WO2008120070A2

WO2008120070A2 - 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) displaying hepatoprotective, hypoammoniemic and detoxic action, a method for its production and a pharmaceutical composition based thereon

Info

Publication number: WO2008120070A2
Application number: PCT/IB2008/000727
Authority: WO
Inventors: Ivan Ivanovych Novik
Original assignee: Aktsionerne Tovarystvo Vidkrytogo Typu <<galychfarm>>; Tovarystvo Z Obmezhenoyu Vidpovidalnistyu <<instytut Fito-Himichnyh Tehnologij>>; Vidkryte Aktsionerne Tovarystvo <<kyivmedpreparat>>
Current assignee: Aktsionerne Tovarystvo Vidkrytogo Typu <<galychfarm>>; Tovarystvo Z Obmezhenoyu Vidpovidalnistyu <<instytut Fito-Himichnyh Tehnologij>>; Vidkryte Aktsionerne Tovarystvo <<kyivmedpreparat>>
Priority date: 2007-03-30
Filing date: 2008-03-17
Publication date: 2008-10-09
Anticipated expiration: 2009-09-30
Also published as: WO2008120070A3; UA86441C2

Abstract

The invention relates to medicine and chemical and pharmaceutical industry, and more particularly to a new physiologically active substance 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) displaying hepatoprotective, hypoammoniemic and detoxic activity, to a method for producing said substance, and to pharmaceutical compositions based thereon. The technical result of implementing the invention is a new physiologically active chemical substance 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) displaying hepatoprotective, hypoammoniemic and detoxic activity, a method for producing the same which method ensures a higher level and a wider spectrum of specific activity while simplifying, and shortening the duration of the manufacturing process, and pharmaceutical compositions based on the new physiologically active substance 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) in various dosage forms the use whereof results in a diversified range of hepatoprotective drugs.

Description

2(HYDROXYETHYL)TRIMETHYLAMMONIUM THIOCTATE (CHOLINE

THIOCTATE) DISPLAYING HEPATOPROTECTIVE, HYPOAMMONIEMIC

AND DETOXIC ACTION, A METHOD FOR ITS PRODUCTION AND A

PHARMACEUTICAL COMPOSITION BASED THEREON

The invention relates generally to medicine and chemical and pharmaceutical industry, and more particularly to a new physiologically active substance 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) which displays hepatoprotective, hypoammoniemic and detoxic activity, to a method for its production and to pharmaceutical compositions based thereon.

The substance of the following structural formula

is a salt of choline base and thioctic acid solvable in water and some polar organic solvents and displays hepatoprotective, hypoammoniemic and detoxic activity.

Regarding hepatology, up to now, there has been remained significant the role of drugs which are used to treat liver and bile duct diseases and of those that protect the liver from a damaging influence of both exogenous and endogenous factors and/or accelerate its natural regeneration (hepatoprotectors). The group of hepatoprotectors includes substances of both vegetable and synthetic origins which exert a differently directed effect on metabolic processes.

The range of domestic drugs which display hepatoprotective, hypoammoniemic and detoxic activities is insufficient and, accordingly, what is relevant is the search for new physiologically active substances which possess such specific activity and the provision, based thereon, of new high-efficacy drugs.

A structural and pharmacological analog to choline thioctate is choline chloride - a choline derivative (2-oxyethyltrimethylammonium hydroxide), which is a lipotropic substance preventing severe liver diseases from occurring during its atheroma. Choline chloride is used in medicine included in hepatoprotective drugs to treat liver diseases. It is also included in combined feed for farm animals. Its disadvantages include an insufficiently high level and limited spectrum of specific activity.

At present, drugs of thioctic (α-lipoic) acid (for instance, Thioctacid) attract close attention to the treatment of liver metabolic diseases. It is well known that thioctic acid plays a substantial role in regulating metabolic processes in the liver. As a coenzyme, it takes part in carbohydrate or protein metabolism, and it is one of the important components of the antioxidant system of organism. Thioctic acid plays an important role in α-keto acid oxidative decarboxylation reactions including pyruvic acid transformation into acetyl-CoA which will enter the Krebs cycle. A complex oxidation process of thioctic acid results in the formation of 3 ATP molecules this increasing significantly a cell energy potential. Thioctic acid activates the oxidation processes of fatty acids in mitochondrions this, in addition provides the cell with energy, reducing the content of substrates therein for triglyceride synthesis and preventing thereby fatty (adipose) degeneration of the liver and hyperlipidemia from developing.

The main reasons for a wide use of thioctic acid to treat liver diseases are as follows:

• Ability of reducing lipid peroxidation (binding free radicals and free tissue iron);

• Participation in fatty acid and acetate oxidation (it prevents fatty steatosis of the liver from developing);

• Participation in α-keto acid decarboxylation (the provision of cell with energy and ketoacidosis development prevention); • Increase in glucose transmembrane transport to the cell (glycogen accumulation and increase in the energy balance of hepatocytes);

• Nitrogen oxide synthesis inhibition by hepatocytes (the prevention and reduction of disorders of the rheological properties and vascular disorders).

The study conducted has shown the prospects of providing a complex physiologically active substance based on choline base and thioctic acid, a further use of which, in particular, as hepatoprotector, improves substantially a patient's quality of life.

Known in the art are hepatoprotective drugs Legalon, Karsil, Siliborum in the form of tablets, capsules, pellets and others which contain as their active ingredients flavonoids extracted from seeds of variegated thistle. These drugs are used to treat acute hepatitis, hepatocirrhosis, in supporting therapy of chronic diseases of the liver. When using these drugs, undesirable side effects in the form of laxative action, allergic reactions of skin tissues are reported. (1)

A known in the art hepatoprotective drug Essentialle contains natural "essential" phospholipids - diglyceride esters of cholinephosphoric acid and unsaturated fatty acids (linolic, linoleic, and others) along with vitamins (pyridoxine, cyanocobalamin, nicotinamide, pantothenic acid). Essentialle is used in the form of 5-mL and 10-mL ampoules which contain respectively 250 mg and 1000 mg of "essential" phospholipids, 2.5 mg and 5 mg of pyridoxine hydrochloride, 10 μg and 15 μg of cyanocobalamin, 25 mg and 100 mg of nicotinamide, 1.5 mg and 3 mg of sodium pantothenate, as well as in the form of capsules which contain 175 mg of "essential" phospholipids, 3 mg of thiamine, 3 mg of riboflavin, 3 mg of pyridoxine hydrochloride, 3 mg of cyanocobalamin, 15 mg of nicotinamide and 3.3 mg of alpha- tocopherol acetate. The drug is useful to treat chronic hepatitis, degeneration and cirrhosis of the liver, as well as to treat liver involvement associated with diabetes, alcoholism and others. Undesirable side effects in the form of dyspeptic disorders are reported. (2)

Known in the art is a hepatoprotective drug, which contains as its active ingredient 2-phenyl-3-carbetoxy-4-dimethylaminomethyl-5-oxibenzofuran hydrochloride (vinboron, phenicaberan) and helps to preserve the liver ravine structure. (3)

Known in the art is a hepatoprotective drug Hepabene in the form capsules, which is a combined drug and contains extracts from medicinal herbs including variegated thistle with at least 22 mg of silibinin per capsule (4).

Known in the art is a hepatoprotective drug Hepabene in the form of infusion solution, which contains a 5% solution of amino acids, which are the most suitable for the organism including arginine (5).

Known in the art is a drug Zixorin, which is used in the form of capsules to treat functional hyperbilirubinemia in patients with chronic diffuse diseases of the liver, to treat jaundice caused by enzymopathies and benign intrahepatic cholestasis (6).

Known in the art is a drag Hepatofalk, which contains choline orotate as its active ingredient. It is used to treat toxic liver disorders (including alcoholic and drug disorders), acute and chronic hepatitis of various ethiology and hepatocirrhosis as a supporting therapy (7).

Known in the art is a drag Lipovitan which contains choline tartrate as its active ingredient and is used to treat toxic liver disorders (8).

Known in the art is a drag Glutarginum (L-arginine L-glutamate) which displays hepatoprotective, hypoammoniemic and detoxic activity and is used in the form of tablets, capsules, powders, injection solutions and infusion solution concentrates (9).

Known in the art is a method for producing arginine amide derivatives, the method comprising: activating N-acylarginine or a salt thereof; the acylation thereby of amines of formula NH₂R in an organic solvent medium at the presence of a base; and liberating the target product, wherein dimethyl formamidinium chloride is used as the activating agent (10).

Known in the art is a method for producing 6-aminocapronic acid glutamate, a substance which displays antifibrinolytic, anti-inflammatory and antimicrobial activity, the method comprising: dissolving glutamic acid and 6-aminocapronic acid in water; adding an activated charcoal to the resulting solution while stirring; filtering the resulting solution of 6-aminocapronic acid glutamate salt; cooling the filtrate for the purpose of further isolating a precipitate; and filtering, washing and drying the resulting crystalline precipitate - 6-aminocapronic acid glutamate salt. A target product yield is 86% (11).

The most similar to a method in accordance with the present invention is the method for producing a water-soluble salt L-arginine L-glutamate, the method comprising: dissolving L-arginine base in water, adding L-glutamic acid, while stirring, to the resulting L-arginine base solution up to pH 6.6 to 6.8; filtering the resulting water solution of L-arginine L-glutamate; introducing the water solution of L-arginine L-glutamate in ethanol till precipitation (at a filtrate wateπethanol ratio of 1:(9.76-10.24); filtering and washing the resulting precipitate of L-arginine L- glutamate; drying L-arginine L-glutamate under vacuum at temperature not higher than 60⁰C for 4 to 5 hours. (12)

The disadvantages of the prototype method include the complexity and a long duration of the manufacturing process, necessity to comply with strict temperature and time conditions.

Accordingly, an object of the present invention is to provide a new physiologically active substance choline thioctate which, displaying hepatoprotective, hypoammoniemic and detoxic activity, possesses a higher level and a wider spectrum of specific activity.

Another object of the present invention is to provide a method for producing a new chemical substance displaying hepatoprotective, hypoammoniemic and detoxic activity, which method, thanks to the sequence of process steps, the selection of process conditions and parameters, enables a substance with a higher level and a wider spectrum of specific activity to be produced while simplifying, and shortening the duration of, the manufacturing process.

Yet another object of the present invention is to provide pharmaceutical compositions based on a new physiologically active substance choline thioctate in various dosage forms. The assigned object is achieved by that, in accordance with the present invention, 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) of formula

displays hepatoprotective, hypoammoniemic and detoxic activity.

The assigned object is achieved by that, in accordance with the present invention, in a method for producing 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate), the method comprises: dissolving a base and an organic acid in polar solvents and further isolating the target product, choline base is used as the base, thioctic acid is used as the organic acid, water and/or ethanol is/are used as the polar solvent(s), and the target product is isolated by removing the solvent by any method known to those skilled in the art.

The assigned object is achieved by that, in accordance with the present invention, a pharmaceutical composition containing an active ingredient and one or more pharmaceutically suitable carriers or solvents contains as the active ingredient 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) of formula

which displays hepatoprotective, hypoammoniemic and detoxic activity.

The technical result of implementing the present invention is the provision of a new physiologically active chemical substance 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) which displays hepatoprotective, hypoammoniemic and detoxic activity, the development of a method for producing the same which method ensures a higher level and a wider spectrum of specific activity while simplifying, and shortening the duration of the manufacturing process, as well as the provision of pharmaceutical compositions based on a new physiologically active substance 2(hydroxyemyl)trimethylammonium thioctate (choline thioctate) in various dosage forms the use whereof results in diversifying the range of hepatoprotective drugs.

Particular examples of preferred embodiments of the invention are the following:

Example 1: To a mixture of 0.8497 g (0.004125 moles) of thioctic acid (calculated with reference to the substance of 100%) in ethanol add 0.5 g (0.004125 moles) of choline base (calculated with reference to the dried substance of 100%) and stir till a clear solution. To produce the target product (choline thioctate), distillate ethanol out of the solution and dry the resulting precipitate under vacuum.

Example 2: To a suspension of 0.8497 g (0.004125 moles) of thioctic acid (calculated with reference to the substance of 100%) in water add 0.5 g (0.004125 moles) of choline base (calculated with reference to the substance of 100%) and stir till a clear solution. To produce the target product (choline thioctate), freeze- drying the solution.

Example 3: Proceed as in Example 2. To produce the target product (choline thioctate), dry the solution by spray drying.

Example 4: To a mixture of 0.5722 g (0.004125 moles) of choline hydrochloride (calculated with reference to the substance of 100%) in ethanol add 0.165 g (0.004125 moles) of sodium hydroxide (calculated with reference to the substance of 100%). Filter a sodium chloride precipitate, add 0.8497 g (0.004125 moles) of thioctic acid (calculated with reference to the substance of 100%) to the filtrate and stir till a clear solution. To produce the target product (choline thioctate), distillate ethanol out of the solution and dry the resulting precipitate under vacuum.

Example 5: To a mixture of 0.8497 g (0.004125 moles) of thioctic acid (calculated with reference to the substance of 100%) in ethanol, add 0.165 g (0.004125 moles) of sodium hydroxide (calculated with reference to the substance of 100%) and mix. To the resulting solution of sodium thioctate, add 0.5722 g (0.004125 moles) of choline hydrochloride (calculated with reference to the substance of 100%). Filter a sodium chloride precipitate. To produce the target product (choline thioctate), distillate ethanol out of the solution and dry the resulting precipitate under vacuum. Example 6: Proceed as in Example 2 at a thioctic acid / choline base molar ratio of 1:1 to 1.2). To the resulting water solution of the target product, add pharmaceutically suitable adjuvants to produce an injection solution by any method known to those skilled in the art.

Example 7: To pharmaceutically suitable adjuvants, add, while stirring, a concentrated water solution of choline thioctate produced in accordance with Example 2, then granulate, then granulate, dry and tablet or capsulate by any method known to those skilled in the art.

In the event that a manufacturer has problems in purchasing the substance choline base, it may be produced in accordance with Examples 4 and 5.

By the method in accordance with the present invention, a new physiologically active substance 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) is produced, which substance displays hepatoprotective, hypoammoniemic and detoxic activity. This is an amorphous, yellowish-colored product (a yield of about 100%) of high purity (a content of main substance is not less than 98.5%), soluble in water and polar organic solvents: C₁₃ H₂₇ NO₃S₂ (molecular weight =309.53)

Elemental Analysis Results

Both interrelation and sequence of the steps of of the method in accordance with the present invention, the selection of conditions and parameters ensure the attainment of the object of the present invention. Thus, the salt-forming reaction in producing choline thioctate occurs the most effectively in both polar solvents because of their specific properties (they perform a solvent function and a reaction medium function concurrently) and reagents - choline base and thioctic acid. It is to be understood that the present invention is not limited by selected ratios of solvents and choline base.

Table 1 Comparative Analysis the Method in Accordance with the Present Invention and the

Prototype Method

The methods to produce choline thioctate by the prototype method failed to produce the target product because of that the output substance dissolves not only in water but also in ethanol, which is used as a salt-forming agent in producing L- arginine L-glutamate. The easiness of performing the salt-forming reaction, as well as its output reagents of a sufficient purity level which are used in an equivalent quantity, ensure a high level of purity and stability of not only the target product but also pharmaceutical compositions when these solutions are used (a step of isolating the target product in a dry form is excluded thereby the manufacturing cycle is simplified significantly). In the method in accordance with the present invention, the temperature conditions of salt-forming do not influence on product purity and are only limited the freezing point and boiling point of solvents which are used as a reaction medium. Only the pre-determined parameters of target product concentration determine a solvent quantity for salt-forming.

In accordance with the present invention, choline thioctate based pharmaceutical compositions are produced in the form of tablets, capsules, powders, injection solutions, infusion solution concentrates, and other dosage forms.

An active substance was selected by means of the pharmacological screening of a number of complex compounds produced and of a comparative pharmacological assessment with respect to Glutarginum (injection dosage form). Preliminary screening studies of pharmacological activity of the complexes were conducted for the existence of their hepatoprotective action.

At the first stage of these studies, the most active compound was revealed based whereon further comparative studies were performed.

In the hepatitis model in rats, which hepatitis was induced by intragastric administration of carbon tetrachloride, there was conducted a comparative study of hepatoprotective activity of those complexes (compositions Nos. 1 to 5) which contain, in 1 niL:

Composition No. 1 : 10.0 mg of choline base + 17.0 mg of α-lipoic (thioctic) acid with 0.4 % of sodium chloride;

Composition No. 2: 14.4 mg of arginine base + 17.0 mg of α-lipoic (thioctic) acid; Composition No. 3: 10.0 mg of choline base + 17.0 mg of α-lipoic (thioctic) acid; Composition No. 4: 9.98 mg of choline base + 10.16 mg of nicotinic acid; Composition No. 5: 9.98 mg of choline base + 12.1 mg of glutamic acid; Drug Berlition commercially available from Berlin-Chemie AG: 133 mg of lipoic (thioctic) acid / 12 mL; Drug Glutarginum for Injection commercially available from Pharmaceutical Company Zdorovye Ltd.: 200 mg / 5 mL (22 mg of arginine + 18 mg of glutamic acid in 1 mL).

The experimental studies were conducted on 48 outbred white rats, both male and female, of 230 to 260 g of body weight. Carbon tetrachloride was administered to the animals in the form of a 50% oil emulsion intragastrically, once, at a dose of 0.8 mL/100 g b.w. Tested drag specimens were administered intravenously into the caudal vein for three days prior to the administration of a toxic agent, as well as for three days following the administration of the toxic agent (to the animals survived. The tested drug specimens were administered to the animals at the following doses:

In this study, the animals were divided into 8 groups of 6 rats in each:

1 - Pathology control;

2 - Pathology + Composition No. 1 at a dose of 0.5 mg/kg;

3 - Pathology + Composition No. 2 at a dose of 0.5 mg/kg;

4 - Pathology + Composition No. 3 at a dose of 0.5 mg/kg;

5 - Pathology + Composition No. 4 at a dose of 0.5 mg/kg;

6 - Pathology + Composition No. 5 at a dose of 0.5 mg/kg;

7 - Pathology + Berlition at a dose of 0.34 mg/kg;

8 - Pathology + Glutarginum at a dose of 0.35 mg/kg. The selection of doses was based on the fact that the animals should receive substances at the same doses close to the doses recommended in clinics for humans.

As a result of the experimental study, it was found that 5 of 6 rats in the pathology control group died on Day 3 to Day 5 after dosing (Table 2) this corresponds to the data, which are known from literature and to our own studies which are the evidence of the development of acute hepatic pathology in rats and death of as a rule 60% to 80% of rats.

Table 2

The administration of test specimens to rats revealed their marked hepatoprotective activity. The highest (100%) effect was observed when composition No. 3 was administered. Composition No. 1 that, in addition to choline and lipoic acid, contained a 0.4 % solution of sodium chloride, exhibited a slightly lower activity (83.3%). The rest of the test compositions and Glutarginum were second to them in specific activity level.

At the same time, the dose of Glutarginum was nearly 3 times higher than the dose of the most active test compound.

As a result of these preliminary experiments, it was found that the most active complex displayed a 100% hepatoprotective activity. In theses experiments, Glutarginum exhibited a 67% effect. It may be, thus, concluded that the physiologically active substance choline thioctate displays a marked hepatoprotective activity, which exceeds that of Glutarginum, which was used at a higher dose. The complex in accordance with the present invention is promising for producing, based thereon, monodrugs or complex drugs of hepatoprotective activity.

Under Power of Attorney V. Mikhailyuk, Patent Attorney

REFERENCES:

1. M. D. Mashkovskiy, Drugs, Torsing, Kharkov (1997), Vol. 1 - 514-516.

2. M. D. Mashkovskiy, Drugs, Torsing, Kharkov (1997), Vol. 1 - 514.

3. The Russian Federation Patent No. 2,211,034, IPC A61K 31/3343, A61P 1/16. Official Bulletin of Rospatent "Inventions, Utility Models" of 27.08.2003.

4. Vidal, Drugs in Russia - Reference Book, Moscow, AstraPharmService (2001) B-132.

5. Vidal, Drugs in Russia - Reference Book, Moscow, AstraPharmService (2001) B-472.

6. M. D. Mashkovskiy, Drugs, Torsing, Kharkov (1997), Vol. 1 - 516.

7. ROTE LISTE 2000, No. 48053.

8. ROTE LISTE 2000, No. 48056.

9. Ukrainian Patent No. 54,880, Cl. C07C229/02, A61K31/195, A61P1/16, A61P39/00, O racial Bulletin "Industrial Property" (2003), No.3, page 2

10. USSR Inventor's Certificate No. 1,266,139. Cl. C07 C 103/187, published on July 03, 19S4.

11. USSR Inventor's Certificate No. 1,248,212. Cl. C 07 C 101/22, A 61 K 31/195, published on July 27, 1983. 12. Ukrainian Patent No. 54,880, Cl. C07C229/02, A61K31/195, A61P1/16, A61P39/00, Official Bulletin "Industrial Property" (2003), No.3, page 3.

Claims

1. 2(hydroxyethyl)trimemylammonium thioctate (choline thioctate) of formula

which displays hepatoprotective, hypoammoniemic and detoxic activity.

2. A method for producing 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate), the method comprising: dissolving a base and an organic acid in polar solvents and further isolating the target product, characterized in that choline base is used as the base, thioctic acid is used as the organic acid, water and/or ethanol is/are used as the polar solvent(s), and the target product is isolated by removing the solvent by any method known to those skilled in the art.

3. A pharmaceutical composition containing an active ingredient and one or more pharmaceutically suitable carriers or solvents, characterized in that said composition contains as the active ingredient 2(hydroxyethyl)trimethylammonium thioctate (choline thioctate) of formula

which displays hepatoprotective, hypoammoniemic and detoxic activity.

Under Power of Attorney V. Mikhailyuk, Patent Attorney