EA031596B1 - Anti-tumour pharmaceutical composition comprising temozolomide, and method for producing the same - Google Patents

Abstract

The invention is related to the field of pharmaceutics, in particular, to an anti-tumour pharmaceutical composition comprising temozolomide, and can be used in medicine, in particular, in oncology. The objective of the invention is provision of an anti-tumour pharmaceutical composition comprising temozolomide that features high efficiency, improved stability, provides high accessibility of tumour cells for the anti-tumour agent, and development of a method for producing such composition. Said objective is attained by provision of a sustained-action anti-tumour pharmaceutical composition for treatment of oncological diseases including temozolomide and pharmaceutically acceptable excipients, characterized in that the composition contains, as pharmaceutically acceptable excipients, polysaccharide phosphates in salt and acidic forms with the following contents of the components, wt.%: temozolomide - 5-20, polysaccharide phosphate - 30-80, polysaccharide phosphates in the acidic form - up to 100.

Description

The invention relates to the field of pharmaceuticals, specifically an antitumor pharmaceutical composition containing temozolomide, and can be used in medicine, in particular in oncology. The objective of the invention is the creation of an antitumor pharmaceutical composition containing temozolomide, which is characterized by high efficiency, has improved stability, provides high accessibility of the antitumor agent to tumor cells, and the development of a method for its preparation. The problem is solved in that the antitumor pharmaceutical composition of prolonged action for the treatment of cancer, containing temozolomide and pharmaceutically acceptable carriers, is characterized in that as a pharmaceutically acceptable carrier, the composition contains polysaccharide phosphates in salt and acid forms with the following components, wt.%: temozolomide - 5-20, polysaccharide phosphate - 30-80, polysaccharide phosphate in acid form - up to 100.

The invention relates to the field of pharmaceuticals, specifically an antitumor pharmaceutical composition containing temozolomide, and can be used in medicine, in particular in oncology.

Temozolomide, 4-methyl-5-oxo-2,3,4,6,8-pentazabicyclo [4.3.0] nona-2,7,9-triene-9-carboxamide of the formula

It is an antitumor alkylating agent and, in the form of oral dosage forms (capsules), is used in the treatment of malignant gliomas of the brain and melanoma and other oncological diseases. Temozolomide penetrates well through the blood-brain barrier, but is characterized by a very short half-life: when administered orally, temozolomide is rapidly absorbed, its maximum plasma concentration is reached after 0.7 hours, and the half-life is 1.8 hours. To maintain an effective concentration of temozolomide, for example, repeated administration of the drug is required in the brain, which is associated with inconvenience for patients and causes a risk of toxic effects.

To overcome these shortcomings, a method has been proposed for the combined treatment of malignant brain tumors [1]. The method includes the surgical removal of tumor tissue, followed by the use of temozolomide incorporated into a gel of highly substituted dextran phosphate. Sterile water is added to the freeze-dried gel, the pH is adjusted to 7.2-7.4, and the resulting mass is introduced into the bed of the removed tumor.

However, adjusting the pH in the operating room does not allow for accuracy, is time-consuming. In addition, as was established by the authors, at a pH of 7.2-7.4, temozolomide decomposes with the formation of dacarbazine impurities A in quantities exceeding permissible.

To implement this and similar methods, it is necessary to create a dosage form that has the necessary not only pharmacological properties, but also corresponds to modern pharmaceutical requirements. Such a composition should be stable during storage, be sterile, have reproducible and marketable technology.

An example of such a composition is a prolonged form of temozolomide in the form of implantable tablets comprising 3-10% temozolomide and a biodegradable polymer material from the number of polyethylene, polypropylene, polyethylene terephthalate, plasticized polyvinyl chloride, crosslinked polyester, polycarbonate, polysulfone, polystyrene, poly (2-pentene) , poly (1,4phenylene), polytetrafluoroethylene and polyanhydrides (mainly polyanhydride obtained by condensation of 3,4-bisf-carboxyphenoxy) propane with sebacic acid in their ratio 20:80 respectively [2]. Upon implantation in the brain of rats, the release of temozolomide from the implant begins after 1 h and lasts for 10 days. Moreover, more than 70 wt.% Temozolomide is released in the first day.

The preparation of such implantable tablets is difficult, requires the use of highly toxic solvents such as chloroform, ethyl acetate, acetone, dichloromethane at the stage of preparation of the polymer material solution. Subsequent spray drying and evaporation of the residual solvent under vacuum cannot guarantee complete removal to an acceptable and safe concentration for the patient. For chloroform, for example, this concentration is very low - not more than 0.006% or 60 ppm. A significant drawback of this drug is the form of the implant in the form of a hard tablet, which can lead to additional trauma to the surgical wound and does not allow the penetration of the cytostatic agent to all remaining tumor cells.

The objective of the invention is the creation of an antitumor pharmaceutical composition containing temozolomide, which is characterized by high efficiency, has improved stability, provides high accessibility of the antitumor agent to tumor cells, and the development of a method for its preparation.

The problem is solved in that the antitumor pharmaceutical composition of prolonged action for the treatment of cancer, containing temozolomide and pharmaceutically acceptable carriers, is characterized in that the composition contains polysaccharide phosphates in salt and acid forms as pharmaceutically acceptable carriers.

Polysaccharides are natural macromolecular compounds and their macromolecules do not contain structural units that could have an undesirable or toxic effect on the body.

It is known that temozolomide stability is achieved at pH 4.5-7.0. However, to achieve this pH range, alkaline, acidic, or buffer solutions are required, which leads to a change in the solubility of temozolomide.

- 1 031596

In the process of research, the authors unexpectedly found out that it is possible to obtain a stable pharmaceutical composition containing temozolomide using a mixture of polysaccharide phosphates in salt and acid forms. Such a composition provides the dissolution of temozolomide in the right amount and maintaining the pH in the right range, ensuring the stability of temozolomide.

The preferred composition is: from 5 to 20 wt.% Temozolomide, from 30 to 80 wt.% Phosphate polysaccharides in salt form and phosphate polysaccharides in acid form to 100 wt.%.

As polysaccharides, dextran, starch, cellulose, and mixtures thereof can be used.

Most preferred is dextran, in particular dextran with a molecular weight of from 40 to 100 kDa, more preferably from 60 to 70 kDa.

The salt form of polysaccharide phosphates is selected from the group consisting of sodium, potassium, magnesium and / or calcium, sodium being most preferred.

According to a preferred aspect of the invention, the weight ratio of temozolomide to polysaccharide phosphates is from 1: 4 to 1:19, most preferably 1: 9.

According to another preferred aspect of the invention, the weight ratio of polysaccharide phosphates in salt form to polysaccharide phosphates in acid form is from 15:85 to 70:30, most preferably 55:35.

According to the present invention also claims a hydrogel for intrathecal use, formed from the claimed composition.

The pharmaceutical composition can be made in any of the pharmaceutically acceptable dosage forms for topical, implantation and oral administration with the addition of any pharmaceutically acceptable excipients: powder, napkin, granules, capsules, tablets, ready-to-use gel, suspension, emulsion, gel powder immediately before use.

An advantageous embodiment of the hydrogel is the mixing of the pharmaceutical composition with sterile water in a ratio of from 1:10 to 1:30, most preferably 1:15.

The problem is also solved by the fact that in accordance with one of the embodiments of the claimed pharmaceutical composition is obtained by a method comprising the formation of a hydrogel by adding water to a mixture of polysaccharide phosphates in salt form and polysaccharide phosphates in acid form;

the introduction of powdered temozolomide into the hydrogel of polysaccharide phosphates in salt form and polysaccharide phosphates in acid form at the following component ratios: from 5 to 20 wt.% temozolomide, from 30 to 80 wt.% polysaccharide phosphates in salt form, the remaining part to 100 wt.% - phosphate polysaccharides in acidic form;

freeze drying the gel to obtain a lyophilisate;

sterilization of powdered lyophilisate by radiation method [3].

In accordance with another embodiment of the claimed pharmaceutical composition is obtained by a method comprising preparing an aqueous solution of temozolomide;

gel preparation by introducing into the solution of temozolomide a dry mixture of polysaccharide phosphates in salt form and polysaccharide phosphates in acid form in the following ratios: from 5 to 20 wt.% temozolomide, from 30 to 80 wt.% polysaccharide phosphates in salt form, the remaining part to 100 wt.% - phosphates of polysaccharides in acidic form;

freeze drying the gel to obtain a lyophilisate;

sterilization of powdered lyophilisate by radiation method.

In accordance with a third embodiment, the inventive pharmaceutical composition is prepared by a process comprising mixing a mixture of polysaccharide phosphates in salt form and polysaccharide phosphates in powder form and powdered temozolomide in the following proportions: from 5 to 20 wt.% Temozolomide, from 30 to 80 wt.% Polysaccharide phosphates in salt form, the remaining part up to 100 wt.% - phosphates of polysaccharides in acid form;

sterilization of powdered lyophilisate by radiation method.

The use of a pharmaceutical composition and a hydrogel formed upon contact of the composition with sterile water or another pharmacologically acceptable liquid (for example, saline or phosphate buffer) is also claimed in the treatment of cancer, in particular intracerebral tumors of grade 2-4, such as oligoastrocytoma, anaplastic astrocytoma , glioblastoma, etc.

The maximum content of temozolomide in the claimed pharmaceutical composition (20 wt.%) Is determined by the solubility of temozolomide in water or another pharmacologically acceptable solvent, added to prepare the implant in the form of a gel.

The lower limit on the content of temozolomide (5 wt.%) Is due to the minimum effectiveness of the claimed pharmaceutical composition.

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The claimed limits on the content in the claimed pharmaceutical composition of polysaccharide phosphate in salt form (from 30 to 80 wt.%) And polysaccharide phosphate in acid form (up to 100 wt.%) Are determined by the increase in the content of 5-aminoimidazole-4-carboxamide hydrochloride (USP Decarbazine Related Compaund A or dacarbazine impurity A), normalized within 0.5 wt.% For the registered medicinal product (temozolomide) [4]. In the claimed pharmaceutical composition, the content of dacarbazine impurity A is less than 0.5 wt.% Up to an amount of 0.01 wt.% Or even an amount not detected by high performance liquid chromatography (HPLC).

The mass ratios of the components of the claimed pharmaceutical composition are selected experimentally, mainly according to the effectiveness of the antitumor effect and storage stability in terms of: Quantitative content of temozolomide, Dacarbazine impurity A, Water absorption, Sterility and are shown in the table.

The composition of the claimed antitumor pharmaceutical composition and storage stability

No. Polysaccharide Saline the form The composition of the pharmaceutical composition, May. % Quality indicators at the time of manufacture Quality indicators after 2.5 years of storage temozo- lomid polysaccharide phosphate in salt form acid polysaccharide phosphate Contents, May. % water- absorption g / g sterile nost Contents, May. % water- absorption g / g sterile nost temozolo mid dacarbazine impurity A temozolomide dacarbazine impurity A one dextran sodium 10 54 36 10 not detected 27 erased. 10 not detected 27 erased. 2 cellulose sodium twenty 56 24 twenty 0.12 28 erased. twenty 0.13 28 erased. 3 dextran magnesium 5 76 24 5 0.18 21 erased. 5 0.18 21 erased. four dextran potassium 10 54 36 10 not detected 27 erased. 10 not detected 27 erased. 5 starch sodium 10 54 36 10 0.28 34 erased. 10 0.28 34 erased. 6 dextran sodium 5 80 fifteen 5 0.22 34 erased. 5 34 erased. 7 dextran calcium 10 54.8 35,2 10 not detected twenty erased. 10 not detected twenty erased. 8 dextran sodium twenty thirty fifty twenty 0.09 24 erased. twenty 0.09 24 erased. 9 starch potassium 5 64.5 30.5 5 0.15 28 erased. 5 0.15 28 erased. 10 starch sodium 10 68.8 21,2 10 0.28 35 erased. 10 0.28 35 erased. eleven dextran sodium 10 64.1 25.9 10 0.08 32 erased. 10 0.08 32 erased.

The invention is illustrated in FIG. 1, 2, where in FIG. 1 shows the postoperative survival of patients with malignant gliomas of the brain after total tumor removal without (control group) and with subsequent implantation (main group) of the claimed antitumor pharmaceutical composition. In FIG. 2 presents the duration of the disease-free period in patients with malignant gliomas of the brain after total removal of the tumor without (control group) and with subsequent implantation (main group) of the claimed antitumor pharmaceutical composition.

Examples of specific performance confirm the possibility of carrying out the invention with a positive effect when using the entire combination of essential features of the invention indicated in its formula.

Example 1. 100.0 g of dextran phosphate obtained by any known method are placed in a reaction vessel, 5 l of distilled water are poured and stirred until a homogeneous suspension is formed. A 10% sodium hydroxide solution was added in portions to the resulting aqueous suspension of dextran phosphate, after adding each portion, the reaction mixture was stirred for 5 minutes and the pH was measured. The addition of a 10% sodium hydroxide solution is completed when the pH of the reaction medium is in the range of 6.0-7.0. The reaction mixture is left for 30 min at room temperature and freeze-dried. The content of dextran phosphate in sodium form and dextran phosphate in acid form in the lyophilisate are 60 and 40 wt.%, Respectively.

10 g of temozolomide are added to 90 g of the lyophilized dried powder obtained from a mixture of dextran phosphate in salt and acid forms, the dry mixture is thoroughly mixed, packaged in 1 g in any suitable sealed package and sterilized by radiation. The resulting pharmaceutical composition contains temozolomide - 10 wt.%, Dextran phosphate in salt form - 54 wt.% And dextran phosphate in acid form - 36 wt.%.

Example 2. 80.0 g of cellulose phosphate obtained by any known method are placed in a reaction vessel, 5 l of distilled water are poured and stirred until a homogeneous suspension is formed. A 10% sodium hydroxide solution is added in portions to the resulting aqueous suspension of cellulose phosphate, and after adding each portion, the reaction mixture is stirred for 5 minutes and the pH is measured. The addition of a 10% sodium hydroxide solution is completed when the pH of the reaction medium is in the range of 6.0-7.0. The reaction mixture is left for 30 minutes at room temperature, mixed thoroughly and freeze-dried. The content of cellulose phosphate in sodium form and cellulose phosphate in acid form in the lyophilisate are 70 and 30 wt.%, Respectively.

To 80 g of the obtained freeze-dried mass from a mixture of cellulose phosphate in saline and acidic

- 031596 molds add 20 g of temozolomide, the dry mixture is thoroughly mixed, placed in molding molds, pressed, the resulting wipes are placed in plastic bags, hermetically sealed and sterilized by radiation. The resulting pharmaceutical composition contains temozolomide - 20 wt.%, Dextran phosphate in salt form - 56 wt.% And dextran phosphate in acid form - 24 wt.%.

Example 3. In a reaction vessel, 100 g of dextran phosphate obtained by any known method are placed, 5 l of distilled water are added, and stirred until a homogeneous suspension is formed. A 10% solution of magnesium hydroxide is added in portions to the resulting aqueous suspension of dextran phosphate, after adding each portion, the reaction mixture is stirred for 5 minutes and the pH is measured. The addition of a 10% solution of magnesium hydroxide is completed when the pH of the reaction medium is in the range of 6.0-7.0. The reaction mixture is left for 30 min at room temperature and freeze-dried. The content of dextran phosphate in magnesium form and dextran phosphate in acid form in the lyophilisate are 80 and 20 wt.%, Respectively.

To 95 g of the lyophilized powder obtained from a mixture of dextran phosphate in salt and acid forms, 5 g of temozolomide is added, the dry mixture is thoroughly mixed, packaged in 1 g in any suitable sealed package and sterilized by radiation. The resulting pharmaceutical composition contains temozolomide - 5 wt.%, Dextran phosphate in salt form 76 wt.% And dextran phosphate in acid form - 19 wt.%.

Example 4. 10 g of temozolomide is dissolved in 3 l of sterile distilled water and 90 g of a mixture containing dextran phosphate in salt form — 60 wt.% And dextran phosphate in acid form — 4% wt. Are added. Stirred until a homogeneous mixture, freeze-dried, the dry mixture is thoroughly mixed, packaged in 1 g in any suitable sealed packaging and sterilized by radiation. The resulting pharmaceutical composition contains temozolomide - 10 wt.%, Dextran phosphate in salt form - 54 wt.%, Dextran phosphate in acid form 36 wt.%.

Example 5. The pharmaceutical composition is prepared analogously to example 1, except that a mixture of starch phosphate in salt form and starch phosphate in acid form is used. The composition of the claimed pharmaceutical composition is shown in the table.

The content of temozolomide and dacarbazine impurity A is determined by HPLC. For the test, a solution is prepared as follows: 0.25 g of the pharmaceutical composition to be tested is added to a 25 ml volumetric flask, 20 ml of dimethyl sulfoxide are added, sonicated for 15 minutes and the volume is adjusted to the mark with the same solvent. The resulting solution was filtered through a membrane filter with a pore size of 0.45 μm. The solution is used freshly prepared.

Chromatography conditions:

a liquid chromatograph equipped with a spectrophotometric detector installed at a wavelength of 254 nm;

stainless steel column 250 * 4.6 mm in size, filled with silica gel octadecylsilyl for chromatography, with a particle size of 5 μm, for example, Nucleodur C18 Gravity.

The speed of the mobile phase is 1 ml / min. The volume of the injected sample is 10 μl.

Mobile phase: 0.846 g / l aqueous solution of sodium hexanesulfonate: methanol: glacial acetic acid in a ratio of 895.5: 100: 4.5.

The sterility of the claimed pharmaceutical composition was determined by standard pharmacopoeial methods.

To determine water absorption, a weighed portion of the claimed pharmaceutical composition was placed in about 50 ml of distilled water and kept for 1 hour. The hydrogel was separated from the excess liquid on a glass filter and placed in a previously weighed box, weighed and dried in a vacuum oven over phosphorus oxide ( V) at a temperature of 323 K to constant weight. The water absorption capacity of the claimed pharmaceutical composition was calculated from the difference in mass of the swollen and dried sample.

The pharmaceutical composition obtained by the method described in any one of examples 1-5 can be used for intrathecal administration, for topical administration, or for the manufacture of other dosage forms for oral administration. The manufacture of oral dosage forms is illustrated by examples 6-8.

Example 6. The pharmaceutical composition obtained as described in any of examples 1-5, in an amount of about 50 mg (calculated on the content of 5 mg of temozolomide) is mixed with excipients lactose - 90 mg, sodium carboxymethyl starch - 7.5 mg, silicon colloidal dioxide 0.2 mg, tartaric acid - 3.0 mg. The resulting mixture is subjected to dry or wet granulation. The resulting granulate is packaged in hard gelatin capsules. The drug is intended for oral administration in a dose of 5 mg of temozolomide.

Example 7. The pharmaceutical composition obtained as described in any of examples 1-5 in an amount of about 500 mg (calculated on the content of 50 mg of temozolomide) is mixed with excipients lactose - 55 mg, sodium carboxymethyl starch - 15 mg, tartaric acid - 10 mg, stearic

- 4 031596 new acid - 14 mg. The resulting mixture is subjected to dry or wet granulation, the resulting granulate is sent to pressing. Get tablets for oral administration with a content of temozolomide 50 mg.

Example 8. The pharmaceutical composition obtained, as described in any of examples 1-5, in the form of a gel in an amount of about 2500 mg (calculated on the content of 250 mg of temozolomide) is mixed with auxiliary substances lactose - 180 mg, sodium carboxymethyl starch - 28 mg, tartaric acid 20 mg and subjected to wet granulation. The granules are dusted with calcium (or magnesium) stearate and packaged in single dose packets. The obtained granulate is used for oral administration in a single dose of 250 mg of temozolomide.

Evaluation of the effectiveness of the claimed antitumor pharmaceutical composition was carried out under clinical conditions in 41 patients (experimental group) with malignant brain tumors Grade II-IV. To 1 g of the claimed pharmaceutical composition containing temozolomide - 10 wt.%, Dextran phosphate in sodium form - 54 wt.% And dextran phosphate in acid form - 36 wt.%, 15 ml of sterile water was added under operating conditions, mixed and left on 20-30 min until a homogeneous dense mass is formed. The resulting mass was implanted in the bed of the removed tumor.

The results presented in FIG. 1 and 2, indicate that the use of the claimed antitumor pharmaceutical composition leads to a significant increase in the average life expectancy of patients with malignant brain tumors and reduces the risk of relapse.

As follows from the table, the claimed pharmaceutical composition in terms of quantitative content of temozolomide, dacarbazine impurity A, water absorption capacity, sterility is stable for 2.5 years of storage.

Information sources.

1. Patent BY 11838, A61K 31/4162, A61K 31/715, publ. 04/30/2009.

2. Patent RU 2322979, A61K 31/4188, A61K 47/30, A61K 9/22, A61P 35/00, publ. 04/27/2008.

3. The State Pharmacopoeia of the Republic of Belarus. - Publishing House: Ministry of Health of the Republic of Belarus, 2006, v. 1, p.671.

4. USP. Draft USP Pending Monograph. Temozolomide.

Claims (14)

CLAIM 1. Antitumor pharmaceutical composition containing temozolomide as an active pharmaceutical ingredient and a pharmaceutically acceptable carrier, characterized in that as the pharmaceutically acceptable carrier the composition contains polysaccharide phosphate and polysaccharide phosphate in acid form. 2. The pharmaceutical composition according to claim 1, characterized in that it contains temozolomide and polysaccharide phosphates in the following amounts: temozolomide - from 5 to 20 wt.%, Polysaccharide phosphate - from 30 to 80 wt.%; polysaccharide phosphate in acid form - the rest up to 100 wt.%. 3. The pharmaceutical composition according to claims 1, 2, characterized in that the phosphates of dextran, starch, cellulose and / or their mixtures are used as polysaccharide phosphates. 4. The pharmaceutical composition according to claims 1 to 3, characterized in that dextran with a molecular weight of from 40 to 80 kDa is used as a polysaccharide. 5. The pharmaceutical composition according to claims 1 to 3, characterized in that the polysaccharide phosphates are sodium, potassium, magnesium, calcium salt and / or their mixture. 6. The pharmaceutical composition according to claims 1-5, characterized in that the mass ratio of temozolomide and the total phosphate content of the polysaccharide is from 1: 4 to 1:19. 7. The pharmaceutical composition according to claims 1-6, characterized in that the mass ratio of temozolomide and the total phosphate content of the polysaccharide is 1: 9. 8. The pharmaceutical composition according to claims 1-7, characterized in that the mass ratio of polysaccharide phosphate and polysaccharide phosphate in acid form is from 30:50 to 80:15. 9. The pharmaceutical composition according to claims 1-8, characterized in that the mass ratio of polysaccharide phosphate and polysaccharide phosphate in acid form is 55:35. 10. A method of obtaining a pharmaceutical composition according to claims 1-9 from a pharmaceutical substance temozolomide and an auxiliary substance, which is a mixture of polysaccharide phosphate and polysaccharide phosphate in acid form, comprising mixing a mixture of polysaccharide phosphate and polysaccharide phosphate in acid form and temozolomide powder and sterilizing the powder composition . 11. A method for preparing a pharmaceutical composition according to claims 1-9 from a pharmaceutical substance temozolomide and an excipient, which is a mixture of polysaccharide phosphate and polysaccharide phosphate in acid form, comprising forming a hydrogel from a mixture of polysaccharide and polysaccharide phosphate in acid form when water is added; introduction of temozolomide powder into a hydrogel from a mixture of polysaccharide phosphate and polysaccharide phosphate in an acid form with - 5 031596 blowing ratios: from 5 to 20 wt.% Temozolomide, from 30 to 80 wt.% Polysaccharide phosphate, the rest up to 100 wt.% Polysaccharide phosphate in acid form; freeze drying the gel to obtain a lyophilisate; sterilization powdered lyophilisate. 12. A method of obtaining a pharmaceutical composition according to claims 1 to 9 from a pharmaceutical substance temozolomide and an excipient, which is a mixture of polysaccharide phosphate and polysaccharide phosphate in an acidic form, comprising preparing an aqueous solution of temozolomide; preparation of the gel by introducing into the solution of temozolomide a dry mixture of polysaccharide phosphate and polysaccharide phosphate in an acidic form in the following ratios: from 5 to 20 wt.% temozolomide, from 30 to 80 wt.% polysaccharide phosphate, the remaining part to 100 wt.% polysaccharide phosphate sour form; freeze drying the gel to obtain a lyophilisate; sterilization powdered lyophilisate. 13. A method of obtaining a pharmaceutical composition according to claims 1-9 of a sterile pharmaceutical substance temozolomide and a sterile excipient, which is a mixture of polysaccharide phosphate and polysaccharide phosphate in an acidic form, by sterile sprinkling them into single-dose containers. 14. The use of the pharmaceutical composition according to claims 1-9 for the treatment of cancer. ABOUT