Products of oxidative decomposition of atorvastatin calcium

FIELD: chemistry.

SUBSTANCE: invention relates to products of oxidative decomposition of atorvastatin calcium, specifically to 4-[6-(4-fluorophenyl)-6-hydroxy-1b-isopropyl-6a-phenyl-1a-phenylcarbamoylhexahydro-1,2-dioxa- 5a-azacyclopropa [a]inden-3-yl]-3-(R)-hydroxybutyric acid, phenylamide 4-(4-fluorophenyl)-2,4-dihydroxy-2-isopropyl-5-phenyl-3,6-dioxabicyclo[3.1.0]hexane-1-carboxylic acid and 4-[1b-(4-fluorophenyl)-6-hydroxy-6-isopropyl-1a-phenyl-6a-phenylcarbamoylhexahydro-1,2-dioxa-5a-azacyclopropa [a]inden-3-yl]-3-(R)-hydroxybutyric acid. The invention also relates synthesis methods thereof, based on oxidation of an atorvastatin salt.

EFFECT: disclosed are products of oxidative decomposition of an atorvastatin salt, which can be used to identify impurities or a product of decomposition of an atorvastatin salt in accordance with approved analytical procedures.

15 cl, 5 tbl, 9 ex

The present invention relates to organic chemistry and to the products of oxidative degradation of atorvastatin calcium, as well as to methods for their preparation. The present invention relates to calcium atorvastatin, which contains almost no products of oxidative degradation, as well as to pharmaceutical compositions containing the specified atorvastatin calcium.

The purity of pharmaceutically active compounds is always an essential factor in ensuring the safety and quality of medicines. As is known in the art, the result of a complex multi-stage process of obtaining pharmaceutically active compounds not only get the desired product, but also impurities, which are related compounds with similar structure. In addition, many pharmaceutically active compounds sensitive to the effects of environmental conditions such as temperature, pH, humidity, light, gases, oxygen, carbon dioxide, reactivity to the environment during processing or storage. These environmental conditions can lead to conversion of pharmaceutically active compounds in the degradation products, which in most cases are less effective than active connection. In addition to a lower effective the particular degradation products also cause adverse side effects, thus, adversely affecting the safety of medicines. Even a very low percentage of impurities or degradation products present in the active compound, can greatly reduce the safety of medicines. In this regard, the great value has the highest possible purity of the input pharmaceutically active compounds. This means that the percentage of the degradation products and impurities present in the pharmaceutically active compound should be the minimum value.

In addition, the pharmaceutical excipients used in pharmaceutical dosage form, also affect the amount of the degradation products and impurities present in pharmaceutically active compounds. Degradation products of pharmaceutical excipients themselves act as reaction centers, initiating the reaction of decomposition of pharmaceutically active compounds in pharmaceutical dosage form.

The sensitivity of various pharmaceutically active compounds to oxidative degradation described in the article K.C. Waterman and others, Stabilization of Pharmaceuticals to Oxidative Degradation, Pharmaceutical Development and Technology, 7, 1, cc.1-32 (2002), which describes possible approaches to the stabilization of pharmaceutically active compounds in terms okoli the nutrient destruction. In the article mentioned above and in other publications discussed the complex mechanism of oxidation in solid pharmaceutical dosage forms, which requires further research, but found that the active compound itself, and more often active compound in pharmaceutical dosage form, can be oxidized. In article S.R. Byrn, etc. (Solid-State Chemistry of Drugs, 2^nded., SSCI, West Lafayette, (1999)) found that atmospheric molecular oxygen interacts with organic crystals and indicated that the reactivity depends on the shape and morphology of the crystals of the active compounds, which determine the permeability of oxygen and its solubility in the crystal lattice, respectively. In some examples, found that reactivity decreases with an increase in the melting temperature, indicating that the inhibition of diffusion of oxygen in compounds with high energy of the crystal lattice.

Up to the present time to prevent or reduce oxidation of the active compound in the pharmaceutical composition of different approaches are used, such as, for example:

1. increasing the concentration of active compound in the pharmaceutical composition if the oxidation caused by the presence of peroxide and metallic impurities in excipients,

2. adding heh atnah agents (for example, citric acid, EDTU, fumaric acid and maleic acid) to remove the metal impurities present in the excipients,

3. the use of pharmaceutical excipients high purity,

4. the use of alternative pharmaceutical excipients or decrease in the quantity of excipients in the pharmaceutical composition, especially if the excipients are caused by oxidation due to their content of impurities peroxides,

5. the use of antioxidants that can prevent or reduce the formation of peroxides in the pharmaceutical composition, however, these antioxidants at the same time not reduce the level already present peroxides. Some well-known antioxidants include:

the initiators of the open circuit (as, for example, thiols and phenols),

- reducing agents with a higher oxidation rate compared to the active compound and thus absorb oxygen present (for example, sulfites and ascorbic acid), and using a combination observed a synergistic effect, for example, the combination of palmitate ascorbic acid and tocopherol),

connection - sinks of peroxides, which decompose peroxides (for example, Fe²⁺) by the method of Fenton, but their use is limited, as is shown techniques, the formation of free hydroxyl radicals, which then initiates reactions involving free radicals and, thus, degradation of active connections

- cyclodextrins, which screens the site of active compounds sensitive to oxidation (K.C. Waterman and others, Stabilization of Pharmaceuticals to Oxidative Degradation, Pharmaceutical Development and Technology, 7, 1, cc.1-32 (2002)).

However, when using particular active compounds cannot provide optimal conditions, and also in modern literature there are only few articles about stabilizing compounds (K.C. Waterman and others, Stabilization of Pharmaceuticals to Oxidative Degradation, Pharmaceutical Development and Technology, 7, 1, s-32 (2002)).

Calcium atorvastatin, poluchalsja Sol (R-(R*,R*))-2-(4-forfinal)-β,δ-dihydroxy-5-(1-methylethyl)-3-phenyl-4-((phenylamino)carbonyl)-1H-pyrrol-1-heptane acid, a known inhibitor of the reductase 3-hydroxy-3-methylglutarylcoenzyme A (HMG-CoA). The specified connection first described in U.S. patent No. 5273995. Ways to get calcium atorvastatin and main intermediate compounds described in U.S. patents№5003080, 5097045, 5103024, 5124482, 5149837, 5155251, 5216174, 5245047, 5248793, 5280126, 5342952 and 5397792.

It is known that inhibitors of HMG-CoA reductase are pharmaceutically active compounds that are sensitive to environmental conditions, pH, humidity, light, temperature, carbon dioxide and oxygen. These compounds are effective t is repitions active compounds designed to ensure dyslipidemias and cardiovascular disease selected from the group comprising dyslipidemia, hyperlipidemia, hypercholesterolemia, atherosclerosis, arteriosclerosis, coronary artery disease, coronary heart disease, etc. associated with the metabolism of lipids and cholesterol. The mechanism of action latinovich compounds is the inhibition of the biosynthesis of cholesterol and other sterols in the liver of humans or animals. These compounds are competitive inhibitors of HMG-CoA reductase or reductase 3-hydroxy-3-methylglutarylcoenzyme A, the enzyme that catalyzes the conversion of HMG-CoA in the mevalonata in the liver of a person or animal, with the specified transformation is an important stage of the biosynthesis of cholesterol in the liver. It is now established that in addition to the above mentioned therapeutic effects statins also have therapeutic actions of another type and, accordingly, are suitable for the treatment of diseases, pathological conditions and disorders that are selected from the group including vascular disorders, inflammatory disease, allergic disease, neurodegenerative disease, malignant disease, viral disease (application WO 0158443), the abnormal state of the bone tissue (application WO 0137876), violations of producing β-am is toidnogo protein precursor, such as Alzheimer's disease or down's syndrome (application WO 0132161).

In modern literature there are practically no data on the reduction of the content of oxidation products in the preparation of atorvastatin and about the identification of the degradation products calcium atorvastatin. Preventing oxidation of the calcium atorvastatin through the process of getting in an inert atmosphere and packaging in an inert atmosphere as described in the application for the grant of a patent Slovenia SI P-200200244. The structure of one of the products of the decomposition of calcium atorvastatin, phenylamide 3-(4-perbenzoic)-2-isobutyryl-3-phenyloxirane-2-carboxylic acid and its formation during photodegradation described in article Hurley .R. and others, Tetrahedron, 49, cc.1979-1984 (1993).

Since the active compounds with a high degree of purification is of great importance, there is a need to identify each impurity or product degradation present in the active compound and/or pharmaceutical compositions. The identification of each impurity or product degradation present in the active compound or pharmaceutical composition is important in respect of such active compounds, for which the sensitivity level of the specified analytical method (for example, IHVR) in determining impurities and/or product degradation differs from the level of justicel the property when determining the active connection. Namely, in such cases, the active compound is considered as pharmaceutically acceptable according to current requirements, although the actual level of impurities or degradation products is outside of the allowed values.

In the present invention according to the existing in the art having offered product calcium atorvastatin with a high degree of purification, characterized by low content of products of oxidative degradation, easiest way to gain and high output due to the identification of three oxidative degradation products present in the calcium atorvastatin and/or containing pharmaceutical compositions.

One object of the present invention relates to new compounds that are products of oxidative degradation calcium atorvastatin the following formulas:

a) compound of formula I

4-[6-(4-forfinal)-6-hydroxy-1-isopropyl-6A-phenyl-1A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxipropionic acid (abbreviated ATV-ceclor),

b) the compound of formula II

phenylamide 4-(4-forfinal)-2,4-dihydroxy-2-isopropyl-5-phenyl-3,6-dioxabicyclo[3.1.0]hexane-1-carboxylic acid (abbreviated ATV-epository),

the connection of the formula III

4-[1b-(4-forfinal)-6-hydroxy-6-isopropyl-1A-phenyl-6A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxipropionic acid (abbreviated ATV-F).

The product of oxidative degradation of calcium atorvastatin described in article Hurley T.R. and others, Tetrahedron, 49, cc.1979-1984 (1993), is characterized by the following formula IV:

,

phenylamide 3-(4-perbenzoic)-2-isobutyryl-3-phenyloxirane-2-carboxylic acid (abbreviated ATV-epoxide).

Oxidative destruction calcium atorvastatin presented in the following diagram:

Compounds of formulas I, II, III and IV of the present invention are present in the drug atorvastatin calcium as a product of oxidative degradation, in this regard, it is important to reduce their number to the minimum value according to the analysis. Such impurities are toxic or in some way have a negative impact on the patient. Due to these reasons it is important to reduce the content of such impurities in the connection to a minimum amount. On the other hand, has a value of detection accuracy level of these impurities and their content should be quantified, for example, using the standards (connected to the I known chemical structure and known analysis method).

The present invention relates to new methods of making compounds of formulas I, II, III and IV of the present invention.

The new compounds of the present invention is produced by oxidation of a solid atorvastatin in salt form (for example, in the form of salts of calcium, sodium, potassium, magnesium or ammonium) in an atmosphere of air or oxygen at elevated temperatures, for example from 40 to 90°C. the Duration of the reaction is from 1 to several days. The oxidation is carried out in a solution of salt of atorvastatin in water, and/or in an organic solvent, and/or mixtures of solvents, such as, for example, acetonitrile, methanol, ethanol, propanol, dichloromethane or methylene chloride, adding hydrogen peroxide or by passing air or oxygen through the solution at a temperature of from about 40 to 90°C. the Solid salt of atorvastatin get by any known method.

The new compounds of the present invention is also obtained by photooxidation of atorvastatin in salt form (for example, in the form of salts of calcium, sodium, potassium, magnesium, ammonium), irradiating the solution of salt of atorvastatin sunlight or artificial sunlight. Salt of atorvastatin get by any known method.

The new compounds of the present invention obtained by the methods described above produce preparative normal-phase or treatment is but-phase chromatography.

In preparative normal-phase chromatography as a stationary phase using silica gel for chromatography or phase on the basis of modified silica gel, for example, contain the following groups: aminopropyl, cyanopropyl, diol or nitrophenyl. The mobile phase contains a mixture of polar alcohol modifier, for example, methanol, ethanol, propanol or acetonitrile, and a non-polar solvent, for example, hexane, dichloromethane, methylcyclohexane, or a combination of more than two of the above solvents.

In preparative reversed-phase chromatography using silica gel containing groups of octadecylsilane or octylsilane. The mobile phase contains a mixture of water with organic or inorganic buffer solution at a concentration in the range from 5 mm to 100 mm and in the range of pH from 2 to 8 in a mixture with one or more organic modifiers selected from alcohols, such as methanol, ethanol and propanol, or acetonitrile.

When allocating new compounds according to the present invention using one or more chromatographic steps. The solvents used in the stages chromatography removed by evaporation and/or lyophilization.

The structure of the new compounds of the present invention, the received and selected by the methods described above is determined by mass spectrometry, and I the cluster magnetic resonance. Ways of identifying patterns and their results are shown in the following examples.

When developing a method for producing stable compositions of calcium atorvastatin established that calcium atorvastatin undergoes degradation upon contact with air or with oxygen. It has been unexpectedly found that the use of various antioxidants, such as, for example, bottled hydroxyanisol, bottled hydroxytoluene, fumaric acid, propylgallate, sodium sulfite, sodium metabisulfite, sodium ascorbate, is not observed to prevent and reduce the formation of oxidative degradation products. Unexpectedly, it was also found that by reducing the oxygen content in the atmosphere can significantly reduce the formation of oxidative degradation products calcium atorvastatin drug calcium atorvastatin or in compositions containing calcium atorvastatin. The observed reduction in the number of products of oxidative degradation is directly proportional to the decrease in oxygen content in the ambient atmosphere. The aforementioned decrease in oxygen content is achieved by replacement of oxygen by an inert gas such as nitrogen or argon, or by reducing the pressure of the atmosphere surrounding the calcium atorvastatin.

Apart from the process of obtaining calcium atorvast is Tina in an inert atmosphere and store it in an inert atmosphere, it is important to record the number of products of oxidative degradation, present in the drug atorvastatin calcium and containing pharmaceutical compositions. To determine the number of unwanted compounds and for accurate quantitative analysis is necessary to obtain the standards of these compounds (i.e., use of compounds of known chemical structure and the appropriate method of analysis). The availability of standards is of particular importance in cases where the sensitivity level for example, analysis GHUR, in the identification of impurities and/or product degradation differs from the level of sensitivity when identifying active compounds. For determination of impurities in pharmaceutically active compounds and pharmaceutical compositions generally used GHUR.

Unexpectedly found that some compounds present in the drug atorvastatin calcium, characterized by a level of sensitivity that is different from the sensitivity level of the calcium atorvastatin, if the detection method GHUR carried out at 250 nm. Namely, the sensitivity level for the new compounds according to the present invention of the formula I is 0,41 for a new connection according to the present invention of formula II level of sensitivity is to 0.72 for a new connection according to the present invention of the formula III, the sensitivity level is 0.48, and the new is th compounds of the present invention of the formula IV level of sensitivity is 1.20 unlike the calcium atorvastatin.

The content of impurities in the active compound and/or pharmaceutical composition is an important factor in the safety of the drug, therefore, should be kept to a minimum content of impurities. Specified is of paramount importance for the degradation products, because their contents in medicine increases during storage of the medicinal product.

Another object of the present invention serves calcium atorvastatin, almost not containing products of oxidative degradation, and pharmaceutical compositions containing the specified atorvastatin calcium, and at least one pharmaceutically acceptable excipient.

In the present invention serves calcium atorvastatin high degree of purification, in which the content of the products of oxidative degradation is less than approximately to 0.29 wt.%.

In the present invention serves calcium atorvastatin high degree of purification, in which the content of ATV-ceclor is less than about 0.09 wt.%.

In the present invention serves calcium atorvastatin high degree of purification, in which the content of ATV-epoxybutane is less than about 0.05 wt.%.

In the present invention serves calcium atorvastatin high degree of purification, in which the content of ATV-F with what is less than about 0.09 wt.%.

In the present invention serves calcium atorvastatin high degree of purification, in which the content of ATV-epoksidine is less than about 0.06 wt.%.

Table 1 below shows the quantity of each product of oxidative degradation present in the drug atorvastatin calcium, depending on a variety of atmospheric conditions in which the conduct of its receipt.

If calcium atorvastatin receive or store in air at room temperature, the formed products of oxidative degradation, which are not formed during the storage of calcium atorvastatin in nitrogen atmosphere.

When comparing the data of table 1 (the sensitivity level for the products of oxidative degradation is 1.0) and table 2 (different levels of sensitivity) is set, the content of these products is significantly different. If the level of sensitivity of 1.00 in the absence of standards of impurities, the measured values for the products of oxidative degradation in the atorvastatin calcium obtained in air or in nitrogen atmosphere, the following values, defined using the more accurate sensitivity levels. The content of oxidative degradation products defined at the level of sensitivity of 1.00, also below the limit of 0.10%, which corresponds to the standard requirements adopted in the pharmaceutical industry. Moreover, when the sensitivity level is above 1.00 contents ATV-epoksidine higher compared with the contents specified at a more precise level of sensitivity.

All the following tests conducted under the following sensitivity levels: 0,41 for ATV-ceclor, 0,72 for ATV-epoxybutane, 0,48 for ATV-F and 1.20 for ATV-epoksidine.

The present invention features a pharmaceutical composition that contains almost pure calcium atorvastatin, which contains less than about 0.6 wt.% products of oxidative degradation and at least one pharmaceutically acceptable excipient.

The present invention features a pharmaceutical composition that contains almost pure calcium atorvastatin, which contains less than about 0.2 wt.% ATV-ceclor and at least one pharmaceutically acceptable excipient.

The present invention features a pharmaceutical composition that contains almost pure calcium atorvastatin, which contains less than about 0.1 wt.% ATV-epoxybutane and at least one pharmaceutically acceptable excipient.

The present invention features the pharmaceutical is Skye composition, containing almost pure calcium atorvastatin, which contains less than about 0.2 wt.% ATV-F and at least one pharmaceutically acceptable excipient.

The present invention features a pharmaceutical composition that contains almost pure calcium atorvastatin, which contains less than about 0.1 wt.% ATV-epoksidine and at least one pharmaceutically acceptable excipient.

The results are shown in tables 3 and 4 suggests that the storage of calcium atorvastatin or a pharmaceutical composition containing atorvastatin calcium in tablet form in air at room temperature within 24 months of the contents of the products of oxidative degradation increases significantly. This effect can be excluded when storing calcium atorvastatin in nitrogen atmosphere.

The pharmaceutical composition of the present invention is administered to the mammal in a pharmaceutical form. Dosage form contains almost pure calcium atorvastatin according to the present invention and at least one pharmaceutically acceptable excipient selected from the group comprising diluents, binders, dezintegriruetsja substances, oil, glidant, flavors, sweeteners, preservatives, dyes, and other excipients used in the preparation of pharmaceutical compositions. The pharmaceutical composition of the present invention is processed in any dosage form, used in the pharmaceutical industry, such as, for example, tablets, dispersible compositions for oral administration, capsules, pellets, granules, etc. In an inert g is for you to use nitrogen or argon for creating inert atmosphere. The pharmaceutical composition can be stored in an inert atmosphere in a blister aluminium/aluminium blister with a coating of homopolymer Al-polychlor-3-veratile/PVC or in bottles.

The pharmaceutical composition of the present invention can be used for the treatment of hypercholesterolemia and hyperlipidemia.

The present invention is illustrated by the following examples without limiting its scope.

Example 1

Receipt and allocation of compounds ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide

Calcium atorvastatin (5 g) was stored in a tightly closed container with a volume of 200 ml in the atmosphere of oxygen at 80°C for 30 days. Thus obtained sample was dissolved in a mixture of 50% acetonitrile/water (vol./about.) and was isolated by preparative chromatography.

Preparative chromatography

Recovery of the products of oxidative degradation was performed by reversed-phase chromatography. To obtain purified compounds was performed two phase chromatography with different mobile phases.

In the first stage separation was performed using a chromatograph for preparative GHUR with column Luna prep 18(2), 10 μm (200 mm×50 mm) and a UV detector at a wavelength of 250 nm. As the two mobile phases used systems a and b: 10 mm solution of ammonium acetate (pH 4.5) and a mixture of 95% acetonitrile/5% tetrahydrofuran (the b./about.) respectively. The flow rate was 140 ml/min, the Elution was performed using the following gradient:

Received four factions, the pH of the first and second fractions were brought to 8-9 potassium hydroxide solution (1 M), pH of the third and fourth fractions brought up to 2-3 with hydrochloric acid (1 M). Fractions were evaporated under reduced pressure. The temperature of the water bath is maintained below 30°C. and the reflux condenser was cooled with water at a temperature of 0°C.

To obtain purified compounds all four fractions were subjected to additional purification.

Purification of the first fraction

The first fraction was purified in the same conditions that were used in the first stage, except the mobile phase a, which used a 10 mm solution of bicarbonate of shumilkin) is. The elution was performed in the following gradient:

Received the first fraction, the pH was brought to 8-9 potassium hydroxide solution (1 M). The fraction was evaporated under reduced pressure in the same way as described for the first stage of separation.

After drying the concentrated fractions were received cleared connection ATV-ceclor, 4-[6-(4-forfinal)-6-hydroxy-1-isopropyl-6A-phenyl-1A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxybutiric acid (170 mg). Chromatographic purity was 97.2 per cent.

Clearing the second faction

The second fraction was purified in the same conditions that were used in the first stage, except that as the mobile phase And used a mixture of 70% 10 mm phosphate buffer solution (pH 7.0)/25% acetonic the Il/5% tetrahydrofuran (about./about./about.) and the following gradient:

Received the first fraction and evaporated under reduced pressure. The concentrated fraction was applied to a column for reversed-phase chromatography, buffer substances washed with water and connect the ATV-F was suirable from the column with a mixture of 80% acetonitrile/20% water (vol./vol.).

After drying the concentrated fractions were received cleared connection ATV-F, 4-[1b-(4-forfinal)-6-hydroxy-6-isopropyl-1A-phenyl-6A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxybutiric acid (185 mg). Chromatographic purity was 97,5%.

Clearing the third faction

The third fraction was purified in the same conditions that were used in the first stage, except that as the mobile phase And used 5 mm Sol the strong acid and the following gradient:

Received the first fraction and evaporated under reduced pressure in the same way as described in the first stage.

After drying the concentrated fractions were received cleared connection ATV-epository, phenylamide 4-(4-forfinal)-2,4-dihydroxy-2-isopropyl-5-phenyl-3,6-dioxabicyclo[3.1.0]hexane-1-carboxylic acid (205 mg). Chromatographic purity was 93.6%.

Cleaning fourth faction

The fourth fraction was purified in the same conditions that were used for the purification of a third faction, except that he used the following gradient:

Received the first fraction and evaporated under reduced pressure in the same way as described in the first stage.

After drying the concentrated fractions were received cleared connection ATV-epoxide, phenylamide 3-(4-perbenzoic)-2-isobutyryl-3-phenyloxirane-2-carboxylic acid (50 mg). Chromatographic purity was 96.2%.

Structure determination of compounds ATV-ceclor

Mass spectrometry

Conditions

Mass high resolution spectra were obtained using a quadrupole time-of-flight mass spectrometer Micromass Q-TOF Ultima Global. Used the ionization electrospray. The source temperature was set equal to 100°C., the temperature of the desolvatation equal to 200°C., the velocity of the gas in the spray was 0 l/h, and the drying gas is 200 l/h Used TOF analyzer W-shaped configuration. The instrument was calibrated using clusters of Na formate. The sample was dissolved in 50% solution of 5 mm ammonium acetate/acetonitrile (about./about.) and served in a mass spectrometer with a constant flow rate of 10 ál/min, the Concentration of the sample solution was 0.05 mg/ml

As an internal standard for high resolution used calcium atorvastatin. In the solution of sample was added to the internal standard, the concentration of 0.01 mg/ml

Registered protonated molecular ion 591,2507 m/z. The calculated elemental composition of C₃₃H₃₆N₂O₇F. the Deviation of calculated values from the measured mass was 0.5 MDA. Compared to the atorvastatin calcium in connection ATV-ceclor identified two additional oxygen atom.

Spectroscopy nuclear magnetic resonance

Conditions

¹H and¹³With NMR spectroscopy was performed using Varian instrument or UNITY INOVA 300 at 300 MHz. The INOVA instrument equipped with a gradient cell pulsed fields for inverse detection.¹H and¹³The NMR spectra were recorded at room temperature.

The samples were dissolved in methanol, chloroform or methanol/chloroform 2:1.

Chemical shifts in ppm million was determined relative to the residual signal of the solvent.

Solvent: CD₃OD (¹H and¹³With NMR-spectra)

Structure:

Structure determination of compounds ATV-epository

Mass spectrometry

Conditions

Mass high resolution spectra were recorded under similar conditions as described for compounds ATV-ceclor.

In the mass spectrum observed adducts of the molecular ion with sodium, 472,1536 m/z, and potassium, 488,1270, in the first case, the calculated elemental composition of C₂₆H₂₄NO₅FNa (the deviation of the calculated mass from the measured values constituted 0,0 MDA), and in the second case, C₂₆H₂₄NO₅FK (deviation of the calculated mass from the measured values was 0.5 MDA).

Protonated molecular ion is not observed due to the rapid removal of water from the molecule (M+H-H_{2 O)⁺=432,1606 m/z, the expected elemental composition of C26H23NO4F. the Deviation of the calculated mass from the measured values was 0.5 MDA. Specified fragmented ion also forms adducts with sodium and potassium.}

Adduct of two molecules of compounds ATV-epoxybutane with sodium observed at 921,3131 m/z, the expected elemental composition of C₅₂H₄₈N₂O₁₀F₂Na. The deviation of the calculated mass from the measured value was 4.4 MDA.

Spectroscopy nuclear magnetic resonance

Conditions

¹H and¹³With NMR-spectra were recorded as described for compounds ATV-ceclor.

Solvent: CDCl₃(¹H NMR)

mix CD₃OD/CDCl₃, 2:1 (¹³With NMR)

Structure:

Structure determination of compounds ATV-F

Mass spectrometry

Conditions

Mass high resolution spectra were recorded as described for compounds ATV-ceclor.

Protonated molecular ion is observed at 591,2507 m/z. The intensity of the molecular ion is much lower than the molecular ion ATV-ceclor. The most intense ion in the MS-spectrum is observed when 573,2406 m/z, which is formed by removing water molecules, the calculated elemental composition for 591,2507 m/z - C₃₃H₃₆N₂O₇F. the Deviation of the calculated mass from the measured values is 1.4 MDA. In the MS-spectrum of compound, there are two additional oxygen atom compared with the atorvastatin calcium.

Spectroscopy of aderno the magnetic resonance

Conditions

¹H and¹³With NMR-spectra were recorded as described for compounds ATV-ceclor.

Solvent: CD₃OD (¹H and¹³With NMR)

Structure:

Structure determination of compounds ATV-epoxide

Mass spectrometry

Conditions

Mass high resolution spectra were recorded as described for compounds ATV-ceclor.

In spectroanalyzer protonated molecular ion 432,1612 m/z, the calculated elemental composition of C₂₆H₂₃NO₄F. the Deviation of the calculated mass from the measured values of 0.1 MDA.

Table 5 shows the spectra MS/MS protonated molecular ion.

Spectroscopy nuclear magnetic resonance

Conditions

¹H and¹³With NMR-spectra were recorded as described for compounds ATV-ceclor.

Solvent: a mixture of CD₃OD/CDCl₃, 2:1 (¹H and¹³With NMR)

Structure:

Example 2

Receipt and allocation of connection ATV-ceclor

A solution of atorvastatin (2 l) was obtained in a mixture of 80% acetonitrile/20% water (vol./about.) at a concentration of 1 mg of atorvastatin in 1 ml of the Solution was placed in a small crucible and kept in the sun for 5 h, then the solution was immediately podslushivaet 1 M potassium hydroxide solution to pH 8-9 and evaporated under reduced pressure until the first signs of turbidity. The temperature of the water bath is maintained below 30°C. and the reflux condenser was cooled with water at a temperature of 0°C.

Then the solution was osvetleni minimal addition of acetonitrile.

Preparative chromatography and the structure determination was carried out similarly to that described in example 1.

After l is utilizacii concentrated fractions were received cleared connection ATV-ceclor (210 mg). Chromatographic purity of the compounds was 96.6 percent.

Example 3

Receipt and allocation of compounds ATV-epository and ATV-epoxide

A solution of atorvastatin (1 l) was obtained in a mixture of 80% acetonitrile/20% water (vol./about.) at a concentration of 1 mg of atorvastatin in ml. of the Solution was placed in a small crucible and kept in the sun for 5 h, then the solution was immediately acidified with 0.5 M phosphoric acid to pH 3.0. The mixture was stirred at room temperature for 2 h and evaporated under reduced pressure to about 1/3 the original volume.

Preparative chromatography and the structure determination was carried out similarly to that described in example 1.

After drying the concentrated fractions were received cleared connection ATV-epository (120 mg). Chromatographic purity of the compounds was 92.6 per cent.

After drying the concentrated fractions were received cleared connection ATV-epoxide (21 mg). Chromatographic purity of the compounds was 95,1%.

Example 4

Receipt and allocation of connection ATV-F

A solution of atorvastatin (800 ml) was obtained in acetonitrile at a concentration of 10 mg of atorvastatin in ml. was Added 12 M sodium hydroxide solution (4 ml) and 30% hydrogen peroxide solution (40 ml). The solution was stirred at 55°C for 5 hours, the Reaction mixture was cooled to room temperature and de who was intervale. The supernatant was evaporated under reduced pressure to about 50 ml, water was discarded and the solid residue was washed with a fresh portion of water. Then the solid residue was dissolved in acetonitrile.

Preparative chromatography and the structure determination was carried out similarly to that described in example 1.

After drying the concentrated fractions were received cleared connection ATV-F (230 mg). Chromatographic purity of the compounds was 98,3%.

Example 5

Calcium atorvastatin received by any known method. The only necessary condition for the entire allocation process calcium atorvastatin is inert atmosphere. In the thus obtained calcium atorvastatin content of each product of oxidative degradation (connection: ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide)defined according to GHUR at a wavelength of 250 nm, less than 0.04%.

Example 6

The atorvastatin calcium obtained by the method described in example 1 was stored in a nitrogen atmosphere at room temperature for 2 years. In the thus obtained calcium atorvastatin content of each product of oxidative degradation (connection: ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide)defined according to GHUR at a wavelength of 250 nm, less than 0.1%.

Example 7

Calcium is atorvastatin, obtained according to the method described in example 1 was stored in air at room temperature for 2 years. In the thus obtained calcium atorvastatin content of the products of oxidative degradation (connection: ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide)defined according to GHUR at a wavelength of 250 nm is 0,856%, 0,636%, 0,905% and 0.741%, respectively.

Example 8

Tablets were obtained from calcium atorvastatin obtained by the method described in example 1, and at least one pharmaceutically acceptable excipient.

In the thus obtained tablets contents oxidative degradation products (compounds: ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide)defined according to GHUR at a wavelength of 250 nm, is 0.11%, 0.07 per cent, of 0.07% and 0.08%, respectively.

The tablets are Packed in blisters and aluminium/aluminum in a nitrogen atmosphere. The blisters kept at room temperature for 2 years. The content of oxidative degradation products (compounds: ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide)defined according to GHUR at a wavelength of 250 nm is 0,18%, and 0.08%, 0,17% and 0.09%, respectively.

Example 9

The tablets obtained as described in example 8, was Packed in blisters and aluminium/aluminum in the air. Received such a way tablets content of products okoli the nutrient degradation (connection: ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide)defined according to GHUR at a wavelength of 250 nm, is 0.13%, and 0.09%, 0.08% and 0,08%, respectively.

The blisters kept at room temperature for 2 years. In the thus obtained calcium atorvastatin content of the products of oxidative degradation (connection: ATV-ceclor, ATV-epository, ATV-F and ATV-epoxide)defined according to GHUR at a wavelength of 250 nm, is 1.75%, and 0.61%, 1,23% and 0.65%, respectively.

1. 4-[6-(4-Forfinal)-6-hydroxy-1-isopropyl-6A-phenyl-1A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxipropionic acid.

2. Phenylamide 4-(4-forfinal)-2,4-dihydroxy-2-isopropyl-5-phenyl-3,6-dioxabicyclo[3.1.0]hexane-1-carboxylic acid.

3. 4-[1b-(4-Forfinal)-6-hydroxy-6-isopropyl-1A-phenyl-6A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxipropionic acid.

4. A method of obtaining a compound selected from the group including: 4-[6-(4-forfinal)-6-hydroxy-1-isopropyl-6A-phenyl-1A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxybutiric acid, phenylamide 4-(4-forfinal)-2,4-dihydroxy-2-isopropyl-5-phenyl-3,6-dioxabicyclo[3.1.0]hexane-1-carboxylic acid, 4-[1b-(4-forfinal)-6-hydroxy-6-isopropyl-1A-phenyl-6A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxilase the strong acid, which is characterized by the fact that the solid salt of atorvastatin oxidize in air or oxygen at elevated temperatures from 40 to 90°C.

5. The method according to claim 4, where the solid salt of atorvastatin is chosen from the group comprising calcium atorvastatin sodium atorvastatin, potassium atorvastatin, atorvastatin magnesium and ammonium atorvastatin.

6. A method of obtaining a compound selected from the group including: 4-[6-(4-forfinal)-6-hydroxy-1-isopropyl-6A-phenyl-1A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxybutiric acid, phenylamide 4-(4-forfinal)-2,4-dihydroxy-2-isopropyl-5-phenyl-3,6-dioxabicyclo[3.1.0]hexane-1-carboxylic acid, 4-[1b-(4-forfinal)-6-hydroxy-6-isopropyl-1A-phenyl-6A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxybutiric acid, in which the oxidation of a salt of atorvastatin carried out in a solution of salt of atorvastatin in water and/or organic solvent and/or in mixtures of solvents selected from the group comprising: acetonitrile, methanol, ethanol, propanol, dichloromethane or methylene chloride, adding hydrogen peroxide or by passing air or oxygen through the solution at a temperature of from about 40 to 90°C.

7. The method according to claim 6, where the salt of atorvastatin is chosen from the group comprising calcium atorvastatin sodium atorvastatin, Kali is atorvastatin, magnesium atorvastatin and atorvastatin ammonium.

8. A method of obtaining a compound selected from the group including: 4-[6-(4-forfinal)-6-hydroxy-1-isopropyl-6A-phenyl-1A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxybutiric acid, phenylamide 4-(4-forfinal)-2,4-dihydroxy-2-isopropyl-5-phenyl-3,6-dioxabicyclo[3.1.0]hexane-1-carboxylic acid, 4-[1b-(4-forfinal)-6-hydroxy-6-isopropyl-1A-phenyl-6A-phenylcarbamoyloxy-1,2-dioxa-5A-azacyclopenta[a]inden-3-yl]-3-(R)-hydroxybutiric acid, which is characterized by the fact that the solution of salt of atorvastatin is irradiated with sunlight or artificial sunlight.

9. The method of claim 8, where the salt of atorvastatin is chosen from the group comprising calcium atorvastatin sodium atorvastatin, potassium atorvastatin, atorvastatin magnesium and ammonium atorvastatin.

10. The method according to claims 4 to 9, which, in addition, includes one or more stages of the selection.

11. The method according to claim 10, where the extraction is chosen from the group comprising preparative normal-phase and reversed-phase chromatography.

12. The method according to claim 11, where for preparative normal-phase chromatography as a stationary phase using silica gel or phase on the basis of modified silica gel, for example, contain the following groups: aminopropyl, cyanopropyl, diol and nitrophenyl.

13. Pic is b-12, where to preparative normal-phase chromatography as a mobile phase, use a mixture of polar alcohol modifier selected from the group comprising methanol, ethanol, propanol and acetonitrile, and a non-polar solvent selected from the group comprising hexane, dichloromethane, methylcyclohexane and any combination thereof.

14. The method according to item 13, where for preparative reverse-phase chromatography using silica gel modified with octadecylsilane or octylsilane.

15. The method according to 14, where for preparative reverse-phase chromatography as a mobile phase, use a mixture of water with organic or inorganic buffer solution in a mixture with one or more organic modifiers selected from the group comprising alcohols and acetonitrile.