RU2614234C2 - PHARMACEUTICAL COMPOSITION BASED ON 3-(4-METHYLIMIDAZOLE-1-YL)IMIDAZO[1,2-b][1,2,4,5]TETRAZINE AS ANTI-TUMOR AGENT - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry and includes pharmaceutical composition, which is solution of pharmaceutically acceptable substance 3-(4-methylimidazole-1-yl)imidazo[1,2-b][1,2,4,5]tetrazine of formula (I) and D-mannitol in water. Composition is used for therapy of malignant tumors in human or animal.

EFFECT: technical result is obtaining stable, long-term stored solution of Substance 1.

1 cl

Description

FIELD OF THE INVENTION

The invention relates to the pharmaceutical industry and includes a pharmaceutical composition comprising a solution of a pharmaceutically acceptable compound 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine of the formula (I ) and D-mannitol in water. The composition is used for the treatment of malignant tumors of humans or animals.

State of the art

According to the World Health Organization, cancer incidence is steadily increasing. Methods of treating cancer are diverse and depend on a number of factors, such as the type, localization and degree of development of the disease, the general condition of the patient. The action of chemotherapeutic drugs is aimed either at killing or displacing cancer cells, or at depriving cells of cell division signals, which ultimately leads to cell death. Currently, for the treatment of cancer, an alkylating antitumor agent Temozolomide - 3,4-dihydro-3-methyl-4-oxoimidazo [5,1-d] [1,2,3,5] tetrazine-8-carboxamide is used, which is heterocyclic an analogue of the drug “imidazotetrazine” being developed. Temozolomide synthesized in the UK in 1984 (Wang Y., Stevens MFG. Synthetic studies of 8-carbamoylimidazo- [5,1-d] -1,2,3,5-tetrazin-4 (3H) -one: A key derivative of antitumour drug temozolomide. Bioorg Med Chem Lett. 1996; 6 (2): 185-188, Patent US 0286088 dated 10.11.2010), has been used in Russia for the past 10 years for the treatment of malignant gliomas. It belongs to the group of alkylating agents of the 2nd generation - imidazotetrazines. After oral administration, it is rapidly absorbed and enters the bloodstream unchanged. Being a small lipophilic molecule, it easily penetrates the blood-brain barrier and accumulates in the tumor tissues in concentrations sufficient to realize its antitumor activity. When it enters the human body, it undergoes spontaneous hydrolysis (without the participation of any enzyme systems), turning into the active metabolite MTIC, and then into the methyldiazonium ion, which alkylates the DNA of tumor cells. Metal groups, joining certain sections of DNA, violate its structure. Ultimately, this leads to numerous DNA breaks, which the cell system of repair is not able to restore, since it is also suppressed by temozolomide. In addition, temozolomide has the ability to inhibit the activity of a number of enzymes responsible for DNA repair in a tumor cell. This is one of the most important mechanisms of action of this drug, which makes it possible to overcome the chemoresistance of tumor cells, which develops quite quickly during treatment with other alkylating agents and, first of all, drugs from the nitrosourea group.

The disadvantage of temozolomide is its extremely low solubility in common solvents such as ethyl acetate, dichloromethane, water, a mixture of organic solvent and water, and in DMSO. Another disadvantage is the instability of the drug, manifested in its sensitivity to light, alkaline media with a pH value of more than 7, and media that contain nucleophilic groups. In addition, the synthesis of temozolomide includes a number of time-consuming and energy-intensive stages, which leads to a high market value of the drug.

An analogue of temozolomide is 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine. Its advantage over temozolomide is a higher cytotoxicity for malignant lymphocyte cultures and a higher complexation constant with an intracellular target, double-stranded DNA. The synthesis of 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine is described in (Izv. AN Ser. Chem., 2011. 961-967). The antitumor use of 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine is described in the patent of the Russian Federation No. 2527258 C1 dated 03/09/2011, but specific compositions are not given. pharmaceutical compositions. The novelty lies in the fact that for the first time we proposed the composition, which allows us to overcome the poor solubility of this substance in water and saline, which previously did not allow its use in the form of injection forms for the treatment of tumors.

Disclosure of invention

This invention relates to a pharmaceutical composition comprising a solution of a pharmaceutically acceptable compound 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine of formula (I) and D-mannitol in water, from which a pharmaceutical preparation can be easily prepared by dilution with aqueous solutions, for example, physiological saline.

The formulation of the pharmaceutical composition of the present invention is a liquid, which, in particular, is an injectable solution, with particular preference for an injection solution (for example, for intravenous, subcutaneous, intramuscular, etc. injections) and intravenous fluids.

The pharmaceutical composition of the present invention can be used in the form of a liquid preparation for the treatment of malignant tumors.

The qualitative component composition of the created dosage form of the injectable drug was selected using the following substances:

1) 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine - active substance;

2) Acetonitrile - the main solvent;

3) D-Mannitol - co-solvent and solution stabilizer;

4) Water for injection is a solvent.

To obtain stable, long-lasting solutions of this component composition, we have developed a method for producing a stable pharmaceutical composition based on 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine.

According to this method, a working solution was prepared containing the compound 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine, acetonitrile, D-Mannitol, introduced into the pharmaceutical composition in as a solubilizer and stabilizer of the solution and water for injection. The concentration of D-mannitol (up to 1%) was selected in an amount sufficient to improve the dissolution of the drug substance and maintain the stability of the final solution over time.

A working solution was prepared from these components. Then acetonitrile was removed from the working solution and the mother liquor was prepared.

Acetonitrile is mixed with water and other solvents; this substance is moderately toxic (toxicity class 3), and although its estimated amount in the final solution is insignificant (up to 1%), the presence of acetonitrile in the injection pharmaceutical composition is undesirable. In this regard, to remove acetonitrile, the resulting working solution was evaporated on a rotary evaporator with vacuum suction at a temperature of +35 - (+ 40) ° С for 40 min and a mother liquor with a known active substance content was obtained. Thus, acetonitrile was excluded from the composition of the mother liquor.

To prepare injectable LF, a measured amount of the mother liquor is adjusted with water for injection to a predetermined concentration.

The composition of the mother liquor

The mother liquor of an injectable pharmaceutical composition based on 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine is a three-component system, with components taken in the following weight ratios (table one):

An example of a method of obtaining a pharmaceutical composition

Materials:

1. 3- (4-Methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine - 20 mg

2. Acetonitrile 200 μl

3. An aqueous solution of D-Mannitol - 5 ml of a 1% solution

Rotary Evaporator: Heildoph, Laborota- 400 effiicient

The laboratory technology for the production of an injectable pharmaceutical composition based on 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine consists of four stages:

The first stage is the preparation of an auxiliary (aqueous solution of D-Mannitol) and a working solution,

The second stage is the preparation of the mother liquor,

The third stage is the preparation of an injection solution,

The fourth stage is sterilization.

The working solution was prepared at room temperature according to the following algorithm.

1) Preparation of an auxiliary 1% solution of D-Mannitol in water for injection.

A portion of D-Mannitol was placed in a conical flask and the amount of water necessary for the formation of a 1% solution was poured.

Stirred until complete dissolution and the formation of a clear yellow solution (1%).

2) A portion of 20 mg of imidazotetrazine in a sterile 20 ml vial was dissolved in 200 μl of acetonitrile. A clear yellow solution forms.

3) 5 ml of D-Mannitol solution was fractionally added to the resulting solution. A clear yellow solution forms.

The resulting working solution was evaporated on a rotary evaporator with vacuum suction at a temperature of +35 - (+ 40) ° C for 40 min and a clear yellow mother liquor was obtained with a content of 20 mg of active ingredient (4 mg / ml).

The resulting mother liquor was stored in a sealed container at low temperatures (+2 - (+ 8) ° C).

Injection preparation

The full volume of the mother liquor was transferred to a 20 ml volumetric flask. 10 ml of water for injection was added and thoroughly mixed without foaming. The solution volume was adjusted to the mark with water for injection. The resulting dosage form is a transparent 0.1% yellow NIK-1277 solution. Sterile vials containing a solution of the injectable pharmaceutical composition 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine were exposed to gamma radiation for 4 hours at 35 kG .

Toxicometric Characterization of Injectable Pharmaceutical Composition 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine. Assessment of acute toxic effects was carried out on white mice of the Balb / c line - males weighing 20-24 grams, obtained from the Central nursery RAMS branch "Stolbovaya".

Clinically healthy animals were injected intravenously with a solution of the injectable pharmaceutical composition 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine in doses of 5 mg / kg (n = 3) 15 mg / kg (n = 3), 25 mg / kg (n = 3). No toxic effects were observed. At a dose of 50 mg / kg (n = 3), one animal died.

Novelty

The novelty lies in the fact that we first proposed the composition to overcome the poor solubility of 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine in water and saline, which previously did not allow its use in the form of injectable forms for the treatment of tumors.

Claims (2)

A pharmaceutical composition for the treatment of malignant tumors containing, as an active component, a pharmaceutically acceptable 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine and two excipients - water and D-mannitol, in the following mass ratio of components: 3- (4-methylimidazol-1-yl) imidazo [1,2-b] [1,2,4,5] tetrazine one D-mannitol 0.005 water 500