EA004882B1 - Sodium-hydrogen-exchanger type 1 inhibitor (nhe-1) - Google Patents

Abstract

1. A compound having the Formula I a prodrug thereof or a pharmaceutically acceptable salt of said compound or of said prodrug with the proviso that [5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine is not included. 2. [5-Cyclopropyl-1-(2-quinolon-5-yl)-1H-pyrazole-4-carbonyl]guanidine, or a pharmaceutically acceptable salt thereof. 3. A method of reducing tissue damage resulting from ischemia or hypoxia comprising administering to a mammal in need of such treatment a therapeutical effective amount of a compound of claim 1, a prodrug thereof or a pharmaceutical acceptable salt of said compound or of said prodrug. 4. A method as recited in claim 3 wherein the tissue is cardiac, brain, liver, kidney, lung, gut, skeletal muscle, spleen, pancreas, nerve, spinal cord, retina tissue, the vasculature, or intestinal tissue. 5. A method as recited in claim 3 wherein the amount of the compound of claim 1, a prodrug thereof or a pharmaceutical acceptable salt of said compound or of said prodrug is about 0.01 mg/kg/day to about 50 mg/kg/day. 6. A method as recited in claim 5 wherein the mammal is a female or male human. 7. A method as recited in claim 6 wherein said tissue is heart tissue. 8. A method as recited in claim 6 wherein said tissue is brain tissue. 9. A method as recited in claim 6 wherein said tissue is liver tissue. 10. A method as recited in claim 6 wherein said tissue is kidney tissue. 11. A method as recited in claim 6 wherein said tissue is lung tissue. 12. A method as recited in claim 6 wherein said tissue is gut tissue. 13. A method as recited in claim 6 wherein said tissue is skeletal muscle tissue. 14. A method as recited in claim 6 wherein said tissue is spleen tissue. 15. A method as recited in claim 6 wherein said tissue is pancreas tissue. 16. A method as recited in claim 6 wherein said tissue is retina tissue. 17. A method as recited in claim 6 wherein the compound is administered prophylactically. 18. A method as recited in claim 6 wherein the compound is administered prior to surgery. 19. A method as recited in claim 6 wherein the compound is administered prior to cardiac surgery. 20. A method as recited in claim 6 wherein the compound is administered during surgery. 21. A method as recited in claim 6 wherein the compound is administered during cardiac surgery. 22. A method as recited in claim 6 wherein the compound is administered within twenty-four hours after surgery. 23. A method as recited in claim 6 wherein the compound is administered within twenty four hours after cardiac surgery. 24. A method as recited in claim 6 wherein the tissue damage resulting from ischemia is ischemic damage and is incurred during organ transplantation. 25. A method as recited in claim 6 wherein the compound is administered to prevent perioperative myocardial ischemic injury. 26. A pharmaceutical composition which comprises an amount of the compound of claim 1, a prodrug thereof or a pharmaceutical acceptable salt of said compound or of said prodrug and a pharmaceutical acceptable carrier, vehicle or diluent. 27. A pharmaceutical composition for the reduction of tissue damage resulting from ischemia or hypoxia which comprises a therapeutical effective amount of the compound of claim 1, a prodrug thereof or a pharmaceutical acceptable salt of said compound or of said prodrug and a pharmaceutical acceptable carrier, vehicle or diluent. 28. A method as recited in claim 6 wherein the compound is administered prior to, during and after surgery. 29. A method as recited in claim 6 wherein the compound is administered prior to, during and after cardiac surgery. 30. A method of producing [5-cyclopropyl-1-(2-quinolon-5-yl)-1H-pyrazole-4-carbonyl]guanidine by administering [5-cyclopropyl-1-(quinolin-5-yl)-1H-pyrazole-4-carbonyl]guanidine to a human.

Description

State of the art

This invention relates to a type 1 (ΝΗΕ-1) sodium hydrogen exchange inhibitor.

Ischemic myocardial damage can be observed both in an outpatient and during surgery and can lead to the development of sudden death, myocardial infarction or congestive heart failure. In medicine, there is a need to prevent or minimize ischemic damage to the myocardium, especially myocardial infarction during surgery. Such treatment is expected to save lives and can shorten hospital stays, improve quality of life, and reduce overall health care costs for high-risk patients.

Pharmacological cardioprotection would reduce the prevalence and progression of myocardial infarction and dysfunction encountered in these surgical operations (during surgery). In addition to reducing myocardial damage and improving myocardial function after ischemia in patients with coronary heart disease, cardioprotection would also reduce the prevalence of heart disease and mortality due to myocardial infarction and dysfunction among “at-risk” patients (those over 65 who does not tolerate physical activity, has coronary artery disease, diabetes mellitus, hypertension), which require extracardial surgery.

The mechanism (s) responsible for myocardial damage observed after ischemia and reperfusion are not fully understood.

A number of publications have reported the use of guanidine derivatives useful for the treatment of, for example, arrhythmias.

In the recently published patent application PCT / 1B 99/00206, published as XV 0 99/43663 September 2, 1999, the description of which is incorporated into this description by reference, discloses a number of inhibitors ΝΗΕ-1, including [5 cyclopropyl-1- (quinolin-5 -yl) -1H-pyrazole-4carbonyl] guanidine. The publication also states that "preferred salts of the immediately preceding compound are mono- or dimesylate salts." One group of preferred compounds, which includes hydroxyquinoline compounds, is described in paragraph 102 of the claims of the published application. Some of these hydroxyquinoline compounds may exist in several tautomeric forms, such as the quinolone form, as described in this patent application. In addition, in the usual manner, the provisional application filed in the United States with serial number 60/162374 was filed October 29, 1999 and relates to crystalline forms of the ΝΗΕ-1 inhibitor described above.

PCT / 1P 97/04650, published June 25, 1998, discloses compounds No. [(substituted five-membered heteroaryl)] guanidine, useful as inhibitors of Να '/ Η' metabolism and therefore effective in the treatment of various diseases such as hypertension, arrhythmia, angina pectoris, myocardial infarction, arteriosclerosis and diabetes complications.

Thus, it is clear that there is a need for compounds for the treatment of myocardial ischemia that occurs during surgery, and the continuation of research in this area.

SUMMARY OF THE INVENTION

This invention relates to a compound of formula I

n a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug, provided that [5-cyclopropyl-1- (quinolin-5-yl) -1H-pyrazole-4-carbonyl] guanidine is not included.

As used herein, the phrase “a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug” includes the condition that [5cyclopropyl-1- (quinolin-5-yl) -1H-pyrazole-4carbonyl] guanidine is not included.

The present invention also relates to a substantially pure [5-cyclopropyl-1- (2-quinolone-5yl) -1H-pyrazole-4-carbonyl] guanidine or a pharmaceutically acceptable salt of said compound.

Alternatively, the above compound is called [5-cyclopropyl-1- (2-quinolone-5yl) -1H-pyrazole-4-carbonyl] guanidine. A preferred salt is a hydrochloride salt, and most preferably a hydrogen monochloride salt.

Another aspect of the present invention is a method of treating a mammal (eg, a human) having a disease or condition mediated by ΝΗΕ-1, administering to the mammal a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for reducing tissue damage (e.g., essentially preventing tissue damage, inducing tissue protection) due to ischemia, comprising administering to a mammal (e.g., a woman or a man) in need of such treatment a therapeutically effective amount of a compound of formula I, its prodrugs or a pharmaceutically acceptable salt of said compound or said prodrug.

Ischemic tissues, individually or in a group, are preferably tissues of the heart, brain, liver, kidney, lung, intestine, skeletal muscle, spleen, pancreas, nerve, spinal cord, retinal tissue, vasculature, or intestinal tract tissue.

It is particularly preferred when the ischemic tissue is cardiac tissue.

It is particularly preferable to administer a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug to prevent ischemic damage to the myocardium during surgery.

It is preferable to administer a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug prophylactically.

Ischemic damage can occur during organ transplantation into either the organ or the patient.

Preferably, a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug is administered before, during and / or shortly after cardiac surgery or extracardiac surgery.

In one aspect of the invention, a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug is applied locally.

A preferred dosage of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug is about 0.01-100 mg / kg / day. A particularly preferred dosage of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug is approximately 0.01-50 mg / kg / day.

Another aspect of the present invention relates to a method for reducing myocardial tissue damage (e.g., essentially preventing tissue damage, inducing tissue protection) during surgery (e.g., coronary artery bypass grafting (SABO) surgery, vascular surgery, percutaneous trans-cavity coronary angioplasty (PTCA) ), by organ transplantation or during other extracardiac surgery), comprising administering to a mammal (e.g., a woman or a man) a therapeutically effective amount of a compound of formula I, th prodrug or pharmaceutically acceptable salt of said compound or of said prodrug.

Another aspect of the present invention relates to a method for reducing myocardial tissue damage (e.g., essentially preventing tissue damage, inducing tissue protection) in patients representing patients with current heart disease (acute coronary syndromes, such as myocardial infarction or unstable angina) or with ischemia brain (eg, “stroke”), comprising administering to a mammal (eg, a woman or man) a therapeutically effective amount of a compound of formula I, a prodrug thereof, or a pharmacist a tactically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a chronic method for reducing myocardial tissue damage (e.g., essentially preventing tissue damage, inducing tissue protection) in patients diagnosed with coronary heart disease (e.g., previous myocardial infarction or unstable angina) or in patients at increased risk of myocardial infarction (e.g. age> 65 and two or more risk factors contributing to coronary heart disease), including administration to a mammal (e.g., female or m zhchine) a therapeutically effective amount of a compound of Formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or of said prodrug.

Another aspect of the present invention relates to a method for the prevention of ischemic damage, comprising the continuous oral administration to a mammal in need of such treatment of a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating cardiovascular diseases, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method of treating arteriosclerosis, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method of treating hypertension, comprising administering to a mammal (e.g., a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating arrhythmia, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating angina pectoris (angina pectoris), comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating heart hypertrophy, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating kidney diseases, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating diabetes complications, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating restenosis, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating proliferative cell diseases, comprising administering to a mammal (e.g., a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating malignant diseases, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating fibrotic diseases, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method of treating glomerular nephrosclerosis, comprising administering to a mammal (e.g., a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating pulmonary fibrosis, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating cerebro-ischemic disorders, comprising administering to a mammal (e.g., a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating myocardial stroke, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating myocardial dysfunction, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating cerebrovascular diseases, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

Another aspect of the present invention relates to a method for treating organ hypertrophy or organ hyperplasia, comprising administering to a mammal (eg, a woman or a man) a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug.

The invention also relates to pharmaceutical compositions that comprise a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug, and a pharmaceutically acceptable carrier, excipient or diluent.

The invention also relates to pharmaceutical compositions for reducing tissue damage due to ischemia, which comprise a therapeutically effective amount of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug, and a pharmaceutically acceptable carrier, excipient or diluent.

By the term “reduction” is meant, in addition to essentially complete prevention, partial prevention or prevention, which is at least greater than that which would result from not using the compound or using a placebo, but less than 100%.

The term “ischemic injury”, as used herein, refers to conditions directly related to a decrease in blood flow to the tissue, for example, due to a blood clot or blockage of blood vessels through which blood supply to the underlying tissues is carried out, which leads, in other words, to a reduced transfer oxygen to such tissue, tissue malfunction, tissue dysfunction and / or necrosis. On the other hand, when the blood flow or perfusion of an organ is adequate in quantity, the amount of oxygen carried by the blood or the average value of organ perfusion can be reduced, for example, in an environment lacking oxygen, so that the oxygen supply to the tissue decreases and tissue malfunctions, dysfunction tissue and / or tissue necrosis.

As used herein, the terms “treatable”, “treat” or “treatment” include preventive (eg, prophylactic) and palliative treatment.

The term “pharmaceutically acceptable salt” refers to non-toxic anionic salts containing anions such as (but not limited to) chloride, bromide, iodide, sulfate, bisulfate, phosphate, acetate, maleate, fumarate, oxalate, lactate, tartrate, citrate, gluconate, methanesulfonate (mesylate) and 4-toluenesulfonate.

Since there is more than one basic moiety, the expression may include polybasic salts (e.g., disols). The expression also refers to non-toxic cationic salts with cations such as (but not limited to) sodium, potassium, calcium, magnesium, ammonium or protonated benzathine (Ν, Ν'dibenzylethylenediamine), choline, ethanolamine, diethanolamine, ethylenediamine, meglamine (Ν methylglucine) , benethamine (Ν-benzylphenethylamine), piperazine or tromethamine (2-amino-2-hydroxymethyl-1,3-propanediol).

The term “prodrug” refers to compounds that are precursors of the drug to be administered, releasing the drug ίη νίνο by some chemical or physiological process (for example, a prodrug brought to physiological pH, or converted to the desired form of the drug by the action of the enzyme). Again, there is a condition that [5-cyclopropyl-1 (quinolin-5-yl) -1H-pyrazole-4-carbonyl] guanidine is not included.

The expression "pharmaceutically acceptable" means that the carrier, diluent, excipients and / or salt must be compatible with the other ingredients of the composition and harmless to the recipient.

As used herein, the terms “reaction inert solvent” and “inert solvent” refer to a solvent or mixture of solvents that do not interact with the starting materials, reagents, intermediates or products in such a way that adversely affects the yield of the desired product.

It is understood that the compound of the present invention may exist in a radioactive label, that is, the compound may contain one or more atoms having an atomic mass or mass number different from the atomic mass or mass number commonly found in nature. The radioisotopes of hydrogen and carbon include, respectively, ² H, ³ H and ¹⁴ C. A compound of the present invention, which contains said radioisotopes and / or other radioisotopes of other atoms, is within the scope of this invention. Radioisotopes of tritium, i.e., ³ N, and carbon-14, i.e., ¹⁴ C, are especially preferred because of the ease of their preparation and detection. A compound of formula I with a radioactive label can usually be obtained by methods known to those skilled in the art. Such compounds with a radioactive label can be obtained by following the procedures described below in the schemes and / or examples, replacing a reagent not labeled with a radioactive label, with a ready-made, available reagent with a radioactive label.

Other features and advantages will be apparent from the description and claims, which describes the invention.

DETAILED DESCRIPTION OF THE INVENTION

The compound of formula I is a metabolite of [5-cyclopropyl-1 - (quinolin-5-yl) -1H-pyrazole-4-carbonyl] guanidine in the human body. It has been identified in human plasma samples obtained after intravenous administration of [5-cyclopropyl-1- (quinolin-5yl) -1H-pyrazole-4-carbonyl] guanidine to humans.

The structure of this compound was independently verified by chemical synthesis. In yet another aspect of the present invention, [5-cyclopropyl-1- (2-quinolon-5-yl) -1Npyrazole-4-carbonyl] guanidine can be prepared by introducing [5-cyclopropyl-1- (quinolin-5-yl) -1Npyrazole -4-carbonyl] guanidine to humans and isolating the desired metabolite from plasma. Alternatively, isolation of the metabolite from the human body is not required, since it is produced by ίη νίνο.

In the General case, the compound of this invention can be obtained by methods similar to those known in the field of chemistry, especially in light of the disclosed method contained in this description. Some methods for preparing the compounds of this invention are further presented by further features of the invention and are illustrated by the following reaction scheme. Other methods are described in the experimental section.

A detailed description of the synthetic aspect of the invention is as follows

According to the above scheme, the compound of formula ΙΑ (obtained as described by Sarrk ΙΌ .; NashShop S.S. GShies. Bos. 1938.60.2104) is dissolved in a protic solvent, such as ethanol, and treated with an appropriate reducing agent, such as dihydrate tin chloride. The resulting mixture was stirred at a temperature of from about 0 to 115 ° C. For about 30 minutes to 24 hours. The resulting mixture was cooled to about 23 ° C and filtered. The resulting solid is purified by a suitable method, such as trituration with hydrochloric acid (e.g., aqueous 1M) at about 100 ° C for about 1 hour, cooled to about 23 ° C and filtered to give the compound of formula II as its hydrochloride salt . Alternatively, a compound of formula II can be isolated from the reaction mixture as a free base, made basic with an inorganic base and extracted with a suitable organic solvent. The compound of formula II can also be isolated as other salts. Other reduction methods by which such reduction can be performed are known to those skilled in the art, such as, for example, catalytic hydrogenation.

The compound of formula II is diazotized in hydrochloric acid and water using sodium nitrite at about 0 ° C. for about 15 minutes to 2 hours. The resulting diazonium salt solution is reduced with a suitable reducing agent, such as tin chloride dihydrate, in hydrochloric acid and water at about 0-23 ° C, for about 0.5-6 hours. The resulting solid is collected by filtration and purified by a suitable method, such as trituration with hydrochloric acid (e.g., aqueous, 1M) at a rate about 23 ° C. for about 30 minutes. The compound of formula III is collected by filtration in the form of its hydrochloride salt.

Alternatively, the compound of formula III can be isolated from the reaction mixture as a free base, made basic with an inorganic base and extracted with a suitable organic solvent. The compound of formula III can also be isolated as other salts. Other reducing agents by which such a conversion can be carried out are known to those skilled in the art.

The compound of formula IV is prepared by methods known to those skilled in the art, such as reacting ди dialkyl acetal, Ν-dialkylformamide, for example, Ν dimethyl acetal, Ν-dimethylformamide, with 3-cyclopropyl-3-oxopropanoic acid ester at approximately 23-115 ° C. for approximately 1-4 hours in the presence or absence of an acid catalyst. The K ¹ and K ² groups represent any suitable alkyl, cycloalkyl, cycloalkylalkyl or arylalkyl group. In addition, two K ² groups can be linked together to form a cyclic fragment.

The compound of formula IV is reacted with a compound of formula III in an alcohol solvent such as ethanol at about 23-115 ° C. for about 15 minutes to 12 hours. When the compound of formula III is used as its hydrochloride salt, it is advantageous to carry out reaction in the presence of an excess of a non-nucleophilic base such as triethylamine. The compound of formula V is isolated by filtration. Alternatively, the compound of formula V can be isolated by other methods, such as concentration, followed by the addition of water and extraction with a suitable organic solvent. Alternatively, the formula V compound may be prepared using the formula III compound and other compounds in lieu of formula IV compounds, such as compounds in which (Β ²⁾ Ν ₂ group is replaced by B ² O group.

The compound of formula V is hydrolyzed with a base, such as sodium or lithium hydroxide, in a suitable solvent, such as water and / or methanol, and / or THF, at about 23 ° C. or at an elevated temperature, such as a reflux temperature in for about 0.5-12 hours. An acid of formula VI is then isolated, for example, by removing organic solvents, acidifying and filtering. Alternatively, a compound of formula VI can be isolated from the reaction mixture by removing organic solvents, acidifying and extracting with a suitable organic solvent.

An acid of formula VI is reacted with guanidine in the presence of a suitable binder. A suitable binder is one that converts a carboxylic acid into a reactive form that forms an amide bond when reacted with an amine.

The binder may be a reagent that affects this condensation during the process in one tank where carboxylic acid and guanidine are mixed together. Typical binding agents are 1- (3-dimethylaminopropyl) 3-ethylcarbodiimide / hydroxybenzotriazole (EOC / HBT) hydrochloride, dicyclohexylcarbodiimide (pCC) / hydroxybenzotriazole (HBT), 2-ethoxy-1-ethoxycarbonyl-i-1,2-din diethylphosphorylcyanide. The reaction is carried out in an inert solvent, preferably an aprotic solvent, at a temperature of about (-) 20 to 50 ° C for about 1-48 hours in the presence of an excess of guanidine as a base. Typical solvents include acetonitrile, dichloromethane, dimethyl sulfoxide, dimethylformamide, chloroform, and mixtures thereof.

The binder may also be a substance that converts a carboxylic acid into an activated intermediate, which is released and / or formed in the first stage and in the second stage allows interaction with guanidine. Examples of such binders and activated intermediates are thionyl chloride or oxalyl chloride to form an acid chloride, cyanurofluoride to form an acid fluoride or alkyl chloroformate such as isobutyl or isopropyl chloroformate, or propanphosphonic acid anhydride for the formation of tertiary phosphoric acid anhydride, or carbonyldiimidazole to form acylimidazole. If the binder is oxalyl chloride, it is preferable to use a small amount of dimethylformamide as a co-solvent with another solvent (such as dichloromethane) to catalyze the formation of the acid chloride. Said activated acid derivative may be reacted with a suitable base by mixing together with the intermediate in a suitable solvent. Suitable solvent / base combinations are, for example, dichloromethane, dimethylformamide or acetonitrile, or mixtures thereof in the presence of an excess of guanidine as the base. Other suitable solvent / base combinations include water or (C1-C5) alcohol or a mixture thereof together with a co-solvent such as dichloromethane, tetrahydrofuran or dioxane, and a base such as sodium, potassium or lithium hydroxide in an amount sufficient to bind the acid released during the reaction. The use of such binders and a suitable choice of solvents and temperatures are known to those skilled in the art or can be readily determined from the literature in light of this description. These and other typical conditions used for the combination of carboxylic acids are described in Noiue \ UeG t. XU, part II, ELUSHEK Eb., S. Tyeppe Ug1ad, 1974, 81idai; M.Vobaizku, Rpps1r1e8 οί Rerybe 8ui1ys818, 8gshdegAeg1ad, Veg1sh 1984; and Tke Rerybez, Lia1u818, 8iu1ke515 aib Vu1odu (eb. E. Sgozz and ^ .Me ^ eη1уΓе ^). Walk 1-5 (Lsabesh 1s Rgezz, ΝΥ 1979-1983).

In one embodiment of the invention, the acid of formula VI is activated with an excess of thionyl chloride at about reflux for about 15 minutes to 3 hours, and the excess thionyl chloride is removed by concentration. The resulting acid chloride is combined with an excess of guanidine hydrochloride and an inorganic base such as sodium hydroxide in tetrahydrofuran and water. The reaction mixture is stirred in the usual manner at 23 ° C or at an elevated temperature, such as a reflux temperature, for about 0.5-6 hours. The compound of formula I is isolated from this reaction mixture in a variety of ways. For example, the reaction mixture is concentrated to remove THF; the aqueous layer was acidified to pH 9 and the solid was collected by filtration. Alternatively, the compound of formula I can be isolated by extraction with an organic solvent. The compound of formula I can be converted to the corresponding hydrochloride salt by treating its solution in methanol with hydrogen chloride in ether and collecting the resulting solid by filtration or concentration. Other salts can be obtained in similar ways.

Those skilled in the art will recognize that it is also possible to directly convert a compound of formula V into a compound of formula I by treating the compound of formula V with an excess of guanidine in an inert solvent such as an alcohol solvent, for example ethanol, or in the absence of a solvent at about 60-150 ° C.

It will be apparent to those skilled in the art that the compound of formula I can exist in several tautomeric forms. All such tautomeric forms are contemplated as part of this invention. For example, all tautomeric forms of the carbonylguanidine moiety of a compound of formula I are included in this invention. All tautomeric forms of the 2-quinolone fragment of a compound of formula I, such as the 2-hydroxyquinoline form, are also included in this invention.

The starting materials and reagents for the compounds described above are also readily available or can be easily synthesized by those skilled in the art using suitable organic synthesis methods.

In addition, when the compound of the present invention forms metabolites, hydrates or solvates, they are also included in the scope of this invention.

Some of the compounds (eg, prodrugs) of this invention are acidic and form a salt with a pharmaceutically acceptable cation. Most of the compounds of this invention are basic in nature and form a salt with a pharmaceutically acceptable anion. All such salts, including disols, are within the scope of this invention and can be prepared by suitable methods. For example, they can be obtained by simple contact with acidic and basic substances, either in an aqueous, or non-aqueous, or partially aqueous medium. Salts are recovered either by filtration, precipitation with a solvent, followed by filtration, evaporation of the solvent, or, in the case of aqueous solutions, lyophilization, as convenient.

A compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug inhibits the sodium / proton exchange system (Να '/ Η') and, therefore, is useful as a therapeutic or prophylactic agent for diseases caused by accelerated sodium / proton transport (Να '/ Η'). for example, with cardiovascular diseases [for example, arteriosclerosis, hypertension, arrhythmias (for example, with ischemic arrhythmia, arrhythmias due to myocardial infarction, arrhythmias after PTCA or after thrombosis, etc.), with angina pectoris, heart hypertrophy, myocardial infarction, heart failure ( for example, in congestive heart failure, acute heart failure, cardiac hypertrophy, etc.), for restenosis after PTCA, shock (for example, hemorrhagic shock, endotoxin shock, etc.)], for renal diseases (for example, for diabetes mellitus, diabetic nephropathy, ischemic acute renal failure, etc.), for organic disorders associated with ischemia or ischemic reperfusion [(for example, for disorders associated with ischemic reperfusion of the heart muscle, for acute renal failure, or for violations caused by surgical treatment, such as coronary artery bypass grafting (CABS), vascular surgery, organ transplantation, extracardiac surgery, or percutaneous transosseous coronary angioplast ка (РТСА)], for cerebrovascular diseases (for example, for ischemic stroke, hemorrhagic stroke, etc.), for cerebro-ischemic disorders (for example, for disorders associated with cerebral infarction, for disorders in the form of complications caused by cerebral hemorrhage, or with cerebral edema). A compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, can also be used as a myocardial protection during coronary artery bypass grafting (CABS) surgery, vascular surgery, during percutaneous transoluminal coronary angioplasty (PTCA), transplantation organs or extracardiac surgery.

A compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, can preferably be used as a myocardial protection agent before, during, or after coronary artery bypass grafting (CABS) surgery, vascular surgery, during percutaneous transverse cavity coronary angioplasty (RTSA), organ transplantation or extracardiac surgery.

A compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug may preferably be used as myocardial protection agents in patients representing patients with current heart disease (with acute coronary syndromes, for example, with myocardial infarction or unstable angina) or with phenomena cerebral ischemia (for example, after a stroke).

A compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, can preferably be used as a means of continuous myocardial protection in patients with diagnosed coronary heart disease (for example, with previous myocardial infarction or unstable angina) or in patients with an increased risk of heart attack myocardium (for example, over 65 years old and two or more risk factors contributing to coronary heart disease).

In addition to the above, a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug is highly remarkable due to its strong inhibitory effect on cell proliferation, for example, proliferation of fibroblast cells and proliferation of smooth muscle blood vessel cells. For this reason, a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug is a valuable therapeutic agent for diseases in which cell proliferation is the primary or secondary cause, and can therefore be used as anti-atherosclerotic agents and as anti long-term complications of diabetes, malignant diseases, fibrotic diseases such as pulmonary fibrosis, liver fibrosis or Brose kidney against glomerular nephrosclerosis, organ hypertrophies or hyperplasias, in particular hyperplasia or hypertrophy of the prostate, pulmonary fibrosis, diabetic complications or recurrent stricture after PTCA, or diseases caused by endothelial cell injury.

The effectiveness of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, as a medicament for treating mammalian diseases (such as humans) described in detail herein, for example, for protecting myocardium during surgery, or protecting myocardium in patients with ongoing heart disease or the phenomena of cerebral ischemia, or for continuous cardioprotection of patients with diagnosed coronary heart disease is demonstrated by actively the compounds of formula I or a pharmaceutically acceptable salt of the compound under typical conditions ίη νίίτο and other preclinical cardio protection tests [see test ίη νίνο in the publication K1esh N. et al., Spsi1a1yup 92: 912-917 (1995); test on an isolated heart in the publication 5> s1y1h V. et al., Sagbuua5siag Kekeages, 29: 260-268 (1995); test for antiarrhythmia in a publication by Wajcake M. et al., Ash.k Pbu8u1., 36: H2430-H2440 (1994); NMR analysis in a publication by Ko1ke et al., ί. T1yugas. SagbyuahsYangd. 112: 765-775 (1996)]. Such tests also provide means by which the activity of a compound of formula I or a pharmaceutically acceptable salt of said compound or said prodrug can be compared with the activities of other known compounds. The results of this comparison are used to determine dosage levels for mammals, including humans, in the treatment of such diseases.

Measurement of human ΝΗΕ1 inhibition activity

Methods for measuring human β-1 activity and inhibitor effectiveness are based on the methods published by Wa18op et al., (Pbu8u1., 24: 6229-6238.1991), where the recovery of intracellular pH mediated by β after intracellular acidification is measured. So fibroblasts stably expressing human ΝΗΕ-1 (SoipShop L. et al., Mo1. Ryagshaso1., 44: 1041-1045 (1993), are plated on collagen-coated 96 well plates (50,000 / well) and grown in culture media (ΌΜΕΜ with a high content of glucose, 10% fetal bovine serum, 50 units / ml penicillin and streptomycin) until confluence Confluent plates are incubated for 30 min at 37 ° C with a pH-sensitive fluorescence probe ΒCΕCΕ (5 μmol; Mo1ci1ag Prgoe, Bademe, OK). Cells loaded with Β6Ε6Ρ are incubated for 30 min at 37 ° C in saturate of media (70 mM choline chloride, 50 mM ΝΗΟ1 ₊ 5 mM KC1, 1 mM MdS1 _2, 1.8 mM SaS1 _2, 5 mM glucose, 10 mM ΗΕΡΕ8, pH 7.5) and then placed in a spectrophotometer | R1ioge8sep1 Ipadshd R1a1e Keabeg (Mo1eci1ag OMse8, CA)]. Β6Ε6Ρ fluorescence is observed using excitation and radiation wavelengths of 485 nm and 525 nm, respectively. Intracellular acidification is initiated by the rapid replacement of the acidic saturating medium with a reducing medium (120 mM No. C1, 5 mM KCl, 1 mM MD1 _2, 1.8 mM SaS1 _2, 5 mM glucose, 10 mM ΗΕΡΕ8, pH 7,5) ± test compound, and observed as a sequential increase fluores ntsii Β6Ε6Ρ, time-dependent. The effectiveness of human ΝΗΕ-1 inhibitors is calculated by the concentration, which reduces the restoration of intracellular pH by 50% (TC ₅₀ ). Under such conditions, the ΝΗΕ amyloride and HOE-642 inhibitors, known from the literature, had a GS50 for человека-1 person of 50 and 0.5 μM, respectively. The compound of formula I showed a 1C ₅₀ value of 200 nM in the above test.

It was previously reported that short periods of myocardial ischemia, accompanied by reperfusion of the coronary artery, protect the heart from subsequent severe myocardial ischemia (Miggu et al., Clinica! 74: 1124-1136, 1986). This phenomenon is known as ischemic preconditioning (preadaptation).

The therapeutic effect of the compounds of the present invention to prevent damage to heart tissue resulting from ischemic stroke can be demonstrated by ίη νίίτο in accordance with the principles presented in the publication bsh and others (Satbyusaks. Kek., 28: 1057-1061, 1994) and specifically disclosed in this description. Cardioprotection, which means a decrease in damage to the myocardium during a heart attack, can be induced pharmacologically by using agonists of adenosine receptors in an isolated, retrograde perfused rabbit heart, used as a model for the preconditioning (pre-adaptation) of myocardial ischemia (Lactius et al. ., 28: 10571061,1994). The ίη νίίτο test described below shows that the compound active during the test can also pharmacologically induce cardioprotection, that is, reduce the volume of myocardial infarction when introduced into an isolated rabbit heart. The effects of the test compound can be compared with ischemic preconditioning (pre-adaptation) and with the effect of the adenosine agonist A1 / A3, ΑΡΝΕΑ (L ⁶ - [2- (4-aminophenyl) ethyl] adenosine), which have been shown to pharmacologically induce cardioprotection in isolated rabbit heart (Lh et al., Sagbyugaks. Kek., 28: 1057-1061,1994). The exact methodology is described below.

The protocol used for these experiments exactly follows the procedure described in the publication of bsh et al., S'agbyugaks. Kek., 28: 1057-1061,1994). New Zealand white rabbit males (3-4 kg) were anesthetized with sodium pentobarbital (30 mg / kg, intravenously). After deep anesthesia was achieved (determined by the absence of an oculomotor reflex), the animal was intubated and ventilated with 100% O ₂ using an artificial lung ventilation apparatus with atmospheric pressure. A left-side thoracotomy was performed, exposing the heart, and a loop (2-0 silk) was loosely attached around the convex branch of the left-sided coronary artery, approximately 2/3 in relation to the apex of the heart. The heart was removed from the chest and quickly (<30 s) installed on the Langendorff apparatus (LapdebogGG). The heart was retrograde perfused with a non-recirculated modified Krebs solution (118.5 mM No. C1, 4.7 mM KC1, 1.2 mM Mg§0 ₄ , 1.2 mM KH ₂ PO ₄ , 24.8 mM

NaHCO3, 2.5 mM CaCl ₂ and 10 mM glucose) at a constant pressure of 80 mm Hg. and a temperature of 37 ° C. During perfusion, a pH of 7.47.5 was maintained, sparging a mixture of 95% O2 / 5% CO2. Heart temperature was carefully controlled using heated saline tanks and a water jacket not only around the tubing of the perfusion system, but also of an isolated heart. Heart rate and pressure in the left ventricle were determined through a latex balloon catheter, which was inserted into the left ventricle and connected via stainless steel tubing to a pressure transducer. The balloon catheter inside the ventricle was inflated to provide a systolic pressure of 80-100 mm Hg. Art. and diastolic pressure <10 mmHg Total coronary blood flow was also continuously monitored using a probe acting in the bloodstream and normalizing heart weight.

The heart was allowed to reach an equilibrium state within 30 minutes, after which the heart should show stable pressure values in the left ventricle within the parameters indicated above. If the heart rate dropped below 180 beats / min at any time up to 30 min of the regional ischemia period, the heart was stimulated (accelerated) at approximately 200 beats / min to continue the experiment. Ischemic preconditioning (preadaptation) was induced by complete cessation of cardiac perfusion (global ischemia) for 5 minutes, followed by reperfusion for 10 minutes. Regional ischemia was provided by compressing the loop around the coronary artery branch. After 30 min of regional ischemia, the loop was released and reperfusion of the heart was performed for an additional 120 min.

Pharmacological cardioprotection was induced by infusion of the test compound at predetermined concentrations, starting 30 minutes before 30 minutes of regional ischemia and continuing until the end of the 120 minute reperfusion period. Hearts into which the test compound was not administered were not subjected to ischemic preconditioning (preadaptation). Compound ΑΡΝΕΑ (500 nM), known from the literature, was perfused through the hearts (into which the test compound was not administered) for a 5-minute period, which ended 10 minutes before 30 minutes of regional ischemia.

At the end of the 120 minute reperfusion period, the loop on the coronary artery was compressed and perfused through the heart with a 0.5% suspension of particles of fluorescent zinc-cadmium sulfate (1-10 μM) (Enke ZaepbPs Comp. (Pa1o Α1ίο, CA)); it stains the entire myocardium, except for the area that is an area of increased risk for the development of a heart attack ("area of increased risk"). The heart was removed from the Langendorff apparatus (LapdspbogHG). blotted dry, wrapped in aluminum foil and stored overnight at -20 ° C. The next day, the heart was cut into 2 mm transverse sections from the apex to the upper part of the ventricles. The sections were stained with 1% solution of triphenyltetrazolium chloride (TTC) in phosphate buffered saline for 20 min at 37 ° C. Then TTC was subjected to interaction with living tissue (containing ΝΆΌ-dependent dehydrogenases), this dye distinguishes living (stains red) tissue and dead tissue (does not stain heart attack tissue). For each left ventricular slice, the infarcted area (unpainted) and the area (area) of increased risk (no fluorescent particles) are calculated using a calibrated image analyzer. To normalize the difference between hearts with ischemic damage in the high-risk area, the data were expressed as the ratio of the area of the heart attack to the area of the “high-risk area” (% ΙΛ / ΛΛΚ.). All data were expressed as mean values ± standard error and statistically compared using the non-parametric Mapi-L1peu criterion with VoiGettos correction for multiple comparisons. Results were considered significant at p <0.05.

The results of the above ίη νίίτο test can be used to demonstrate that the compounds of this invention induce significant cardioprotection compared to the control group.

The compound of formula Ι, its prodrug or a pharmaceutically acceptable salt of said compound or said prodrug can be administered by any method that delivers the compound of formula Ι, its prodrug or pharmaceutically acceptable salt of said compound or said prodrug, preferably to the desired tissue (for example, liver tissue and / or hearts). Such methods include oral, parenteral, intraduodenal routes of administration of the drug, etc. In general, a compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, is administered in single (for example, once daily) or in multiple doses, or as a continuous infusion, for example, in isotonic saline.

A compound of Formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug is useful, for example, to reduce or minimize damage by acting directly on any tissue that may be susceptible to ischemic / reperfusion injury (e.g., heart, brain, lung, kidney, liver, intestine, skeletal muscle, net net), due, for example, to the mediation of ΝΗΕ-1, as a result of an ischemic event (for example, myocardial infarction). The active compound is therefore useful for prophylactic use in order to prevent (e.g., in the future or prophylactically), reduce or delay tissue damage (e.g., myocardial tissue) in patients at increased risk of ischemia (e.g., myocardial ischemia).

In general, a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug according to this invention is administered orally or parenterally (for example, intravenously, intramuscularly, subcutaneously or intramedullary). Topical application may also be indicated, for example, when the patient suffers from gastrointestinal upsets or whenever the drug is best applied to the surface of a tissue or organ, as determined by the hospital staff doctor.

The amount and timing of administration of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug will, of course, depend on the subject being treated, the severity of the disease, the route of administration and the indications prescribed by the doctor. Thus, due to patient-to-patient variability, the dosages below are recommendations, and the physician may select the dosage of the drug to achieve a treatment that the physician considers appropriate for the patient. Given the level of treatment required, the physician must balance a number of factors, such as the patient’s age, the presence of a pre-existing disease, and the presence of other diseases (for example, cardiovascular disease).

Typically, an amount of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, is effective that is effective against ischemia. A preferred dosage is about 0.01-100 mg / kg / day of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug according to this invention. A particularly preferred dosage is about 0.01-50 mg / kg / day of a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug according to this invention.

In one route of administration, a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, can be administered only prior to heart surgery (e.g., within 24 hours before surgery), during or after heart surgery (e.g., during 24 hours after surgery) when there is a risk of myocardial ischemia. In a particularly preferred method, a saturation dose (loading dose) of about 1-300 mg is infused over a period of about 1 minute to 1 hour before surgery, followed by a continuous infusion of about 1-100 mg / kg / day to maintain the preoperative , operating and postoperative periods, including, for example, from about 2 to 7 days of postoperative treatment. The compound of the present invention can also be administered on a continuous daily basis.

A compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug is usually administered in the form of a pharmaceutical composition comprising a compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug, together with a pharmaceutically acceptable excipient, carrier or diluent. Thus, a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug, can be administered in any suitable dosage form: oral, parenteral, rectal, or transdermal.

The pharmaceutical composition for oral administration may be in the form of solutions, suspensions, tablets, pills, capsules, powders, and the like. Tablets containing various excipients, such as sodium citrate, calcium carbonate and calcium phosphate, are used along with various disintegrants, such as starch, preferably potato or tapioca starch, and some complex silicates, together with binding agents, such as polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricants such as magnesium stearate, sodium lauryl sulfate and talc are often used to make tablets. Solid compositions of a similar type are also used as the contents of soft and hard gelatin capsules; preferred materials in this regard also include lactose or milk sugar, as well as high molecular weight polyethylene glycols. When aqueous suspensions and / or elixirs are required for oral administration, the compounds of this invention can be combined with various sweeteners, flavoring agents, coloring agents, emulsifiers and / or suspending agents, as well as with such diluents as water, ethanol, propylene glycol, glycerin and the like. their various combinations.

For parenteral administration, solutions can be used, for example, in sesame or peanut oil or in aqueous propylene glycol, as well as sterile aqueous solutions of the corresponding water-soluble salts. In such aqueous solutions, if necessary, a suitable buffer and a liquid diluent can be introduced, first of all providing isotonicity with a sufficient amount of saline or glucose. Such aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal injections. In this regard, all used sterile water products can easily be obtained by standard methods well known to specialists in this field.

For transdermal (e.g. topical) administration, dilute sterile aqueous or partially aqueous solutions are prepared (usually with a concentration of about 0.1-5%), in other words, similar to the above parenteral administration solutions.

Methods for preparing various pharmaceutical compositions with a specific amount of active ingredient are known, or will be apparent in the light of this description, to those skilled in the art. For example, for the preparation of pharmaceutical compositions, see Publication RiattaseiIsa1 8c1spes5. Musk RiYMnpd Sotrapu, Eakeg, Ra., 15th edition (1975).

The pharmaceutical compositions of the invention may contain, for example, 0.0001-95% of a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug. In any case, the composition or composition to be administered will contain an amount effective to treat the disease / condition of the subject being treated.

A compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug will in any case be administered in a suitable composition. The following formulation examples are for illustration only and should not be construed as limiting the scope of the present invention.

In the compositions below, the “active ingredient” is a compound of formula I, a prodrug thereof, or a pharmaceutically acceptable salt of said compound or said prodrug.

Composition 1. Gelatin capsules.

Hard gelatin capsules are prepared using the following ingredients:

Ingredient Amount, mg / capsule

Active ingredient 0.25-100

Starch, ΝΡ 0-650

Flowing starch powder 0-50

Liquid silicone 350 centistokes 0-15

A tablet formulation is prepared using the following ingredients: Formulation 2. Tablets.

Ingredient Amount (mg / tablet) Active Ingredient 0.25-100

Microcrystalline cellulose 200-650

Silica fuming 10-650

Stearic acid (81eaga1 aeI) 5-15

The components are mixed and compressed to form tablets.

Alternatively, tablets, each containing 0.25-100 mg of active ingredients, can be formulated as follows.

Composition 3. Tablets.

Ingredient

Active ingredient

Starch

Cellulose, microcrystalline Amount, mg / tablet 0.25-100 45 35

Polyvinylpyrrolidone (10% solution in water) 4

Sodium Carboxymethyl Cellulose 4.5

Magnesium Stearate 0.5

Talc 1

The active ingredient, starch and cellulose are passed through a No. 45 mesh US sieve and mixed thoroughly. A solution of polyvinyl pyrrolidone is mixed with the resulting powders, which are then passed through a No. 14 mesh US sieve. The granules thus obtained are dried at 50-60 ° C. and passed through a No. 18 US sieve. Sodium carboxymethyl cellulose vine, magnesium stearate and talc, previously passed through a No. 60 US sieve, are then added to the granules, which, after mixing, are pressed onto a tablet machine to make tablets.

Suspensions, each of which contains 0.25-100 mg of the active ingredient in a 5 ml dose, are prepared as follows

Composition 4. Suspensions.

Ingredient Number, mg / 5 ml Active ingredient 0.25-100 Sodium Carboxymethyl Celluloea fifty Syrup 1.25 Benzoic acid solution 0.10 Corrigent In dost. count Dye In dost. count Purified water Up to 5 ml

The active ingredient is passed through a No. 45 mesh US sieve and mixed with sodium carboxymethyl cellulose and syrup until a homogeneous paste is formed. The benzoic acid solution, flavor and color are diluted with a portion of water and added with stirring.

Amount (wt.%) 0.25 25.75

74.00

Then add a sufficient amount of water to obtain the desired volume.

For aerosol, a solution is prepared containing the following ingredients.

Composition 5. Aerosol.

Ingredient

Active ingredient Ethanol

Propellant 22 (chlorodifluoromethane)

The active ingredient is mixed with ethanol and the mixture is added to a portion of the propellant 22, cooled to 30 ° C, and transferred to a filling device. The required amount is then fed into a stainless steel container and diluted with the remaining propellant. Then the valve blocks are attached to the container.

Suppositories are prepared as follows.

Composition 6. Suppositories.

Ingredient Amount (mg / suppository)

Active ingredient 250

Glycerides of saturated 2000 fatty acids

The active ingredient is passed through a No. 60 mesh US sieve and suspended in saturated fatty acid glycerides previously melted using the minimum heat required. The mixture is then cast into a suppository mold with a nominal capacity of 2 g and allowed to cool.

The composition for intravenous use is prepared as follows.

Composition 7. Intravenous solution.

Ingredient Amount

Active ingredient 25 mg

Isotonic saline 1000 ml

A solution of the above ingredients is administered intravenously to the patient.

General experimental techniques

¹ H NMR spectra (300 and 400 MHz) were recorded on a Wapai IiNu + 300 or 400 spectrometer (Wapai Co., Pa1o L11o, SL) equipped with two Raman channels, indirect detection and pulsed field gradients (only along the оси axis). Usually, the spectra were obtained at a temperature close to room temperature (21 ° C), and standard programs of self-blocking and auto-adjustment were used for the installed samples. Chemical shifts of the signals were expressed in parts per million towards a weak field, relative to trimethylsilane. The peak shapes were designated as follows: s, singlet; d, doublet, t, triplet, sq, quadruplet; m, multiplet; sh.s, wide singlet. Resonances designated as exchangeable in a separate NMR experiment, when the sample was shaken with several drops of E ₂ O in the same solvent, were not detected. Mass spectra of chemical ionization at atmospheric pressure (APC1M8) were obtained on a Είδοηκ P1aGogt II or M1stotakk спект spectrometer (M1go8ta88, Mapseyeg, I.K.). When describing the intensity of chlorine or bromine-containing ions, the assumed ratio of intensities was observed (approximately 3: 1 for ³⁵ C1 / ³⁷ C1-containing ions and 1: 1 for ⁷⁹ Vg / ⁸¹ Vg-containing ions), M is based on ³⁵ C1 and ⁷⁹ Vg. In some cases, only representative peaks of ¹ H NMR and APC1M8 are provided.

Chromatography on a column was carried out either with Wakeg 8Scca Cl1 silica gel (40 μm) (S.W. Wakeg, RYShrKitd, N.1.), Or with 8Shca Ce1 60 silica gel (EM 8c1epse5, Bjota, N.1.) On glass columns or on flash columns 12.40 or 75 (Vu1ade) (Ciag1o11e8uSe, UA) under reduced nitrogen pressure. Radial chromatography was performed using a chromatotron (Sytota1o1top), (Natlkop, Pa1o A11o, CA). Unless otherwise specified, commercially available reagents were used. Dehydrated dimethylformamide, 2-propanol, methanol, dimethyl sulfoxide, 1,2-dichloroethane, tetrahydrofuran, toluene, dichloromethane and other solvents for the reaction were supplied A16ps11 Sjet1ea1 Sotrapu (Mi ^ aikee, Khuscheoscch).

Microanalyses were carried out in the analytical laboratory 8c11 \\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\ n: n <s <= -> The terms "concentrated in vacuum" and "removed in vacuum" refer to the removal of solvent under reduced pressure on a rotary evaporator with a solution temperature of less than 90 ° C. The reactions carried out at “0-20 ° C” or “0-25 ° C” were carried out with the initial cooling of the vessel in an isolated ice bath, which allowed raising the temperature to room temperature for several hours. The abbreviation "min" and "h" means "minutes" and "hour", respectively.

Example 1. 5-amino-2quinolone hydrochloride.

A solution of 5-nitro-2-quinolone (12.37 g, 65 mmol) (prepared as described by Sarrk S.; NutShop C.8. 1.At.Syet.8oc. 1938.60.2104) in ethanol (176 ml) under nitrogen atmosphere was treated with 8pC1 ₂ -2H ₂ O (73.4 g, 325 mmol). The heterogeneous reaction mixture was allowed to mix at 23 ° C for 2 hours and heated at the boiling point under reflux for 2 hours. The resulting heterogeneous mixture was cooled to 23 ° C and filtered. The solid was triturated with HCl (1 M, aq., 78 ml, 78 mmol) at reflux for 1 h, cooled to 23 ° C and filtered to give 10.08 g of the desired product (78% yield).

Ή NMR (400 MHz, DMSO-66) δ 6.42 (d, 1 = 10, 1H), 6.71 (d, 1 = 7.6, 1H), 6.81 (d, 1 = 8, 1H ),

7.27 (d, 1 = 8, 1H), 8.04 (d, 1 = 10, 1H), 11.71 (br s, 1H).

APC1M8 161 [M + 1] ⁺ .

Example 2. 5-Hydrazino-2-quinolone hydrochloride.

To a solution of 5-amino-2-quinolone hydrochloride (10.08 g, 51.2 mmol) in HCl (concentrated, 42.7 ml) and H ₂ O (18.2 ml) at 0 ° C was added dropwise a NaNO solution ₂ (3.53 g, 51.2 mmol) in H2O (26.4 ml) while maintaining the temperature below 5 ° C. The resulting red suspension was allowed to mix at 0 ° C for 1 h.

In a separate three neck round bottom flask equipped with mechanical stirrer, cooled to 0 ° C slurry 8pS1 ₂ -2H ₂ O (25.41 g, 112.6 mmol) in HC1 (concentrated, 34.2 mL) and H ₂ O (93 9 ml). A red suspension of a diazonium salt, prepared as described above while maintaining the temperature below 5 ° C., was added dropwise to the suspension. The reaction mixture was heated to 23 ° C and allowed to stir for 3 hours. The resulting suspension was filtered. The collected solid was triturated with HC1 (1 M aqueous, 61.4 ml, 61.4 mmol) at 23 ° C for 30 minutes. The solid was collected by filtration to obtain 7.88 g of the desired product (yield 73%).

'|| NMR (400 MHz, DMSO-66) δ 6.43 (d, 1 = 10, 1H), 6.63 (d, 1 = 8, 1H), 6.88 (d, 1 = 8, 1H),

7.38 (t, 1 = 8, 1H), 7.98 (d, 1 = 10, 1H), 8.97 (br s, 1H), 10.40 (br s, 1H), 11, 77 (br s, 1H).

APC1M8 176 [M + 1] ⁺ .

Example 3. Methyl 5-cyclopropyl-1- (2-quinolon-5-yl) -1H-pyrazole-4-carboxylate.

A solution of methyl 3-cyclopropyl-2- (dimethylaminomethylene) -3-oxopropanoate (7.85 g, 39.8 mmol) in EUN (257 ml) was treated under nitrogen with triethylamine (7.77 ml, 55.8 mmol), then 5-hydrazino-2quinolone hydrochloride (7.88 g, 37.2 mmol). The resulting heterogeneous reaction mixture was heated at reflux for 4 hours and cooled to 23 ° C. A pale yellow solid precipitated was collected by filtration to obtain 5.06 g of the desired product (yield 44%).

'H NMR (300 MHz, DMSO-66) δ 0.59-0.73 (m, 4H), 1.87 (m, 1H), 3.79 (s, 1H), 6.54 (d, 1 = 10, 1H), 7.17 (d, 1 = 10, 1H), 7.34 (d, 1 = 7.5, 1H), 7.49 (d, 1 = 8, 1H), 7.66 (t, 1 = 8, 1H), 8.08 (s, 1H), 12.10 (s, 1H).

APC1M8 310 [M + 1] ⁺ .

Example 4. 5-Cyclopropyl-1- (2-quinolone-5yl) -1H-pyrazole-4-carboxylic acid.

Suspension of methyl 5-cyclopropyl-1- (2quinolon-5-yl) -1H-pyrazole-4-carboxylate (5.06 g, 16.4 mmol) in THF (81.8 ml) and MeOH (40.9 ml) treated with LUN (1 M aqueous, 81.8 ml, 81.8 mmol) and heated at the boiling temperature under reflux for 2.5 hours. The resulting heterogeneous mixture was cooled to 23 ° C. MeOH and THF were removed in vacuo. The resulting suspension was cooled to 0 ° С and acidified to pH 3-4 using НС1 (concentrated). The mixture was filtered. The solid was dried to give 4.94 g of the desired product (yield 100%).

Ή NMR (400 MHz, DMSO-bb) δ 0.58 (d, 1 = 4.4, 2H), 0, b4 (d, 1 = 8, 2H), 1.82 (m, 1H), b, 50 (d, 1 = 10, 1H), 7.12 (d, 1 = 10, 1H), 7.27 (d, 1 = 7, b, 1H),

7.44 (d, 1 = 8.4, 1H), 7, b0 (t, 1 = 8, 1H), 7.97 (s, 1H), 12.03 (br s, 1H), 12, 38 (br s, 1H).

ARS1MZ 29b [M + 1] ⁺ .

Example 5. [5-Cyclopropyl-1 (2-quinolon-5-yl) -1H-pyrazole-4-carbonyl] guanidine hydrochloride.

A solution of 5-cyclopropyl-1- (2-quinolone-5yl) -1H-pyrazole-4-carboxylic acid (13.27 g, 45.0 mmol) in ZOS1 ₂ (b0 ml) was heated at reflux for 1 h. Excess ZOS1 ₂ was removed in vacuo. The residue was treated with dry (dehydrated) toluene and concentrated in vacuo. A solution of the solid residue in THF (50 ml) was treated with a solution of guanidine hydrochloride (15.47 g, 1b2 mmol) in No. 1OH (2 M, aqueous, 1b2 ml, 324 mmol) in a nitrogen atmosphere. The resulting mixture was stirred at 23 ° C. for 2 hours. Then, the reaction mixture was concentrated in vacuo to remove THF. The aqueous layer was cooled to 0 ° C and acidified to pH 9. The precipitated pale yellow solid was collected to give 11.41 g of the free base of the desired compound. A solid solution in MeOH (700 ml) was treated with HC1 (1M in ether, 84.9 ml, 84.9 mmol). The volume of solvent was reduced in vacuo. The resulting pale brown precipitate was collected to give 8.49 g (51% yield). (Additionally, the product in the form of a free base can be obtained by extraction of the aqueous layer using ESAc-THF (1: 1)).

Ή NMR (400 MHz, DMSO-bb) δ 0.51 (m, 2H), 0.70 (dd, 1 = 2, 8.4, 2H), 1.92 (m, 1H), b, 53 ( dd, 1 = 1, b, 10, 1H), 7.17 (d, 1 = 10, 1H), 7.32 (d, 1 = 7, 1H), 7.48 (d, 1 = 8.4 , 1H), 7, b4 (t, 1 = 8.2, 1H), 8.34 (br.s, 2H), 8, b0 (br.s, 3H), 11, b4 (s, 1H), 12.10 (s, 1H).

АРС1МЗ 337 [M + 1] ⁺ .

Example b. Identification of [5-cyclopropyl-1- (2-quinolon-5-yl) -1H-pyrazole-4-carbonyl] guanidine in human plasma samples by liquid chromatography (LC) / mass spectroscopy (MS) / MS and LC / NMR.

5-cyclopropyl-1 - (quinolin-5yl) -1H-pyrazole4-carbonyl] guanidine monohydrate monohydrate was intravenously administered to a man (men and women). Human plasma samples were pooled and precipitated with a double volume of acetonitrile. After centrifugation, the supernatant was concentrated in a water bath in an atmosphere of Ν ₂ at 40 ° C.

Aliquots (100 μl or 50 μl each) of concentrated samples were added to a BS / MZ / MZ instrument equipped with a (4 x b x 150 mm) Kx-C8 column, analyzed on a PE Zs1ekh AP1 2000 mass spectrometer or Είηηί ^ αη BS'O and tested on a UV detector at 254 nm. The eluent from the column was separated so that 5% of the eluent was directed to a mass spectrometer, and the remaining amount was to a UV detector. Analyzes were performed using a flow rate of the mobile phase of 1 ml / min, the mobile phase consisting of 5 mM ammonium formate (pH 3.0) as solvent A and acetonitrile as solvent B. The B / A ratio gradient included values from 5/95 up to 40 / b0 for a 20 minute period after 3 min of the initial retention period. Under such conditions, an LC metabolite was detected with a retention time of 11.8 minutes ([5-cyclopropyl-1- (quinolin-5-yl) -1H-pyrazole-4-carbonyl] guanidine was detected at 13.7 minutes). The difference in 1b mass units between [5cyclopropyl-1- (quinolin-5-yl) -1H-pyrazole-4carbonyl] guanidine (t / ζ 321) and metabolite (t / ζ 337) from human plasma suggests that it is formed by oxidation . The ion spectrum of the product after BS / MZ / MZ of the protonated metabolite at t / ζ 337 ([M + H] ⁺ ) indicated the presence of fragment ions at t / ζ 278, 2b0, 250, 23b, 223 and 208.

Salt was removed from the above concentrated sample by extraction of the solid phase. The sample was concentrated to approximately 5-10 μg metabolite. 50 μl of this sample was added to a BS / NMR instrument equipped with a (4 × b × 150 mm) Column Kx-C8 column, analyzed on a Vgikeg NMR spectrometer (500 MHz) and checked on a UV detector at 235 nm. The analyzes were performed using a flow rate of the mobile phase of 1 ml / min, and the mobile phase consisted of 15 mM deuterated aqueous formic acid with pH 3.3, which was dehydrated in I ₂ O, as solvent C, and acetonitrile-bz, as a solvent I. The I / C gradient included values from 5/95 to 20/80 over a 17 min period after a 3 min period of initial retention. The required metabolite was recorded in the NMR sample, using the method of stopping the flow at peak at 13, b min. The proton NMR spectra and the POP spectra of the human metabolite contained two doublets (b, b4 and 7.41 ppm), which were associated with each other, but not with any other peaks. No peak at 8.92 ppm toward the weak field, which is present in [5-cyclopropyl-1- (quinolin-5-yl) -1Npyrazol-4-carbonyl] guanidine, it was assumed that the 2-position of the quinoline ring corresponds to the oxidation site. Based on these data, the metabolite structure was determined as [5cyclopropyl-1- (2-quinolon-5-yl) -1H-pyrazole-4 carbonyl] guanidine. Comparison of the ¹ H LC / NMR spectrum of the metabolite and the synthetic standard of [5-cyclopropyl-1 - (2-quinolon-5-yl) 1H-pyrazole-4-carbonyl] guanidine hydrochloride obtained as described in Example 5 confirmed the established structure.

Claims (30)

CLAIM 1. The compound of formula I, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug, provided that [5-cyclopropyl-1 - (quinolin-5-yl) -1H-pyrazole-4-carbonyl] guanidine is not included. 2. [5-Cyclopropyl-1 - (2-quinolon-5-yl) -1 Npyrazol-4-carbonyl] guanidine or a pharmaceutically acceptable salt thereof. 3. A method of reducing tissue damage due to ischemia or hypoxia, comprising administering to a mammal in need of such treatment a therapeutically effective amount of a compound according to claim 1, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug. 4. The method according to claim 3, where the tissue is tissue of the heart, brain, liver, kidney, lung, intestine, skeletal muscle, spleen, pancreas, nerve, spinal cord, tissue of the retina, vasculature, or tissue of the intestinal tract. 5. The method according to claim 3, where the amount of the compound according to claim 1, its prodrug or a pharmaceutically acceptable salt of said compound or said prodrug is from about 0.01 to 50 mg / kg / day. 6. The method according to claim 5, where the mammal is a woman or man. 7. The method of claim 6, wherein said tissue is heart tissue. 8. The method of claim 6, wherein said tissue is brain tissue. 9. The method of claim 6, wherein said tissue is liver tissue. 10. The method of claim 6, wherein said tissue is kidney tissue. 11. The method of claim 6, wherein said tissue is lung tissue. 12. The method of claim 6, wherein said tissue is intestinal tissue. 13. The method of claim 6, wherein said tissue is skeletal muscle tissue. 14. The method of claim 6, wherein said tissue is spleen tissue. 15. The method according to claim 6, where the specified tissue is tissue of the pancreas. 16. The method of claim 6, wherein said tissue is retinal tissue. 17. The method according to claim 6, where the compound is administered prophylactically. 18. The method according to claim 6, where the compound is administered before surgery. 19. The method of claim 6, wherein the compound is administered prior to heart surgery. 20. The method according to claim 6, where the compound is administered during surgery. 21. The method according to claim 6, wherein the compound is administered during heart surgery. 22. The method according to claim 6, where the compound is administered within 24 hours after surgery. 23. The method according to claim 6, where the compound is administered within 24 hours after heart surgery. 24. The method according to claim 6, where the tissue lesion due to ischemia is an ischemic lesion and occurs during organ transplantation. 25. The method according to claim 6, where the compound is administered to prevent ischemic damage to the myocardium during surgery. 26. A pharmaceutical composition that contains a compound according to claim 1, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug and a pharmaceutically acceptable carrier, excipient or diluent. 27. A pharmaceutical composition for reducing tissue damage due to ischemia or hypoxia, which comprises a therapeutically effective amount of a compound according to claim 1, a prodrug thereof or a pharmaceutically acceptable salt of said compound or said prodrug, and a pharmaceutically acceptable carrier, excipient or diluent. 28. The method according to claim 6, where the compound is administered before, during and after the operation. 29. The method according to claim 6, where the compound is administered before, during and after heart surgery. 30. Method for the production of [5-cyclopropyl-1- (2-quinolon-5-yl) -1H-pyrazol-4-carbonyl] guanidine by the administration of [5-cyclopropyl-1 (quinolin-5-yl) -1H-pyrazol-4 -carbonyl] guanidine to humans.