Method of producing a2a-adenosine receptor agonist and polymorphic modifications thereof

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

 

This application is a partial continuation of patent application U.S. serial No. 11/701699, filed February 2, 2007, which claimed priority from provisional patent application U.S. serial No. 60/801857, filed may 18, 2006, and provisional patent application U.S. serial No. 60/765114, filed February 3, 2006, the complete disclosure of which is incorporated into this description by reference.

The technical field to which the invention relates

The present invention relates to a method of large-scale obtain the agonist receptor And2A-adenosine, and also belongs to a polymorphic modifications of this compound and to methods for selection of specific polymorphic modifications.

The level of technology

Adenosine is a natural nucleoside, which exerts its biological properties through interaction with the family of adenosine receptors, known as A1And2AAnd2Band a3, each of which modulate important physiological processes. One of the biological properties of adenosine is its action as a coronary vasodilator, with the specified property is a result of interaction with the receptor And2Aadenosine. It was found that this effect of adenosine is useful for visualization of the heart when necessary in order to expand the coronary artery before application of the radiopharmaceutical (e.g., thallium 201) and by observation thus obtained images to determine the presence or absence of coronary artery disease. The advantage of this technology lies in the fact that they have avoided the more traditional method of inducing coronary sosudorasshiratmi through exercises on running "track", which is certainly undesirable for the patient with coronary disease.

However, the use of adenosine has several shortcomings. Adenosine has a very short half-life in humans (less than 10 seconds), and has all the properties associated with agonists And1And2AAnd2Band a3the receptors. Thus, the selective agonist of the receptor And2Aadenosine would provide an excellent way to exercise coronary sosudorasshiratmi, especially agonist with a longer half-life and fewer side effects.

The class of compounds having these desirable properties, is disclosed in U.S. patent No. 6403567, the full disclosure of which is hereby incorporated by reference. In particular, it is shown that one connection, disclosed in this patent, (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamic represents you ocollection agonist receptor And 2A-adenosine and is now undergoing clinical trials as a coronary vasodilator, suitable for cardiac imaging.

Given the heightened interest in this and similar compounds, it became desirable discovery of new synthesis methods, which provide a convenient method of obtaining large quantities of substances with good yield and high purity. The patent, which discloses the desired compound (U.S. patent No. 6403567), provides several ways to obtain a connection. However, although these methods are suitable for synthesis on a small scale, all the synthesis methods disclosed in the patent, use protective group, which is undesirable for synthesis on a large scale.

In addition, we discovered that the desired product (that is, (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamic) can exist in at least three different crystalline forms, the most stable of which is a monohydrate. This polymorphic modification is stable under stress conditions relative humidity up to its melting point. Accordingly, it is desirable that the final product produced by the new synthesis, was obtained in the form of a stable monohydrate.

The INVENTION

Thus, the present invention is to provide a convenient synthesis for large-scale production (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate and its polymorphic modifications, preferably in the form of its monohydrate. Accordingly in a first aspect the invention relates to a method for obtaining compounds of formula I:

The formula I

including

contacting the compounds of formula (3):

with methylamine.

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

In the second embodiment, the product is isolated as a pure monohydrate by dissolving the product in a solvent, for example dimethyl sulfoxide, add purified water, filter formed in this way the suspension, washing the contents of the filter with water and then with ethanol and drying solids, which is under vacuum at a temperature not exceeding 40°C.

In the second aspect of the invention relates to a method for connection fo the formula (3):

including

contacting the compounds of formula (2):

with ethyl-2-formyl-3-oxopropionate.

In one embodiment, the reaction is performed in ethanol at a temperature of approximately 80°C., with about 1.1 molar equivalents of ethyl-2-formyl-3-oxopropionate.

In the third aspect of the invention relates to a method for obtaining compounds of formula (2):

including

contacting the compounds of formula (1):

with hydrazine.

You can apply 14,3-16,7-fold molar excess of hydrazine and the reaction can be conducted at a temperature of approximately 60-65°C. In one embodiment, the hydrazine is first heated to approximately 60-65°C, followed by addition of the compounds of formula (1). The compound of the formula (2) can be distinguished by (a) cooling the reaction mixture to about 40°C, (b) adding 4,2-4,9 mass equivalent amount of water at the same time maintaining the temperature at approximately 40°C, (C) cooling the mixture to about 10°C and maintain at this temperature for at least about 1 hour, (d) filtering, (e) washing the contents of the filter with water and then ethanol, and (f) drying the solids, which will is seeking under vacuum at a temperature which does not exceed 30°C, for at least 12 hours.

The above method is suitable for large-scale synthesis of the desired product, which is provided with a good yield, despite the presence in the final product one minor impurities. It is shown that this mixture is immutable intermediate compound of the formula (2); that is, the compound of the formula:

Although this impurity can be removed from the final product by crystallization, it was decided to look for an alternative synthesis, which has all the advantages of the above synthesis, but does not give the compound of the formula (2) as an impurity in the final product.

Thus, in the fourth aspect of the invention relates to a method of 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 by contacting the compounds of formula (4):

with methylamine.

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

In another embodiment, the reaction is carried out at a temperature around the 2.5-to 7.5°C.

In yet another embodiment, the product is isolated as a pure monohydrate by dissolving the product in a solvent, for example dimethyl sulfoxide, add purified water, filtering the thus obtained suspension, washing the contents of the filter with water and then with ethanol and drying solids, which remains under vacuum at a temperature not exceeding 40°C. In some other embodiments, implementation of the product can be identified as pure monohydrate by (a) degassing the reaction mixture under vacuum at not more than 35°C to remove excess methylamine, (b) drop the vacuum and cooling to 0-5°C for approximately from 15 minutes to one hour, (C) filter formed thereby suspensions, (d) washing the contents of the filter with water and then ethanol and (e) drying the solid substance, which remains under vacuum at a temperature not exceeding 40°C.

In some embodiments, the implementation of (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamic additionally cleaned to provide monohydrate forms by (i) dissolving the compound in a solvent such as dimethyl sulfoxide is heated to approximately 78-88°, (ii) filtering any solid impurities from the solution, (iii) washing the additional p is storyteller, (iv) adding the solution to purified water, which is supported at approximately 78-88°C, thus forming a suspension, (v) mixing the suspension, (vi) cooling the suspension, (vii) filtering, (viii) washing the contents of the filter with water and then ethanol, and (ix) drying the solids, which remain under vacuum at a temperature not exceeding 40°C.

In the fifth aspect of the invention relates to a method of synthesis of compounds of formula (4):

comprising contacting compounds of the formula (2):

with excess ethyl-2-formyl-2-oxopropionate, preferably about 2-10-fold molar excess, more preferably approximately 5-10-fold excess. In one embodiment, the reaction is performed in ethanol at a temperature of approximately 80°C. Ethyl-2-formyl-3-oxopropionate present in 5-10-fold excess.

In an additional embodiment, also provides an additional method of synthesis of compounds of formula (4), comprising contacting the compound of formula (2)

with excess ethyl-2-formyl-3-oxopropionate in the presence of acid. The reaction takes place by boiling under reflux, and, as a rule, is carried out in ethanol. The reaction may proceed as in the presence and in Otsu is the result of HCl as catalyst. Apply up to 0.1 molar equivalent, preferably about 0.05 molar excess of HCl and approximately 5-10-fold molar excess, preferably approximately 6,8-7,5-fold molar excess of ethyl-2-formyl-3-oxopropionate. The product of this reaction can be distinguished by (a) cooling the reaction mixture after completion of the reaction to about 10°C, (b) filtering (C) washing the contents of the filter with ethanol and (d) drying the solids, which remains under vacuum at a temperature not exceeding 40°C.

Definition and basic settings

Figure 1 presents1H NMR spectrum monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate (form A).

Figure 2 shows thermal analysis monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate.

Figure 3 shows x-ray diffraction pattern monohydrate (1-{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 diffractogram (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate, form C.

Figure 5 shows x-ray diffractogram (1-{9-[(4S,2R,3R,5R)-3,4-Digue is droxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate, form than form A.

In the context of the present description, the following terms and expressions are generally intended to determine the following values, unless otherwise stated.

“Optional” or “optionally” means that consistently described event or circumstance may or may 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 1, which is sufficient for effective treatment, as defined below, when administered to a mammal in need of such treatment. therapeutically effective amount will vary depending on the subject and conditions of the disease to treat, mass and age of the subject, the severity of the conditions of the disease, the route of administration and the like, which can be easily determined by conventional specialist.

The term “treatment” or “treating” means any treatment of a disease in a mammal, including:

(i) preventing the disease, i.e. an obstacle for the development of clinical symptoms;

(ii) inhibiting the disease, i.e. suspension of clinical symptoms; and/or

(iii) the relief of illness is evania, that is, the regression of clinical symptoms.

As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion medium, coating, antimicrobial and antifungal agents, isotonic and preventing absorption agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except in those cases where any conventional medium or agent is incompatible with the active ingredient, its use in therapeutic compositions is assumed. You can also enter in the composition of the additional active ingredients.

The term “polymorphs” is intended to include amorphous modifications 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.

Discovered that this compound can exist in at least three different crystalline forms referred to in this document as form A, form B, form C and an amorphous product.

Form And: This polymorphic modification 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 and water is, or from a polar solvent, such as water and dimethyl sulfoxide. It is shown that the form is a monohydrate and is the most stable of the various polymorphic modifications at ambient temperatures. It is stable in a relatively wet stress States up to its melting point.

Form: This polymorphic modification get by evaporation under vacuum 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 temperatures. X-ray analysis of the crystals was significantly different from any other polymorphic modifications (see figure 4), but to determine its composition was difficult because the x-ray analysis gave disordered broad peaks and polymorphic modification contained various amounts of water. Found that it is difficult with a high probability to reproduce the receipt of this polymorphic modifications.

Form: This polymorphic modification 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 the crystals was significantly different from any other polymorphic modifications (see IG). It is shown that polymorphic modification With is an unstable hydrate, which at elevated temperatures desolvation in an unstable form.

Amorphous material: This polymorphic modification get through heating polymorphous modifications of Forms And at a temperature up to 200°C. This polymorphic modification is unstable in the presence of atmospheric moisture, forming various hydrates.

Methods of analysis of forms a, b, C, and amorphous material

Powder x-ray diffraction

Analysis by powder x-ray diffraction (XRPD) was performed on a Shimadzu XRD-6000 powder x-ray diffractometer using Cu Kα radiation. The instrument was equipped with an x-ray tube with fine focus and the voltage and current in the tube was set to 40 kV and 40 mA. The divergence and scattering slits were set at 1 and the receiving slit was set at 0.15 mm Diffracted radiation was determined NaI scintillation detector. Used theta-two theta continuous scan at 3°/min (step 0.4 sec/0,02°) from 2.5 to 40°2θ. To check the settings used silicon standard. Data were collected and analyzed using XRD-6000 v. 4.1 software.

Analysis by powder x-ray diffraction (XRPD) also performed using Inel XRG-3000 difr is Chimera, equipped with a CPS (sensitive with a curved position detector 28 intervals of 120°. Calibration was performed using a silicon reference standard. The voltage and current in the tube was set to 40 kV and 30 mA. Monochrome slit was set at 5 mm close to 80 μm. The samples were placed on a holder for samples of aluminum with silicon liner or glass capillaries XRPD-quality. Each capillary was mounted in the goniometer head, which was brought into effect to provide torsional capillary during data collection. Received real-time data using Cu-Kα radiation at a resolution of 0.03°2θ. Typical data collected over a period of 300 seconds. The damages XRPD drawings shows only data points within the interval 2.5 to 40°2θ.

Thermal analysis

Thermogravimetric (TG) analysis was performed on a TA Instruments 2050 or 2950 thermogravimetric analyzer. Calibration standards were Nickel and AlumelTM. The samples were placed in a tray of samples of aluminum, was inserted into the TG furnace and accurately weighed. The samples were heated in nitrogen at a rate of 10°C/min or 300 or 350°C. until otherwise approved, the mass of the samples before analysis balanced at 25°C in the TGA furnace.

Analysis by the method of differential scanning calorimetry (DSC) was performed on a TA Instruments WPPT is erentielles scanning calorimeter 2920. Accurately weighed samples were placed or tortuous pallets or sealed pallets that contain the puncture, to allow for pressure relief. Each sample was heated in nitrogen at a rate of 10°C/min or 300 or 350°C. as the calibration standard used metal indium. The temperature was held in the high jump.

Infrared spectroscopy

Infrared spectra were obtained on Magna 860®Fourier transformed infrared (FT-IR) spectrophotometer (Nicolet Instrument Corp.), equipped with Ever-Glo mid/far IR source, a beam splitter with an extended interval of bromide of potassium and detector of deuterated triglycine sulfate (DTGS). Until otherwise approved, for a sample survey was used Spectra-Tech, Inc. accessory for diffuse reflectance. Each range is 256 jointly added scans at a spectral resolution of 4 cm-1. Obtaining samples for compounds was placed in microcosm and the alignment material using frosted glass slides. The installation of the basic data were obtained using centered reflector in place. The spectra represent the set of data with a single beam sample dataset with background single beam. is librescu wavelength of the instrument was performed using polystyrene.

NMR spectroscopy

1H NMR spectra in solution phase was obtained at ambient temperature on the model of the spectrometer Bruker AM-250, when operating by 5.87 T (Larmor frequency:1N=250 MHz). Data time interval was obtained using the width of the ripple 7.5 PS and time of collection in 1,6834 seconds on the spectral window at 5000 Hz. Collected a total of 16384 data points. Between transition States have used the time delay 5 seconds relaxation. Each set of data typically consisted of 128 together averaged transient States. Spectra were processed using GRAMS132 A1 software, version 6.00. The weakening of the free-induction (FID) was zeroed to four times the number of data points and exponentially multiplied by a factor of line broadening, equal to 0.61 Hz before Fourier transformation. In1N spectra conducted an internal assignment of signals to tetramethylsilane was (0 ppm), which was added as an internal standard.

Alternative NMR analysis was carried out as described in example 4.

Analysis of sorption/desorption of moisture

The data of sorption/desorption of moisture collected on a VTI SCA-100 sorption Analyzer pair. The data of sorption and desorption were collected in the range from 5% to 95% relative humidity (RK) with 10% intervals relative humidity by purging with nitrogen. As the calibration standard is used in the sodium chloride (NaCl) and polyvinylpyrrolidone (PVP). Criteria of equilibrium used for analysis was less than 0,0100% mass change for 5 minutes with a maximum time of equilibrium in 180 minutes, if the criterion of mass has not been reached. Put on the schedule data has not been adjusted for initial moisture content.

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 shown in reaction scheme I. All reactions are usually carried out in a nitrogen atmosphere.

A REACTION SCHEME I

Stage 1 - Getting 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 about 45-55°C. When the reaction is finished, the product of formula (2) was isolated by dilution with proton solvent in which the compound of the formula (2) has limited the NUU solubility, for example ethanol or isopropanol. The mixture is stirred for about 1-5 hours and then filtered. The solid is purified by stirring with water, filtering and washing with water and then with isopropanol and drying under vacuum and take it to the next stage without purification.

Stage 2 - Getting formula (3)

The compound of the formula (2) is then converted into the compound of the formula (3) by reaction with about 1-1,2 molar equivalents of ethyl-2-formyl-3-oxopropionate. The reaction is performed in proton solvent, preferably ethanol, at approximately the boiling temperature under reflux for about 2-4 hours. After cooling to approximately 0°C. the solid is filtered off, washed with cold ethanol and dried under reduced pressure. The product of formula (3) is taken to the next stage without purification.

Stage 3 - the final product

The final product obtained from the compounds of formula (3) by reaction with methylamine, preferably aqueous methylamine. The reaction is carried out at approximately room temperature for approximately 4 hours. The product of formula 1 allocate the traditional way, for example by filtering, washing the solids with cold ethanol and drying under reduced pressure./p>

Obtain the original substance

As starting substances in stage 1 apply (4S,2R,3R,5R)-2-(6-amino-2-globulin-9-yl)-5-(hydroxymethyl)oxolan-3,4-diol. This connection is available in the market.

In as the starting material in stage 2 is used ethyl-2-formyl-3-oxopropionate. It is available on the market, or it can be obtained as shown in reaction scheme II.

A REACTION SCHEME II

Ethyl-3,3-diethoxypropionate 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 distinguish the traditional way, for example by addition of water and extraction of the impurities traditional solvent, for example tert-butylmethylamine ether, acidification of the aqueous phase, for example, hydrochloric acid, followed by extraction with a solvent, such as dichloromethane, and removal of solvent from the dried extract under reduced pressure. Ethyl-2-formyl-3-oxopropionate purified by distillation under reduced pressure.

The preferred method 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 shown in reaction scheme III.

A REACTION SCHEME III

Stage 1 - Getting formula (2)

As before, the compound of the formula (2) is produced from compound of formula (1) by reaction with hydrazine monohydrate in the absence of solvent. However, in this method, 14,3-16,7-fold excess of hydrazine monohydrate is first heated to approximately 60-65°C, followed by addition of the compounds of formula (1). The temperature of the support at around 60-65°C during the interaction, which takes to complete approximately 1 to 3 hours. When the level of residual compounds of formula (1) in the mixture is not more than approximately 0,10%, the reaction mixture is cooled to approximately 40°C. the temperature of the support, while slowly add water. Once added approximately 4,2-4,9 mass equivalence, the mixture is cooled to about 10°C and maintained at this temperature for a period of not less than 1 hour.

Then the product produce by filtration, washed with water and then washed with absolute ethanol. The solid is dried under vacuum at up to 30°C for not less than 12 hours and then take to the next stage without additional purification.

Stage 2 - Getting formula (4)

The compound of the formula (2) is then converted into the compound of the formula (4) by reaction with an excess of ethyl-2-formyl-3-oxoprop is ionata, for example 2-10-fold excess, preferably approximately 5-10-fold, ideally approximately 6,8-7,5-fold excess. The reaction can be conducted under the same conditions as the conditions described for obtaining the compounds of formula (3) in reaction scheme (I).

An alternative to the reaction mixture, you can also add about 0.05 molar equivalents of acid, such as HCl. Reactions provide an opportunity to continue at the boiling point under reflux for about 2-4 hours, until the level of residual compounds of the formula (2) will not be not more than 0.50% and the amount of any of compounds of formula (3), which may be formed, will not amount to no more than 2.5%. After cooling to about 10°C., the solid is filtered off, washed with absolute ethanol and dried under vacuum at up to 40°C to remove residual ethanol. The product of formula (4) is taken to the next stage without purification.

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

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

Accordingly, although the compound of the formula (4) is represented only as a derivative (2E) alkene, the term “compound of formula (4)is intended to include both the case where it is the only (2E) - isomer, and a case where the main part of the product is a (2E) isomer and also present a minor part (2Z) - isomer. The transformation of compounds of formula (4) in the final product by reaction with methylamine as described in stage 3, flows in the same way, if the compound of the formula (4) as (2E) isomer or as a mixture of (2E) - isomer and (2Z) - isomer.

Stage 3 - the final product

The final product obtained from the compounds of formula (4) by reaction with methylamine, preferably aqueous methylamine. The reaction is carried out initially at approximately 0-5°C. for about 8 hours, preferably in the reactor under pressure, followed by raising the temperature to 50-60°C. after approximately 1 hour and maintaining the temperature for 15-30 minutes.

Alternative first methylamine placed in a pressure vessel and cooled to approximately 2.5 and 7.5°C, and then add the compound of the formula (4), while maintain the temperature. The reaction proceeds up until the level of residual compounds of the formula (3) will not be less than approximately 0,10%.

After C is the conclusion of the product emit the traditional way, for example, by degassing under vacuum at not more than approximately 35°C to remove excess methylamine. Then the vacuum is lowered and the mixture is cooled to about 0-5°C. and maintained at temperature for 15 minutes to one hour followed by filtration. The solid, thus obtained, was washed with water and then with ethanol and dried under reduced pressure at a temperature of not more than approximately 40°C.

This method provides a (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 polymorphic modification can further purify by dissolving in dimethyl sulfoxide (DMSO), filtration of the solution of any solid impurities, rinsing additional DMSO and deposition monohydrate from the solution by adding to water. This method is particularly effective when a solution of DMSO and water is heated to approximately 78-88°C and maintained at this temperature with stirring for approximately 1 hour. After stirring, the suspension is slowly cooled to approximately 20°C. Then produce the final product by filtration, washed with purified water and then ethanol, and dried as described previously.

EXAMPLE 1

Getting ethyl-2-formyl-3-oxopropionate

Three or chetyrehosnuju round bottom flask, equipped with a wand magnetic stirrer, a thermocouple, a thermometer with digital display, inlet and outlet for gas and an additional funnel was purged with argon. In the additional funnel was loaded ethyl-3,3-diethoxypropionate (64,5 g) in tetrahydrofuran. In the reaction flask was loaded sodium hydride (of 21.2 g of 60%dispersion) followed by tetrahydrofuran. The contents of the flask was cooled to 0-5°C in a bath with ice was 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. bath was Removed with ice and the contents allowed to warm to ambient temperature. Consumption ethyl-3,3-diethoxypropionate was monitored by TLC analysis. The reaction was suppressed by adding ice water (10,6 about.), was extracted three times with methyl tert-butyl ether (5,4 about. each) and discard the organic layers. The aqueous phase was acidified with 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 over sodium sulfate. The solvent was removed under reduced pressure and the residue was distilled under vacuum to provide ethyl-2-formyl-3-oxopropionate, 27,92 g, yield 70%.

EXAMPLE 2

A.Getting 2-hydrazinophthalazine (2)

Flask, equipped with a mechanical stirrer, an inlet and outlet for gas and thermocouple, was purged with argon. Added hemihydrate 2-chlorobenzene (53,1 g) followed by the addition of hydrazine monohydrate (134 g). The mixture was stirred under heating at 40-45°C for 2 hours. The reaction was monitored by TLC analysis. Upon completion of the reaction, the heat source 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. Solid was transferred into a clean flask, equipped with a mechanical stirrer, was added water (300 ml). The suspension was stirred at room temperature for 18 hours and the solids were isolated by filtration. The filter cake was washed with ice water (300 ml) followed by washing with ice ethanol (300 ml). The solid was dried under reduced pressure to obtain 2-hydrazinophthalazine (41,38 g, 81.4 per cent of the output of 99.3% purity).

CenturyAlternative getting 2-hydrazinophthalazine (2)

The vessel for the reaction containing hydrazinehydrate (258 g, 250 ml)was heated to 40-50°C. To the warm mixture in parts was added to the hemihydrate 2-chlorides is on (100 g), maintaining the temperature between 45-55°C. the Temperature was maintained at this value for two hours and then was added deionized water (500 ml) over a period of 30 minutes, maintaining the temperature at 45-55°C. the Mixture is then gradually cooled to 0-5°C over a period of 3 hours, then stirred at this temperature for an additional 30 minutes. The solid was then filtered and washed with cold (2-5°C) deionised water (200 ml) followed by washing with ethanol (400 ml). The solid was dried under vacuum for 12 hours to obtain 2-hydrazinophthalazine.

C.Alternative getting 2-hydrazinophthalazine (2)

In the vessel for the reaction load hydrazinehydrate (1285 g). The solution is heated to approximately 62°C and add 2-chloramination (500 g) maintaining the temperature of approximately 62°C. the Mixture support at 62°C for at least 2 hours, until the level of residual 2-chlorobenzene in a mixture is equal to not more than 0,10%. The mixture is cooled to approximately 40°C and the mixture was checked to confirm the presence of solids. Slowly add water (2275 g), while maintaining a temperature of approximately 40°C. the Mixture is cooled to about 10°C and maintained for at least 1 hour. The product distinguish filtering, promyvayut (1195) and then absolute ethanol (1885). The product is dried under vacuum at a temperature up to 30°C for at least 12 hours to obtain 2-hydrazinophthalazine.

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-2-formyl-3-oxopropionate (23,93 g to 0.17 mol) were placed in a flask equipped with a mechanical stirrer, an inlet and outlet for gas and reflux. To the flask was added 2-propanol followed by the addition of 2-hydrazinophthalazine (44,45 g, 0.15 mol). The mixture was heated at the boil under reflux with stirring for 2-4 hours, monitoring the reaction by TLC analysis. When the reaction was deemed complete, the heat source was removed and the mixture was cooled to room temperature. The suspension was cooled with stirring in a bath with ice for 1.5-2 hours. The solids were isolated by vacuum filtration and washed with ice-cold 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.

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-Digue is droxy-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, monitoring the reaction by HPLC analysis. The main amount of excess methylamine was removed under reduced pressure and the remaining mixture was cooled to 0°C for 2 hours. The solid was filtered off, washed with ice cold ethanol fortress 200 and was 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-yl)-N-methylcarbamate in the form of its monohydrate.

The structure of the substance was confirmed using1H NMR (see figure 1 and below). Thermal analysis (see figure 2) presented the results coinciding with the presence of one water molecule. Received the XRD pattern (figure 3).

Spectra1The h and13C-NMR was obtained as follows. Two sample obtained above was weighed and dissolved in d6-DMSO - 5.3 mg used for1H-spectra and 20.8 mg used for13With spectra. All spectra were recorded at ambient temperature on the spectrometer JEOL Eclipse+400 c operating frequency of 400 MHz for1The h and 100 MHz for13C.

Designation13With the shift (millions of shares) 1N shift (millionths)Multipletness, splitting (Hz)
2150,5 or 150,3-
4156,4-
4A117,9-
6140,0to 8.41
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
4'85,73,96m
5'61,53,67, 3,57m
5'-OH-5,02D., of 5.7
A140,98,07d, 0,8
B120,2-
Cof 129.68,95d, 0,8
Dof 161.7-
E25,6was 2.76d, 4,6
NH2-to 7.77users
NH-8,35kV, 4,6

EXAMPLE 5

And 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 (4)

A mixture of 2-hydrazinophthalazine (100 g, 0.34 mol), ethyl-2-formyl-3-oxopropionate (242 g, 1.7 mol) and absolute ethanol were loaded into the reactor and the mixture was heated at the boil under reflux for 2 hours. When the reaction was deemed complete, the heat source was removed and the mixture was gradually cooled to 5-10°C. over a period of 3 hours. The suspension was stirred for 30 minutes at this temperature and the mixture was filtered. The solid is washed with cold (5-10°C.) absolute ethanol and then dried under vacuum at a temperature which did not exceed 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.

CenturyAlternative 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 (4)

In the vessel for the reaction load 2-hydrazinophthalazine (450 g) and add absolute ethanol (11376 g), HCl (7.47 g) and ethyl-2-formyl-3-oxopropionate (1557). The mixture is heated at the boil under reflux, and from it draw samples up until the level of residual 2-hydrazinopyridazine in the mixture will not be more than 0.50%, the level of the compounds of formula (3) will not be more than 2.5%. The mixture is slowly cooled to CA is approximately 10°C. The product, a compound of the formula (4), isolated by filtration and washed with absolute ethanol (5121 g). The product is dried under vacuum at 40°C up until the residual ethanol will not be more than 5000 ppm 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 of the product of example 5A gave the following results: C, 48,75%; N, A 4.86%; N, 18,05%; Oh, 27,57. Theoretical values: C, 49,72%; N, 4,74%; N, Of 18.45%; Oh, 27,09. The analysis results are within the limits of experimental error for the hemihydrate of the desired product (48,89%; N, 4,81%; N, 18.1 Percent; Oh, 28,12).

Spectra1The h and13C-NMR was obtained as follows. 20.2 mg of the compounds of formula (4) was dissolved in ~0.75 ml DMSO-d6and spectra were obtained at ambient temperature spectrometer JEOL ECX-400 c operating frequency of 400 MHz for1The h and 100 MHz for13C. Chemical shifts attributed to DMSO solvent, 2,50 MD1N and 39.5 ppm for13C.

RESULTS

Chemical shifts for1H and13Given in table 1. Two isomers in a ratio of ~60/30 was observed in the spectra of1N and13With identified as a major and minor in the table.

Atomand131N chemical shift (millionths)Multipletnessb, splitting (Hz)
21 (primary)192,49,96d, 3,6
21 (minor)187,69,83
22 (minor)167,1--
22 (primary)165,2--
15 (minor)161,8--
15 (primary)of 161.7--
6 (primary)153,1--
6 (minor)152,9--
2 (minor)149,4- -
2 (primary)149,3--
19 (minor)148,0which 9.22d, 13,0
4 (minor)147,9--
4 (primary)147,8--
19 (primary)147,59,26d, 12,4, d, 3,6
8 (primary)144,98,87
8 (minor)144,7cent to 8.85
12143,18,20-8,23m
14 (minor)132,89,20d, ~0,7
14 (primary)to 132.69,12 d, ~0,7
5 (primary)120,7--
5 (minor)120,6--
13of 116.7--
20 (minor)107,2--
20 (primary)106,1--
1' (primary)87,96,07d, 5,3
1' (minor)87,9the 6.06d, 5,3
4'85,8as 4.02kV, a 3.9
2' (minor)74,1to 4.62kV ~5,4
2' (primary)74,1br4.61kV ~5,4
3'70,14,22kV, 4,2
5'61,03,62, to 3.73m
23, 1660,3-60,84,25-4,39m
17, 24a 14.1-14,21,28-1,38m
18 (primary)-12,51d, 12,4
18 (a minor)-11,47d, 13,0
2'-OH (primary)-5,63d, 6,1
2'-OH (minor)-5,62d, 6,1
3'-OH-and 5.30d, 5,1
5'-OH-5,08t, 5,5

what was confirmed, the compound of the formula (4) is a mixture of two isomers:

EXAMPLE 6

A.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)

Aqueous 40% solution of methylamine (1300 ml) was placed in a reactor under pressure, cooled to 0-5°C was added the product of example 5A (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, tracking the completion of the reaction. Upon completion the mixture was heated, keeping the temperature between 50 and 60°C for 1 hour, and then cooled to less than 30°C for 1 hour. When the temperature was below 30°C, the mixture was degirolami, using a 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 at 100-150 mm Hg. The vacuum is then interrupted and replaced with nitrogen, keeping the temperature at 0-5°C for at least 30 minutes. The solid product was then filtered off, 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, obtaining (1-{9-[(4S,2R,3R,5R)-3,-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate in the form of its monohydrate.

CenturyAlternative 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)

Vessel working under pressure, load a solution of 47% methylamine (10080 g). The solution is cooled and added the compound of the formula (4)as obtained in example 5B above (600 g) while maintaining the temperature rate of 5°C. the Mixture is stirred at a temperature around 5°C up until the level of residual compounds of the formula (3) in the mixture shall not be less than 0.10%. The reaction mixture Tegaserod under vacuum at not more than 35°C to remove excess methylamine. The vacuum drop and the mixture is cooled to 2.5°C and incubated for at least 30 minutes the Product is further isolated by filtration, washed with water (not less than 9000 g) and then absolute ethanol (no less than 2964 g). The product is dried under vacuum at temperatures up to 40°C. until the residual ethanol will be not more than 5000 ppm of obtaining the crude (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.

Spectra1The h and13C-NMR was obtained as follows. Two samples of substances obtained in example 6A, was weighed and dissolved in d6-DMSO - 5.3 mg used for1H-spectra and 20.8 mg used for13With spectra. All spectra reg is there when the ambient temperature spectrometer JEOL Eclipse +400 c operating frequency of 400 MHz for1The h and 100 MHz for13C.

Designation13With the shift (millionths)1N shift (millionths)Multipletness, splitting (Hz)
2150,5 or 150,3-
4156,4-
4A117,9-
6140,0to 8.41
7a150,5 or 150,3-
1'86,95,94d, 6,2
2'73,7to 4.62m
2'-OH-5,50 d, 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., of 5.7
A140,98,07d, 0,8
B120,2-
Cof 129.68,95d, 0,8
Dof 161.7-
E25,6was 2.76d, 4,6
NH2- to 7.77users
NH-8,35square, 4,6

Elemental analysis of the product gave the following results: C, 43,96%; N, 4,94%; N, Of 27.94%; Oh, 27,57%. Theoretical values: C, 44,12%; N, 4,94%; N, 27,44%; Oh, 27,09%. The analysis results are within the limits of experimental error for monohydrate.

EXAMPLE 7

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

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 obtained as in example 4 (100 g)in dimethylsulfoxide (300 ml) was filtered through a prefilter 0.6 to 0.8 microns and 0.2 microns filter to remove any solid impurities. The filtrate is then slowly added over 1 hour to deionised water (1 liter) with stirring, and the thus obtained suspension was stirred for at least 1 hour. The solid was filtered, washed with deionized water (2×1 l) and dried under vacuum for at least 1 hour.

The dried product is then re-suspended in deionized water (1.5 l) for at least 2 hours, was filtered, washed with deionized water (1 liter) followed the industry is some absolute ethanol (750 ml). The purified product was dried under vacuum at a temperature not more than 40°C for at least 12 hours from the receipt monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate free from any impurity 2-hydrindantin.

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

The solution monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate (400 g) was dissolved in DMSO (1320 g) and the solution is filtered through a series of successive filters of 0.6-0.8-micron and 0.2-micron. To flush the system filters apply additional DMSO (880 g). The solution is slowly added to purified water (5000 g) while maintaining the temperature level equal to 83°C. the Product begins to crystallize during the addition, and the suspension is stirred at 83°C for approximately 1 hour. The mixture is slowly cooled to 20°C. the Product emit filtered and washed with purified water (8000 g).

Solids are loaded into the vessel and add purified water (6000 g). The suspension is stirred for about 1 hour. The product distinguish filtration and washed with purified water (4000 g) and then absolute ethanol (3160 g). The product is dried under in the cosmology vacuum at temperatures up to 40°C as long until the content of residual water will be not more than 5.5%, and the residual ethanol not more than 2000 ppm with getting monohydrate (1-{9-[(4S,2R,3R,5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-6-aminopurin-2-yl}pyrazole-4-yl)-N-methylcarbamate.

1. 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 contacting the compounds of formula (4):

with aqueous methylamine at a temperature of approximately 2,5-7,5°C.

2. The method according to claim 1, where the reaction is carried out in a sealed reactor under pressure.

3. The method according to claim 2, where the product emit through
(a) degassing under vacuum at a temperature not exceeding 35°C to remove excess methylamine,
(b) flushing the vacuum and cooling to 0-5°C for approximately 15 minutes to an hour,
(c) filtering educated so suspension
(d) washing the contents of the filter with water and then ethanol and
(e) drying the remaining solids under vacuum at a temperature not exceeding 40°C.

4. The method according to claim 3, where 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-methylcarbamic additionally cleaned by
(i) dissolving the dried solids, poluchennogo the stage (e) according to claim 3 in a solvent, such as dimethylsulfoxide,
(ii) filtering any solid impurities from the solution,
(iii) washing with additional solvent, such as dimethylsulfoxide,
(iv) adding the solution to purified water, which is supported at a temperature of approximately 78-88°C with the formation of so suspension
(v) mixing the suspension,
(vi) cooling the suspension,
(vii) filtering
(viii) washing the contents of the filter with water and then ethanol and
(ix) drying the remaining solids under vacuum at a temperature not exceeding 40°C.

5. The method according to claim 4, where the residual water content in the final product is not more than 5.5%, and the residual ethanol is not more than 2000 ppm.

6. The method of obtaining the compounds of formula (4):

including the interaction of the compounds of formula (1)

from 14.3-16,7-fold molar excess of hydrazine hydrate at a temperature of approximately 60-65°C To produce the compounds of formula (2)

followed by contacting the compounds of the formula (2) with excess ethyl-2-formyl-3-oxopropionate optionally in the presence of acid.

7. The method according to claim 6, where the stage of contacting the compounds of formula (2) is carried out in ethanol.

8. The method according to claim 6 where the use of acid, representing the Wallpaper HCl in the amount of up to 0.1 molar equivalent.

9. The method of claim 8, where the reaction is carried out by boiling under reflux.

10. The method according to claim 6, which use approximately 5-10-fold molar excess of ethyl-2-formyl-3-oxopropionate.

11. The method according to claim 10, where the use of approximately 6,8-7,5-fold molar excess of ethyl-2-formyl-3-oxopropionate.

12. The method according to claim 11, where the end product produce by
(a) cooling the mixture obtained after completion of the reaction, up to approximately 10°C,
(b) filtering
(c) washing the contents of the filter with ethanol and
(d) drying the remaining solids under vacuum at a temperature not exceeding 40°C.

13. The method of obtaining the compounds of formula (2):

including
the interaction of the compounds of formula (1)

from 14.3-16,7-fold molar excess of hydrazine hydrate at a temperature of approximately 60-65°C.

14. The method according to item 13, where the hydrazine hydrate are initially heated to a temperature of about 60-65°C. followed by the addition of compounds of formula (1).

15. The method according to item 13, where the compound of the formula (2) was isolated by
(a) cooling the reaction mixture to about 40°C,
(b) adding 4,2-4,9 mass equivalents of water while maintaining a temperature of approximately 40°C,
(c) cooling the mixture to a temperature of approximately 10°C and maintain the Oia such temperature for at least 1 h,
(d) filtering
(e) washing the contents of the filter with water and then ethanol and
(f) drying the remaining solids under vacuum at a temperature not exceeding 30°C, for at least 12 hours



 

Same patents:

FIELD: medicine, pharmacology, bioorganic chemistry, pharmacy.

SUBSTANCE: invention relates to the effective using amount of β-L-2'-deoxynucleoside of the formula (I) or (II) used in manufacturing a medicinal agent used in treatment of hepatitis B, pharmaceutical compositions containing thereof, and methods for treatment of hepatitis B. Proposed agent shows the enhanced effectiveness in treatment of hepatitis B.

EFFECT: enhanced and valuable medicinal properties of agent.

83 cl, 6 tbl, 11 ex

The invention relates to certain oxipurinol the nucleosides, compounds related data oxipurinol the nucleosides, acyl derivatives and compositions that contain at least one of these compounds

The invention relates to novel acyl derivatives of guanosine formula I, inosine formula II, xanthosine formula III, deoxyinosine formula IV, deoxyguanosine formula V, inosine - 2',3'-(acyclic)dialcohol formula VI or pharmaceutically acceptable salts

The invention relates to O6substituted derivatives of guanine, method of their production and to their use for the treatment of tumor cells

The invention relates to a method for obtaining enriched beta-anomer nucleoside of the formula I, where T is fluorine and R is the corresponding nucleoside described in paragraph 1 of the formula

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 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: 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 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: 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.

EFFECT: valuable medicinal properties of compounds.

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

The invention relates to medicine and provides substances that are effective against tumors and viruses, for which conventional anti-tumor agents and antiviral agents exhibit only insufficient effects, and have cancerostatic action and antiviral effects on different tumor immune

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

Up!