CS228104B2 - Preparation of esters of nitric acid and n-/2-hydroxyethyl/nicotinamides - Google Patents

Abstract

PURPOSE:N-(2-hydroxyethyl) nicotinic acid amide nitric acid ester or its salt I.

Description

The present invention relates to a process for the preparation of a nitric acid ester of N- (2-hydroxyethyl) nicotinamide of the following formula.

^ СОННЩСНОЩ

This compound can be used as a pharmaceutical ingredient since it has been found to be useful in the treatment of circulatory disorders.

Some nicotinic acid derivatives or nicotinamide dorivatives are already known in the art, such as Japanese Patent Application Publication No. 1624/1976, U.S. Patent 3,092,634, U.S. Patent No. 3 168,438 and "Mie Medical Journal", Vol. 16 (3), pp. 207-211, 1967.

The above Japanese Patent Application Publication No. 1624/1976 describes nicotinic acid ester derivatives and the preparation of these derivatives by reacting 1-nicotinyl-glycerin or 1-nicotinyl-2,3-isopropylidene glycerin with pumped nitric acid to give 1? -r ^ i ^ k ^ c ^ t ^ i ^ yy ^^^ - ^ g ^^ lyceininir 2,3-di2

Nitrate, and this compound is then converted to a dioxane addition compound having coronary vasodilatory effects.

U.S. Pat. Nos. 3,092,634 and 4,168,438 disclose the preparation of bis, nitrate esters of N, N-bis - (.beta.-hydroxyethyl) -totinamide, which also exhibits coronary-vasodilatory effects by the reaction of dtethanclaminonitrate with nicotinic acid chloride.

However, the above-mentioned compounds are characterized by short-term effects or adverse effects with respect to blood pressure or heart activity, and therefore they cannot be used as medicaments to stimulate blood circulation, for example in ischemic heart disease. Under these conditions, it seems highly desirable to develop some more suitable drug.

In "Mie Medical Journal", Vol. 16 (3), pp. 207-211, 1967, there is an article relating to 2-iquinotinamide thiol. This compound is referred to herein as an anti-tumoral compound only, and no significant pharmacological effect is reported in this journal.

SUMMARY OF THE INVENTION It is an object of the present invention to provide a process for the preparation of such compounds which can be used in the treatment of circulatory diseases, which compounds have a greater effect compared to the compounds used hitherto.

The compounds of the present invention can be represented by the following formula (I)

CONHCH it _{Ch l of} (I)

The essence of the process for the preparation of this nitric acid ester of N- (2-hydroxyethyl) nickel; According to the invention, the compound of formula (I) is reacted by reacting a compound of formula (II)

COOH (II) or a reactive derivative thereof on the carboxyl group with a compound of formula III

NH ₂ —CH ₂ CH ₂ —ONO ₂ (III) or a reactive derivative of this compound on an amino group, in an aqueous medium or in an organic solvent, at a temperature ranging from –10 to +50 ° C for 0.5 hours to 4 hours hours, and optionally in the presence of an inorganic base, an amine compound or an amide accelerator, to form a compound of Formula I.

In a preferred embodiment of the process of the invention, the reactive derivative of the compound of formula II is an acid halide, active amide or active ester thereof. The reaction is preferably carried out at a temperature ranging from 0 to 10 ° C and the organic solvent is preferably selected from benzene, toluene, tetrahydrofuran, diethyl ether, dioxane, dimethylformamide, chloroform, methylene chloride, acetonitrile, acetone, carbon tetrachloride or ethyl ster steric acid. acetic. It is also preferred to use as the inorganic base a compound selected from the group consisting of potassium acetate, sodium acetate, potassium carbonate, sodium carbonate, sodium hydroxide, calcium octaine or calcium carbonate, and amine compounds preferably used in this process are pyridine, triethylamine, dimethylaniline or picoline.

The compounds prepared by the process of the present invention are novel compounds which exhibit superior effects in the treatment of circulatory diseases, such as coronary vasodilation, antihyportensive, antiarrhythmic, ainticoagulatory, and peripheral vasodilatory activity over prior art compounds. Thus, the compounds of the present invention may be used in the treatment of ischemic heart disease as anti-hypertension drugs, as anticoagulant drugs, as anti-rhythmic drugs and as peripheral vasodilators, including cerebral vasodilators and renal vasodilators.

Alternatively, a reactive derivative of a compound of formula III, prepared by reacting a compound of formula III with phosphorus trichloride, methylchlorophosphite or ethylchlorophosphite, and a compound of formula II, starting from room temperature to room temperature, is used as the starting compound. the reflux temperature of the solvent used for a time ranging from 0.5 hours to 3 hours in a neutral solvent or in an organic basic solvent, and optionally in the presence of an amine compound. In this embodiment, a solvent selected from the group consisting of benzene, toluene, xylene, dioxane or teitna is used as the neutral solvent, hydrofuran is selected from the group consisting of pyridine, triethylamine, dimethylaniline and picoline and the organic base solvent is selected from the group consisting of pyridine, triethylamine, dimethylaniline and picoline.

In another alternative embodiment, a compound selected from the group consisting of N, N'-dicyclohexylcarbodiimide, N-cyclohexyl-N'-morpholinethylcarbodiimide, N, N'-diethylcarbodiimide, diphenylketene-N-cyclohexylimine, pentamethylene ketene-N-cyclohoxylimine, triethylphosphorite, ethyl polyphosphate and isopropyl polyphosphate.

In this embodiment, the reaction is carried out at a temperature ranging from room temperature to the reflux temperature of the solvent used, and preferably in an inert solvent selected from the group consisting of benzene, toluene, tetrahydrofuran, chloroform, methylene chloride and acetonitrile.

In carrying out the process of the invention, the above-mentioned functional derivatives of the compounds of formula II on the carboxyl group may, as already mentioned, be acid halides, acid anhydrides, active amides, active esters and the like. Such commonly used derivatives are, for example, acid chlorides, acid azides, acid anhydrides such as anhydrides derived from the two-moles of the compound of formula II and anhydrides thereof with other acids such as dialkyl phosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, benzylphosphoric acid, halogenated phosphoric acid, dialkylphosphoric acid, sulfuric acid, thiosulphuric acid, sulfuric acid, alkylcarbonic acid, fatty acids such as pivalic acid, valeric acid, isovaleric acid, 2-ethylbutanoic acid or trichloroacetic acid, or aromatic carboxylic acids such as benzoic acid, amides such as amides with imidazole, imidazole in the 4-position, dimethylpyrazole, triazole or tetrazole, and esters such as cyanomethyl ester, 4-niirophenyl ester, 2,4-dinitrophenyl ester, t ^ lchlorfenylester, pentachlorophenyl methanaulfonylifenylester, íenylazofenylester, phenylthio ester, 4-nit'rofenylthioester p-kresylthioesíer, carbo-xymethylithioester, pyranylester, pyridylester, quinolin-8-y ^] ^^^ _S -cis tic t «r Ν esters, Ν-dimethylhydroxylamine, 1-hydroxyl-2- (1H) -pyridine, N-hydroxysuccinimide or N-hydroxyphthalimide.

The process for the preparation of the compounds of the formula I according to the invention can be practiced, for example, by reacting a reactive derivative of the compound of the above formula II on a carboxyl group with a compound of the above formula III in order to condense these compounds at temperatures ranging from - 10 to 50 ° C, preferably in the range from 0 to 10 ° C, for a time ranging from 0.5 hours to 4 hours. Various solvents such as water, benzene, toluene, tetrahydrofuran, diethyl ether, dioxane, dimethylformamide, chloroform, methylene chloride, acetonitrile, acetone, carbon tetrachloride, ethyl acetate and the like can be used for the above reaction. Inorganic basic compounds such as alkali or alkaline earth metal hydroxides, carbonates or octains such as sodium acetate, sodium carbonate, potassium acetate, potassium carbonate, sodium hydroxide, calcium acetate, calcium carbonate may be used as accelerators for the above reaction. or amine compounds such as pyridine, triethylamine, dimethylaniline, picoline or the like can be used.

The above condensation reaction between a compound of formula II and a compound of formula III whose amine group has been activated by phosphorus trichloride, ethyl chlorophosphite, methyl chlorophosphite or the like can be carried out in a conventional manner at a temperature ranging from ambient to reflux temperature of the solvent. is used for a given reaction, for a time ranging from 0.5 to 3 hours. The solvent commonly used in the present invention is a neutral solvent such as benzene, toluene, xylene, dioxane or tetrahydrofuran, or a basic solvent such as pyridine, triethylamine, dimethylamine, dimethyyaniline or picoline. When a neutral solvent is used in this embodiment, it is preferable to add an amine compound such as pyridine, triethylamine, dimethylaniline, picoline or the like.

In another embodiment of the process of the invention, the compound of formula II may be reacted with the compound of formula III in an inert solvent in the presence of an amide-forming agent, for example an imide compound such as N, N'-dicyclohexylcarbodiimide, N-cyclohexyl- N, N-morpholinoethylcarbodiimide, N, N'-diethylcarbodiimide or the like, mine compounds such as di-phenylketene-N-cyclohexylimine, pentamethylene ketene-N-cyclohexylimine or the like, or in the presence of a phosphate or phosphioate such as triethylphosphorothiethane, triethylphosphorothiethane, ,. isopropyl polyphosphate or the like, at temperatures ranging from room temperature to the reflux temperature of the solvent used, for a time ranging from 1 to 5 hours. Various inert solvents such as benzene, toluene, tetrahydrofuran, chloroform, dioxane, acetonitrile and dimethylformamide can be used for this reaction. ,

The compounds prepared by the process of the present invention of formula I may be formulated in conventional manner into pharmaceutical compositions in the form of tablets, granules, powders, capsules, suspensions, parenteral injections, suppositories or the like. For preparing tablets, powders, granules or capsules filled with powder or granules, the compounds prepared by the process of the invention may be mixed with one or more pharmaceutical carriers such as lactose, starch, mannitol, kaolin, crystalline cellulose, talc, carbonate, calcium, magnesium stearate or the like.

In the preparation of soft capsules filled with a liquid preparation, the compounds prepared according to the invention are dissolved in oil, and these compounds can also be suspended in acacia gum or in an aqueous solution of sucrose, adjusting the pH. On the other hand, it should be noted that the compounds prepared by the process of the invention may be mixed with mannitol to prepare parenteral injections.

The compounds prepared by the process of the present invention may be used in any form with respect to pharmaceutical compositions, in an amount sufficient to exert an effect of the compounds in the treatment or prevention of circulatory disorders, but in such an amount as it will not cause adverse effects when the compositions are administered. A unit amount in a form such as a tablet or capsule may comprise the ordinarily active compound in an amount ranging from 5 to 20 milligrams when the composition is administered orally. When the composition is administered parenterally, a unit dose, such as in the form of a lamp, contains an amount of active compound in the range of 1 to 10 milligrams.

It goes without saying that the immediate dose varies, which of course depends on the individual patient's condition, and this dose must be determined; separately for each patient. However, the dangerous and commonly used dose, in terms of the amount of active compound, is between 10 and 100 milligrams, preferably 10 to 60 milligrams of active compound per day for an adult patient when administered orally. and generally, the dose is 1 to 100 milligrams, preferably 1 to 50 milligrams per day for an adult patient when parenteral injection is involved.

The accompanying figures illustrate the effects of the compounds prepared by the process of the present invention compared to the compositions used so far, and a detailed discussion of the results will be given in the following section describing the performance of the compounds.

Figure 1 is a graph of the percent increase in coronary blood flow when the compound prepared according to Example 1 or nitroglycerin as a comparator is administered imtravenously according to Experiment 1.

Figure 2 is a graph showing the duration of coronary blood flow increase when the compound was prepared according to Example 1 and the compound and nitroglycerin as comparator were administered intravenously according to Experiment 1.

Figure 3 is a graph showing the change in left-lung venitricular tension when the compound prepared according to Example 1 of the invention or nitroglycerin as a comparator is administered intravenously according to Experiment 1.

Figure 4 is a graph showing the change in cardiac activity when the compound prepared according to Example 1 of the invention or nitroglycerin as comparator is administered intravenously according to Experiment 1.

5, 6, 7 are graphs showing changes in electrocardiograms when the compound prepared according to Example 1 of the present invention is administered intravenously.

Figure 8 is a graph depicting changes in systemic blood pressure when a compound prepared according to Example 1 of the present invention or nitroglycerin as reference is administered intravenously according to Experiment 1.

Figures 9 and 10 are graphs showing the changes in systemic blood pressure when the compound prepared according to Example 1 of the invention is administered intravenously according to Experiment 1.

Figures 11 and 12 are graphs showing changes in aortic blood flow when the compound prepared according to Example 1 of the invention is administered intravenously.

Figure 13 is a test image showing the effect of the compound of Example 1 according to the invention on the platelet coagulation caused by the adenosine phosphate of Experiment 7.

Figure 14 is a test image showing the effect of the compound prepared by the process of the present invention on the platelet coagulation caused by the collagen of Experiment 7.

Experiment 1

The acute toxicity (LD ₅₀ ) of the compound of Example 1 was determined in female and male SD rats (4 weeks old) to which the compound was administered orally or intravenously. The LD ₅₀ dose in both male and female cases ranged from 1200 to 1300 milligrams per kilogram body weight for oral administration, and for intravenous administration ranged from 800 to 1000 milligrams per kilogram animal weight .

Experiment 2

Growth-crossed dogs that were anesthetized by intravenous administration of sodium pentobarbiitain were subjected to oxygen-assisted thoracotomy using a Bird's respirator, and then various physical phenomena were determined and determined as follows.

1) Coronary blood flow (CBF)

The electromagnetic flowmeter probe was attached to the circumflex branch or the beginning of the forward descending branch of the left coronary artery.

2) Coronary perfusion pressure (СВР)

A fine teter connected to the pressure transducer was placed on the distal side of the probe in the circumflex branch of the left coronary artery.

3) Aortic blood flow [AOBF]

An electromagnetic flow meter probe was inserted at the onset of the aorta.

4) Left ventricular tension (LVT)

The extensometer was sewn into the front wall of the left ventricle.

5) Systemic blood pressure (SBP)

This pressure corresponds to the pressure in the marginal areas of the body (hands, feet, etc.), during which the catheter connected to the pressure transducer was inserted into the right femoral artery.

6) Heart activity (HR)

Heart rate was measured with a cardiotachometer using pulse pressure as a trigger pulse.

7) Electrocardiogram (RCG)

The electrocardiogram was recorded using a single-pole conduit on the heart surface using a set of different electrodes on a portion of the anterior wall of the left ventricle.

In addition to the above experiments 1-7, several of the test animals were subjected to renal blood flow (RBF) and -femoral blood flow (FBF) measurements, with the electromagnetic flow probe attached to the left renal artery and left femoral. artery.

The test compound prepared according to Example 1 was dissolved in physiological saline or distilled water and then administered intravenously, orally or sublingually. In order to compare with the effects of the test compounds, initroglycerin as a control drug was administered in the same manner.

Experiment results:

I) Intravenous substance

a) Changes in coronary blood flow:

The diiassolic coronary flow started to increase 10 to 20 seconds after intravenous administration of the test compounds at a dose of 10 µg / kg. The symmetrical coronary flow rate increased at doses of test compound from 250 pg / kg or more, accompanied by a transient decrease in flow rate immediately after administration of the test compound. Mean coronary blood flow showed a sustained increase after administration of test compounds.

The increasing coronary blood flow is shown in Figures 1 and 2, which is expressed as the maximum percentage change in blood flow and duration of action over the values before administration of the test compound. Figure 1 shows the dose of the test compound in pg / kg body weight by the x-axis, administered intravenously. The value given here corresponds to X + SD (N = 5), with X representing the number of animals tested, i.e. 5. The maximum percentage change in blood flow is plotted on the y-axis. The hatched values correspond to nitroglycerin as a comparative substance - and empty - The comparison of the effects of the compound prepared according to the invention with nitroglycerin before and after administration of the compound and nitroglycerin, respectively, is indicated by a solid line over the respective rectangles.

Figure 2 shows the duration of action of the compound prepared according to Example 1-a nitroglycerin in a dose-dependent manner. The x-axis represents the dose of the compound prepared as described in Example 1 of α-nitroglycerin in µg / kg of weight administered intravenously. The meaning of X + SD (N = 5) is the same as in Fig. 1.

The y-axis shows the duration of the effect (i.e., the effect on coronary blood flow) in minutes. Again, there is a comparison of both the compound prepared by the method of Example 1 and the nitroglycerin in terms of duration of action before and after administration.

Administration of the compound of the present invention at doses of 10 µg / kg resulted in a substantial increase in coronary blood flow, and at doses above 500 µg / kg (intensive administration), an increase in coronary In this latter case, the duration of the increased flow rate was nearly 3 hours The effect of the compound prepared by the process of the invention is considerable with respect to the effect of nitroglycerin as a comparator, especially in terms of duration of action.

b) Change in left ventricular tension.

FIG. 3 is a graphical representation of the change in left ventricular tension; In this graph, the dose of the compound of Example 1 and Example 1 is plotted on the x-axis. nitroglycerin in pg / kg body weight, given as an intravenous dose and the meaning of the X + SD symbol (N = 5) is the same as in Figure 1. The y-axis shows the percentage change. The hatched values refer to nitro / lycerin and the open rectangles refer to the compound of the invention.

The left-sided ventricular tension showed a slight decrease after intravenous administration of the compound of Example 1 at doses of 50 ₍ µg / kg or greater. This decrease was almost equivalent to that observed with nitroglycerin at doses over the test compound. or less even at doses of 500 µg / kg (intravenously).

c) Change in heart activity.

A graph depicting changes in cardiac activity is shown in Figure 4. Again, the dose administered in .mu.g / kg - (intravenous administration) is plotted on the x-axis, with X + + SD (N = 5) having the same meaning as in As shown in Figure 4, cardiac activity shows almost no change when administering the compound prepared according to Example 1 of the invention at doses up to 20 µg / kg, while at at doses of 50 µg / kg or greater (intravenous administration) there is a slight dose-dependent decrease up to 17% at doses of about 500 µg / kg.

The values for the compound prepared according to Example 1 are shown by an empty rectangle, while the hatched values are for nitroglycerin, which are given as comparative. Cardiac activity increased with administration of nitroglycerin at doses of 10 µg / kg or greater (again intravenous administration).

d) Change in electrociardiogram.

5 to 7 are graphs showing changes in electrocardiograms for the compound prepared according to Example 1 of the invention administered intravenously. FIG. the x-axis plot of the dose of the compound of the invention in µg / kg administered intravenously, with the symbol X + SD [N - = 5] having the same meaning as in Fig. 1. refers to heart beat, or cardiac activity. As the heart rate increases, the PP interval becomes shorter. From the graph of FIG. 5, it is apparent that the PP intervals were prolonged in a dose-dependent manner by intravenous administration of the compound of Example 1 at. dosages greater than 50 µg / kg.

In Figure 6, the X-axis dose of the compound of Example 1 in µg / kg (compound administered intravenously) is again plotted, with the X + SD symbol [N = 5] as previously defined, and the y-axis plotting the percentage change in PQ interval This PQ interval means conduction of excitation from the atrium to the ventricular ventricle If this PQ interval is of unsuitable length, there will be an irregularity in the conduit.A chart in Figure 6 shows that the PQ interval shows almost no change at doses up to 1000 / zg / kg administered intravenously.

In Fig. 7, the dose of the compound of Example 1 in µg / kg (compound administered intravenously) is again plotted on the x-axis, with the symbol X + SD (N = 5) having the same meaning as above. percentage change of QT _C interval. If we assume that QRST means excite the ventricle, and QT interval is a waveform excitation of the heart chamber. Symbol QTc then represents the QT interval corrected for heart rate. the graph in Fig. 7 shows that QTc intervals prolonged, depending on the dose used, the compounds prepared according to Example 1 of the present invention at doses greater than 50 µg / kg (intravenous administration).

e] Change in systemic blood pressure

Figure 8 is a graph depicting changes in systemic blood pressure when the compound prepared according to Example 1 of the invention or nitroglycerin is administered intravenously with results referring to Experiment 1. The x-axis shows the dose of the administered compound of Example 1 or nitroglycerin in µg / kg (intravenous), wherein the values for nitroglycerin are scored and empty boxes are applied to the compound of the invention [prepared according to Example 1]. The symbol X + SD (N-5) is as previously defined. . The y-axis shows the percentage change in systemic blood pressure.

In FIGS. 9 and 10, the x-axis dose of the compound prepared according to the invention in µg / kg is always plotted. intravenously, with the symbol X +. SD (N = 5) is as previously defined. Figure 9 shows the maximum percentage change in systemic pressure on the y-axis, and Figure 10 shows the duration of this reduced blood pressure, in minutes, on the y-axis.

It can be seen from the graphs of Figures 8-10 that said systemic blood pressure exhibited a substantial dose-dependent decrease in intravenous administration of the test compound of the present invention at doses greater than 50 µg / kg. The effect of the test compound of the present invention on systemic blood pressure was not stronger than that of nitroglycerin as a comparative, but the duration of the effect caused by the test compound was greater than that exhibited by nitroglycerin.

f) Change in aortic blood flow.

Changes in aortic blood flow when administered with the compound of the invention (intravenous) are shown in the graphs of Figures 11 and 12. Both of these figures show the x-axis doses of the compound of the present invention in µg / kg (intravenous) ], the meaning of the X + SD symbol (N = 5) being the same as above The y-axis shows the maximum percentage change in this aortic flow, while the y-axis shows the duration of the effect in Fig. 12 in minutes.

The graphs of Figures 11 and 12 show that aortic blood flow showed a slight dose-dependent increase in the intravenous administration of the compound prepared according to Example 1 of the invention, at doses ranging from 10 µg / kg. Percentage rise. at a dose of 500 µg / kg it reached a maximum of 40 ° / o, but the duration of this effect was not significantly greater compared to the degree of increase in coronary blood flow.

g) Change in renal and femoral blood flow.

Renal and femoral blood flow increased despite the decrease in systemic blood pressure by intravenous administration of the compound of Example 1 at doses ranging from 100 to 250 yg / kg. The increase in renal and femoral blood flow was less than coronary blood flow in terms of the degree of increase in flow and duration of action.

From the above test results, it was confirmed that the compound of the present invention had a greater effect on coronary blood flow than that of nitroglycerin as a comparative, and in addition, the compound of the present invention caused a decrease in blood pressure, decreased heart rate and decreased cardiac tension, which was not excessive at high doses.

In addition, the fact that the compound of the present invention does not interfere with excitatory conduction in the heart, as shown by the non-prolongation of the PQ interval of the electrocardiogram, suggests that the compounds of the present invention are very useful as therapeutic agents in ischemic heart disease.

The compounds of the present invention may be used as antihyperteinous drugs or peripheral vasodilators as they exhibit a long lasting and gentle anitihypertensive effect and a dilatory effect on femoral and renal vessels, as well as a beneficial effect on ischemic heart disease.

II) Oral and sublingual administration

The compound prepared according to Example 2 was sublingually administered to dogs in the form of tablets containing said compound in an amount ranging from 1 to 10 mg. About 2 minutes after administration, coronary blood flow increased significantly. The compound of the invention obtained by the procedure of Example 2 showed almost no change in systemic blood pressure, cardiac tension and aontal flow at a dose of 10 mg. Conversely, sublingual administration of tablets containing the hydrochloride of the active compound of the present invention showed no significant increase in coronary blood flow. When the compounds of the invention, prepared as described in Examples 1 or 2, either in free form or in hydrochloride form, were administered intraduodally at doses of 50 µg / kg or greater, a prolonged increase in coronary blood flow was observed.

It is also apparent from the test results that the compounds of the invention may be used in the form of various preparations, such as parenteral injection, capsules, tablets, granules or powder for oral or sublingual administration. Conversely, nitroglycerin, as a comparator, is not absorbed through the intestinal walls, and therefore administration in this manner is not possible and only sublingual administration is possible.

Experiment 3

Growth-crossing dogs that were anesthetized by intravenous administration of sodium pentobarbital in an amount ranging from 30 to 40 mg / kg were subjected to thoracotomy under the influence of oxygen using a Bird's respirator. A catheter was then inserted in the proximal direction at the beginning of the left coronary artery branch and a contrast agent was injected into the catheter.

The morphological changes in the left coronary artery were filmed using 35 mm kinofilm before and after injection containing the compound of the invention. From the performed analyzes, it was found that coronary heat was greatly dilated by intravenous administration of a compound of the invention at doses of 100 µg / kg or greater.

Experiment 4

The same procedure as in Experiment 2 was carried out in this case, with the increased crossbreeding of the dogs being anesthetized by intravenous administration of sodium pentobarbital. The left anterior descending or circumflex branch of the coronary pulse was mechanically contracted or occluded on the distal side of the portion on which the electromagnetic flow meter probe was applied to experimentally induce a cardiac ischemic event, and the effect of the compound of Example 1 of this invention against this effect was examined.

With incomplete withdrawal, coronary blood flow was moderate. enlarged by the effect of the compound of Example 1 administered intravenously at doses of 100 µg / kg or less, while at doses of 250 µg / kg or more, there was a slight decrease.

After administration of the compounds of the present invention, a more preferred ST increase in the electrocardiogram in the cardiac ischemic portion is achieved, while at the same time regeneration of the ventricular tension in the ischemic portion is observed. After complete occlusion, the compound of the present invention was not observed to improve ST increase or ventricular tension at the midpoint of the ischemic portion, but improvement at the edge point of the ischemic particles was noted

Similarly, when sublingual tablets containing 10 mg of the compound prepared according to Example 2 in free form were administered, the coronary blood flow increased as well as the ST elevation in the electrocardiogram and ventricular tension was reduced.

Experiment 5

I) The guinea pig nipple muscle was isolated and placed in an organic bath which was filled with an oxygenated Tyrode solution (composition: calcium 1.8 mM, potassium 2.7 mM) at 30 ° C. One end of said muscle was attached to an organic bath and the other end was connected to a strain gauge by a fiber.

The effect of the compound prepared according to Example 1 on muscle contraction strength was observed by subjecting the muscle to electrical stimulation (20 V, 5 msec, 1 Hz) using an Ag-AgCl electrode. The calcium ion or isoproterenol antagonist activity of the compound of the present invention was also investigated.

From the test results, it was found that contraction of the nipple muscle that was induced by electrical stimulation was inhibited by the use of a compound of the invention at a concentration greater than 1 µg / ml, and that the calcium ion showed a contradictory effect to this inhibitory effect. On the other hand, when using isoproterenol at a concentration of 0.08 µg / ml, the nipple muscle exhibited excessive excitation, with two or more irregular contractions observed for each individual electrical stimulation. At this stage, the addition of a compound of the invention to said solution at a concentration of more than 2 µg / ml inhibited excessive excitation.

II) In this embodiment, a spiral strip was isolated from the dog's coronary artery, which was placed in an organic bath formed with a free calcium ion lock solution. One end of the tape was attached to an organic bath and the other end was connected to a strain gauge. In this way, the effect of the compound prepared according to Example 1 on the tape contraction caused by polarization of the solution by the addition of 43 mM potassium ions and 1 mM calcium ions was examined.

The results of the tests showed that the contraction caused by depolarization with potassium ions was inhibited by the compound prepared according to the invention, and this inhibitory effect was reduced by the addition of calcium ions.

III) The effect of calcium ions on the increase in coronary blood flow was examined using anesthetized dogs that were anesthetized with sodium pentobarbital in the same manner as in Experiment 2, Embodiment I. The increase in coronary flow induced by administration of the compound of this invention of the invention was inhibited by a larger dose of calcium ions.

IV) The guinea pig (Taenia coli) of the guinea pig was suspended in an organic bath consisting of Tyrode's solution to examine the effect of the compound prepared according to Example 1 on the spontaneous contraction and on the contraction induced by potassium ion depolarization.

The test results show that spontaneous contraction and potassium ion depolarization induced by the contraction was inhibited by the use of a compound of the invention at a concentration of 2 µg / ml, the inhibitory effect being reduced by the addition of calcium ions.

Analysis of the results showed that the compound of the present invention showed a contradictory effect to the effect of calcium ions, with an inhibitory effect on cardiac contraction, a dog-like coronary artery smooth muscle inhibitory effect, and a guinea pig taenia coli contraction inhibitory effect caused by the compounds of the present invention. was opposed to the effect of the added calcium ions. It should also be noted that it was also. It has been found that the compounds of the present invention exhibit an antiarrhythmic effect, since the compounds of the present invention have been shown to inhibit excessive nipple muscle irritation induced by isoproterenol administration.

Attempt 8

According to this experiment, insufficient blood flow was induced such that the dog's coronary artery was experimentally loaded, for example by contracting the coronary artery for a period of time, leaving the artery in this state. Accordingly, peripheral blood pressure and peripheral blood flow were periodically repeated, and spontaneous fluctuations repeated at intervals of several minutes, accompanied by an ST increase in the electrocardiogram. This phenomenon was very similar to the seizure in one variant of angina pectoris in a clinical case.

Periodic fluctuations were found to be caused by periodic spasm in the coronary artery contracted. In this case, therefore, the effect of the compounds of the present invention on this variant of the angina pectoris model was investigated.

The results of the tests showed that periodic fluctuations in coronary blood pressure and coronary flow can be inhibited by intravenous administration of the compound of Example 1 in an amount of 250 g / kg or greater. That is, the effect of a compound of the above periodic fluctuation in blood flow. The effective duration of this inhibitory action ranges from 25 to 40 minutes.

As mentioned above, the compounds of the present invention are effective in treating angina pectoris.

Attempt 7

In this experiment, it was examined whether or not the compound prepared according to Example 1 exerted an anti-diagnostic effect on platelets, in particular an anti-embolic effect.

1. Preparation of the test solution

1.1 Platelet rich plasma treatment [PRP solution].

Rabbit blood, which contained 38% sodium citrate (blood to citriconate ratio 9: 1), was subjected to centrifugation at 100 rpm for 10 minutes with the liquid above. sediment was used as a PRP solution.

1.2 Treatment of the adenosine diphosphate solution (ADP solution).

Adenosine diphosphate was dissolved in physiological saline at a concentration of 100 µg / ml.

1.3 Preparation of the collagen suspension.

Fifty milligrams of collagen, obtained from beef [Slunaliy], was suspended in five million liters of saline, and homogenized in a glass homogenizer for five minutes, then centrifuged at 500 rpm for 5 minutes. for five minutes, and the supernatant was used as a collagen suspension.

1.4 The compound of Example 1 was dissolved in physiological saline to obtain a test solution.

2. Measurement method

The measurement was performed on a conventional aggregometer.

A 0.5 ml PRP solution and a 0.025 ml test solution or a 0.025 ml physiological saline or a physiological saline solution as a 0.025 ml control solution were placed in two tubes and then incubated at 37 ° C. C for 2 minutes. Next, 0.025 milliliters of ADP solution or 0.0.25 milliliters of collagen suspension was added to each tube, and the effect of the compound of the invention on platelet aggregation induced by ADP or collagen was examined.

The effect of the compound of Example 1 on platelet coagulation caused by adenosine diphosphate according to Experiment 7 is shown in Figure 13, while Figure 14 shows a test image showing the effect of the compound of the invention on platelet coagulation caused by collagen. Both graphs show the x-axis time in minutes and the у-axis the degree of platelet aggregation. The two upper curves relate to different concentrations of the adenosine diphosphate or collagen solution used, i. 2.5. 10 ~ ³ M and 1.0. 10 ³ M, and the lowest curve indicates a control solution, i.e., 10 µM. physiological saline.

As shown in FIGS. 13 and 14, the initial aggregation rate induced by the ADP solution in the test compound was the same as that of the sample, but the test compound accelerated the dissociation of aggregated platelets in the course of the course. On the other hand, the test compound prevented the initial collagen-induced aggregation, with the highest level of aggregation less than that of the controls. From the above results it can be seen that the inhibitory effect of the compound of the present invention on platelet aggregation has been confirmed, and the compound of the present invention has also been found to be useful as an anticoagulant.

Attempt 8

In the same manner as in Experiment 2 (5), systemic blood pressure was measured when the test compounds were administered intravenously. The results are shown in the following table.

Table

Test compound

Dose [/ zg / kg]

Percent change

S. B. P.

Duration (min)

Example 1 compound

Compound of Example 8 Nitroglycerin

Comparative test:

In this test, a comparison of nitric acid ester and N- (2-hydroxyethyl) nicotinamide for pharmacological activity with prior art dinitroxydiethylamide nicotinate nitrate as claimed in U.S. Pat. No. 3,036,074 was made.

Try:

In this experiment, the same procedure as described in Experiment 2, points 1, 5 and 6 was performed, with dogs being anesthetized by intravenous administration of 30-40 mg / kg sodium pentobarbital, and a change in coronary blood flow (CBF) was noted in this experiment. , systemic blood pressure (SBP), and cardiac activity, following administration of a compound of the present invention or a comparative compound. In addition, the effects of the compound of the present invention and the comparator on the change in coronary blood flow during coronary artery contraction * and after angina pectoris induced artificially were measured according to the procedures of Experiments 4 and 6.

Results:

a) Change in coronary blood flow (CBF):

The increase in CBF and the duration of this increase in CBF after administration of the compound of the present invention or the comparator is shown in Fig. 1 and Fig. 2.

Intravenous administration of a compound of the invention at a dose of 20 µg / kg showed a significant increase in coronary blood flow. In addition, the effect of the compound of the present invention on the increase in coronary blood flow was significantly superior to that of the comparator at different doses.

Similarly, intravenous administration of a compound of the present invention at 100 µg / kg caused a 20 minute duration of action, while the comparative intravenous duration was significantly shorter. It is clear from the above that the differences in the effects of both substances are significant.

b) Change in systemic blood pressure (SBP).

As shown in Figure 3, the degree of systemic blood pressure drop induced by administration of a compound of the present invention is lower than that of the comparative compound.

c) Change in heart rate (HR)

As shown in Fig. 4, cardiac activity showed almost no change when the compound of the present invention was administered, while it increased significantly when the comparator was administered.

d) Change in coronary blood flow after coronary artery contraction

As shown in Fig. 5, the coronary flow rate increased by intravenous administration of the compound of the present invention, and a decrease in this value was observed when the comparator compound was administered.

e) Effects of a compound of the invention and a comparator on various angina pectoris conditions

Intravenous administration of a compound of the invention at a dose greater than 150 µg / kg had an inhibitory effect on abnormal change in electrocardiogram (ST-increase) and periodic fluctuations in coronary blood flow. In contrast, no inhibitory effect was observed by intravenous administration of the comparator even at doses of 300 µg / kg.

Discussion:

It is generally known that, as far as the pharmacological activity is concerned, anti-anginal drugs are required to increase coronary blood flow over longer periods of time, while avoiding substantial changes in cardiac activity and blood pressure. In these aspects, the compounds of the present invention, as can be seen from the results of tests a), b) and c) above, are much better in terms of effect as anti-anginal drugs compared to the comparator.

Furthermore, in various experiments using pathologic models, and in particular in tests using different models of angina pectoris induced by coronary artery contraction, the compound of the present invention was found to exhibit excellent effects not observed with the comparator. .

In the following examples, the process for preparing the compounds of the invention is characterized in detail.

Example 1

To a mixture of 5 grams sodium bicarbonate, 15 milliliters of water, 1.69 grams of monoethanolamine nitrate, and 20 milliliters of chloroform, 2.5 grams of nicotinyl chloride hydrochloride were slowly added over 10-30 minutes with stirring at a temperature ranging from 0 to 5 ° C. Stirring was continued for an additional 30 minutes and then the chloroform layer was separated. The remaining aqueous layer was extracted with chloro228104 form and the extract was combined with a separate chloroform layer. The organic layer thus obtained was washed with a solution of potassium carbonate in water, dried over anhydrous sodium sulfate and finally evaporated to dryness under reduced pressure. The residue was dissolved in a 1: 1 mixture of ether and isopropanol (1: 1) and hydrogen chloride was bubbled through the solution with continuous cooling to give 2.35 grams of N- (2-hydroxyethyl) -nicotinamide hydrochloride nitrate. Recrystallization from ethanol gave colorless needle crystals with a melting point of 132 ° C.

Analysis for C ₈ H ₁₀ N ₃ O ₄ Cl:

Calculated:

% C, 38.80;% H, 4.07;% N, 16.96; Found:

% C, 38.89;% H, 4.02;% N, 16.72.

IR spectrum (cm ^-1 ):

NH, 3255, C = 0, 1669, ONO2 _; 1640.

Example 2

To a solution consisting of 1.69 grams of monoethanolamine nitrate in 20 ml of pyridine was slowly added 2.5 grams of nicotinyl chloride hydrochloride over a period of 10 to 30 minutes with stirring at 5 T. Then the reaction mixture was stirred for another 30 minutes. minutes and then evaporated to dryness. The residue was dissolved in chloroform and the solution was washed with aqueous sodium bicarbonate solution. The organic layer was separated, dried over anhydrous sodium sulfate and finally evaporated to dryness under reduced pressure. The residue was chromatographed on silica gel, eluting with benzene / ethanol (5: 1). The eluent was evaporated to a semi-solid which was crystallized from diethyl ether to give 1.97 g of nitric acid ester. and N- (2-hydroxyethyl) -nicotinamide.

Recrystallization of the crystals thus obtained from a mixture of diethyl ether and ethanol gave colorless needles having a melting point ranging from 92 to 93 degrees Celsius.

Example 3

In this example, a solution consisting of 10 grams of nitric acid ester hydrochloride and N- [2-hydroxyethyl) -nicotinamide, was neutralized in water with an aqueous solution of sodium bicarbonate. The solution was extracted with chloroform, and the extract was dried over anhydrous sodium sulfate and then evaporated to dryness under reduced pressure. The residue was crystallized from diethyl ether to give 7 grams of nitric ester and N- (2). Recrystallization from a mixture of isopropanol and diethyl ether gave colorless needle crystals having a melting point of 93 ° C.

IR spectra ^[cm-1 "):

NH, 3250, ONO2, 1630.

Example 4

In this example, to a mixture of 1.69 grams of monoethanolamine nitrate in 5 mL of water, 3.8 mL of 35% potassium carbonate solution and 17 mL of tetrahydrofuran, 3 grams of isonicotinyl chloride hydrochloride and 11.2 mL · 35% aqueous potassium carbonate solution were added. during 30 minutes with stirring at a temperature ranging from 0 to 5 degrees Celsius. The reaction mixture was stirred for an additional 15 minutes and the organic layer was separated.

The obtained aqueous layer was extracted with tetrahydrofuran and the extract was combined with a previously separated organic layer. The combined organic layers thus obtained were dried over anhydrous sodium sulfate and the solution was filtered. The solvent was evaporated in vacuo and crystallization of the residue gave ^L es er nitric acid and N- (2-hydroxyethyl) -isonicotinamide.

The product thus obtained was dissolved in diethyl ether and treated dropwise with a mixture of diethyl ether and fuming nitric acid under cooling to give 1.5 g of nitric ester and N- [2-hydroxyethyl) isonicotinamide nitrate. Recrystallization from isopropanol gave crystals of m.p. 150 ° C (dec.).

Analysis for C ₈ H ₁₀ N ₄ O 7:

Calculated:

% C, 35.04;% H, 3.68;% N, 20.43; Found: ·

H, 3.59; N, 20.38.

IR spectrum ^[cm-1]:

NH, 3280, C-O, 1671, ONO ₂ , 1625.

Example 5

3 grams of nicotinyl chloride hydrochloride and 11.2 milliliters of 35% solution were added to a mixture of 1.83 grams of nitric acid ester and isopropanolamine nitrate in 5 milliliters of water, 3.8 milliliters of 35% aqueous potassium carbonate solution and 17 milliliters of tetrahydrofuran. potassium carbonate over 30 minutes with stirring at a temperature ranging from 0 to 5 ° C. The reaction mixture was 222310

The mixture was stirred for an additional 15 minutes and the resulting organic layer was separated.

The aqueous layer was extracted with tetrahydrofuran and the extract was combined with a previously separated organic layer. The combined extracts were dried over anhydrous sodium sulfate and filtered. Evaporation of the solvent under reduced pressure and crystallization of the residue gave the nitric acid ester of N- (2-hydroxypropyl) nicotinamide.

The product was dissolved in acetone and hydrogen chloride was bubbled through the solution to give 1.5 grams of nitric acid ester and N- (2-hydroxypropyl) nicotinamide hydrochloride as colorless crystals. The melting point of the product was 161 ° C (dec.).

Analysis for C9H12N3O4Cl:

Calculated:

% C, 41.31;% H, 4.62;% N, 16.06; Found:

% C, 41.38;% H, 4.60;% N, 16.11.

IR spectrum (tin- ¹ ):

NH, 3230, C = 0, 1672, ONO ₂ , 1620.

Example 6

The procedure was similar to that in Example 4, in which nicotinyl chloride hydrochloride was reacted with a nitric acid ester and a propanolamine nitrate to give the nitric acid ester and N- (3-hydroxypropyl) nicotinamide hydrochloride. The yield was 75 ° / o, the melting point of the product thus obtained was 127 ° C (dec.).

Analysis for C5H12N3O4Cl:

Calculated:

% C, 41.31;% H, 4.62;% N, 16.06; Found:

% C, 41.40;% H, 4.53;% N, 16.15.

IR spectrum (cm ^-1 ):

NH, 3235, C = O, 1672, ONO ₂ , 1617.

Example 7

The procedure was similar to that described in Example 4, wherein isonicotinyl chloride hydrochloride was reacted with a nitric acid ester and a propanolamine nitrate to give the nitric acid N- and 3-hydroxypropyl) isonicotinamide hydrochloride. The yield was 72 ° / o and a melting point of 125 ^q C (dec).

Analysis for C 8 H 11 N 3 O 4 Cl:

Calculated:

S:% C, 41.31;% H, 4.62;% N, 16.06; Found:

% C, 41.40;% H, 4.58;

IR spectrum [cm ^-1 ]:

NH, 3230, C = O, 1672, ONO ₂ , 1615.

Example

The procedure was similar to that in Example 4, in which picolinyl chloride hydrochloride with monoethanolammonium nitrate salt nitrate was reacted to give an ester of nitric acid and N- (2-hydroxyethyl) -picolinamide to yield 78 g. % and the melting point of the product thus obtained was in the range of 55 to 56 ° C.

Analysis for CyHgN4O / 1

Calculated:

45.50% C, 4.29% II, 19.89% N; Found:

H, 4.10; N, 19/17.

IR spectrum (cm ¹ ) ·

NH, 3362, C = O, 1665, ONO ₂ , 1620.

Example 9

The procedure was similar to that in Example 4, wherein the nicotinyl chloride hydrochloride was reacted with 2,3-dihydroxypropylamine to give N- (2,3-hydroxypropyl) nicotinamide dinitrate as a pale yellow oil.

Example 10

To 40 ml of a pyridine solution containing 2.2 grams of ethanolamine nitrate was added 15 ml of a pyridine solution containing 1.37 grams of phosphorus trichloride under cooling with water. After stirring for 1 hour at room temperature, 2.5 g of nicotinic acid was added to the resulting mixed solution, and the solution was allowed to react for 48 hours with stirring at 40 ° C.

After completion of the reaction, pyridine was filtered off under reduced pressure and then extracted with ethyl acetate, further washed with sodium bicarbonate as an aqueous solution. The pH of the organic layer was adjusted to 1 by the addition of 2N hydrochloric acid solution, and the aqueous layer thus formed was separated and basified by the addition of aqueous potassium carbonate solution and then extracted with ethyl acetate. The organic layer was dried over sulfate

Compound prepared according to Example 2 20 mg lactose 176 mg magnesium stearate 4 mg total 200 mg / sodium capsule and then concentrated under reduced pressure. Ethyl ether was added to obtain the nitric acid ester of N- (2-hydroxy) nicotinamide. Yield 20%. Recrystallization from a mixed solution of acetone and water gave crystals of the product, m.p. 92 DEG C. (dec.).

Example 11

Pharmaceutical preparations:

a) Sublingual tablets

Compound prepared according to example 2 5 mg lactose 19.7 mg mannitol 25 mg magnesium stearate 0.3 mg total mg / tablet

c) Hard capsules

The above ingredients - were uniformly mixed in the above - amounts, each capsule weighing 200 mg and designated no. 3, and further filled with a mixture on a filling machine so that each capsule weighed 250 mg.

(d) Granules

Compound prepared according to Example 2 10 mg lactose 710 710 corn-starch paste 280 ms

The ingredients were uniformly mixed in the above amounts and the resulting mixture was formed into tablets each having a diameter of 5 mm and weighing 50 mg.

overall

1000 mg / bp

b) Tablets for internal use

The above ingredients - were uniformly mixed and kneaded - and then the mixture was granulated to form granules, each granule having a diameter of 1 mm.

Compound prepared according to

e) Parenteral injection of Example 1 10mg lactose, 5mg corn starch 20mg crystalline cellulose 225mg magnesium stearate 0.5mg

Compound prepared according to Example 2 mannitol overall

-mg mg mg / dose total

100 mg / tablet

The above ingredients were uniformly mixed in the above amounts and the mixture was formed into tablets each having a diameter of 7 mm and weighing 100 mg.

The above ingredients were dissolved in 1 ml of distilled water and the solution was sterilized - and then filtered. The solution thus prepared was a unit dose, freeze-dried and finally sealed for use as a parenteral injection. The freeze-dried mixture was dissolved in 1 mL of distilled water and could be used as a parenteral injection.

Claims (13)

A process for the preparation of a nitric acid ester of N- (2-hydroxyethyl) nicotinamide of the formula I CONHCHCCHiONOz (II) or a reactive derivative thereof on a carboxyl group with a compound of formula III, characterized in that the compound of formula II is reacted -CH2CH2-NH ₂ ONO 2 (III) or with a reactive derivative thereof, · to an amino group in an aqueous medium or in an organic solvent at a temperature ranging from -10 to +50 ^o C for 0.5 hour to 4 hours, optionally in the presence of an inorganic base, an amine compound or an amide-forming agent, to form a compound of formula I. 2. A process according to claim 1, wherein the reactive derivative of the compound of formula (II) is an acid halide, active amide or active ester thereof. 3. The process of claim 1 wherein the reaction is carried out at a temperature in the range of from 0 to 10 ° C. 4. A process according to claim 1, wherein the organic solvent is selected from the group consisting of benzene, toluene, tetrahydrofuran, diethyl ether, dioxane, dimethylformamide, chloroform, methylene chloride, acetone, trieth, acetone, carbon tetrachloride or ethyl acetate. 5. A process according to claim 1 wherein the inorganic base is a compound selected from the group consisting of potassium acetate, sodium acetate, potassium carbonate, sodium carbonate, sodium hydroxide, calcium acetate or calcium carbonate. 6. The process of claim 1, wherein the amine compound is a compound selected from the group consisting of pyridine, triethylamine, dimethylaniline or picoline. 7. The process of claim 1 wherein the starting compounds are a reactive derivative of a compound of formula III, which is prepared by reacting a compound of formula III with phosphorus trichloride, methylchlorophosphite or ethylchlorophosphoric acid, and a compound of formula II. at a temperature ranging from room temperature to the reflux temperature of the solvent used for a time ranging from 0.5 hour to 3 hours in a neutral solvent or in an organic basic solvent, and optionally in the presence of an amine compound. 8. The process of claim 7, wherein the neutral solvent is a solvent selected from the group consisting of benzene, toluene, xylene, dioxane or tetrahydrofuran. 9. The process of claim 7 wherein the amine compound is selected from the group consisting of pyridine, triethylamine, dlmethylamine and picoline. 10. The process of claim 7, wherein the organic basic solvent is selected from the group consisting of pyridine, triethylamine, dimethylaniline, and picoline. 11. The method of claim 1 wherein the amide accelerator is a compound selected from the group consisting of N, N'-dicyclohexylcarbodiimide, N-cyclohexyl-N'-morpholinethylcarbodiimide, N, N'-diethylcarbodiimide, diphenylketene-N. -cyclohexylimine, pentamethylenketene-N-cyclohexylimine, triethylphosphite, ethyl polyphosphate and isopropyl polyphosphate. 12. The process of claim 11 wherein the reaction is carried out at a temperature ranging from room temperature to the reflux temperature of the solvent used. 13. The process of claim 11 wherein the reaction is carried out in an inert solvent selected from the group consisting of benzene, toluene, tetrahydrofuran, chloroform, methylene chloride, and acetonitrile.