Crystals of taxane derivatives and method for their preparing

FIELD: organic chemistry of natural compounds, medicine, oncology.

SUBSTANCE: invention relates to a novel crystalline form of (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)ethylideneoxy]-5,20-epoxy-1-hydroxytax-11-ene-13-yl-(2R,3S)-3-(tert.-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate that shows the diffraction picture of roentgen rays in powder with characteristic peaks at diffraction angles (θ)= 6.2^o, 10.3^o, 10.7, 11.4^o and 12.0, and a method for its preparing. Method involves carrying out the crystallization step by using organic solvent chosen from group consisting of ketone type solvent, nitrile solvent type and their mixture, or mixture of said solvent and water. Also, invention relates to an antitumor agent based on the prepared crystalline form. Invention provides the stable quality of a medicinal agent based on its lower hygroscopicity.

EFFECT: improved and valuable properties of compounds.

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Technical area

This invention relates to new crystals derived taxane with antitumor activity, and the way they are received.

The rationale for creating theinventions

It is assumed that Taxol, a compound exhibiting antitumor activity, which is based on the inhibition of depolymerization of microtubules during cell division, effectively in clinical practice as an antitumor agent with a mechanism of action different from the action mechanism of conventional anticancer agents. Different types of derivatives of Taxol, respectively, have been described in publications. For example, the described compounds, in which the Deputy entered in the pyridine ring of the side chain, located in the 13th position for the suppression of the modification (transformation) of these compounds during metabolism (international publication WO 01/27115). In particular, the compound described in example 7 of the above-mentioned international publication, (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate hardly metabolized by human liver microsomes and is assumed to be an antitumor agent that can be administered orally.

That is for the connection, described in example 7 of the above-mentioned international publication, the method of obtaining this compound is described in example 9 of the publication, as in example 7. Example 9 describes the target connection, provided by operations similar to those described in stage 4 in example 8". With regard to the explanation that discusses the fact that the solvent in the extract of the reaction mixture containing the target compound was one stripped off, the residue is purified using chromatography on silica gel and recrystallized from a mixture of water and ethanol to obtain a solid substance, which, as defined, the crystals of the target compound. However, no physico-chemical assessment is not given in examples 7 and 9 of the above-mentioned international publication, which confirms the fact that the above mentioned compound was isolated in the form of crystals.

Description of the invention

The authors of this invention have repeated most of the experiments relating to the method of example 9 of the above-mentioned international publication. The authors ultimately concluded that to obtain a crystalline substance by vapor deposition using a mixture of water with ethanol as solvent, it is absolutely impossible, and the target substance was unexpectedly obtained in the form of oil or amorphous substan the AI. In view of these circumstances, the authors of this invention have conducted various studies with the aim of obtaining the above-mentioned compounds in the form of a crystalline substance, and the authors were first successfully obtained crystals of the above compound using a solvent other than a mixture of water and ethanol. The authors also found that there are at least two types of polymorphism of crystals of the above-mentioned compounds, and received a mixture of two types of crystals depending on conditions such as the type of solvent for the crystallization of the crystals.

From the point of view of continuous product applications, stability, quality, etc. as an active ingredient of a medicinal product, it is desirable to use a crystalline, not amorphous substance, and it is desirable that the crystalline substance represented one type of crystals. Due to these reasons, the authors of this invention have conducted various studies to obtain the same type of crystals of the above-mentioned compounds and as a result discovered that crystalline substance (substance) of the above-mentioned compounds consisting of one type of crystals can be obtained by using acetone, mixtures of acetone and water or in a mixture of acetonitrile and water as the organic solvent used for crystallizes the above compounds. This invention was created on the basis of these discoveries.

The invention thus allows to obtain crystals of (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate, which show characteristic peaks at angles of diffraction (2θ), equal to 6.2°, 10,3°, 10,7°, 11,4° and 12.0°in the film x-ray diffraction powder.

This invention also provides a method of obtaining the above-mentioned crystals, which includes the step of crystallization with the use of an organic solvent selected from the group consisting of a solvent type, ketone, solvent type nitrile and mixtures thereof or mixtures of these organic solvents and water. Preferably organic solvents selected from the group consisting of a ketone solvent type, solvent type, nitrile, and mixtures thereof, can be used acetone, or acetonitrile and a mixture of acetone and water or a mixture of acetonitrile and water as the preferred solvent for crystallization. When using the above-mentioned mixture of organic solvent and water, the water content in the above-mentioned mixture is preferably 60% by the Assos or less more preferably in the range from 40 to 50% by mass.

In accordance with a particularly preferred embodiment of the above-mentioned method of performing the above method is provided, when a mixture of acetone and water containing from 40 to 50% water by weight, or a mixture of acetonitrile and water containing from 40 to 50% water by weight, is used as the crystallization solvent in an amount of from 20 to 25 wt. parts by weight of the compound, the compound crystallizes at a temperature in the range of values from 0 to 45°and the resulting crystals are dried under reduced pressure with stirring at a temperature in the range from 30 to 60°C.

This invention, moreover, is an antitumor agent containing as an active ingredient the above-mentioned crystal; antitumor agent in the form of a pharmaceutical composition, which contains as an active ingredient the above-mentioned crystal and one or more kinds of pharmaceutical additives; the use of the above-mentioned crystal for producing the above-mentioned antitumor agent; and a method of therapeutic treatment on the tumor (treatment), which includes an introduction to the patient a therapeutically effective amount of the above-mentioned crystal. This invention also provides IP is the use of the above-mentioned crystal for the production of a medicinal product, containing as the active ingredient (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate.

A brief explanation of the figures

Figure 1 shows the diffraction pattern x-ray powder crystal of the present invention (β crystals).

Figure 2 shows the diffraction pattern x-ray powder mixture of polymorphic crystals (α crystals), which differ from the crystals of the present invention, and crystals according to this invention (β crystals).

Figure 3 shows the curve of thermogravimetric/differential thermal analysis (TG/DTA) of the crystals of the present invention (β crystals).

Figure 4 shows the curve of thermogravimetric/differential thermal analysis (TG/DTA) of the mixture of polymorphic crystals (α crystals), which differ from the crystals of the present invention, and crystals of the present invention (β crystals).

Figure 5 shows the curve of thermogravimetric/differential thermal analysis (TG/DTA) of amorphous solids.

Fig.6 shows the results of measurement of the nature of the absorption of moisture by the crystals of the present invention (β crystals and amorphous substance, measured relative to the additional humidity from 30 to 90%.

The best way to embodiments of the invention

(1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate is a compound represented by the following formula, and can be easily obtained by qualified technicians in this area in accordance with the method described in example 7 international publication WO 01/27115.

(In the formula, Me represents a methyl group, AC represents an acetyl group, Bz represents benzoyloxy group, and BOC represents tert-butoxycarbonyl group.)

The crystals of this invention are crystals (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate, which has characteristic peaks at angles of diffraction (2θ), equal to 6.2°, 10,3°, 10,7°, 11,4° and 12.0°in the film x-ray diffraction powder (these crystals are also referred to as "β crystals" in the description). Due to the fact that the diffraction angles (2θ) in the film x-ray diffraction powder generally have an experimental error in the range of the values less than 5%, each of the above-mentioned diffraction angles should be identified as multiple series of values representing the error less than 5%. Therefore, crystals with diffraction angles of the peaks of x-ray diffraction powder, identical to the above-mentioned angles with the experimental error less than 5% of the crystals as well as crystals having identical specified angles of diffraction peaks fall within the scope of this invention.

The crystals of this invention can be obtained by using the compounds, following the method described in example 7 international publication WO 01/27115, by crystallization from a solvent in which the organic solvent is selected from the group consisting of a solvent type, ketone, solvent type nitrile and mixtures thereof or mixtures of the above-mentioned organic solvent and water. As for the solvent type ketone, acetone, methyl ethyl ketone and the like, they can be used, and acetonitrile and the like can be used as solvent type nitrile. Among these solvents, preferred are acetone and acetonitrile.

When used for crystallization in a solvent mixture of organic solvent and water, the water content in the above-mentioned mixture is preferably 60% by m the SSE or less more preferably from 40 to 50% by weight. As for the mixture of organic solvent and water, a mixture of acetone and water and a mixture of acetonitrile and water is preferred. Especially preferred solvent for crystallization is a mixture of acetone and water containing from 40 to 50% water by weight, or a mixture of acetonitrile and water containing from 40 to 50% water by weight. For crystal deposition mentioned above, the connection can be directly dissolved in a mixture of acetone and water, a mixture of acetonitrile and water and the like. An alternative connection can be dissolved in a solvent such as acetone or acetonitrile, and then the solution with the corresponding water content can be added to conduct crystallization.

The amount of solvent used in crystallization, is not specifically limited. The majority is from about 5 to 50 parts by weight, preferably from about 20 to 25 parts by weight, of the weight of the above-mentioned compounds. The crystallization temperature is not particularly limited. The temperature is preferably, for example, in the interval from 0 to 45°C. the Time of crystallization mainly from about 8 hours to 1 day. The crystals after crystallization can be collected by filtration and then dried in the usual way to obtain crystals of this izaberete the Oia. When the crystals are dried, preferably to reduce sharply the temperature of the crystals. Drying is preferably carried out in air or under reduced pressure with stirring at a temperature in the range from room temperature up to 60°C. the Drying is preferably conducted at a temperature in the range from 30 to 60°C. If the temperature of drying is low, the crystals can be contaminated with crystals of other crystalline forms of (α crystals). Do the crystals of the present invention the crystallization and drying under simple control based on the information or not, each of the diffraction angles of the obtained crystal x-ray diffraction powder is identical to any of the above-mentioned diffraction angles. Figure 1 shows the diffraction pattern x-ray powder crystal of the present invention (β crystals), and figure 2 shows the diffraction pattern for the mix α and β crystals. The crystalline substance of this invention has higher stability than amorphous substance. For example, as shown in the examples, the hygroscopic crystalline substance (substance) of the present invention is lower than that of amorphous substances that detects high stability of the crystalline substance. In addition, the increased stability Krista is symbolic substance of this invention can also be confirmed using other methods, such as the experiment of irradiation of light.

Using (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate as an antitumor agent described in more detail in international publication WO 01/27115, and the crystal of the present invention can be used as an active ingredient the above-mentioned antitumor agent. A full description of international publication WO 01/27115 included in this invention in the form of links. Referring to the description of the above-mentioned international publication, qualified professionals may use pharmaceutical preparation containing a crystal of the present invention as an active ingredient of the antitumor agent. The crystals of this invention can also be used for the production of a medicinal product containing the aforementioned compound as an active ingredient. For example, the crystal of this invention can be used for the production of injection or solutions, dosage forms in the form of solutions.

Pharmaceutical preparation containing a crystal of the present invention as an active ingredient, preferably provided in the form of pharmaceutical compositions that contain Asa crystals of the present invention as an active ingredient and one or more pharmaceutical additives. Route of administration of a drug of the present invention is not strictly regulated, and the drug can be administered orally or parenterally. (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate, when it is used as an active ingredient of the drug of the present invention, is characterized the fact that exerts its anticancer effects even in oral administration. Thus, oral administration is preferred.

Examples of the pharmacologically and pharmaceutically acceptable additives used in the production of the above-mentioned pharmaceutical compositions, include, but are not limited to excipients, loosening agents or loosening additives, binding agents, lubricating agents, film coatings, dyes, diluents, basic materials, dissolving agents or dissolving additives, isotonic agents, pH regulators, stabilizers, dispersing agents, substances that enhance the adhesiveness, and the like.

The examples of the preparation of suitable dosage forms for oral administration include, for example, tablets, powders, granules, capsules, and so p is such. The examples of the preparation of suitable dosage forms for parenteral administration include, for example, injection, fusion drips, suppositories, inhalation forms, plasters and the like. Among them are capsules and tablets are preferred. For preparation suitable for oral administration forms can be used, for example, excipients such as glucose, lactose, D-mannitol, starch and crystalline cellulose; loosening agents or loosening additives such as carboximetilceluloza, starch and calcium carboxymethyl cellulose; binders such as hydroxypropylcellulose, hypromellose, polyvinylpyrrolidone and gelatin; lubricants such as magnesium stearate and talc; film agents such as hypromellose, sucrose, polyethylene glycol and titanium oxide; bases such as vaseline, liquid paraffin, polyethylene glycol, gelatin, kaolin, glycerin, purified water and tallow, as pharmacologically and pharmaceutically acceptable additives.

The dose of the drug of the present invention is not strictly regulated and may be appropriately selected depending on various conditions, such as tumor type, age, weight, symptoms and characteristics of the patient, and the like. The drug is usually injected in number, var is IRowset from 0.5 mg to 50 mg, preferably from about 1 mg to 20 mg per 1 m²the surface area of the body.

Examples

Further, the invention is specifically explained and illustrated using examples. However, the scope of the invention is not limited to these examples.

Example 1

(1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate (30.0 g, 34 mol) is added to the acetone (45 ml) and dissolved by stirring under heating on a water bath at about 45°C. To the solution is added dropwise water (30 ml) at about 45°under stirring and after adding stirred for 2 hours. Then the water bath is cooled to approximately 23°and the solution is stirred over night. The precipitated crystals are collected by filtration and washed with a mixture of acetone and water (30 ml)containing 60% water.

The crystals are dried under reduced pressure, equal to about 600 mm RT. century, when about 60°With stirring for 3 hours. Then the crystals 1.5 hours then dried under reduced pressure, equal to about 150 mm, RT. Art., and 1 hour at a pressure of approximately 30 mm RT. Art., receiving 27 g (90%) of white crystals.

The obtained crystals are subjected to analysis x-ray diffraction powder. The result was characteristically observed the peaks at angles of diffraction (2θ ), equal 6,18°, 10,30°that is 10.68°, 11,38° and 11,96°.

The results of the analysis of x-ray diffraction powder is shown in figure 1. In addition, the results of thermogravimetric analysis and differential thermal analysis (TG/DTA) of the crystals is shown in Fig. 3.

Example 2

(1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate (30.0 g, 34 mol) is added to the acetone (45 ml) and dissolved by stirring under heating on a water bath at about 45°C. To the solution is added dropwise water (80 ml) at about 45°under stirring and after adding stirred for 2 hours. Then the water bath is cooled to approximately 23°and the solution is stirred over night. The precipitated crystals are collected by filtration and washed with a mixture of acetone and water (80 ml)containing 60% water.

The crystals are dried under reduced pressure, equal to about 60 mm RT. century, when about 60°receiving 27 g (90%) of white crystals.

The obtained crystals are subjected to analysis x-ray diffraction powder. In the observed characteristic peaks, similar to those that were obtained in example 1.

Example 3

(1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)this is identix]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate (2.5 g, 2.8 mol) is added to the acetone (38 ml) and stirred under heating on a water bath at about 45°C. To the solution is added dropwise water (25 ml) at about 45°under stirring and after adding continue stirring for 2 hours. Then the water bath is cooled to approximately 23°and the solution is stirred over night. The precipitated crystals are collected by filtration and washed with a mixture of acetone and water (25 ml)containing 60% water.

The crystals are dried under reduced pressure, equal to about 600 mm RT. century, when 60°With stirring for 3 hours. Then the crystals are dried in air at room temperature (about 19° (C)to give 2.3 g (92%) of white crystals.

The obtained crystals are subjected to analysis x-ray diffraction on the sample powder. In the observed characteristic peaks that are close to those obtained in example 1.

Example 4

(1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate (2.5 g, 2.8 mol) are added to acetonitrile (5 ml) and stirred while heating on a water bath at about 45°C. To the solution, then add water (3 ml) at about 45°and leave to stand at room temperature for 1 hour. Then the solution is left is aout to stand at about 5° With over 22 hours. The precipitated crystals are collected by filtration and washed with cold mixture of acetonitrile:water (1:2.5 ml).

The crystals are dried under reduced pressure at 40°C for 22 hours, getting 0,37 g (74%) of white needle-shaped crystals.

The obtained crystals are subjected to analysis x-ray diffraction powder. In the observed characteristic peaks that are close to those obtained in example 1.

Example 5

Was measured moisture absorption of the crystals of the present invention (β crystals and amorphous material at a relative humidity of from 30 to 90%. Were used samples weighing each about 20 mg, and the relative humidity was changed from 30%in increments of 10% using a microbalance (automatic device for measuring the absorption of moisture). Temperature measurement was equal to 25°C. Quantitative change was determined when the sample was given the change to 0.03% or less, for 30 min (maximum aging time 180 minutes). The results are shown in Fig.6. Amorphous substance with high humidity absorbs more moisture by 1% in comparison with the crystals of the present invention.

Industrial applicability

This invention provides a crystalline substance consisting of one type of crystal compound useful as an antitumor agent. Applied the e crystals of the present invention help to provide a stable quality of the medicinal product.

1. Crystals of (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate that demonstrate the diffraction pattern x-ray powder with characteristic peaks at the angles of diffraction (2θ) is equal to 6.2°, 10,3°, 10,7°, 11,4° and 12.0°.

2. The method of producing crystals according to claim 1, which includes the step of crystallization with the use of an organic solvent selected from the group consisting of a solvent type, ketone, solvent type nitrile and mixtures thereof or mixtures of the specified organic solvent and water.

3. The method according to claim 2, in which an organic solvent chosen from the group consisting of a solvent type, ketone, solvent type nitrile and mixtures thereof is an organic solvent selected from the group consisting of acetone, acetonitrile, and mixtures thereof.

4. The method according to claim 2, in which the crystallization was carried out using a mixture of acetone and water or in a mixture of acetonitrile and water.

5. The method according to claim 4, in which a mixture of acetone and water, or a mixture of acetonitrile and water contains from 40 to 50% water by weight.

6. The method according to any of claim 2 to 5, further comprising a stage of drying the crystals obtained by crystallization at a temperature of from 30 to 60�B0; C.

7. The antitumor agent comprising as an active ingredient a crystal according to claim 1.

8. The crystal according to claim 1, used for getting medicines containing (1S,2S,3R,4S,5R,8R,9S,10R,13S)-4-acetoxy-2-benzoyloxy-9,10-[(1S)-2-(dimethylamino)utilizandose]-5,20-epoxy-1-hydroxydec-11-EN-13-yl(2R,3S)-3-(tert-butoxycarbonylamino)-3-(3-fluoro-2-pyridyl)-2-hydroxypropionate, as the active ingredient.