl nitrate solution was completely added to the isatin solution, the mixture was stirred at room temperature for 0.5 hours. Then it was poured into cold water and stirred for another 0.5 hours. The precipitate formed was filtered off, washed with a large amount of cold water, and dried in air.

In the next step, if necessary, alkylation of the obtained isatins (III) was carried out. For this, a solution of isatin (III) in an anhydrous solvent was prepared, argon was blown through the solution for 15 minutes. Then, sodium hydride was added to the reaction mixture. The solution was stirred at room temperature for 0.5 hours, until the evolution of hydrogen ceased. Then, the alkylating agent R1Hal was added to the reaction mixture and the reaction mixture was stirred for 7 days (168 hours) at room temperature. Then, the solvent was removed in a vacuum of a rotary evaporator, the obtained solid residue was washed with cold water and extracted with ethyl acetate or chloroform. The organic extracts were combined, dried over anhydrous sodium sulfate. The solvent was removed in a vacuum rotary evaporator.

The following are examples of specific examples of the performance of the claimed compounds.

Example 1. Obtaining 5-methoxy-isatin

A solution of 14.7 g (0.089 mol) of chloral hydrate, 84 g of sodium sulfate, 9.1 g (0.074 mol) of p-anisidine, 25 ml of 6N hydrochloric acid and 18.5 g (0.27 mol) were placed in a 1 L round-bottom flask. ) hydroxylamine hydrochloride in 0.5 l of water. After that, the reaction mixture was stirred at a temperature of 60 ° C for a day, after which it was cooled to room temperature. The precipitate was filtered off, washed with a small amount of cold water to a neutral pH and dried in air. Received 2- (hydroxyimino) -N- (4-methoxyphenyl) acetamide in 59% yield (45.8 g).

Then, in a three-necked flask with a volume of 250 ml, equipped with a thermometer and a mechanical stirrer, 105 ml of 85% sulfuric acid was placed. Maintaining a temperature in the range of 55 - 70 ° C, 25 g (0.13 mol) of 2- (hydroxyimino) -N- (4-methoxyphenyl) acetamide were gradually added in equal portions with an interval of 2 minutes, after which they were kept at a temperature for another 10 minutes 80 ° C. The solution was cooled to room temperature and poured into a glass containing 1300 g of ice, the precipitate was filtered off, washed with a small amount of cold water until the pH of the washings was neutral. The resulting precipitate was dried in air. 5-methoxyisatin was obtained in 88% yield (21.78 g).

Mp (exp.) = 201-205 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 3.71 (s, 3H); 6.80 (d, 1H, J = 8.23); 7.00 (s, 1H); 7.17 (dd, 1H, J = 1.79, J = 6.03); 10.81 (s, 1H).

Example 2. Obtaining 1-methyl-5-bromisatin

The synthesis was carried out analogously to example 1, using 0.074 mol of p-bromoaniline, a mol of 2- (hydroxyimino) -N- (4-bromophenyl) acetamide was obtained in 66% yield (mp. (Exp.) = 172 ° C). Using 0.084 mol of 2- (hydroxyimino) -N- (4-bromophenyl) acetamide, 5-bromisatin was obtained in 96% yield (mp. (exp.) = 252 ° C).

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 6.82 (d, 1H, J = 8.3), 7.37 (d, 1H, J = 8.3) 7.46 (s, 1H); 10.76 (s, 1H).

Then 5 g (0.022 mol) of 5-bromisatin was dissolved in 40 ml of THF, 0.64 g (0.024 mol) of sodium hydride was added to it. The reaction mixture was stirred 30 minutes until the evolution of hydrogen ceased, then 2.7 ml (0.044 mol) of methyl iodide was added and left at room temperature with stirring for 168 hours. At the end of the reaction, the excess THF was removed in a vacuum of a rotary evaporator, the resulting brown precipitate was washed with cold water and extracted with ethyl acetate. The organic extracts were combined, dried over anhydrous sodium sulfate, and the solvent was removed in a vacuum of a rotary evaporator. The yield of 1-methyl-5-bromisatin was 95% (5.1 g). Mp (exp.) = 153-154 ° C.

Example 3. Obtaining 1-benzylisatin

The synthesis was carried out analogously to example 2, using 0.074 mol of aniline, obtaining 2- (hydroxyimino) -N-phenylacetamide with a yield of 74% (mp (exp.) = 173 ° C) using 0.404 mol of 2- (hydroxyimino) -N -phenylacetamide, obtaining isatin with a yield of 88% (mp. (exp.) = 201 ° C) using 0.038 mol of isatin and 0.042 mol of benzyl bromide, obtained 1-benzyl isatin with a yield of 97%.

Mp (exp.) = 117 ° C.

¹ H NMR (CDCl ₃ , δ, ppm, J, Hz): 4.92 (s, 2H); 6.78 (d, J = 8.09, 1H); 7.08 (t, J = 7.58, 1H); 7.32 (m, 5H); 7.48 (t, J = 7.6, 1H); 7.58 (t, J = 7.33, 1H).

Example 4. Obtaining 5-nitroisatin

520 ml of concentrated sulfuric acid and 52.09 g (0.352 mol) of isatin were placed in a 1-liter three-necked round-bottomed flask equipped with a mechanical stirrer and a thermometer. The reaction mixture was cooled in an ice bath to 0 ° C and 35.55 g (0.352 mol) of potassium nitrate was slowly added in small portions with stirring while maintaining the temperature of the solution in the range of 0-5 ° C. After complete addition, cooling was stopped, stirred at room temperature for 0.5 hours, then the contents of the flask were slowly poured into a glass containing 1.5 kg of finely chopped ice with stirring. The yellow precipitate of 5-nitroisatin was filtered off, washed with a small amount of cold water to a neutral pH and dried in air. The yield was 47% (32.16 g).

Mp (exp.) = 253 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 7.09 (d, 1H, J = 8.6); 8.2 (d, 1H, J = 2.3), 8.44 (dd, 1H, J = 8.54, J = 2.5); 11.67 (s, 1H).

Example 5. Obtaining 5,7-dinitroiztin

5 g (0.034 mol) of Isatin was dissolved in 50 ml of concentrated sulfuric acid with constant stirring. The resulting solution was cooled to 0 ° C, then 7 g (0.068 mol) of potassium nitrate were added in small portions, keeping the temperature in the range of 0-15 ° C. After complete addition, the mixture was heated to 55 ° C and kept for 2.5 hours at this temperature, after which it was poured into 700 g of ice. The resulting solution was extracted with diethyl ether (3 times 200 ml), dried over anhydrous sodium sulfate and the excess solvent was removed in vacuo. Thus, 5,7-dintroisatin was obtained with a yield of 46% (3.7 g).

Mp (exp.) = 208-209 ° C.

NMR ¹ H (DMSO-d ₆ , δ, ppm, J, Hz): 7.09 (s, 1H); 8.2 (s, 1H); 11.67 (s, 1H, NH).

Example 6. Obtaining 5-methoxy-7-nitroisatin

Acetyl nitrate was prepared as follows: in a round bottom flask, 0.34 ml (0.0030) of concentrated nitric acid was slowly added dropwise to 2.97 ml of acetic anhydride. Then, the resulting solution was slowly added dropwise to a solution of 0.5 g (0.0028) of 5-methoxyisatin in 2 ml of acetic anhydride, preventing the mixture from heating. After complete instillation, the reaction mixture was stirred for 0.5 hours. Then it was poured into 18 ml of cold water and stirred for another 0.5 hours. The precipitate formed was filtered off, washed with a small amount of cold water, and dried in air. Thus, 5-methoxy-7-nitroisatin was obtained in a yield of 57% (0.35 g).

Mp (exp.) = 230-235 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 3.86 (s, 3H), 7.58 (d, 1H, J = 2.15), 7.33 (d, 1H, J = 2.40), 11.55 (s, 1H).

Example 7. Obtaining 5-methoxy-4-nitroisatin

In a 100 ml three-necked flask equipped with a thermometer and a mechanical stirrer, 65 ml of concentrated sulfuric acid was placed. Maintaining the temperature in the range of 55–70 ° C, 13.9 g (0.072 mol) of 2- (hydroxyimino) -N- (4-methoxyphenyl) acetamide were gradually added in equal portions with an interval of 2 min, after which they were kept at a temperature for another 10 min 80 ° C. The reaction mixture was cooled in an ice bath to 0 ° C and 7.23 g (0.072 mol) of potassium nitrate was slowly added in small portions with stirring, keeping the solution at a temperature of 0-5 ° C. After complete addition, cooling was stopped, stirred at room temperature for 0.5 hours, then the contents of the flask were slowly poured into a glass containing 1.5 kg of finely chopped ice with stirring. The precipitated precipitate of 5-methoxy-4-nitroisatin was filtered off, washed with a small amount of cold water to a neutral pH and dried in air. The yield was 10% (1.59 g).

T. pl. (exp.) = 238-240 ° C (decomp.).

NMR ¹ H (DMSO-d ₆ , ppm, J, Hz): 3.86 (s, 3H), 7.06 (d, 1H, J = 8.72), 7.57 (d ., 1H, J = 8.72), 11.15 (s, 1H).

¹³ C NMR (DMSO-d ₆ , δ, ppm): 57.88; 109.43; 115.62; 123.97; 138.13; 143.74; 145.39; 159.19; 180.27.

IR (liquid paraffin, cm ^-1 ): 972 (COC), 1089, 1282, 1351 (NO ₂ ), 1379, 1441 (CH ₃ ), 1540, 1598, 1636 (NO ₂ ), 2847 (O-CH ₃ ) 3262 (NH).

Mass spectrum (electron impact, 70 eV), m / z (I,%): 222 (78, M +), 178 (40), 120 (77), 80 (47), 65 (57), 53 (33 ), 30 (100), 15 (54).

Example 8. Obtaining 5,6,7-trimethylisatin

The synthesis was carried out analogously to example 1, using 0.074 mol of 2,3,4-trimethylaniline, received 5,6,7-trimethylisatin with a yield of 35%.

¹ H NMR (CDCl ₃ , δ, ppm, J, Hz): 1.91 (s, 3H), 2.28 (s, 3H), 2.60 (s, 3H), 7 52 (s, 1H); 10.23 (s, 1H).

Example 9. Obtaining (5-methoxy-2-oxoindol-3-ylidene) acetonitrile

5.3-methoxyisatin in an amount of 10.3 g (0.058 mol) was placed in a 100 ml round-bottom flask, dissolved in 30 ml of 1,4-dioxane, then a solution of 4.94 g (0.058 mol) of cyanoacetic acid and 9.68 was added with stirring ml (0.07 mol) of triethylamine in 5 ml of 1,4-dioxane. Stirring of the reaction mixture was carried out for 4 hours, after completion of the reaction, 30 ml of concentrated hydrochloric acid was added to the mixture. The reaction mixture was left at room temperature for 4 days. The precipitated crystals were filtered off, washed with cold water and dried in air. Received (5-methoxy-2-oxoindolin-3-ylidene) acetonitrile in 53% yield (7.8 g).

Mp (exp.) = 290-292 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 3.73 (s, 3H); 6.47 (s, 1H); 6.79 (d, J = 8.6, 1H); 7.03 (dd, J = 2.5, 8.6, 1H); 7.35 (d, J = 2.3, 1H); 10.65 (s, 1H).

Example 10. Obtaining (1-methyl-5-bromo-2-oxoindolin-3-ylidene) acetonitrile

After this, the synthesis was carried out analogously to example 9, using 0.021 mol of 1-methyl-5-bromisatin, (1-methyl-5-bromo-2-oxoindolin-3-ylidene) acetonitrile was obtained in 89% yield.

T. pl. (exp.) = 194-196 ° C.

¹ H NMR (CDCl ₃ , δ, ppm, J, Hz): 3.23 (s, 3H), 6.38 (s, 1H), 6.75 (d, J = 8, 33, 1H), 7.57 (d, J = 8.30, 1H), 8.16 (s, 1H).

Example 11. Obtaining (1-benzyl-2-oxindol-3-ylidene) acetonitrile

Then the synthesis was carried out analogously to example 9, using 0.032 mol of 1-benzylisatin, (1-benzyl-2-oxoindolin-3-ylidene) acetonitrile was obtained in 82% yield.

M.p. (exp.) = 154-156 ° C.

¹ H NMR (CDCl ₃ , δ, ppm, J, Hz): 4.93 (s, 2H); 6.39 (s, 1H); 6.75 (d, J = 7.83, 1H); 7.1 (t, J = 7.58, 1H); 7.33 (m, 6H); 8.09 (d, J = 7.58, 1H).

Example 12. Obtaining (5-nitro-3-hydroxy-2-oxindol-3-ylidene) acetonitrile

To a solution of 10.4 g (0.054 mol) of 5-nitroisatin in 25 ml of 1,4-dioxane was added a solution of 4.61 g (0.054 mol) of cyanoacetic acid and 9 ml of triethylamine in 10 ml of 1,4-dioxane, the mixture was vigorously stirred at room temperature for 4 hours. Then 25 ml of concentrated hydrochloric acid was added, the reaction mixture was concentrated in a vacuum of a rotary evaporator. The resulting oil was dissolved in 10 ml of ethyl acetate and poured into a glass containing 300 ml of cold water with stirring. The precipitate was filtered off, washed with a little cold water and dried in air. (5-Nitro-3-hydroxy-2-oxoindolin-3-yl) acetonitrile was obtained in 16% yield (2.05 g).

Mp (exp.) = 160-165 ° C.

NMR ¹ H (DMSO-d ₆ , δ, ppm, J, Hz): 2.97 (d, J = 16.7, 1H), 3.12 (d, J = 16.6, 1H ), 3.56 (s, broad OH), 6.94 (s, 1H), 7.05 (d, J = 8.8, 1H), 8.20 (d, J = 8 6, 1H); 8.32 (s, 1H); 11.19 (s, 1H).

Example 13. Obtaining (5,7-dinitro-3-hydroxy-2-oxindol-3-yl) acetonitrile

The synthesis was carried out analogously to example 12, using 0.013 mol of 5,7-dinitroisatin, (5,7-dinitro-3-hydroxy-2-oxoindolin-3-yl) acetonitrile was obtained in a yield of 60%.

Mp (exp.) = 155-159 ° C.

NMR ¹ H (DMSO-d ₆ , δ, ppm, J, Hz): 3.29 (s, 2H), 3.49 (s broad, 1H), 8.62 (d, 1H, J = 2.14), 8.84 (d, 1H, J = 2.14), 12.04 (s, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 25.67; 70.68; 116.88; 122.43; 124.42; 130.72; 135.17; 141.84; 143.92; 177.57.

IR (liquid paraffin, cm ^-1 ): 1320 (NO ₂ ); 1550 (NO ₂ ); 1750 (C = O); 2280 (CN); 1950-3500 (OH); 3250 (C (O) NH); 3480 (C (O) NH).

Elemental analysis: found (%): C 43.48; H 2.49; N, 19.24; calculated for C ₁₀ H ₆ N ₄ O ₆ (%): C 43.18; H 2.17; N, 20.14.

Example 14. Obtaining (5-methoxy-7-nitro-3-hydroxy-2-oxindol-3-yl) acetonitrile

In a 10 ml round bottom flask equipped with a reflux condenser, a solution of 0.5 g (0.0023 mol) of 5-methoxy-7-nitroisatin and 0.191 g (0.0023 mol) of cyanoacetic acid in 6 ml of ethanol in the presence of 0.025 ml ( 0.0003 mol) of piperidine for 3 hours. Then the reaction mixture was cooled to room temperature, the excess solvent was removed in a vacuum rotary evaporator. The obtained solid residue was washed several times with cold water, dried in air. (5-Methoxy-7-nitro-3-hydroxy-2-oxoindolin-3-yl) acetonitrile was obtained in 71% yield (0.422 g).

Mp (exp.) = 125-130 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 3.10-3.20 (m, 2H), 3.84 (s, 3H), 6.98 (br.s, 1H), 7.51 (d, 1H, J = 2.53), 7.54 (d, 1H, J = 2.53), 11.23 (s, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 26.08; 56.62; 71.17; 106.60; 117.11; 120.44; 131.35; 132.41; 135.23; 154.89; 177.26.

IR (liquid paraffin, cm ^-1 ): 1019 (COC), 1292, 1306, 1316 1535, 1590 (NO ₂ ), 1742 (C = O), 2277 (CN), 2847, 2936, 2978 (CH), 3224 (NH), 3334 (OH).

Mass spectrum (electron impact, 70 eV), m / z (I,%): 263 (74, M +), 223 (100), 193 (14), 168 (12), 148 (21), 122 (9 ), 106 (5), 79 (4), 44 (11), 30 (11).

Example 15. Obtaining (5-nitro-3-hydroxy-2-oxindol-3-yl) acetic acid

In a round-bottom flask equipped with a reflux condenser, a solution of 1.08 g (0.0056 mol) of 5-nitroisatin and 0.631 g (0.0061 mol) of malonic acid in 1.5 ml of pyridine in the presence of 0.1 ml (0.0014 mol) was boiled ) piperidine for 24 hours, then the reaction mixture was cooled to room temperature. The precipitate was filtered off, washed with a solution of hydrogen chloride in 1,4-dioxane, then with cold water, and dried in air. Thus, (5-nitro-3-hydroxy-2-oxoindol-3-yl) acetic acid was obtained in a yield of 49% (0.691 g).

T. pl. (exp.) = 215-216 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 3.05 (d, 1H, J = 16.51), 3.15 (d, 1H, J = 16.51) 6.40 (s. Broad, 1H), 7.00 (d, 1H, J = 8.68), 8.20 (dd, 1H, J = 8.62, J = 2.38), 8, 27 (d, 1H, J = 2.32), 10.99 (s, 1H).

¹³ C NMR (DMSO-d ₆ , δ, ppm): 66.80; 72.66; 110.11; 120.18; 127.01; 132.97; 142.46; 149.94; 170.83; 178.92.

Mass spectrum (HRMS-ESI): 253.0455, C ₁₀ H ₉ N ₂ O ₆ (M + H), calculated: 252.1803, C ₁₀ H ₈ N ₂ O ₆

Elemental analysis: found (%): C 46.99; H 3.47; N, 10.90; calculated for C ₁₀ H ₈ N ₂ O ₆ (%): C 47.63; H 3.20; N, 11.11.

IR (liquid paraffin, cm ^-1 ): 1339 (NO ₂ ); 1540 (NO ₂ ); 1656 (C = O); 1681 (C = O); 2686-3232 (OH); 3060 (C (O) NH).

Example 16. Obtaining (5-methoxy-4-nitro-3-hydroxy-2-oxindol-3-yl) acetic acid

In a round-bottom flask equipped with a reflux condenser, a solution of 1 g (0.0045 mol) of 5-methoxy-4-nitroisatin and 0.47 g (0.0045 mol) of malonic acid in 7 ml of pyridine in the presence of 0.05 ml (0 , 0007 mol) of piperidine for 15 hours, then the reaction mixture was cooled to room temperature, 5 ml of a solution of hydrogen chloride in 1,4-dioxane was added. The solution was condensed in a vacuum of a rotary evaporator, then the obtained solid residue was washed with a solution of hydrogen chloride in 1,4-dioxane and water, dried in air. Thus, (5-methoxy-4-nitro-3-hydroxy-2-oxoindol-3-yl) acetic acid was obtained in a yield of 70% (0.89 g).

Mp (exp.) = 240-245 ° C.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 2.83 (d, 1H, J = 16.63), 2.99 (d, 1H, J = 16.63) 3.80 (s, 3H), 6.48 (broad s, 1H), 6.94 (d, 1H, J = 8.56), 7.16 (d, 1H, J = 8.56), 10.54 (s, 1H), 12.28 (br s, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 42.07; 57.40; 73.66; 112.54; 114.71; 123.64; 137.34; 141.93; 145.52; 170.31; 177.51.

Mass spectrum (electron impact, 70 eV), m / z (I,%): 282 (9, M +), 255 (2), 223 (5), 164 (2), 146 (5), 134 (2 ), 120 (3), 84 (84), 56 (46), 44 (100), 29 (47), 15 (46).

Example 17. Obtaining (5,7-dinitro-2-oxindol-3-ylidene) acetic acid

In a round-bottom flask equipped with a reflux condenser, a solution of 1.02 g (0.0043 mol) of 5.7-dinitroisatin and 0.45 g (0.0043 mol) of malonic acid in 10 ml of pyridine in the presence of 0.1 ml (0.0014) was boiled mol) of piperidine for 60 hours, then the reaction mixture was cooled to room temperature. The precipitate was filtered off, washed with a solution of hydrogen chloride in 1,4-dioxane, then with cold water, and dried in air. Thus, (5,7-dinitro-2-oxoindol-3-ylidene) acetic acid was obtained in a yield of 53% (0.64 g).

T. pl. (exp.) = 198-200 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 7.22 (s, 1H), 8.94 (s, 1H), 9.46 (s, 1H), 11, 75 (s. Broad, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 119.06; 122.37; 126.74; 128.08; 137.74; 140.14; 140.64; 160.99; 165.79.

Elemental analysis: found (%) C 43.15; H 2.03; N, 15.10; calculated for C ₁₀ H ₅ N ₃ O ₇ (%) C 43.02; H 1.81; N, 15.05.

IR (liquid paraffin, cm ^-1 ): 1323, 1339, 1358 (NO ₂ ); 1524, 1549 (NO ₂ ); 1623 (C = O); 1669 (C = O); 3269 (C (O) NH); 3522 (OH).

Example 18. Obtaining (5-bromo-2-oxoindol-3-ylidene) acetic acid

The synthesis was carried out analogously to example 17, using 2.57 g (0.011 mol) of 5-bromisatin, boiling was carried out for 35 hours, (5-bromo-2-oxoindole-3-ylidene) acetic acid was obtained in 21% yield.

T. pl. (exp.) = 275-280 ° C (decomp.).

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 6.98 (s, 1H), 7.31 (d, J = 8.21, 1H), 7.70 (d ., J = 9.04, 1H), 8.42 (s, 1 H), 12.21 (s, 1H).

¹³ C NMR (DMSO-d ₆ , δ, ppm): 114.58; 117.91; 118.39; 125.97; 128.74; 133.84, 138.99; 139.58; 161.24; 166.69.

IR (liquid paraffin, cm ^-1 ): 1011 (C-Br), 1617 (NH, NC = O), 1644 (C = O), 1710 (C = O), 3060 (NH), 3375 (OH).

Example 19. Obtaining 2- (5-methoxy-2-oxoindolin-3-ylidene) malonic acid monoethyl ester

In a 250 ml round bottom flask, a solution of 3 g (0.017 mol) of 5-methoxyisatin and 3.5 ml (0.021 mol) of malonic acid diethyl ether in 100 ml of ethyl alcohol in the presence of 0.5 ml (0.005 mol) of piperidine was boiled for 15 hours . Then the reaction mixture was cooled to room temperature and poured into a glass containing 500 ml of cold water. The mixture was left for 4 days, after which the obtained crystals were filtered off, washed with cold water and dried in air. Thus, 2- (5-methoxy-2-oxoindolin-3-ylidene) malonic acid monoethyl ester was obtained in 69% yield (3.39 g).

T. pl. (exp.) = 240-245 ° C.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.27 (t, J = 7.20, 3H); 4.23 (q, J = 7.09, 2H); 3.83 (s, J = 7.58, 3H); 6.84 (d, J = 9.42, 1H); 7.16 (d, J = 8.97, 1H); 7.28 (s. 1H).

Elemental analysis: found (%) C 57.66; H 4.46; N, 4.83; calculated for C ₁₄ H ₁₃ NO ₆ (%): C 57.73; H 4.50; N, 4.81.

IR (liquid paraffin, cm ^-1 ): 1620 (C = O), 1650 (C = C), 3510 (OH).

Example 20. Obtaining 2- (5-bromo-2-oxoindolin-3-ylidene) malonic acid monoethyl ester

The synthesis was carried out analogously to example 19, using 0.0133 mol of 5-bromisatin, 2- (5-bromo-2-oxoindolin-3-yl) malonic acid monoethyl ester was obtained in 67% yield.

T. pl. (exp.) = 180-185 ° C.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.55 (t, J = 10.10 3H); 4.36 (q, J = 7.08, 2H); 7.45 (dd, J = 2.28, J = 8.97, 1H); 7.73 (dd, J = 2.14, J = 8.33, 1H); 8.31 (d, J = 1.89, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 24.20; 25.65; 39.79; 40.21; 41.70; 46.82; 114.74; 120.19; 127.36; 130.80; 134.24; 138.59; 140.50.

Mass spectrum (HRMS-ESI): 341.3231, C ₁₃ H ₁₁ BrNO ₅ (M + H); calculated: 340.1262, C ₁₃ H ₁₀ BrNO ₅ .

IR (liquid paraffin, cm ^-1 ): 1640 (C = O), 1660 (C = C), 3530 (OH).

Example 21. Obtaining monoethyl ether (5-nitro-2-oxindol-3-ylidene) malonic acid

The synthesis was carried out analogously to example 19, using 0.016 mol of 5-nitroisatin, monoethyl ether (5-nitro-2-oxindol-3-ylidene) malonic acid was obtained in 81% yield.

Mp (exp.) = 260-265 ° C.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.72 (t, J = 6.88, 3H); 4.74 (q, J = 7.12, 2H); 7.19 (dd, J = 2.71, J = 8.56, 1H); 8.21 (dd, J = 3.01, J = 7.55, 1H); 9.15 (d, J = 8.66, 1H).

Elemental analysis: found (%): C 50.38, H 3.09, N 9.10, calculated for C ₁₃ H ₁₀ N ₂ O ₇ (%): C 50.59; H 3.29; N, 9.15.

IR (liquid paraffin, cm ^-1 ): 1350 and 1540 (NO ₂ ), 1640 (C = O), 1660 (C = C), 3510 (OH).

Example 22. Obtaining ethyl ester (5-methoxy-2-oxindol-3-ylidene) acetic acid

2 g (0.0069 mol) of 2- (5-methoxy-2-oxoindolin-3-ylidene) malonic acid monoethyl ester and 16 ml of 2-ethoxyethanol were added to a 100 ml round bottom flask equipped with a reflux condenser with a calcium chloride tube. The mixture was boiled for 4 hours with stirring. After completion of the reaction, the mixture was cooled to room temperature, the excess solvent was removed in a vacuum of a rotary evaporator, the resulting oil was dissolved in a small amount of ethyl acetate, and passed through a layer of silica gel. Excess solvent was removed in a vacuum rotary evaporator. Thus, ethyl acetate (5-methoxy-2-oxoindolin-3-ylidene) acetic acid was obtained in 33% yield (0.56 g).

Mp (exp.) = 230-240 ° C.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.13 (t, J = 7.07, 3H); 4.25 (q, J = 4.14, 2H); 5.65 (s, 1H); 6.97 (dd, J = 6.84, J = 13.41, 1H); 7.93 (dd, J = 3.48, J = 10.33, 1H); 8.12 (d, J = 3.15, 1H); 10.54 (s, 1H).

IR (liquid paraffin, cm ^-1 ): 1640 (C = O), 1680 (C = C).

Example 23. Obtaining ethyl ester (5-nitro-2-oxindol-3-ylidene) acetic acid

The synthesis was carried out analogously to example 22, using 0.0023 mol of monoethyl ether (5-nitro-2-oxindol-3-ylidene) malonic acid, ethyl ester (5-nitro-2-oxindol-3-ylidene) acetic acid was obtained with a yield of 26 %

T. pl. (exp.) = 175-180 ° C.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.25 (t, J = 7.08, 3H); 4.27 (q, J = 4.55, 2H); 5.77 (s, 1H); 7.05 (dd, J1 = 14.90, J2 = 8.71, 1H); 8.11 (dd, J1 = 9.47, J2 = 2.27, 1H); 8.61 (d. J = 2.02, 1H); 11.05 (s., 1H).

Elemental analysis: found (%) C 54.66, H 3.67, N 10.42, calculated for C ₁₂ H ₁₀ N ₂ O ₅ (%) C 54.97, H 3.84, N 10.68.

IR (liquid paraffin, cm ^-1 ): 1520 (NO ₂ ), 1610 (C = O), 1630 (C = C).

Example 24. Obtaining 2- (5,6,7-trimethyl-2-oxindol-3-ylidene) -2-carbamide-acetic acid

The synthesis was carried out analogously to example 21, using 0.0059 mol of 5,6,7-trimethylisatin and 0.0059 mol of malonamide, boiling was carried out for 50 hours, 2- (5,6,7-trimethyl-2-oxindole-3- ylidene) -2-carbamide-acetic acid with a yield of 28%.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.83 (s, 3H), 2.30 (s, 3H), 2.60 (s, 3H), 4 11 (s, 2H); 7.63 (s, 1H); 10.45 (s, 1H).

Example 25. Obtaining (5,6,7-trimethyl-2-oxindol-3-ylidene) acetmide

The synthesis was carried out analogously to example 22, using 0.0059 mol of 2- (5,6,7-trimethyl-2-oxindol-3-ylidene) -2-carbamide-acetic acid, (5,6,7-trimethyl-2- was obtained oxindol-3-ylidene) acetmide in 40% yield.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.83 (s, 3H), 2.30 (s, 3H), 2.60 (s, 3H), 4 11 (s, 2H), 4.22 (s, 2H), 6.69 (s, 1H), 7.63 (s, 1H), 10.45 (s, 1H).

Example 26. Obtaining (5-methoxy-2-oxoindol-3-yl) acetonitrile

2.54 g (0.01 mol) of (5-Methoxy-2-oxoindolin-3-ylidene) acetonitrile was dissolved in 40 ml of ethyl acetate and 10 ml of 3N hydrochloric acid. Then added 1.2 g (0.02 mol) of zinc dust, there was a significant heating of the reaction mixture. At the end of the reaction was judged using thin layer chromatography. After termination of the reaction, the organic layer was separated, the aqueous layer was extracted with ethyl acetate. The organic extracts were combined, washed with cold water, dried over anhydrous sodium sulfate, and the solvent was removed in a vacuum of a rotary evaporator. Received (5-methoxy-2-oxoindolin-3-yl) acetonitrile with a yield of 57% (1.09 g).

Mp (exp) = 179-180 ° C.

¹ H NMR (CDCl ₃ , δ, ppm, J, Hz): 2.77 (dd, J = 17.0, J = 9.1, 1H); 3.10 (dd, J = 17.0, J = 4.8, 1H); 3.70 (dd, J = 8.6, J = 4.3, 1H); 3.81 (s, 3H); 6.89 (dd, J = 8.6, J = 1.2, 1H); 7.02 (s, 1H); 7.20 (d, J = 8.6, 1H); 8.45 (s, 1H, NH).

Example 27. Obtaining (1-methyl-5-bromo-2-oxoindolin-3-yl) acetonitrile

The synthesis was then carried out analogously to 26, using 0.0013 mol of (1-methyl-5-bromo-2-oxoindolin-3-ylidene) acetonitrile, and (1-methyl-5-bromo-2-oxoindolin-3-yl) acetonitrile was obtained yield 72%.

Mp (exp.) = 167-169 ⁰ С

¹ H NMR (CDCl ₃ , δ, ppm, J, Hz): 2.73 (dd, J = 16.5, J = 8.09, 1H); 3.10 (dd, J = 16.8, J = 8.09, 1H); 3.96 (s, 3H); 3.68 (dd, J = 8.59, J = 4.29, 1H); 6.77 (d, J = 8.26, 1H); 7.49 (d, J = 8.24, 1H); 7.61 (s, 1H).

Example 28. Obtaining (1-benzyl-2-oxoindolin-3-yl) acetonitrile

The synthesis was carried out analogously to example 26, using 0.0077 mol (1-benzyl-2-oxoindolin-3-ylidene) acetonitrile, (1-benzyl-2-oxoindolin-3-yl) acetonitrile was obtained in a yield of 60%.

Mp (exp.) = 151-153 ° C.

¹ H NMR (CDCl ₃ , δ, ppm, J, Hz): 2.76 (d, J = 15.9, 1H); 2.96 (d, J = 15.9, 1H); 3.10 (dd, J = 16.67, J = 4.8, 2H); 3.68 (t, J = 4.55, 1H); 4.88 (d, J = 10.1, 2H); 6.73 (d, J = 7.83, 1H); 7.03 (t, J = 7.33, 1H); 7.22 (m, 5H); 7.44 (d, J = 7.33, 1H).

Example 29. Obtaining (5-acetamido-3-hydroxy-2-oxindol-3-yl) acetonitrile

A solution of 0.5 g (0.0021 mol) of (5-nitro-3-hydroxy-2-oxoindol-3-yl) acetonitrile and 0.2 ml (0.023 mol) of acetic anhydride in 7 ml of acetic acid was placed in a round bottom flask, equipped with a reflux condenser. Then, 1 g (0.008 mol) of zinc dust was quickly sprinkled with a considerable heating of the reaction mixture, and the solution was boiled for 2 hours. After the reaction mixture was cooled to room temperature, concentrated in a vacuum of a rotary evaporator. The resulting dry residue was dissolved in 20 ml of a mixture of methanol: ethyl acetate (1: 1) and passed through a layer of silica gel (R _f = 0.7). The resulting solution was collected, the excess solvent was removed in vacuo. The yield of (5-acetamido-3-hydroxy-2-oxindol-3-yl) acetonitrile was 45% (0.23 g).

Mp (exp.) = 123-128 ° C.

NMR ¹ H (DMSO-d6, δ, ppm, J, Hz): 2.01 (s, 3H), 2.80 (d, J = 16.7, 1H), 2.96 ( d, J = 16.4, 1H); 6.61 (s, broad 1H); 6.77 (d, J = 8.3, 1H); 7.50 (d, J = 8.6, 1H); 7.69 (s, 1H), 9.83 (s, 1H), 10.39 (s, 1H).

Example 30. Obtaining (5,7-bis (acetamido) -3-acetoxy-2-oxindol-3-yl) acetonitrile

The synthesis was carried out analogously to example 29, using 0.003 mol (5,7-dinitro-3-hydroxy-2-oxoindolin-3-yl) acetonitrile and 0.058 mol of acetic anhydride, (5,7-bis (acetamido) -3-acetoxy-was obtained 2-oxindol-3-yl) acetonitrile in 79% yield.

Mp (exp.) = 156-158 ^° C.

¹ H NMR Spectrum (DMSO-d ₆ , δ, ppm, J, Hz): 1.90 (s, 3H), 2.01 (s, 3H), 2.04 (s, 3H), 2 90 (d, 1H, J = 16.7), 3.02 (d, 1H, J = 16.6), 7.56 (s, 1H); 7.76 (s, 1H), 9.49 (s, 1H), 9.96 (s, 1H), 10.1 (s, 1H).

¹³ C NMR (DMSO-d ₆ , δ, ppm): 21.81, 23.77, 24.24, 26.75, 72.79, 112.72, 115.72, 117.39, 121 86, 129.55, 130.99, 134.72, 168.54, 168.98, 173.29, 176.62.

Mass spectrum (HRMS-ESI): 345.1193, C ₁₆ H ₁₇ N ₄ O ₅ (M + H) _, calculated: 344.3220, C ₁₆ H ₁₆ N ₄ O ₅ .

Example 31. Obtaining (5-methoxy-7-acetamido-3-acetoxy-2-oxindol-3-yl) acetonitrile

The synthesis was carried out analogously to 29, using 0.00057 mol (5-methoxy-7-nitro-3-hydroxy-2-oxoindolin-3-yl) acetonitrile and 0.0014 mol of acetic anhydride, (5-methoxy-7-acetamido- was obtained 3-acetoxy-2-oxindol-3-yl) acetonitrile in 76% yield.

Mp (exp.) = 185-190 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 1.83 (s, 3H), 2.05 (s, 3H), 2.94 (d, 1H, J = 16.55), 3.05 (d, 1H, J = 16.55), 3.70 (s, 3H), 6.89 (d, 1H, J = 1.89), 7.25 (d, 1H, J = 2.27), 9.83 (s, 1H) 10.51 (br s, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 22.95; 23.95; 26.60; 55.97; 72.88; 107.08; 109.24; 117.43; 123.01; 126.80; 131.72; 155.37; 169.03; 176.51.

IR (liquid paraffin, cm ^-1 ): 742 (CH ₂ ), 1043 (COC), 1466 (CH ₂ ), 1558 (NH), 1614 (NH), 1723 (C = O), 1727 (C = O) , 1738 (C = O), 2258 (CN), 2840, 2938, 2963 (CH), 3000-3650 (NH, NH).

Mass spectrum (electron impact, 70 eV), m / z (I,%): 275 (16), 257 (21), 233 (3), 215 (26), 193 (13), 164 (18), 127 (14), 110 (3), 84 (5), 43 (100), 15 (45).

Example 32. Obtaining (5-acetamido-3-acetoxy-2-oxoindol-3-yl) acetic acid

1 g (0.0039 mol) of (5-Nitro-3-hydroxy-2-oxoindol-3-yl) acetic acid was dissolved in 7.5 ml of acetic acid in the presence of 0.92 ml (0.0098 mol) of acetic anhydride was boiled within 0.5 hours. Then the reaction mixture was cooled to room temperature, passed through a layer of silica gel (eluent acetic acid). The solvent was removed in a vacuum rotary evaporator. Received (5-acetamido-3-acetoxy-2-oxoindol-3-yl) acetic acid with a yield of 79% (0.95 g) (mp. (Exp.) = 232-235 ° C).

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.99 (s, 3H), 2.71 (s, 1H), 6.67 (d, 1H, J = 8.34 ), 7.41 (d, 1H, J = 8.46), 7.46 (s, 1H), 9.79 (s, 1H), 10.06 (s, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 22.34; 24.24; 42.94; 73.73; 109.51; 116.45; 120.25; 132.78; 133.91; 138.46; 168.26; 174.88; 178.84.

Elemental analysis: found (%): C 54.39; H 4.68; N, 10.35; calculated for C ₁₂ H ₁₂ N ₂ O ₅ (%): C 54.55; H 4.58; N, 10.60.

IR (liquid paraffin, cm ^-1 ): 1203, 1225 (CO); 1554 (NH); 1610 (C = O); 1714 (C = O); 2592-3400 (OH), 3099 (NH).

Example 33. Obtaining (5-methoxycarbonylamino-2-oxindol-3-yl) acetonitrile

In a 100 ml round bottom flask, a solution of 1 g (0.0047 mol) (5-nitro-2-oxindol-3-ylidene) acetonitrile with 3.4 g (0.052 mol) of zinc was prepared in 10 ml of 1,4-dioxane. To this solution, 30 ml of a solution of hydrogen chloride in 1,4-dioxane were slowly added dropwise, while a significant heating of the reaction mixture was observed. After complete instillation, stirring of the solution was continued until the evolution of hydrogen ceased (2-3 hours). The solution was purified from the precipitate by filtration, and the solvent was removed in a vacuum of a rotary evaporator. The dry residue was dissolved in 15 ml of tetrahydrofuran, 2.3 ml (0.023 mol) of piperidine and 0.72 ml (0.0093 mol) of methyl chloroformate were added. The reaction mixture was boiled for 1.5 hours in a round bottom flask equipped with a reflux condenser, then cooled to room temperature, the excess solvent was removed in a vacuum of a rotary evaporator. The resulting dry residue was washed with 1,4-dioxane. The precipitate was separated by filtration, the excess solvent was removed in a vacuum rotary evaporator. Thus, (5-methoxycarbonylamino-2-oxindol-3-yl) acetonitrile was obtained in 20% yield (0.23 g).

Mp (exp.) = 145-150 ° C.

¹ H NMR (DMSO-d ₆ , δ, ppm, J, Hz): 3.00 (dd, 1H, J = 6.19, J = 17.05), 3.17 (dd, 1H, J = 5.56, J = 17.05), 3.64 (s, 3H), 3.80 (t, 1H, J = 5.56), 6.79 (d, 1H, J = 8.21), 7.24 (d, 1H, J = 7.96), 7.54 (s, 1H), 9.51 (s, 1H), 10.50 (s, 1H).

NMR ¹³ C (DMSO-d ₆ , δ, ppm): 18.21; 42.06; 51.98; 109.99; 116.27; 118.64; 119.62; 128.00; 134.03; 138.33; 154.67; 176.75.

IR (liquid paraffin, cm ^-1 ): 1069 (CO), 1240 (CO), 1463 (CH ₃ ), 1683 (C = O), 1719 (C = O), 2263 (CN), 2848 (O-CH ₃ ), 3108 (NH), 3305 (NH).

Mass spectrum (electron impact, 70 eV), m / z (I,%): 245 (100, M +), 205 (47), 186 (15), 173 (49), 131 (10), 119 (11 ), 91 (5), 77 (3), 59 (14), 15 (10).

Example 34. Obtaining monoethyl ester of 2- (5-methoxy-2-oxoindolin-3-yl) malonic acid

A solution of 0.1 g (0.00034 mol) of 2- (5-methoxy-2-oxoindolin-3-ylidene) malonic acid monoethyl ester in 200 ml of ethyl alcohol was hydrogenated in a H-Cube Research Complex flow reactor, pressure H ₂ 1 atm., Room temperature, Pd / C catalyst 10%, flow rate 1 ml / min. After complete hydrogenation, the solvent was removed in a vacuum of a rotary evaporator. Thus, 2- (5-methoxy-2-oxoindolin-3-yl) malonic acid monoethyl ester was obtained in 88% yield (0.87 g).

T. pl. (exp.) = 220-225 ° C.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 0.87 (t, J = 7.20, 3H); 3.87 (s, 2H); 4.27 (q, J = 7.03, 2H); 7.05 (d, J = 6.70, 1H); 4.81 (d, J = 8.34, 1H); 6.68 (dd, J = 5.65, J = 7.83, 1H); 7.08 (d, J = 2.65, 1H); 7.19 (dd, J = 2.65, J = 8.46, 1H); 10.83 (s, 1H).

Elemental analysis: found (%): C 57.03, H 5.13, N 4.66; calculated for C ₁₄ H ₁₅ NO ₆ (%): C 57.34, H 5.16, N 4.78.

IR (liquid paraffin, cm ^-1 ): 1640 (C = O).

Example 35. Obtaining (5-trifluoroacetamido-3-trifluoroacetoxy-2-oxoindol-3-yl) acetonitrile

The synthesis was carried out analogously to example 29, using 0.0025 mol (5-nitro-3-hydroxy-2-oxindol-3-yl) acetonitrile and 0.0063 mol of trifluoroacetic anhydride, (5-trifluoroacetamido-3-trifluoroacetoxy-2-oxoindole was obtained -3-yl) acetonitrile in 55% yield.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 3.56 (d, J = 17.06, 1H), 3.68 (d, J = 17.06, 1H) 7.46 (dd, J = 3.29, J = 8.84, 1H), 8.59 (d, J = 2.15, 1H), 8.64 (dd, J = 2, 02, J = 8.97, 1H), 10.45 (s, 1H), 11.03 (s, 1H).

Example 36. Obtaining (5-bromo-3-hydroxy-2-oxindol-3-yl) acetic acid

The synthesis was carried out analogously to example 17, using 0.033 mol of 5-bromisatin and 0.033 mol of malonic acid, boiling was carried out for 6 hours, (5-bromo-3-hydroxy-2-oxindol-3-yl) acetic acid was obtained in a yield of 47%.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 2.94 (d, J = 16.04, 1H), 3.01 (d, J = 16.17, 1H) 7.26 (dd, J = 1.51, J = 8.09, 1H), 7.32 (dd., J = 2.65, J = 7.84, 1H), 8.01 (d ., J = 2.02, 1H), 10.96 (s, 1H).

Example 37. Obtaining (5-methoxycarbonylamino-3-methoxycarboxy-2-oxindol-3-yl) acetic acid

The synthesis was carried out analogously to example 33, using 0.0012 mol of (5-nitro-3-hydroxy-2-oxindol-3-yl) acetic acid and 0.003 mol of methyl chloroformate, (5-methoxycarbonylamino-3-methoxycarboxy-2-oxindole- 3-yl) acetic acid with a yield of 13%.

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 3.37 (d, J = 16.06, 1H), 3.44 (d, J = 16.17, 1H) 3.55 (s, 3H), 3.80 (s, 3H), 7.52 (dd, J = 2.02, J = 8.84, 1H), 7.60 (dd, J = 2.78, J = 8.71, 1H), 8.42 (d, J = 1.16, 1H), 10.45 (s, 1H), 10.96 (s, 1H) .

Example 38. Obtaining (5,6,7-trimethyl-2-oxindol-3-yl) acetamide

The synthesis was carried out analogously to example 26, using 0.001 mol (5,6,7-trimethyl-2-oxindol-3-ylidene) acetamide, (5,6,7-trimethyl-2-oxindol-3-yl) acetamide was obtained with a yield of 15 %

¹ H NMR (DMSO-d ₆ , ppm, J, Hz): 1.83 (s, 3H), 2.30 (s, 3H), 2.60 (s, 3H), 2 96 (q, J = 1.78, 1H), 3.15 (q, J = 2.02, 1H), 4.11 (s, 2H), 4.55 (t, J = 1 96, 1H), 6.69 (s, 1H), 7.63 (s, 1H), 10.45 (s, 1H).

The study of the biological activity of new derivatives of melatonin.

Table 1 shows the data on anti-glaucoma activity. The ability of melatonin and its synthesized analogues to reduce IOP was evaluated on normotensive chinchilla rabbits, males, weighing about 2 kg. For all investigated substances, a concentration of 0.1% (mass / volume) was used, which is 4.3 μm for melatonin. All substances were dissolved in a 0.05 M phosphate buffer solution, pH 7.4, containing 1% DMSO (volume). A portion was dissolved in DMSO and then diluted to the desired concentration with a buffer solution.

The solutions were instilled using an automatic dispenser of 50 μl in both eyes. IOP was measured before instillation and then for 6 hours at intervals of 1 hour using a Tonovet automatic veterinary blood pressure monitor (Icare, Finland). The hypotensive effect of each substance was evaluated in a group of 3 animals (6 eyes). To introduce corrections for normal diurnal IOP fluctuations, IOP was simultaneously measured in 3 intact animals (6 eyes). In the next experiment, animals were taken after a break of at least 7 days. Since the rate of decrease in IOP in different animals of IOP is different, during the study we focused on the maximum decrease in IOP in each eye, and not on the average decrease at a specific time point.

The affinity studies of the proposed compounds of the general formula (I) with respect to the MT ₁ and MT ₂ subtypes of the melatonin receptor and the in vivo study of antiglaucoma activity were carried out.

Affinity studies were conducted by competitive inhibition using [125] iodine-melatonin as a radioligand on CHO-K1 cells containing recombinant human MT ₁ and MT ₂ melatonin receptors. The binding of new compounds has been studied compared to melatonin.

Table 1

Compound, (1 mg / ml) Formula Max IOP reduction, mmHg (t _max.ff , h) Melatonin

2.41 (2)

3.75 (2)

2.88 (5)

4.25

1.52

5.65 (5)

2.00 (2)

2.51 (3)
Normalization of ophthalmotonus after 28 hours.

4.8 (3)

3.06 (2)

1.75 (4 h)

2.12 (3 h)

4.16 (2 h)

4.0 (2 h) Timolol (5 mg / ml) 2.9 (2 h)

Claims (35)

1. Derived indolinone of General formula (I)

or a pharmaceutically acceptable salt thereof, where a means

R1 is H, benzyl, allyl, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkylcarbonyl, C ₆ aryloxycarbonyl, benzoyl, C ₁ -C ₆ alkyloxycarbonyl, C ₁ -C ₆ alkylcarbonyl, m means 1-3, R2 is CN, NHC (O) C ₁ -C ₅ alkyl, NHC (O) C ₃ -C ₆ cycloalkyl, NHC (O) OC ₁ -C ₅ alkyl, NHC (O) C ₁ -C ₅ haloalkyl, COR6, where R6 is OH, OC ₁ -C ₅ alkyl, NH ₂ , NH (C ₁ -C ₅ alkyl), wherein R3 is H, OH, OC ₁ -C ₅ alkyl, C ₁ -C ₅ alkyl (CO) O -, C ₁ -C ₅ alkyl O (CO) O-, C ₁ -C ₅ haloalkyl (CO) O-, or R2 is halogen as the sole substituent, C ₁ -C ₅ alkyl, C ₁ -C ₅ alkoxy, wherein R3 is C ₁ -C ₅ alkoxy, C ₁ -C ₅ alkyl (CO) O-, C ₁ - C ₅ alkyl O (CO) O-, C ₁ -C ₅ haloalkyl (CO) O-, R4 is CN; COR6, where R6 is OH, OC ₁ -C ₅ alkyl; or C (O) N (C ₁ -C ₅ alkyl) ₂ , except when a) R1 = H, R2 = 5-NHAc, R3 = OH, R4 = CN, m = 1, b) R1 = H, Me, R2 = 5-NHAc, R3 = H, R4 = CN, m = 1. 2. The indolinone derivative of formula (I) according to claim 1, used as ligands MT ₁ , MT ₂ and MT ₃ (enzyme quinone reductase 2) subtype of melatonin receptors. 3. An indolinone derivative of formula (I) according to claim 1, having inhibitory activity against MT _{3 of a} subtype of the melatonin receptor (quinone reductase 2 enzyme). 4. The indolinone derivative of formula (I) according to any one of paragraphs. 1, 2 or 3 to obtain a medicinal product for the treatment and prevention of ophthalmic diseases. 5. An indolinone derivative of formula (I) according to claim 1 for use in the treatment of glaucoma. 6. The use of the indolinone derivative of the general formula (I)

or its pharmaceutically acceptable salt as ligands MT ₁ , MT ₂ and MT ₃ (enzyme quinone reductase 2) subtype of melatonin receptors, where a means

R1 is H, benzyl, allyl, C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkylcarbonyl, C ₆ aryloxycarbonyl, benzoyl, C ₁ -C ₆ alkyloxycarbonyl, C ₁ -C ₆ alkylcarbonyl, m means 1-3, R2 is CN, NHC (O) C ₁ -C ₅ alkyl, NHC (O) C ₃ -C ₆ cycloalkyl, NHC (O) OC ₁ -C ₅ alkyl, NHC (O) OC ₃ -C ₆ cycloalkyl, NHC ( O) C ₁ -C ₅ haloalkyl, NHC (O) OC ₁ -C ₅ haloalkyl, COR6, where R6 is OH, OC ₁ -C ₅ alkyl, NH ₂ , NH (C ₁ -C ₅ alkyl) or N (C ₁ -C ₅ alkyl) ₂ , in which R3 means H, OH, OS ₁ -C ₅ alkyl, C ₁ -C ₅ alkyl (CO) O-, C ₁ -C ₅ alkyl O (CO) O-, C ₁ - C ₅ haloalkyl (CO) O-, or R2 is halogen as the sole substituent, C ₁ -C ₅ alkyl, C ₁ -C ₅ alkoxy, wherein R3 is C ₁ -C ₅ alkoxy, C ₁ -C ₅ alkyl (CO) O-, C ₁ - C ₅ alkyl O (CO) O-, C ₁ -C ₅ haloalkyl (CO) O-, R4 is CN; COR6, where R6 is OH, OC ₁ -C ₅ alkyl; or C (O) N (C ₁ -C ₅ alkyl) ₂ , except when a) R1 = H, R2 = 5-NHAc, R3 = OH, R4 = CN, m = 1, b) R1 = H, Me, R2 = 5-NHAc, R3 = H, R4 = CN, m = 1. 7. The use of claim 6 for the manufacture of a medicament for the treatment and prevention of ophthalmic diseases. 8. The use of claim 7, wherein the ophthalmic disease is glaucoma. 9. The use as an active ingredient of the compound of formula (I) according to claim 1 for the preparation of a pharmaceutical composition suitable for the treatment and prevention of an ophthalmic disease. 10. The use of claim 9, wherein the ophthalmic disease is glaucoma. 11. A pharmaceutical composition for the treatment of ophthalmic diseases, containing an effective amount of a compound of formula (I) according to claim 1. 12. The pharmaceutical composition according to claim 11, wherein the ophthalmic disease is glaucoma. 13. The pharmaceutical composition according to claim 11, which is an eye drop.