RU2659160C1 - Bortezomib lyophilizate production method and the bortezomib containing pharmaceutical composition in the form of stable lyophilizated product produced by said method - Google Patents

Abstract

FIELD: technological processes.

SUBSTANCE: invention relates to the bortezomib lyophilizate (bortezomib substance) production process. Bortezomib lyophilizate is produced by successive production first of the mannitol aqueous solution with its concentration in the solution of 10–20 mg/ml; then the bortezomib aqueous solution production in the resulting mannitol aqueous solution with the bortezomib concentration of 1.0–2.5 mg/ml. Next, performing the product lyophilic drying in the freeze-drying chamber in several steps under certain temperature conditions, with the freeze drying steps certain duration. Producing the stable bortezomib lyophilizate with the product high purity of up to 99.85 wt.%. Also disclosed is the pharmaceutical composition containing, as an active ingredient, the freeze-dried bortezomib lyophilizate, and purified water or physiological saline as the pharmaceutically acceptable carriers.

EFFECT: process according to the invention enables production of the stable and highly purified target product – bortezomib lyophilizate, as well as ability to control all the process stages and their implementation conditions.

5 cl, 2 ex

Description

The present invention relates to a method for producing a lyophilisate of bortezomib and a lyophilisate of bortezomib, in particular obtained by the above method, as well as a pharmaceutical composition comprising this lyophilisate.

Bortezomib is a modified dipeptidylboronic acid with the chemical name [(1R) -3-methyl-1 - ([((2S) -3-phenyl-2 [(pyrazin-2-ylcarbonyl) amino] propanoyl] amino) butyl] boronic acid, also known in the art as “N- (2-pyrazine) carbonyl-L-phenylalanine-L-leucine boronic acid.” Bortezomib has the following structural formula

Bortezomib is known to have a solubility lower than 1 mg / ml in 0.9% (w / v) sodium chloride. It is marketed under the trade name Velcade® Injection. It is used in a lyophilized form, which after restoration of moisture content forms a solution consisting of mannitol ester in equilibrium with bortezomib. Bortezomib is known to be the first therapeutic protease inhibitor, in particular, a reversible inhibitor of chymotrypsin-like 26S proteasome activity in mammalian cells. It is approved in the United States for the treatment of recurrent multiple myeloma and lymphoma from mantle cells.

Pharmaceutical compositions of boronic acid compounds obtained by lyophilization of an aqueous mixture comprising a boronic acid compound and a compound comprising at least two hydroxyl groups are disclosed in WO 2002/059130 A1 and WO 2002/059131 A1. WO 2010/039762 A2 obviously discloses stable sugarless pharmaceutical compositions of bortezomib.

To ensure sufficient dissolution of bortezomib for lyophilization, WO 2002/059130 A1 uses water and approximately 40% tert-butanol as a co-solvent. However, the use of a large number of expensive organic cosolvents, which can be purchased in the distribution network with a limited degree of purification, for example, to approximately 99.9%, is disadvantageous. Typically, co-solvent impurities remain in the lyophilized product, as well as a significant amount of residual co-solvent. In addition, the use of a large number of solvents is disadvantageous because the removal of the solvent in vacuum during lyophilization requires a large amount of time and energy.

As described in documents WO 2010/089768 A2, 12/08/2010, and WO 2010/039762 A2, 04/08/2010 A2, the preparation of bortezomib is associated with significant problems regarding stability and solubility, in particular due to the low solubility of bortezomib in water or 0.9% (v / v) sodium chloride solution.

So, the existing technology for the preparation of bortezomib lyophilisate involves the dissolution of the substance in tert-butanol. After the substance is dissolved, water and mannitol are added to the solution. Tert-butanol is selected for reasons of subsequent lyophilization of the solution and as a solvent for the substance (US 2002188100, 12/12/2002).

It was also found that in the substance of bortezomib several of its crystalline forms may be present at the same time, as a result, when dissolved in water mannitol, part of the substance does not dissolve, and visually the solution may remain cloudy, despite an increase in the temperature of the solution and pH.

However, there are ways to convert such a substance into a form convenient for dissolution.

For this, you can, for example, choose a solvent for crystallization of the substance bortezomib, from which the desired water-soluble form, for example, methanol, crystallizes, another option involves dissolving the substance in methylene chloride, and evaporating the solution on a rotary evaporator, after which a water-soluble amorphous form of bortezomib.

Bortezomib has been found to have a pH-dependent dissolution in water, dissolving much more easily at high pH values.

This approach is implemented, for example, in patent WO 2010/089768 A2, 08/12/2010, where the dissolution of the substance bortezomib is carried out in an environment with a pH of from 6.8 to 8.4, preferably from 7.9 to 8.1. However, it turned out that an increase in the pH of the solution over 6.5-6.7 subsequently leads to a significant increase in impurities in the dosage form.

In patent application WO 2010/089768, A2, when preparing a solution of the substance bortezomib, to improve solubility, heating is carried out to 40-50 °.

In this document WO 2010/089768, in order to overcome this stability problem, the preparation of the compositions includes bortezomib and tromethamine, with a pH ranging from 6.8 to 8.4. The composition is obtained by mixing bortezomib and tromethamine and adding water with continuous stirring. The solution is usually heated to 45 ° C-50 ° C to form a clear solution. It is described that the obtained lyophilisate restores moisture content in 48 seconds. However, heating bortezomib, which, according to the description, has low chemical resistance, is disadvantageous, since heating usually increases the rate of decay. In addition, restoring the moisture content of the obtained lyophilisate is not preferred.

Document WO 2011/110286 also refers to the problem of intrinsic instability of bortezomib, in particular its aqueous solutions. To overcome the stability problem of bortezomib solutions, substantially non-aqueous liquid compositions or compositions are provided in which bortezomib is formulated with a Lewis heterobifunctional base as a donor compound. However, non-aqueous compositions are disadvantageous for the preparation of pharmaceutical compositions, in particular for injection.

In another patent source EA 021179, 04/30/2015, a method for producing a lyophilisate of bortezomib or a salt or derivative thereof is disclosed, which comprises the steps of:

a) preparing an aqueous solution comprising a compound containing at least two hydroxyl groups separated by at least two connecting atoms in a chain or ring, or a compound comprising a hydroxyl group and an amino group separated by at least two connecting atoms in a chain or ring, wherein the solvent system consists of water;

b) dissolving bortezomib or its salt or derivative in the aqueous solution obtained in step a);

c) lyophilizing the aqueous solution obtained in step b), optionally after diluting the above solution with water.

Bortezomib or a salt or derivative thereof (hereinafter “bortezomib”) used in accordance with the method of the present invention is known to those skilled in the art. The synthesis of bortezomib is described, for example, in documents US 6617317 or WO 2005/097809 A2.

A compound having at least two hydroxyl groups separated by at least two connecting atoms in a chain or ring, as used herein, also referred to as a “dihydroxyl compound”, means any compound having two or more hydroxyl groups in which two hydroxyl groups are separated at least two connecting atoms. A compound having a hydroxyl group and an amino group separated by at least two connecting atoms in a chain or ring, as used herein, also referred to as an “amino hydroxyl compound”, means any compound having at least one hydroxyl group and at least one amino group separated at least two connecting atoms. The connecting atoms of the dihydroxyl compound or the amino hydroxyl compound may be in a chain or ring, and the chain or ring may include carbon atoms and optionally one or more heteroatoms, for example, N, S or O. Examples of dihydroxyl compounds are sugars, pinanediol, pinacol, perfluoropinacol , ethylene glycol, diethylene glycol, catechol, 1,2-cyclohexanediol, 2,3-butanediol, 1,2-butanediol and glycerol. In a preferred embodiment, the dihydroxyl compounds are sugars, more preferably monosaccharides or disaccharides. Examples of suitable sugars are glucose, sucrose, fructose, trehalose, mannitol and sorbitol. In a preferred embodiment, the sugar used is reduced sugar, commonly known as a sugar alcohol such as mannitol, sorbitol, erythritol or pentaerythritol, most preferably D-mannitol.

Examples of aminohydroxyl compounds are hydroxyl amino acids, aminoalcohols and aminohydroxyl thiols, preferably 2-aminoethanol, series, 2-amino-1,3-propanediol, tromethamine, cysteine or cysteinol (2-amino-3-hydroxy-propan-1- thiol). In a preferred embodiment, the amino hydroxyl compound is 2-aminoethanol, 2-amino-1,3-propanediol, cysteine or cysteinol.

The solvent system used to prepare the aqueous solution of the dihydroxyl compound or the amino hydroxyl compound in step a) is an aqueous solvent system comprising water as a solvent. It is contemplated that up to 10 wt.% Of a co-solvent, for example, an organic co-solvent, or up to 5 wt.%, Or up to 1 wt.% Of the co-solvent can also be used in the solvent system. According to the known invention, the solvent system essentially consists of water, that is, only up to 0.1 wt.%, More preferably up to 0.05 wt.%, In particular up to 0.01 wt.% Or less, are present in the solvent system substances other than water. Typically, purified water is used, preferably highly purified water (according to Ph. Eur.). Thus, with the exception of water, other cosolvents in the solvent system are present in small amounts.

According to a known method for producing a lyophilisate of bortezomib or a salt or derivative thereof, an aqueous solution of a dihydroxyl compound or an aminohydroxyl compound in water is prepared in step a), in a preferred embodiment, until the dihydroxyl compound or aminohydroxyl compound is completely dissolved. Typically, the concentration of the dihydroxyl compound or amino-hydroxyl compound in the aqueous solution prepared in step a) is at least about 10 mg / ml, preferably at least about 20 mg / ml, more preferably at least about 25 mg / ml, in particular at least about 30 mg / ml, for example at least about 50 mg / ml. The upper limit of the concentration of the dihydroxyl compound or the amino hydroxyl compound can reach the solubility limit of the corresponding compound in water, typically up to about 200 mg / ml, preferably up to about 150 mg / ml, in particular up to about 100 mg / ml. A typical concentration range of the dihydroxyl compound or amino hydroxyl compound in the solution prepared in step a) is from about 10 to about 200 mg / ml, preferably from about 15 to about 180 mg / ml, more preferably from about 20 to about 150 mg / ml, in particular from about 25 to about 100 mg / ml, for example from about 50 to about 100 mg / ml.

According to a known method for producing a lyophilisate of bortezomib or a salt or derivative thereof, in step b), bortezomib or its salt or derivative, in particular bortezomib free acid, is dissolved in the aqueous solution obtained in step a). In a preferred embodiment, the bortezomib solution is dissolved in the aqueous solution obtained in step a) at a temperature lower than about 30 ° C, preferably at room temperature, that is, about 20 ° C, or lower, more preferably about 15 ° C or lower, in particular approximately 10 ° C or lower, for example, approximately 5 ° C. In a preferred embodiment, the solution obtained in step a) is cooled to an appropriate temperature before adding bortezomib. Bortezomib can be used in solid form for dissolution. In one preferred embodiment, bortezomib is suspended in water, typically in a weight ratio of water to bortezomib from 10: 1 to 1: 1, in a preferred embodiment, from 2: 1 to 1: 1, and the suspension is added to the solution obtained in step but). The amount of bortezomib in the solution in step b) is preferably such that the molar ratio of the dihydroxyl compound or aminohydroxyl compound to bortezomib is from about 0.5: 1 to about 100: 1, more preferably from about 5: 1 to about 100: 1 in particular from about 10: 1 to about 100: 1, for example from about 10: 1 to 20: 1.

The amount of bortezomib in the solution in step b) is preferably such that the weight ratio of the dihydroxyl compound or aminohydroxyl compound to bortezomib is from about 0.5: 1 to about 100: 1, more preferably from about 5: 1 to about 100: 1 in particular from about 10: 1 to about 100: 1, for example from about 10: 1 to 20: 1.

In one embodiment of the known method, the solution before lyophilization may include one or more pharmaceutically acceptable excipients, carriers, solvents, excipients, salts, buffers, stabilizers or solubilizers, as is known to those skilled in the art. In a preferred embodiment, during lyophilization of the solution, none of the additional excipients or adjuvants is present in the aqueous solution, except for bortezomib and the dihydroxyl compound.

The lyophilization step c) is carried out by a method known to those skilled in the art. Typically, the solution to be lyophilized is prepared to achieve complete dissolution. After this stage, a second stage is carried out, at which the temperature is significantly reduced, for example, to a level of from about -80 to -20 ° C, in a preferred embodiment, to a level of from about -40 to -15 ° C, and a vacuum is usually applied approximately 100-200 mTorr. The reduced temperature and vacuum are generally maintained from about 5 to about 24 hours.

After this, the temperature is usually raised to a level of from about 10 to about 25 ° C, for example about 20 ° C, while maintaining or increasing the vacuum, for example to 20-50 mTorr, as a rule, until the residual solvent is substantially removed.

In one embodiment of the known method, a bortezomib solution comprising a dihydroxyl compound or an amino hydroxyl compound obtained in step b) is used in the lyophilization step c) without further dilution, that is, without adding additional water before lyophilization. Thus, in this embodiment, the solution to be lyophilized has the same concentration of the dihydroxyl compound or the amino hydroxyl compound as in step a) as described above. In a preferred embodiment, before subjecting the solution obtained in step b) to lyophilization, the above solution is diluted by adding water. Preferably, sufficient water is added to reduce the concentration of the dihydroxyl compound or amino hydroxyl compound to about 20 mg / ml or lower, in the preferred embodiment, about 15 mg / ml or lower, in particular about 10 mg / ml or lower, before exposure to the diluted solution lyophilization in step c).

The known invention also relates to a lyophilisate obtained by the method according to this invention. The resulting lyophilisate is characterized by a low amount of impurities, in particular degradants of bortezomib, and rapid restoration of moisture content.

In addition, the known invention relates to a lyophilisate of bortezomib or its salt or derivative, which are free from impurities from a co-solvent and do not contain a co-solvent.

The known invention also describes a pharmaceutical composition comprising a lyophilisate according to the present invention, in particular a lyophilisate obtained in a known manner. The pharmaceutical composition may also include appropriate excipients and adjuvants known to those skilled in the art.

This known technical solution according to EA 021179 can be selected as the closest analogue (prototype) of the claimed invention.

The technical task of the claimed invention is the creation of a new method for producing a lyophilisate of a boron acid compound of bortezomib or its salt or derivative, which provides a more stable (highly stable) and highly purified target product - bortezomib lyophilisate and provides control over all stages of the process and the conditions for their implementation.

Thus, in accordance with the technical task, the technical result achieved is a controlled process for obtaining a stable and highly purified final target product - bortezomib lyophilisate, and its increased efficiency when used in the pharmaceutical composition as an active active substance. Moreover, upon receipt of it by the method according to the claimed invention, organic solvents are not used at all; upon receipt of the bortezomib lyophilisate (bortezomib substance) by the claimed method as a solvent in the preparation of mannitol solutions, only water is used to dissolve bortezomib. The exclusion of an organic solvent during the dissolution of mannitol, bortezomib is one of the significant advantages of the claimed method compared to the above patent EA 021179, which provides for the use of an organic solvent, which reduces the safety of the process for producing bortezomib lyophilisate, and also increases unnecessary impurities.

As mentioned above, when describing the prior art, it is known that the dissolution of bortezomib in water and, in particular, in an aqueous solution of mannitol, is easier and faster at high pH values from 6.8 to 8.4.

In the technical solution claimed in this invention, the pH is maintained at the stages of dissolution of mannitol in water and bortezomib in an aqueous solution of mannitol, respectively, a pH in the range from 4.0 to 5.0 and a pH from 5.0 to 6.5.

Obtained by the claimed method, bortezomib (lyophilisate bortezomib) is characterized by a very low content of impurities, in particular, degradants of bortezomib and rapid restoration of moisture content in an aqueous solution, for example, in an aqueous solution of sodium chloride, in particular, a 0.9-1.0% solution of chloride sodium (saline).

So, the technical task and the technical result are achieved by the claimed method of obtaining the lyophilisate of bortezomib, including the following stages:

(a) preparing an aqueous solution of mannitol until it is completely dissolved with its concentration in a solution of 10-20 mg / ml at 0-60 ° C for 0.5-5.0 hours and maintaining the pH from 4.0 to 5.0.

(b) dissolving the substance of bortezomib in the resulting aqueous solution of mannitol for 1.0 to 5.0 hours to obtain a concentration of bortezomib in the resulting solution of 1.0-2.5 mg / ml at 15-30 ° C. while maintaining the pH of the solution in a pH range in the range from 5.0 to 6.5;

(c) sterilizing filtration of the solution from step (b) using filtration under vacuum or filtration under a pressure of 0.1-0.6 MPa and dosing it into vials;

(d) freeze-drying the product from step (c) in a freeze-drying chamber in several stages for 24-72 hours, with primary drying being carried out first in the temperature range (-30 ° C) - (- 20 ° C), increasing the temperature on the shelf in the chamber every 30-60 minutes at 0.5-5.0 ° C / h, thereby performing cycles of gradual rise in temperature and holding at a given temperature until the temperature reaches (-5 ° C) - (- 2 ° C)

- then carry out exposure at this temperature for 0.5-5.0 hours, then set the drying temperature in the range from 0 to (-1 ° C), maintain at this temperature for 5-30 hours to remove about 90 wt.% water from the product;

- then carry out a gradual increase in temperature with a rise speed of 0.5-5.0 ° C / h until the temperature of the shelves of the freeze-drying chamber reaches 25-30 ° C for 5-25 hours and a vacuum of 1-150 m / Torr;

(f) filling the freeze-drying chamber with inert gas, corking the bottles and filling them with caps.

Moreover, the regulation and control of the pH values of the solutions in stages (a) and (b) during the implementation of the method according to the invention is carried out using alkali metal hydroxide NaOH (sodium hydroxide) or KOH (potassium hydroxide), for example, by fractional addition of an alkali solution.

Upon receipt of the lyophilisate of bortezomib, the substance bortezomib is used, which is the free acid of bortezomib, or its salt, or its derivative (hereinafter “bortezomib”).

As a result of numerous experiments, it was found that three parameters during the implementation of the method, such as time, temperature and pH value, were established to achieve the set technical result. In 1-5 hours, and mainly in 3 hours, bortezomib dissolves in an aqueous solution of mannitol at a pH of from 5.0 to 6.5. The ideal pH value is 5.7-6.0, the higher the pH, the better and faster the dissolution of bortezomib in an aqueous solution of mannitol, but we, in the course of work, revealed a dependence at pH above 6.5 during the storage process, the formation of impurities bortezomib in a dosage form (lyophilisate). Correction and maintenance of pH was carried out by fractional addition of a NaOH solution until a predetermined pH value was reached. The dependence of the dissolution of bortezomib in a mannitol solution is also known, and in particular, the higher the temperature, the better and faster its dissolution occurs, but it is also known that with increasing temperature, the degradation of bortezomib also leads to the formation of impurities; temperature not higher than 25 ° C, on average about 22-23 ° C. The dissolution of bortezomib in a mannitol solution was carried out in an inert gas medium - nitrogen.

Another invention claimed in this application, included in the claimed group of inventions, is a pharmaceutical composition containing bortezomib in the form of a lyophilized product (lyophilisate) obtained by the method described above, which is one of the inventions of the claimed group, in therapeutically effective amounts and pharmaceutically acceptable carriers.

In this case, preferably the pharmaceutical composition contains a lyophilisate of a fully water-soluble crystalline form of the substance bortezomib.

The claimed pharmaceutical composition as pharmaceutically acceptable carriers contains purified water, physiological saline (0.9-1.0% aqueous solution of sodium chloride).

The claimed pharmaceutical composition may additionally contain various auxiliary target additives, such as buffers (citric acid, acetic acid, tartaric acid), antioxidants (e.g., butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, α-tocopherol), hydrophilic antioxidants (sodium EDTA, thioglycerol, preservatives (phenol, trimerozal, chlorobutanol, etc., for example, in a concentration of from 0.001 wt.% and up to less than 5% wt./mass. of the total composition) and other commonly known recall nutritional supplements (fillers, cryoprotectants, lipoprotectors (e.g. amino acids), polymers (gelatin, albumin), or mixtures thereof in acceptable amounts.

Obtained by the claimed method, the lyophysilate of bortezomib has increased stability over a long period of time (at least 2-36 months).

The claimed pharmaceutical composition can be used in therapeutically effective amounts to obtain (or used) in the treatment of malignant tumors, (cancer).

The active active pharmaceutical ingredient - Bortezomib is a highly selective reversible inhibitor of the activity of the 26S proteasome, is a modified boronic acid.

The proteasome 26S is present in the nucleus and cytosol of all eukaryotic cells and is a key component that catalyzes the breakdown of the main proteins involved in cell life cycle management. Bortezomib inhibits the chymotrypsin-like action of the proteasome, causes inhibition of proteolysis and leads to apoptosis.

Myeloma cells are almost 1000 times more susceptible to apoptosis caused by bortezomib than normal plasma cells.

The main factor explaining the ability of a bortezomib proteasome inhibitor to kill myeloma cells is its ability to block NF-kB activation. In normal cells, NF-kB (which exists as the p50-p65 dimer) is bound to the inhibitory protein LkB, which keeps it inactive in the cytosol. Some tumors contain activated forms of NF-kB, and the proteasome plays an important role in this activation, as it catalyzes the proteolytic generation of the NF-kB p50 subgroup from the inactive p150 precursor and the destruction of the LkB inhibitory protein. Activated NF-kB, penetrating the nucleus, helps the cell survive and proliferate. By inhibiting the proteasome and, therefore, inhibiting the activation of NF-kB, bortezomib helps to reduce the number of antiapoptotic factors, inflammatory molecules, cell adhesion molecules (which allow connective cells to attach to bone marrow cells) and cytokines (which stimulate myeloma cell growth).

Bortezomib causes a slowdown in the growth of human tumors in many experimental models, including multiple myeloma.

The claimed method for producing a lyophilisate of bortezomib allows to obtain a lyophilisate of bortezomib, which has a purity of approximately 99.80-99.85 wt.%.

The pharmaceutical composition is used in the form of a solution for infusion or injection (after reconstitution).

Obtained by the claimed method, the lyophilisate bortezomib is contained in the pharmaceutical composition in an amount of from about 0.01 to 99.0 wt.% Of the total weight of the composition.

The following are examples to illustrate the claimed invention without limiting the claimed invention.

Example 1

Preparation of a solution of lyophilisate of the substance bortezomib per 1000 vials.

In the prepared water (purified water) for injection (≈2000 g) in a glass reactor, mannitol (≈35 g) is added with stirring until it is completely dissolved to obtain a mannitol concentration in solution of 10 mg / ml at 0 ° C for 4-5 hours and the pH of the solution in the range of pH = 4-5.

Then, the substance of bortezomib (hereinafter “bortezomib”) is added to the resulting mannitol solution (with a concentration of 10 mg / ml) and it is dissolved for 1 h to obtain a solution of bortezomib with a concentration in the solution of 1.0 mg / ml at 15 ° C and at maintaining the pH of the solution in the range from 5.0 to 6.5 in a nitrogen nitrogen gas environment to obtain a bortezomib concentration of 1.0 mg / ml.

Sterilized filtration of the resulting solution of bortezomib in an aqueous solution of mannitol (under vacuum) and the subsequent dosage of the solution in vials are carried out.

Next, freeze drying of the obtained solution of bortezomib in the freeze drying chamber is carried out in several stages for 24 hours, subject to the following conditions of freeze drying at each stage:

- first carry out the initial drying of the solution at -20 ° C, increasing the temperature on the shelves of the freeze-drying chamber every 30-35 minutes by 2.0 ° C / h, thereby performing alternating cycles of gradual temperature rise and holding at this temperature until the temperature is reached to (-2 ° C).

Carry out exposure at this temperature for 3 hours. Next, set the drying temperature on the shelves of the chamber 0 ° C, maintain at this temperature for 30 hours to remove about 90 wt.% Water from the product.

Next, the drying temperature is gradually increased until the temperature on the shelves of the freeze-drying chamber reaches 25 ° C with a temperature rise rate of 5 ° C / h for 5 hours and a vacuum of 1-10 m / torr.

The freeze-drying chamber is filled with inert gas (nitrogen), the bottles are corked and sealed with caps.

A bortezomib lyophilisate is obtained with a product purity of ≈99.80 wt.%.

Example 2

The process of obtaining a lyophilisate of bortezomib is carried out analogously to example 1, but with varying process conditions within the ranges of temperature, time and other conditions given in the claims:

- prepare an aqueous solution of mannitol until it is completely dissolved with a solution concentration of 20 mg / ml (at 60 ° C for 0.5 h at pH = 5.0);

- dissolve the substance of bortezomin in the resulting aqueous solution of mannitol for 3.0-5.0 hours to obtain a solution with a concentration of bortezomib in it of 2.5 mg / ml at 23-25 ° C, the pH of the solution in the range from 5.7 to 6 5 and in an inert atmosphere of nitrogen gas;

- carry out sterilization filtration of the solution under a pressure of 0.1-0.6 MPa and dosing it into bottles.

While maintaining the pH in these ranges is carried out by adding, if necessary, NaOH or KOH.

Freeze drying of the resulting bortezomib solution is carried out for 72 hours under the following conditions:

- initial drying at (-30 ° C) with increasing temperature every 60 minutes by 5 ° C / h until the temperature is reached (-5 ° C);

- exposure at this temperature for 0.5 hours;

- set the temperature on the shelves of the chamber to 0 ° C and maintain at this temperature for 0.5 hours until about 90 wt.% water is removed from the product.

The temperature is gradually increased to 30 ° C over 25 hours (with a temperature rise rate of 0.5 ° C / h for 25 hours in a vacuum of 150 m / Torr (or possibly 100 m / Torr). The chamber is filled with an inert gas (nitrogen); cork the bottles and fill them with caps.

Get the lyophilisate bortezomib with a purity of 99.85 wt. %

Bortezomib lyophilisate restores moisture quickly within 10-15 seconds after mixing it with an aqueous solution, in particular, a 0.9% wt sodium chloride solution (physiological solution) at room temperature, at about 20-23 ° C, I get essentially a pharmaceutical composition based on lyophilisate bortezomib.

The pharmaceutical composition may additionally contain various auxiliary additives (antioxidants, preservatives, buffers, etc., as well as mixtures thereof in acceptable amounts known to specialists).

Thus, the claimed method allows to obtain bortezomib lyophilisate with a high degree of purity, which has increased stability with a concentration of bortezomib in the lyophilisate of 1.0-2.5 mg / ml, which ultimately provides a pharmaceutical composition containing bortezomib lyophilisate with increased stability and efficiency at using her.

Claims (13)

1. A method of obtaining a lyophilisate of bortezomib, comprising the following stages: (a) preparing an aqueous solution of mannitol until it is completely dissolved with its concentration in a solution of 10-20 mg / ml at 0-60 ° C for 0.5-5.0 hours and a pH in the range from 4.0 to 5.0 ; (b) dissolving the substance of bortezomib in the resulting aqueous solution of mannitol for 1.0 to 5.0 hours to obtain a concentration of bortezomib in the resulting solution of 1.0-2.5 mg / ml at 15-30 ° C. while maintaining the pH of the solution in a pH range from 5.0 to 6.5 and in an inert nitrogen gas atmosphere; (c) sterilizing filtration of the solution from step (b) using filtration under vacuum or filtration under a pressure of 0.1-0.6 MPa and dosing it into vials; (d) freeze-drying the product from step (c) in a freeze-drying chamber in several stages for 24-72 hours, wherein - first carry out primary drying in the temperature range (-30 ° C) - (-20 ° C), increasing the temperature on the shelf in the chamber every 30-60 minutes by 0.5-5.0 ° C / h, thereby cycles of a gradual rise in temperature and holding at a given temperature until a temperature is reached (-5 ° C) - (-2 ° C), - then carry out exposure at this temperature for 0.5-5.0 hours, then set the drying temperature in the range from 0 to (-1 ° C), maintain at this temperature for 5-30 hours to remove about 90 wt.% water from the product; - then carry out a gradual increase in temperature with a rise speed of 0.5-5.0 ° C / h until the temperature of the shelves of the freeze-drying chamber reaches 25-30 ° C for 5-25 hours and a vacuum of 1-150 m / Torr; (f) filling the freeze-drying chamber with inert gas, corking the bottles and filling them with caps. 2. The method of producing bortezomib lyophilisate according to claim 1, characterized in that the regulation and maintenance of the pH values of the solutions in stages (a) and (b) of the method, if necessary, is carried out by adding alkali metal hydroxide NaOH or KOH to achieve the desired pH value . 3. A method for producing a bortezomib lyophilisate according to claim 1, characterized in that the bortezomib free acid or its salt or its derivative is used as the substance of bortezomib. 4. A pharmaceutical composition containing bortezomib in the form of a stable lyophilized product obtained by the method according to claim 1 in therapeutically effective amounts and, as pharmaceutically acceptable carriers, it contains purified water or physiological saline. 5. The pharmaceutical composition according to claim 4, characterized in that it further comprises auxiliary target additives, such as buffers, antioxidants, preservatives, as well as their acceptable mixture.