Method of producing a2a-adenosine receptor agonist and polymorphs thereof

FIELD: chemistry.

SUBSTANCE: present invention relates to methods for large-scale production of a A2A_adenosine receptor agonist, particularly a monohydrate of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide: . The invention also discloses methods of producing intermediate products used to produce said monohydrate, and directly the monohydrate of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide.

EFFECT: novel methods of producing 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide, which enable to obtain large amounts of the end product with good output and high degree of purity.

15 cl, 6 ex, 5 dwg

 

This application claims the priority of provisional patent application U.S. series No. 60/801857, filed may 18, 2006, and provisional patent application U.S. series No. 60/765114, filed February 3, 2006, the full contents of which are incorporated by the reference.

The scope of the invention

The present invention relates to a method of large-scale obtain agonist A2A-adenosine receptor, and also relates to polymorphs of this compound and methods for selecting a particular polymorph.

The level of technology

Adenosine is found in the nature of a nucleoside, which exerts its biological properties through interaction with the family of adenosine receptors, known as A1, A2A, A2Band A3that regulate important physiological processes. One of the biological effects of adenosine is to act as a coronary vasodilatator; this effect is a consequence of the interaction with A2A-adenosine receptor. It is shown that this property of adenosine applicable as an aid for x - ray and CT of the heart when the coronary arteries expand before administration of contrast medium (for example, Tali 201), and thus, when viewing pictures obtained in this way, you can determine the nalitch is e or absence of coronary artery disease. The advantage of this technology is that it allows you to avoid the most traditional way of inducing coronary dilation of the vessels by means of physical exercise on a treadmill that is extremely undesirable for the patient with coronary disease.

However, the introduction of adenosine has several shortcomings. Adenosine has a very short half-life in humans (less than 10 seconds), and also has all the effects associated with agonists A1, A2A, A2B, and A3the receptors. Thus, the use of selective agonists A2A-adenosine receptor could provide the best way of producing coronary vasodilation, in particular agonist with a longer half-life and fewer side effects or their absence.

The class of compounds which have these desired properties, has been described in U.S. patent No. 6403567, a full description of which is thus included as a reference. In particular, it is shown that the connection described in this patent, (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamic, is a highly selective agonist A2A-adenosine receptor, and is currently undergoing clinical trials as coronary shall vasodilatator, used in cardiac graphy.

Given the increased interest in this and similar compounds, we need to search for new methods of synthesis, which provide a convenient method of obtaining large quantities of substances with a good yield and a high degree of purification. The patent, which describes the desired compound (U.S. patent No. 6403567) provides several ways to obtain this connection. However, although these methods are suitable for the synthesis of small quantities, the synthetic methods described in the patent, the use of protective groups, which are undesirable for large-scale synthesis.

Additionally, it was revealed that the obtained product, which is (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate) exists, at least in three different crystalline forms, the most stable of which is a monohydrate. This polymorph stable under stress conditions relative humidity up to its melting temperature. Thus, it is desirable that the final product created by the new synthesis, was obtained as a stable monohydrate.

The INVENTION

Thus, the objective of this invention is to provide a suitable way to obtain large-scale

Formula I,

including

the interaction of the compounds of formula (3)

with methylamine.

In one embodiment, the method of chemical reaction is carried out in an aqueous solution of methylamine, initially at a temperature of about 0-5°C, followed by warming to about 50-70°C. Alternatively, a chemical reaction carried out as described above, but in a sealed reaction the reactor under pressure.

In the second embodiment, the way the product is isolated in the form of pure monohydrate by dissolving the product in a solvent, such as dimethylsulfoxide, with the addition of pure water, filtering the suspension thus formed, washing the contents of the filter with water, then with ethanol, and drying the solid residue which remains, in vacuum at a temperature not exceeding 40°C.

In the second aspect of the invention relates to compounds of formula (3)

including

the interaction of the compounds of formula (2)

with ethyl-2-formyl-3-oxopropionate.

In one embodiment, the method p is an action carried out in ethanol, at a temperature of approximately 80°C, with about 1.1 molar equivalents of ethyl ester of 2-formyl-3-oxopropionate acid.

In the third aspect of the invention relates to compounds of formula (2)

including

the interaction of the compounds of formula (1)

with hydrazine.

The above synthesis suitable for large-scale synthesis of a desired product, which provides a good yield, although in the final product seen one small admixture. It is shown that this impurity is constant intermediate compound of the formula (2); it is represented by the following structural formula:

Although this impurity can be removed from the final product by crystallization, it was decided to continue the search for alternative synthesis, which would have all the advantages of the above-described synthesis, but do not form compounds of the formula (2) in the form of impurities in the final product.

Thus, in the fourth aspect of the invention relates to a method of obtaining (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate through interaction of the compounds of formula (4)

with methylamine.

In one of the embodiments with the person of the chemical reaction is carried out in an aqueous solution of methylamine, initially at a temperature of about 0-5°C, followed by heating to approximately 50-70°C. Alternatively, a chemical reaction carried out as described above, but in a sealed reactor under pressure.

In the second embodiment, the product is isolated in the form of pure monohydrate by dissolving the product in a solvent, such as dimethylsulfoxide, with the addition of pure water, filtering the suspension thus formed, washing the contents of the filter with water, then with ethanol, and drying the solid residue which remains, in vacuum at a temperature not exceeding 40°C.

In the fifth aspect of the invention relates to a method for obtaining compounds of formula (4)

including the interaction of the compounds of formula (2)

with excess ethyl ester of 2-formyl-3-oxopropionate acid, preferably from about 2-10 fold excess, more preferably 5-10 fold excess.

In one of the embodiments the reaction is carried out in ethanol, at a temperature of approximately 80°C. Ethyl ester of 2-formyl-3-oxopropionate acid is present in 5-10 fold excess.

Definitions and General options

Figure 1 is1H NMR spectrum monohydrate (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxol the h-2-yl]-6-aminopurin-2-yl}-pyrazole-4-yl)-N-methylcarbamate (Form A).

Figure 2 depicts thermal analysis monohydrate (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}-pyrazole-4-yl)-N-methylcarbamate.

Fi shows x-ray diffraction monohydrate (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}-pyrazole-4-yl)-N-methylcarbamate.

Figure 4 shows x-ray diffraction monohydrate (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}-pyrazole-4-yl)-N-methylcarbamate form B.

Figure 5 shows x-ray diffraction monohydrate (l-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}-pyrazole-4-yl)-N-methylcarbamate form C compared with form A.

Used in the present description, the following words and phrases, as a rule, have the meanings which are explained below, except if the context in which they are used indicates otherwise.

"Optional" or "optionally" means that further described event or circumstance may occur or not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur.

The term "therapeutically effective amount" refers to the amount of the compounds of formula I that is sufficient to be effective, as defined below, when making an introduction to a mammal who is in need of this impact. Therapeutically effective amount largely depends on the subject and state the condition being treated, the weight and age of the subject, the severity of the condition of the disease, the route of administration and the like, and can be easily determined by the expert in this field.

The term "therapy" or "therapeutic" means any treatment of a disease in a mammal, including:

(i) preventing the disease, which provides the non-development of clinical symptoms of the disease;

(ii) inhibiting the disease, which provides a delay in the development of clinical symptoms; and/or

(iii) the cure of the disease, which provides the regression of clinical symptoms.

As used herein, "pharmaceutically suitable carrier" includes any and all solvents, dispersants, coatings, antibacterial and antifungal substances, isotonic agents and substances that prevent the absorption and the like, the Use of such media and agents in pharmaceutically active substances is well known in this field. Except, if any suitable substance or environment is incompatible with the active ingredient, it is assumed their use in therapeutic compositions. D. the additional active ingredients may also be included in the composition.

Assume that the term "polymorph" includes non-crystalline forms and solvate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate.

It was found that this compound is stable when the existence of at least three different crystalline forms, referred to herein as Form A, Form B, Form C and amorphous state.

Form A: This polymorph can be obtained by crystallization (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate of proton solvents, such as ethanol or mixtures of ethanol/water or polar solvent, such as dimethylsulfoxide/water. It is shown that the Form is A monohydrate and most stable of the various polymorphs at ambient temperature. It is tolerant to stress conditions relative humidity up to its melting temperature.

Form B: This polymorph get through vacuum evaporation of a solution of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate in triptoreline at ambient temperature. X-ray analysis of these crystals clearly differed from any other polymorphs (see figure 4), but it was difficult to determine its structure is s, since x-ray analysis gave an unordered fuzzy peaks, and polymorph contained various amounts of water. It was found that the receipt of this polymorpha significantly difficult to reproducibly.

Form C: This polymorph get through suspension (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate in acetonitrile over a long period of time at 60°C. X-ray analysis of these crystals clearly differed from any other polymorphs (see figure 5). It is shown that polymorph C is an unstable hydrate, which at elevated temperatures, desolvated in an unstable form.

Amorphous substance: This polymorph get through heating polymorph Form A at temperatures up to 200°C. This polymorph unstable in the presence of atmospheric moisture, forming an unstable hydrate.

Methods of analysis of Forms A, B, C, and Amorphous substance

X-ray diffraction powder

Analysis of x-ray powder diffraction (XRPD) was performed on x-ray powder diffractometer Shimadzu XRD-6000 using Cu Kα radiation. The instrument was equipped with a finely focused x-ray tube, and the voltage and current were set at 40 kV and 40 mA, respectively. The divergence and scattering slits were set H1" and the receiving slit was set at 0.15 mm Diffracted radiation was analyzed by a NaI scintillation detector. Continuous scanning theta-two theta was carried out at 3°/min (0.4 sec/a 0.02° step) from 2.5 to 40° 2θ. To verify instrument settings used silicon standard. Data were collected and analyzed using software XRD-6000 W.

Analysis of x-ray powder diffraction (XRPD) was also performed using diffractometer Inel XRG-3000, equipped with a detector CPS (non-linear position-sensitive) with 28 ranking of 120°. Calibration was performed using a standard silicon sample. The voltage and current in the tube were set at 40 kV and 30 mA, respectively. Monochromatic slit was set at 5 mm by 80 mm. The samples were placed in aluminum sample holder with silicone paste or glass capillaries for XRPD. Each capillary was mounted on the lid of the goniometer, which moved to ensure rotation of the capillaries during data collection. Data in real-time collected using Cu Kα radiation at a resolution of 0.03° 2θ. Typically, data is collected over a period of 300 seconds. Graphics XRPD samples showed only the experimental points in the limit of 2.5 to 40° 2θ.

Thermal analysis

Thermogravimetric (TG) analysis was performed on the equipment TA 2050 or 2950 termor mimeticism analyzer. Calibration standards were Nickel and Alumel™. The samples were placed in an aluminum pan for the samples was inserted into the TG of the heating installation and carefully balanced. The samples were heated in nitrogen at 10°C/min to 300 or 350°C. Unless otherwise stated, the mass of the samples was balanced at 25°C in TGA wind cabinets before analysis.

Analyses of differential scanning calorimetry (DSC) was performed on the equipment TA differential scanning calorimeter 2920. Carefully balanced, the samples were placed in a tight container or in hermetically closed containers that contained pores, in order to reduce the pressure. Each sample was heated in nitrogen at 10°C/min to 300 or 350°C. indium Metal was used as a calibration standard. Temperature recorded in the area of maximum heat transfer.

Infrared spectroscopy

Infrared spectroscopy was performed on an infrared spectrophotometer, Fourier transform (FT-IR) Magna 860® (Nicolet Instrument Corp.), equipped middle/far IR source Ever-Glo, the advanced level of the beam splitter potassium bromide and deuterated tropicanahotel detector (DTGS). Unless otherwise specified, sampling was applied instrument diffuse reflection Spectra-Tech, Inc. (Collector™). Each spectrum consisted of 256 complementary sitiveni the spectral resolution of 4 cm -1. Get sample for the connection consisted of placing the sample in a microcapsule and alignment of the substance on the plane frosted glass. Establishing baseline was performed by adjusting the mirror in position. The spectrum was represented by a single index of the sample to single-beam indicator baseline. Calibration wavelengths in the device carried out with the use of polystyrene.

NMR Spectroscopy

Range phase solution1H NMR was carried out at ambient temperature spectrometer AM-250 model Bruker working at by 5.87 T (operating frequency:1H=250 MHz). The data interval was obtained using pulse width of 7.5 ps and exposure time 1,6834 second spectral window of 5000 Hz. Were collected in total, 16384 experimental points. Reducing the exposure time of 5 seconds was used between the transition States. Each set of data, usually consisted of a 128 average transient States. The spectrum was processed using the software 132 GRAMS Al, version 6,00. The free induction signal (FID) was zeroed four times the number of coordinates data and exponentiale was multiplied by the expansion factor of the spectral line 0,61 Hz before Fourier transformation. Spectra1Η compared with tetramethylsilane was (0 ppm), which was added to the quality of the ve internal standard.

Alternatively, the NMR analysis was performed as described in Example 4.

Analysis of sorption/desorption of water vapor

The data of sorption/desorption of water vapor collected on steam sorption analyzer VTI SGA-100. The data of sorption and desorption were collected in the range from 5% to 95% relative humidity (RK) at 10% RH intervals by injecting nitrogen. As the calibration standards used sodium chloride (NaCl) and polyvinylpyrrolidone (PVP). Criteria of equilibrium used for analysis were less than 0,0100% mass change within 5 minutes, with a maximum time of balance 180 minutes, if the criterion weight did not meet. Image data based upon the initial moisture content have not changed.

Nomenclature

The structure of the compound (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate is the following:

Synthesis of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate

One way for large-scale synthesis of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate represented in reaction scheme I.

A REACTION SCHEME I

One hundred is the s 1 - Obtain formula (2)

The compound of the formula (2) is produced from compound of formula (1) by reaction with hydrazine monohydrate in the absence of solvent. The reaction is carried out at a temperature of approximately 40°C plus/minus 5°C. When the reaction is complete, the product of formula (2) is recovered by mixing with proton solvent in which the compound of the formula (2) has limited solubility, for example, ethanol or isopropanol. The mixture is stirred for about 1-5 hours, and then filtered. The solid residue purified by mixing with water, filtering and washing with water, then with isopropanol and dried under vacuum, and move on to the next stage without purification.

Stage 2 - Getting formula (3)

The compound of the formula (2) is then transferred to the compound of formula (3) through reaction with approximately 1-1,2 molar equivalent of ethyl ester of 2-formyl-3-oxopropionate acid.

The reaction is performed in proton solvent, preferably ethanol, at the boiling point under reflux for about 2-4 hours. After cooling to approximately 0°C. the solid precipitate filtered off, washed with cold ethanol and dried under reduced pressure. The product of formula (3) is transferred to the next stage without purification.

Stage 3 - the final product

End ol the product obtained from the compounds of formula (3) through reaction with methylamine, preferably with aqueous methylamine. The reaction is carried out at room temperature for approximately 4 hours. The product of formula I produce using conventional methods, for example by filtration, washing the precipitate with cold ethanol and drying under reduced pressure.

Obtain the original substance

(4S,2R,3R,5R)-2-(6-amino-2-globulin-9-yl)-5-(hydroxymethyl)oxolan-3,4-diol is used as the starting material in stage 1. This compound is commercially available.

Ethyl ester of 2-formyl-3-oxopropionate acid used as the starting material in stage 2. It is commercially available, or can be obtained as shown in reaction scheme II.

A REACTION SCHEME II

Ethyl ester of 3,3-diethoxypropionate acid interacts with ethyl formate in the presence of a strong base, preferably sodium hydride. The reaction is carried out at approximately 0-5°C for approximately 24 hours. The product produce by conventional means, for example by addition of water and extraction of the impurities in the usual solvents, such as tert-butylmethylamine ether, acidification of the aqueous phase, for example, hydrochloric acid, followed by extraction with a solvent, such as dichloromethane, and evaporation of the solvent from the dried extract at Pont the leaders introduce pressure.

The preferred method of large-scale synthesis of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate represented in reaction scheme III.

Reaction scheme III

Stage 1 - Getting formula (2)

The compound of the formula (2) are obtained from the compounds of formula (1) by reaction with hydrazine monohydrate in the absence of solvent. The reaction is carried out at a temperature of about 45-55°C plus/minus 5°C. When the reaction is over, the product of formula (2) is recovered by mixing with proton solvent in which the compound of the formula (2) has limited solubility, such as ethanol or isopropanol. The mixture is stirred for about 1-5 hours and then filtered. The solid residue purified by mixing with water, filtering and washing with water, then ethanol or isopropanol and drying under vacuum, and move on to the next stage without purification.

Stage 2 - Getting formula (4)

The compound of the formula (2) is then transferred to the compound of formula (4) using the reaction with an excess of ethyl ester of 2-formyl-3-oxopropionate acid, for example 2-10 fold excess, preferably approximately 5-10 fold excess. The reaction is performed in proton solvent, for example ethanol is, at the boiling point under reflux for about 2-4 hours. After cooling to approximately 0°C, the solid precipitate is filtered, washed with cold ethanol and dried under reduced pressure, and the product of formula (4) is transferred to the next stage without purification.

The compound of the formula (4) are depicted as alkinoos derivative (2E), because it is the major isomer formed in this reaction. However, it should be noted that in this reaction may also be formed of a significant number alkinoos derived (2Z), structural formula

named as ethyl(2Z)-3-({9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-oxolan-2-yl]-2-[4-(etoxycarbonyl)pyrazolyl]purin-6-yl}amino)-2-formylpropyl-2-ENOAT.

Thus, although the compound of the formula (4) are presented only as alkenone derivative (2E), it is assumed that the term "compound of formula (4)means that the connection is only isomer (2E) or connection, the bulk of which is the isomer (2E), and there is a small part of the isomer (2Z). The transformation of compounds of formula (4) in the final product by reaction with methylamine as described in stage 3, is carried out in the same way, if the compound of the formula (4) in the form of isomer (2E) or in the form of a mixture Isom the RA (2E) and isomer (2Z).

Stage 3 - the final product

The final product obtained from the compounds of formula (4) through reaction with methylamine, preferably with aqueous methylamine. Initially, the reaction is carried out at approximately 0-5°C for 8 hours, preferably in the reactor under pressure, followed by raising the temperature to 50-60°C for about 1 hour, and maintaining the temperature for 15-30 minutes Product produce using conventional methods, for example by cooling to 0-5°C and leaving a vacuum for approximately 1 hour, thereby removing methylamine. The vacuum is removed, and the remaining contents left at 0-5°C for at least 30 minutes, followed by filtration. The residue, thus obtained, was washed with water, then with ethanol and dried under reduced pressure.

This process provides (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamic in the form of its monohydrate. This polymorph can further purify by dissolving in dimethyl sulfoxide, filtering any solid impurities from the solution and deposition monohydrate from the solution by adding water.

EXAMPLE 1

Obtaining the ethyl ester of 2-formyl-3-oxopropionate acid

Three or chetyrehmetrovuyu rugroden flask, equipped with magnetic stirrer, thermocouple, digital thermometer, inlet and gas outlet and an additional tube was purged with argon. Ethyl ester of 3,3-diethoxypropionate acid (64,5 g) in tetrahydrofuran was loaded into an additional funnel. Sodium hydride (21,2 g 60% colloidal solution) was loaded into the reaction flask followed by tetrahydrofuran. The contents of the flask was cooled to 0-5°C in an ice bath and added ethyl formate (257 g). The mixture was cooled to 0-5°C and the contents of the additional funnel was added dropwise, maintaining an internal temperature less than 5°C. the Ice bath was removed and the contents allowed to warm to ambient temperature. Consumption ethyl ester of 3,3-diethoxypropionate acid was controlled by TLC analysis. The reaction was stopped by adding ice-cold water (10.6 volumes) and was extracted three times with methyl tert-butyl ether (5.4 volume each) and remove the organic layers. The water phase was added concentrated hydrochloric acid to pH 1 to 1.5. The acidified aqueous layer was extracted three times with dichloromethane and the combined organic layers were dried with sodium sulfate. The solvent was removed under reduced pressure, and the residue was distilled in vacuo to obtain ethyl ester of 2-formyl-3-oxopropionate acid, 27,92 g, 70% yield.

EXAMPLE 2

A.Receiving the s 2-hydrazinophthalazine (2)

The reaction flask, equipped with a mechanical stirrer, inlet and gas outlet and a thermocouple, was purged with argon. Added hemihydrate 2-chloroadamantane (53,1 g), followed by the addition of hydrazine monohydrate (134 g). The mixture was stirred under heating to 40-45°C for 2 hours. The course of the reaction was observed using TLC analysis. When the reaction has ended, the heating element was removed and added ethanol (800 ml). The mixture was stirred for 2 hours at ambient temperature, then the precipitate was collected by filtration. The filter cake was washed with ethanol and dried under reduced pressure for 30 minutes. Sediment was transferred to a clean reaction flask equipped with a mechanical stirrer, and added water (300 ml). The suspension was stirred at room temperature for 18 hours and the precipitate was isolated by filtration. The filter cake was washed with ice water (300 ml), then washed ice ethanol (300 ml). The precipitate was dried under reduced pressure to obtain 2-hydrindantin (41,38 g, 81.4 per cent yield, 99.3% of the pure product).

B.Alternative getting 2-hydrazinophthalazine (2)

The reaction vessel containing hydrazine hydrate (258 g, 250 ml)was heated to 40-50°C. To a mixture of warm portions added to the hemihydrate of chlorobenzene (100 g), maintaining the temperature is between 45 and 55°C. The temperature was maintained at this level for two hours and then added deionized water (500 ml) over 30 minutes, maintaining the temperature at 45-55°C. the mixture is Then gradually cooled to 0-5°C for 3 hours, then stirred at this temperature for the next 30 minutes. The precipitate was then filtered and washed with cold (2-5°C) deionized water (200 ml), then washed with ethanol (400 ml). The precipitate was dried under vacuum for 12 hours to obtain 2-hydrindantin.

EXAMPLE 3

Getting ethyl-1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-carboxylate (3)

Ethyl ester of 2-formyl-3-oxopropanoic acid (23,93 g to 0.17 mol) were placed in a reaction flask equipped with a mechanical stirrer, inlet and gas outlet and a reflux condenser. In the reaction flask was added 2-propanol, then added 2-hydrindantin (44,45 g, 0.15 mol). The mixture was heated to boiling point under reflux with stirring for 2-4 hours, while monitoring the progress of the reaction by TLC analysis. When it was decided that the reaction has ended, the heating element was removed and the mixture was cooled to room temperature. The suspension was cooled with stirring in an ice bath for 1.5 to 2 hours. The precipitate was isolated group is a rotary vacuum filtration and washed with ice 2-propanol. The product, ethyl-1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-carboxylate, was dried under reduced pressure to constant weight. Output 54,29 g pure product (by HPLC) 96,6%.

EXAMPLE 4

Obtain 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate

A mixture of ethyl-1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-carboxylate (46.4 g) and methylamine (40% in water, 600 ml) was stirred at ambient temperature for approximately 4 hours, while monitoring the progress of the reaction by TLC analysis. Most of the redundancy of methylamine was removed under reduced pressure and the remaining mixture was cooled at 0°C for 2 hours. The solid was filtered off, washed with ice-ethanol fortress 200, and dried under reduced pressure to obtain 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-carboxamide in the form of its monohydrate, 36,6 g pure product 99,6%.

Structure of the substance was confirmed1H NMR (see figure 1 and below). Thermal analysis (see figure 2) presented the results corresponding to the presence of one water molecule. Received the profiles of x-ray powder diffraction (figure 3).

1H and13C NMR spectra were obtained in the following way. Two sample obtained above was weighed and dissolved in d6-DMSO - 5.3 mg used for1H spectrum, and 20.8 mg used for13C spectrum. All spectra were obtained at ambient temperature spectrometer JEOL Eclipse*400, producing 400 MHz to1H and 100 MHz for13C.

Label13C shift(ppm)lH shift(ppm)Frequency, splitting, Hz(Hz)
2150,5 or 150,3-
4156,4-
4a117,9-
6140,0to 8.41s
7a150,5 or 150,3-
1'86,95,942'73,7to 4.62m
2'-OH-5,50D, 6,2
3'70,54,17m
3'-OH-5,23D, 4,7
4'85,73,96m
5'61,53,67, 3,57m
5'-OH-5,02D, 5,7
A140,98,07D, 0,8
In120,2-
of 129.68,95D, 0,8
Dof 161.7 -
E25,6was 2.76D, 4,6
NH2-to 7.77br s
NH-8,35Q, 4,6

Cleaning monohydrate 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-carboxamide

The solution of the monohydrate of 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-carboxamide (100 g) in dimethyl sulfoxide (300 ml) was filtered through a 0.6 to 0.8 micron prefilter and 0.2 micron pre-filter to remove any solid impurities. The filtered substance is then slowly over a period of 1 hour was added to deionized water (1 liter) with stirring, and thus stirring the suspension for a period of not less than 1 h the Solid precipitate was filtered, washed with deionized water (2×1 l) and dried in vacuum for a period of not less than 1 hour. The dried product is then mixed to obtain a slurry with deionized water (1.5 l) for a period of not less than 2 hours, filtered, and washed with deionized water (1 liter), then washed absolutely the m ethanol (750 ml). The purified product was then dried under vacuum at a temperature not more than 40°C for not less than 12 hours from the receipt of the monohydrate of 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-carboxamide, free from any admixture of 2-hydrindantin.

EXAMPLE 5

Getting ethyl-(2E)-3-({9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-oxolan-2-yl]-2-[4(etoxycarbonyl)pyrazolyl]purine-6-yl}amino)-2-formylpropyl-2-enoate.

A mixture of 2-hydrindantin (100 g, 0.34 mol), ethyl ester of 2-formyl-3-oxopropanoic acid (242 g, 1.7 mol) and absolute ethanol were loaded into the reaction flask, and the mixture was heated to boiling under reflux for 2 hours. When it was decided that the reaction has ended, the heating element was removed and the mixture was gradually cooled to 5-10°C for 3 hours. The suspension was stirred for 30 minutes at this temperature, and the mixture was filtered. The precipitate was washed with cold (5-10°C) absolute ethanol and then dried under vacuum at a temperature not exceeding 40°C to obtain ethyl(2E)-3-({9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-oxolan-2-yl]-2-[4(etoxycarbonyl)pyrazolyl]purine-6-yl}amino)-2-formylpropyl-2-enoate.

Elemental analysis gave the following results: C, 48,75%; H, a 4.86%; N, 18,05%; O, 27,57. In theory the s: C, 49,72%; H, 4,74%; N, OF 18.45%; O, 27,09. Within experimental errors, the analysis corresponds to the hemihydrate of the expected product (C, 48,89%; H, 4,81%; N, 18.1 Percent; O,28,12).

1H and13C NMR spectra were obtained in the following way. 20.2 mg of the compounds of formula (4) was dissolved in ~0.75 ml DMSO-d6, and spectra were obtained at ambient temperature spectrometer JEOL ECX-400 NMR producing 400 MHz to1H and 100 MHz for13C. Chemical shifts were relative to the solvent DMSO, to 2.50 ppm for1H and 39.5 ppm for13C.

RESULTS

Chemical shifts1H and13C are listed in Table 1. Two isomers with a ratio of ~60/30 were found in the range of1H and in the spectrum of13C, marked in the table as main and side.

Atoma13C Chemical shift (ppm)lH Chemical shift (ppm)Frequency, splitting, Hz
21 (primary)192,49,96d, 3,6
21 (side)187,69,83S
22(side) 167,1--
22(primary)165,2--
15 (side)161,8--
15(primary)of 161.7--
6(primary)153,1--
6(side)152,9__
2(side)149,4__
2(primary)149,3--
19(side)148,0which 9.22d, 13,0
4(side)147,9--
4(primary) 147,8--
19(primary)147,59,26d, 12,4, d, 3,6
8(primary)144,98,87s
8 (side)144,7cent to 8.85s
12143,18,20-8,23m
14(side)132,89,20d,~0,7
14(primary)to 132.69,12d,~0,7
5(primary)120,7--
5(side)120,6--
13of 116.7--
20(side) 107,2--
20(primary)106,1--
1' (primary)87,96,07d, 5,3
1' (side)87,9the 6.06d,5,3
4'85,8as 4.02q,a 3.9
2'(side)74,1to 4.62q,~5,4
2'(primary)74,1br4.61q,~5,4
3'70,14,22q,4,2
5'61,03,62, to 3.73m
23, 1660,3-60,84,25-4,39m
17, 241,1-14,2 1,28-1,38m
18(primary)-12,51d, 12,4
18 (side)-11,47d, 13,0
2'-OH (primary)-5,63d, 6,1
2'-OH (side)-5,62d, 6,1

Confirmed that the compound of the formula (4) is a mixture of two isomers:

EXAMPLE 6

Obtain 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate from compound (4)

40% Aqueous solution of methylamine (1300 ml) was placed in a reactor under pressure, cooled to 0-5°C and added the product of Example 5 ethyl-(2E)-3-({9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)-oxolan-2-yl]-2-[4-(etoxycarbonyl)pyrazolyl]purine-6-yl}amino)-2-formylpropyl-2-ENOAT (100 g). The mixture was stirred at 0-5°C for at least 8 hours, monitoring the reaction to completion. When was the reaction is completed, the mixture was heated, keeping the temperature between 50 and 60°C for 1 hour, and then cooled to less than 30°C during the time period of 1 hour. When the temperature was below 30°C, the mixture was degirolami by the pressure of 100-150 mm Hg, allowing the temperature to drop to 0-5°C. the Mixture was stirred at 0-5°C for at least 1 hour, maintaining the pressure of 100-150 mm Hg. Then the vacuum was replaced with nitrogen, keeping the temperature between 0-5°C for at least 30 minutes. Then the solid product was filtered, washed with water (3×500 ml), then with absolute ethanol (625 ml). The product was dried under vacuum, not allowing the temperature to exceed 40°C, to obtain 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate in the form of its monohydrate.

1H and13C NMR spectra were obtained in the following way. Two sample obtained above was weighed and dissolved in d6-DMSO - 5.3 mg used for1H spectrum, and 20.8 mg used for13C spectrum. All spectra were obtained at ambient temperature spectrometer JEOL Eclipse*400, producing 400 MHz to1H and 100 MHz for13C.

Label13C shift (ppm)1H shift(ppm)Frequency,splitting, Hz
2150,5 or 150,3-
4156,4-
4a117,9-
6140,0to 8.41s
7a150,5 or 150,3-
1'86,95,94D, 6,2
2'73,7to 4.62m
2'-OH_5,50D, 6,2
3'70,54,17m
3'-OH-5,23D, 4,7
4'85,7 3,96m
5'61,53,67, 3,57m
5'-OH-5,02D, 5,7
A140,98,07D, 0,8
In120,2-
of 129.68,95D, 0,8
Dof 161.7-
E25,6was 2.76D, 4,6
NH2-to 7.77br s
NH-8,35Q,4,6

Elemental analysis gave the following results: C, 43,96%; H, 4,94%; N, of 27.94. Theoretical: C, 44,12%; H, 4,94%; N, 27,44%; O, 27,09. The analysis corresponds to the monohydrate within their the values of the experiment.

1. The way large-scale obtain monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate

including the interaction of the compounds of formula (3)

with an aqueous solution of methylamine at an initial temperature of about 0-5°C followed by heating to about 50-70°C., and the reaction is carried out in a sealed reactor under pressure.

2. The method according to claim 1, additionally including the selection of the final product (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate in the form of pure monohydrate by:
(a) dissolving the product in a solvent,
(b) adding purified water,
(c) filtering the suspension obtained in this way,
(d) washing the contents of the filter with water followed by rinsing with ethanol and
(e) drying the solid residue which remains under vacuum at a temperature not exceeding 40°C.

3. The method according to claim 2, where the solvent used in stage (a)is dimethyl sulfoxide.

4. The method of obtaining the compounds of formula (3)

including the interaction of the compounds of formula (2)

with about 1.1 molar equivalent of ethyl ester of 2-formyl-oxopropanoic acid in a solvent.

5. The method according to claim 4 where the solvent is ethanol.

6. The method according to claim 5, where the reaction is carried out at a temperature of approximately 80°C.

7. The method of obtaining monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate by reacting the compounds of formula (4)

with an aqueous solution of methylamine at an initial temperature of about 0-5°C followed by heating to about 50-70°C. in a sealed reactor under pressure.

8. The method according to claim 7, additionally including the selection of the final product (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate in the form of pure monohydrate by:
(a) dissolving the product according to claim 7 in a solvent,
(b) adding purified water,
(c) filtering the suspension obtained in this way,
(d) washing the contents of the filter with water followed by rinsing with ethanol and
(e) drying the solid residue which remains under vacuum at a temperature not exceeding 40°C.

9. The method of claim 8, where the solvent used in stage (a)is dimethyl sulfoxide.

10. The method of obtaining the compounds of formula (4)

including the interaction of the compounds of formula (2)

with CA is approximately 2-10-fold molar excess of ethyl ester of 2-formyl-3-oxopropionate acid in a solvent.

11. The method according to claim 10, where the solvent is ethanol.

12. The method according to claim 11, where the reaction is carried out at a temperature of approximately 80°C.

13. The method according to claim 10, where approximately 5-10-fold molar excess of ethyl ester of 2-formyl-3-oxopropionate acid interacts with the compound of the formula (2).

14. Monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate, which is in crystalline form.

15. Monohydrate at 14, where the crystalline form is characterized by the following H NMR spectrum:

73.7
13With the shift (ppm)1H shift (ppm)Frequency, splitting, Hz (Hz)
150.5 or 150.3-
156.4-
117.9-
140.08.41s
150.5 or 150.3-
86.95.94D 6.2
4.62m
-5.50D 6.2
70.54.17m
-5.23D 4.7
85.73.96m
61.53.67, 3.57m
-5.02D 5.7
140.98.07D, 0.8
120.2-
129.68.95D, 0.8
161.7-
25.62.76D 4.6
-7.77br s
-8.35Q 4.6



 

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FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing a monohydrate of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide through contact of a compound of formula

with aqueous methylamine at temperature equal to approximately 2.5-7.5°C. The invention also relates to a method of producing an intermediate compound of formula (4): involving reaction of a compound of formula (1) with 14.3-16.7-fold molar excess hydrazine hydrate at temperature equal to approximately 60-65°C to obtain the corresponding hydrazine of formula (2), followed by contact between the compound of formula (2) and excess ethyl-2-formyl-3-oxopropionate, optionally in the presence of an acid.

EFFECT: method enables to obtain, in a single step, a crystalline compound in form of a monohydrate and also exclude undesirable impurities of the compound of formula 2 in the end product owing to use of intermediate product 4.

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SUBSTANCE: nucleic base (e.g. uracil, cytosine, adenine, guanine, hypoxanthine, xanthine or similar) reacts with perfluoroalkyl halide in the presence of sulphoxide, peroxide and an iron compound to obtain a perfluoroalkyl-substituted nucleic base.

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FIELD: chemistry.

SUBSTANCE: invention relates to phosphoramidite derivatives of general formula where Bx denotes adenine, guanine, cytosine, thymine or uracil, where the amine group of adenine, guanine and cytosine can be optionally protected by a protective group selected from acetyl and phenoxyacetyl; R1 is a substitute of general formula in which R11, R12 and R13 are identical or different, and each denotes hydrogen or alkoxy; R2a and R2b are identical or different, and each denotes alkyl; and WG1, WG2 denote a cyano group. The invention also pertains to a multistep method of producing the said compounds. The invention also relates to intermediate compounds of the said method, namely: an intermediate ether compound of general formula where L is a halogen or a C1-C5alkylthio group; WG1 is a cyano group; an intermediate compound of general formula where Bx denotes adenine, guanine, cytosine, thymine or uracil, where the amine group of adesine, guanine and cytosine can be optionally protected by a protective group selected from an acetyl group and a phenoxyacetyl group; and WG1 denotes a cyano group; an intermediate compound of general formula where Bx is as described above; R1 is a substitute of general formula (2); an intermediate compound of general formula where Bx is as described above; A is a silicon-containing substitute of general formula or where R6 denotes alkyl and WG1 denotes a cyano group. The invention also relates to a method of producing an oligonucleotide of general formula where each B independently denotes adenine, guanine, cytosine, uracil or thymine; each R independently denotes H or hydroxyl and at least one of R denotes hydroxyl; Z denotes H or a phosphate group; and n is an integer between 1 and 100, involving steps A-G, characterised by use of said phosphoramidite derivatives as a monomer compound of nucleic acid at step B.

EFFECT: high yield.

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EFFECT: antilipolytic effect of compounds.

30 cl, 7 dwg, 31 ex

FIELD: chemistry.

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EFFECT: high degree of purity with high output.

1 ex

FIELD: chemistry.

SUBSTANCE: method implies that suspension 2-amino-6-azido-9-(2,3,5-tri-O-acetyl-β-D-ribofuranozile)purine in 60% anhydrous hydrogen fluoride solution of pyridine is diazotizied with tert-butylnitrite during 1 hour at (-18) - (-22)°C. Reaction mixture is decomposed with cut ice. Reaction product is purified by, flash-chromatography on aluminum oxide. Then produced 2-fluorine-6-azido-9-(2,3,5-tri-O-acetyl-β-D-ribofuranozile)purine is hydrogenated at air pressure in 10% acetic acid solution of absolute ethyl acetate with 10% palladium on carbon solution occurrence during 18 hours. Reaction product is purified in acetonitrile solution by flash-chromatography on aluminum oxide at 50-55°C and crystallized from alcohol.

EFFECT: production of compound of high purity with high output.

2 ex

FIELD: chemistry.

SUBSTANCE: invention applied for relates to process of obtaining 2,6- dichlor-9-(2,3,5-tru-O-acetyl-β-D-ribofuranozyl) purine and may be used in organic chemistry and pharmaceutical industry. The process involves conduction of 2,6- dichlor-9-(2,3,5-tru-O-acetyl-β-D-ribofuranozyl) purine with tret-butyl nitrite in the methylene chloride medium at (-18)-(-22)°C during 2 hours in presence of pyridine hydrochloride and phosphorus oxychloride followed by decomposing the reaction mixture with chipped ice, and cleansing of the target product in methylene chloride with flash-chromatography on silica gel.

EFFECT: obtaining of substance with high grade of purity and high output by simplified technology.

1 ex

FIELD: chemistry.

SUBSTANCE: this invention covers method of production of 2-chloroadenosine and may be used in organic chemistry and pharmaceutical industry. The method includes ammonolysis of 2.6-di-chloro-9-(2,3,5-tri-O-acetyl-(β-O-ribofuranozyl)purine in absolute ethyl acetate saturated with ammonia at 0°C during 3 days with further hydrolysis of obtained 5'-0-acetyl-2-chloro-adenosine with 20% ammonia solution in methanol at 20°C during 6 hours, isolation of desired product from the reaction mixture by boiling in mixture of chloroform and methanol, their volumetric ratio 3:1, and purification by crystallization from water.

EFFECT: production of substance with high purity.

1 ex

FIELD: organic chemistry, medicine.

SUBSTANCE: invention relates to compound of the formula (I) wherein each among R represents independently hydrogen atom, (C1-C6)-alkyl, (C3-C7)-cycloalkyl, phenyl or phenyl-(C1-C3)-alkyl; X and X' represent -CH2OH, -CO2R2, -OC(O)R2, -CH2OC(O)R2 or C(O)NR3R4 wherein R2, R3 and R4 represent independently hydrogen atom (H), (C1-C6)-alkyl substituted optionally with one-three (C1-C6)-alkoxy-groups, (C1-C6)-alkylthio-groups, halogen atoms, hydroxy-, amino-, mono-(C1-C6)-alkyl)-amino-, di-(C1-C6)-alkyl)-amino-group; Z and Z' represent independently (C1-C6)-alkyl broken optionally with one-three sulfur atoms (S) or non-peroxide oxygen atom (O), or they absent; n = 1-3; or to its pharmaceutically acceptable salt. Compounds are agonists of adenosine A2A-receptors and can be used for inhibition of inflammatory response or inflammation treatment.

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56 cl, 1 tbl, 21 dwg, 37 ex

The invention relates to nucleoside analogs of formula (1) in which R1represents H or a group protecting the hydroxyl, R2represents H, a group protecting the hydroxyl group of phosphoric acid, a protected group, phosphoric acid or a group of the formula P(R3R4in which R3and R4are the same or different and represent a hydroxyl group, a protected hydroxyl group, alkoxygroup, allylthiourea, cyanoacetylurea, amino group, substituted alkyl group; And represents alkylenes group containing from 1 to 4 carbon atoms, and a represents a substituted purine-9-ilen group or substituted 2-oxopyrimidine-1-ilen group containing at least one Deputy, selected from hydroxyl groups, protected hydroxyl groups, amino groups, protected amino groups, alkyl groups

FIELD: organic chemistry, medicine.

SUBSTANCE: invention relates to compound of the formula (I) wherein each among R represents independently hydrogen atom, (C1-C6)-alkyl, (C3-C7)-cycloalkyl, phenyl or phenyl-(C1-C3)-alkyl; X and X' represent -CH2OH, -CO2R2, -OC(O)R2, -CH2OC(O)R2 or C(O)NR3R4 wherein R2, R3 and R4 represent independently hydrogen atom (H), (C1-C6)-alkyl substituted optionally with one-three (C1-C6)-alkoxy-groups, (C1-C6)-alkylthio-groups, halogen atoms, hydroxy-, amino-, mono-(C1-C6)-alkyl)-amino-, di-(C1-C6)-alkyl)-amino-group; Z and Z' represent independently (C1-C6)-alkyl broken optionally with one-three sulfur atoms (S) or non-peroxide oxygen atom (O), or they absent; n = 1-3; or to its pharmaceutically acceptable salt. Compounds are agonists of adenosine A2A-receptors and can be used for inhibition of inflammatory response or inflammation treatment.

EFFECT: valuable medicinal properties of compounds.

56 cl, 1 tbl, 21 dwg, 37 ex

FIELD: chemistry.

SUBSTANCE: this invention covers method of production of 2-chloroadenosine and may be used in organic chemistry and pharmaceutical industry. The method includes ammonolysis of 2.6-di-chloro-9-(2,3,5-tri-O-acetyl-(β-O-ribofuranozyl)purine in absolute ethyl acetate saturated with ammonia at 0°C during 3 days with further hydrolysis of obtained 5'-0-acetyl-2-chloro-adenosine with 20% ammonia solution in methanol at 20°C during 6 hours, isolation of desired product from the reaction mixture by boiling in mixture of chloroform and methanol, their volumetric ratio 3:1, and purification by crystallization from water.

EFFECT: production of substance with high purity.

1 ex

FIELD: chemistry.

SUBSTANCE: invention applied for relates to process of obtaining 2,6- dichlor-9-(2,3,5-tru-O-acetyl-β-D-ribofuranozyl) purine and may be used in organic chemistry and pharmaceutical industry. The process involves conduction of 2,6- dichlor-9-(2,3,5-tru-O-acetyl-β-D-ribofuranozyl) purine with tret-butyl nitrite in the methylene chloride medium at (-18)-(-22)°C during 2 hours in presence of pyridine hydrochloride and phosphorus oxychloride followed by decomposing the reaction mixture with chipped ice, and cleansing of the target product in methylene chloride with flash-chromatography on silica gel.

EFFECT: obtaining of substance with high grade of purity and high output by simplified technology.

1 ex

FIELD: chemistry.

SUBSTANCE: method implies that suspension 2-amino-6-azido-9-(2,3,5-tri-O-acetyl-β-D-ribofuranozile)purine in 60% anhydrous hydrogen fluoride solution of pyridine is diazotizied with tert-butylnitrite during 1 hour at (-18) - (-22)°C. Reaction mixture is decomposed with cut ice. Reaction product is purified by, flash-chromatography on aluminum oxide. Then produced 2-fluorine-6-azido-9-(2,3,5-tri-O-acetyl-β-D-ribofuranozile)purine is hydrogenated at air pressure in 10% acetic acid solution of absolute ethyl acetate with 10% palladium on carbon solution occurrence during 18 hours. Reaction product is purified in acetonitrile solution by flash-chromatography on aluminum oxide at 50-55°C and crystallized from alcohol.

EFFECT: production of compound of high purity with high output.

2 ex

FIELD: chemistry.

SUBSTANCE: invention pertains to the method of obtaining 2-amino-6-azido-9-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)purine and can be used in organic chemistry and pharmaceutical industry. The method lies in that, 2-amino-6-azido-9-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)purine and sodium azide interact in the presence of the above mentioned tetrametylammonium chloride boiled for 4 hours in absolute acetonitrile. The obtained compound is cleaned by elution of benzol. The residue is dissolved in chloroform and the desired product is separated during precipitation using hexane.

EFFECT: high degree of purity with high output.

1 ex

FIELD: chemistry.

SUBSTANCE: in compound of formula (I): , R1 represents C1-4-alkoxy C3-6cycloalkyl optionally substituted with atom of halogen, hydroxyl, trifluoromethyl, optionally substituted with halogen atom 5-6-member heterocyclyl, in which heteroatoms are selected from oxygen, optionally substituted with halogen atoms phenyl or optionally substituted with halogen atoms 5-6-member heteroaryl, in which heteroatoms are selected from nitrogen and/or sulfur; R2 represents hydrogen or trifluoromethyl; R3 represents hydrogen, optionally substituted with atom of halogen, C3-6cycloalkyl, optionally substituted with atom of halogen, trifluoromethyl, C1-4-alkyl phenyl, optionally substituted with atom of halogen, trifluoromethyl, C1-4-alkoxy heterocyclyl, which has in ring 1-2 heteroatoms, selected from nitrogen, oxygen or sulfur, or optionally substituted with C1-4-alkyl 5-6-member heterocyclyl, which has in ring 1-2 heteroatoms, selected from nitrogen or oxygen, R4 and R5 independently represent hydrogen; X represents covalent bond or lower alkylene; X1 represents covalent bond or lower alkylene, Y represents covalent bond or lower alkylene, optionally substituted with hydroxy or cycloalkyl; and Z represents -C=C-, -R6C=CR7- or -CHR6CHR7-, where R6 and R7 in each position represent hydrogen or lower alkyl.

EFFECT: antilipolytic effect of compounds.

30 cl, 7 dwg, 31 ex

FIELD: chemistry.

SUBSTANCE: invention relates to phosphoramidite derivatives of general formula where Bx denotes adenine, guanine, cytosine, thymine or uracil, where the amine group of adenine, guanine and cytosine can be optionally protected by a protective group selected from acetyl and phenoxyacetyl; R1 is a substitute of general formula in which R11, R12 and R13 are identical or different, and each denotes hydrogen or alkoxy; R2a and R2b are identical or different, and each denotes alkyl; and WG1, WG2 denote a cyano group. The invention also pertains to a multistep method of producing the said compounds. The invention also relates to intermediate compounds of the said method, namely: an intermediate ether compound of general formula where L is a halogen or a C1-C5alkylthio group; WG1 is a cyano group; an intermediate compound of general formula where Bx denotes adenine, guanine, cytosine, thymine or uracil, where the amine group of adesine, guanine and cytosine can be optionally protected by a protective group selected from an acetyl group and a phenoxyacetyl group; and WG1 denotes a cyano group; an intermediate compound of general formula where Bx is as described above; R1 is a substitute of general formula (2); an intermediate compound of general formula where Bx is as described above; A is a silicon-containing substitute of general formula or where R6 denotes alkyl and WG1 denotes a cyano group. The invention also relates to a method of producing an oligonucleotide of general formula where each B independently denotes adenine, guanine, cytosine, uracil or thymine; each R independently denotes H or hydroxyl and at least one of R denotes hydroxyl; Z denotes H or a phosphate group; and n is an integer between 1 and 100, involving steps A-G, characterised by use of said phosphoramidite derivatives as a monomer compound of nucleic acid at step B.

EFFECT: high yield.

7 cl, 1 dwg, 21 ex

FIELD: chemistry.

SUBSTANCE: nucleic base (e.g. uracil, cytosine, adenine, guanine, hypoxanthine, xanthine or similar) reacts with perfluoroalkyl halide in the presence of sulphoxide, peroxide and an iron compound to obtain a perfluoroalkyl-substituted nucleic base.

EFFECT: high cost effectiveness as an intermediate compound for producing medicinal agents.

15 cl, 6 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing a monohydrate of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide through contact of a compound of formula

with aqueous methylamine at temperature equal to approximately 2.5-7.5°C. The invention also relates to a method of producing an intermediate compound of formula (4): involving reaction of a compound of formula (1) with 14.3-16.7-fold molar excess hydrazine hydrate at temperature equal to approximately 60-65°C to obtain the corresponding hydrazine of formula (2), followed by contact between the compound of formula (2) and excess ethyl-2-formyl-3-oxopropionate, optionally in the presence of an acid.

EFFECT: method enables to obtain, in a single step, a crystalline compound in form of a monohydrate and also exclude undesirable impurities of the compound of formula 2 in the end product owing to use of intermediate product 4.

15 cl, 7 ex, 5 dwg

FIELD: chemistry.

SUBSTANCE: present invention relates to methods for large-scale production of a A2A_adenosine receptor agonist, particularly a monohydrate of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide: . The invention also discloses methods of producing intermediate products used to produce said monohydrate, and directly the monohydrate of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide.

EFFECT: novel methods of producing 1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazol-4-yl)-N-methylcarboxamide, which enable to obtain large amounts of the end product with good output and high degree of purity.

15 cl, 6 ex, 5 dwg

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