HUT75093A - New anti-radical pharmaceutical compositions and process for producing them - Google Patents

Abstract

The invention relates to free radical production-inhibitory pharmaceutical compositions containing as active ingredient 3-methyl-7-ethyl-9 alpha -(4-chlorobenzoyloxy)-3,7-diazabicyclo[3,3,1]-nonane prepared in a known manner or a therapeutically acceptable salt thereof together with fillers, diluents and other formulation-promoting additives known in se and commonly used in the preparation of pharmaceutical compositions. Furthermore, the invention relates also to a process for the preparation of these compositions, to their use for the preparation of pharmaceutical compositions having free radical production inhibitory effect and for a method for treating clinical conditions in patients induced by oxidative stress or weakening or exhaustion of the endogenous antioxidant protection of the organism.

Description

Date of filing: January 1, 1995? ..

50P3

The present invention relates to 3-methyl-7-ethyl-9a- (4-chlorobenzoyl-oxy) -3,7-diazabicyclo [3,3,1] nonane, or a pharmaceutically acceptable salt thereof, which is an active ingredient of the invention. pharmaceutically acceptable salts thereof, and pharmaceutically acceptable excipients, diluents, and other excipients which are conventional in the preparation of pharmaceutical compositions, and a process for their preparation.

THE/

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SERVICE INVENTION

Oj, pharmaceutical compositions having a root-producing effect and a process for their preparation

RICHTER Gedeon Chemical Factory Co., Budapest

The inventors:

dr. PELLIONISNÉ dr. Margit Paróczai, 45 % dr. Karpak Egon, 30 % dr. Katalin REGÖS-SZABÓ, 10 % Budapest, dr. Erzsébet RÖTH, 15 % Pécs.

Date of filing: 1995.01 ....

BACKGROUND OF THE INVENTION The present invention relates to a novel root body having an effect of 1 <£ £ V **.

as an active ingredient of the known 3-METHYL-1-7 1-9a- ethyl (4-l'S'r _oj,? K-1 ox i o'í _Z) -3,7-diazabicyclo [3,3,1Jnonánt (the hereinafter referred to as bisaramyl) or a pharmaceutical composition containing a pharmaceutically acceptable salt thereof, and a process for the preparation thereof.

Bisaramil, its preparation and its action are described in Hungarian Patent Application No. 184,960. In the specification, the class of compounds containing bisaramil is described as having antiarrhythmic activity, supported by activity data.

If the pacemaker stimulates the functional units on the appropriate conduction pathways in the heart in time, the coordinated contraction of the atria and ventricles will result in full functioning of the heart pump.

If the heart's pacing or pacing system is damaged, arrhythmias of different types and severities can occur. Some arrhythmias occur in healthy individuals without sequelae, but can also lead to life-threatening conditions, in which medication is essential. The cited patent describes such therapeutic use of bisaramil.

Another very significant group of heart diseases is related to coronary heart disease. Coronary artery obstruction or stress-induced coronary spasm, which is the cause of ischemic cardiovascular damage, and its consequences are among the leading factors in the mortality statistics of developed countries. For patients after myocardial infarction, the course of the disease is decisively influenced by the size of the dead area. Thus, therapeutic efforts aimed at reducing mortality are both aimed at preventing infarction and reducing the extent of infarction. ¹

In the living organism and inorganic environment, fragments of the electron system, uncompensated by enzymatic and non-enzymatic means, are formed continuously, so-called free radicals, which are eliminated by antioxidant type compounds (reduced glutathione, ascorbic acid, vitamin E, etc.) and enzymes. (superoxide dismutase, catalase, glutathione peroxidase). In the case of malfunctioning of the body, free radicals initiate a chemical chain reaction called lipid peroxidation and primarily cause irreversible damage to the membrane structure of the cellular, non-lipid-rich lipid-rich membrane. N.Y. Acad. Sci., 393, 1-22 (1982)].

In the early 1980s, studies exploring the role of reactive oxygen species in various pathological processes such as oxygen toxicity, inflammation, and ischemia-induced tissue reactions became known. [Can. J. Physiol. Pharmacol., 60, 1346-1352. (1982)].

The importance of the role of the superoxide anion (or superoxide radical) of formula 02 ", formed by uptake of an electron from the oxygen molecule by enzymes, was first demonstrated in ischemic conditions after intestinal ischemia [Gas t roent e ro 1, .-29.

(1981)].

During ischemia, the enzymatic and non-enzymatic antioxidant systems normally present in tissues and cells can no longer counteract the increase in free radical production, as they are rapidly utilized, inactivated, or degraded in the oxygen deficient state.

It is now well-known that free radical damage is associated with the development of a number of chronic health disorders of various etiologies, such as pulmonary dilatation, cardiovascular, and inflammatory and autoimmune diseases, diabetes, gastric ulcer, liver injury, and cancer.

White blood cells play an important role in free radical damage. Namely, ischemia promotes the attachment of white blood cells to endothelial cells of the vascular wall and then to the vascular wall. During this process, toxic oxygen radicals are produced which kill the cells and release enzymes that break down the structures that have not yet been destroyed.

The present invention is based on the newly discovered anti-root-producing effect of the known bisaramil, which is not surprising in any way due to the previously known effect of bisaramil on the cardiac pacing and excitatory system.

The following assays demonstrate the inhibitory effect of bisaramil on root production:

1. Investigation of the root-producing ability of isolated white blood cells

Venous blood from healthy mongrel dogs was prepared as a rich suspension in leukocytes. Superoxide radical production was achieved by the addition of forbo1-myristate acetate (4β, 9α, 12β, 13α, 20-pentahydroxylglyl-1,6-dien-3-one-12β-πιΐ cross-13-acetate) to the suspension. We stimulated. Addition of a phorbol myristate acetate superoxide residue to a cell suspension of polymorphic neutrophilic granulocytes. Meanwhile, the Fe ³⁺ ions in the ferricithochrome C complex are reduced to Fe ²⁺ . The resulting Fe ²⁺ -containing pink ferrocytochrome C complex allows spectrophotometric monitoring of the reaction at 550 nm. The formation of the pink ferrocytochrome C complex is proportional to the amount of superoxide radicals formed. When measuring the ability to generate the root, various amounts of the test substance were first added to the suspension with bisaramyl monohydrochloride for 5 minutes at 37 ° C and the suspension was stimulated with the radical. Measurements were taken in the next liver, after which the results were obtained.

1._t á b 1 áz a t

Percentage change in white blood cell production by bisaramil Treatment

Concentration [µg / ml]

Relative root production L%]

Kont ro1 1 Bisarami1

100

The data in the table show that bisaramil is able to inhibit the root-producing activity of isolated white blood cells in a dose-dependent manner.

To demonstrate that the known antiarrhythmic activity of bisaramil does not result in newly recognized free radical inhibitory activity, some of the commercially available and clinically considered antiarrhythmic drugs have been similarly tested as described above. The results are shown in Table 2.

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Leukocyte Table 2 1 tablespoon root % change in known antia rhythmic drugs Treatment Concentration Relative root production [Μg / ml 1 Γ 1% Contro 1 1 - 100 L i doca in 10 165 50 124 75 140 procainamide 10 113 50 78 75 85 sotalol 10 94 50 76 75 85 amiodarone 10 141 50 169 75 206 Verapami1 10 144 50 96 75 85

Chemical names of the above active substances:

Lidocaine = α- (diethyl-1-amino) -2,6-dimethyl-acetamide

Procainamide = 4-amino-N - [(2-diethyl-1-amino) -ethyl] -benzamide

Sotalol = N- {4- [1-hydroxy-2 - (<1-methylethyl> amino) ethyl] phenyl} methanesulfonamide

Amiodarone = [2-Butyl-1-benzofuran-3-yl] - {4- [2- (diethyl-amino) -ethoxy] -3,5-diiodophenyl} -ketone

Verapamil = 5 - {[2- (3,4-dimethoxyphenyl) ethyl] methyl 1-amino} -2- (3,4-dimethoxyphenyl) -2-isopropylvaleronitrile.

From the data in the table, it can be seen that the known antiarrhythmic drugs do not inhibit the production of white blood cells by the root, and in some cases even stimulate them.

In order to demonstrate that the in vivo inhibitory effect on toxic oxygen atoms can be measured in vivo, the following animal study was performed:

2. Examination of physical performance in mice

The swimming physical test was used to determine maximum physical performance. This consisted of placing 32 g of LATI CFLP male mice in a pool filled with 20 ° C water which was bubbled by air bubbling. To make swimming difficult, the animals had a tail weight of 2 g. Maximum performance was measured from the time the mice were placed in the water to the end of the swim movement. Mice were tested once daily for 2 days to exclude good or poorly floating animals. Experimental groups were selected from animals with a mean value. One group contained 10 mice. Treatment was with bisaramyl monohydrochloride dissolved in physiological saline solution through the stomach tube 1 hour before the swimming test. The control group received only the same volume of saline.

Muscle performance is limited by the amount of available energy, the accumulation of lactic acid and other metabolites, and the increased lipid peroxidation due to relative hypoxia, which leads to the formation of toxic oxygen radicals [Biochem. Biophys. Gap. Commun., 107, 1198-1205. 1981]). In our studies, bisaramil has been shown to significantly increase muscle performance in a dose-dependent manner, even at single doses (Table 3). With once-a-day, repeated treatment, the increase in performance is even more evident (Table 4).

Table 3

The change of maximum swimming time is due to a single bisarami1 hand 1 and

Dose [mg / kg] swimming time [sec] change r%] before treatment after treatment 3 66.6 78.0 + 17.1 10 70.8 101.0 + 42.7 20 65.4 109.2 + 67.0 T 4 b 1 á z a t

The change of the maximum swimming time is 7 daily bisarami1 hands 1 and effect a

Dose [mg / kg] change in swimming time [sec] before treatment on day 7 of treatment_f% 1

7x10 66.0 105.0 +59.1 control 70.2 75.6 +7.7

The following study was conducted to confirm that the increase in physical performance was due to the antioxidant effect of lipid peroxidation.

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The physiological antioxidant protection of the body was reduced to be ionic sulfoximinine (BS) [L- (S-n-butyl) -homocysteine-SR sulfoximin]. As a result of BS treatment, the amount of reduced glutathione (GSH), which provides protection against oxidative stress, is reduced in the body, which may also manifest itself in a decrease in physical performance. Indeed, we found that GSH depletion induced by BS reduced physical performance by nearly 20% (Table 5). Bisaramil treatment after GSH depletion not only restored the original performance but also significantly enhanced it.

Table 5

Changes in maximal swimming time after a single bisarami1 treatment after GSH depletion

Group swimming time [sec] change before treatment after treatment_Γ% 1

GSH depletion 64.8 52.8 -18.5 GSH depletion + 30 mg / kg po · HC1 bisarami1 67.8 112.2 + 65.5 In summary, the test eredményekbő1 it is visible

that bisaramil, due to its potent antioxidant effect, can be used in all conditions where oxidative stress is the underlying cause of pathological events or when the endogenous antioxidant defense of the body is weakened or depleted.

Thus, the therapeutic use may be in the following diseases, disorders: bronchial asthma, spastic bronchitis, pulmonary fibrosis, Parkinson's disease, stroke, head trauma, rheumatoid arthritis, arteriosclerosis, hypertension, thalassemia, myocardial infarction, hemorrhagic shock, ischemic bowel, kidney and cerebrovascular disease, toxic liver damage, cataract, aging, cancer.

. The bisaramil and acid addition salts thereof are formulated into pharmaceutical compositions in admixture with conventional non-toxic inert solid or liquid carriers and / or excipients suitable for enteral or parenteral administration. The pharmaceutical compositions may preferably contain from 5 to 200 mg of active ingredient per unit dosage form and may be achieved by administering one or more dosage units per day. The daily dose

It may range from 5 to 200 mg, but depending on the patient's condition and responsiveness, the severity of the disease being treated, the physician may vary both upwards and downwards based on his / her expertise. Suitable excipients in the formulations include water, gelatin, lactose, lactose, starch, pectin, magnesium stearate, stearic acid, talc, vegetable oils such as peanut oil and olive oil, and the like. It can be used. The active ingredient may be in the form of conventional pharmaceutical compositions such as, for example, solid, e.g., round or square tablets, dragees, capsules such as gelatin capsules, pills, suppositories, and the like. in the form of. The amount of solid carrier can vary over a wide range, preferably about 50 mg to about 1 g. The compositions may optionally contain conventional pharmaceutical excipients such as preservatives, stabilizers, wetting agents, emulsifying agents, and the like. may also be included. They are prepared by conventional techniques such as screening, mixing, granulating and pressing the components in the case of solid compositions. The formulations may also be subjected to other standard pharmaceutical techniques, such as sterilization, to form solutions for injection (injection).

The following examples illustrate the process of the invention without limiting it.

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Preparation of 3-Methyl-7-ethyl-9c- (4-chlorobenzoyloxy) -3,7-diazabicyclo [3.3.11] nonane and its monohydrochloride # 18.4 g (0.1 mol) of 3-methyl-7- Ethyl 3,7-diazabicyclo [3.3.1] nonan-9a-ol is dissolved in 80 ml of anhydrous pyridine and 22.7 g (0.13 mol) of A solution of chlorobenzene-1-chloride in 80 ml of anhydrous pyridine is added dropwise. The reaction mixture was stirred at room temperature for 3 hours and then concentrated in vacuo to remove solvent. The residue was dissolved in water (120 ml), acidified to pH 2 with concentrated hydrochloric acid and extracted with ether (50 ml) to remove non-basic substances. The aqueous phase was basified with potassium carbonate and extracted with chloroform (3 x 60 mL). The combined chloroform phases are washed with 20 ml of saturated sodium chloride solution, dried over anhydrous magnesium sulfate, filtered and washed twice with 5 ml of chloroform each time. The filtrate was evaporated to solvent-free. The residue was crystallized from diisopropylether. 27.45 g (85%) of 3-methyl-7-ethyl-9a- (4-chlorobenzoyloxy) -3,7-diazabicyclo [3.3.1] nonane are obtained. Mp: 74.5-75.5 'C.

From the base obtained above, a monohydrochloride salt is prepared in a known manner with an equivalent amount of hydrochloric acid. After recrystallization from isopropyl alcohol, it melts at 240-242 ° C.

2. p é 1 d a

200 mg tablets with 50 mg active ingredient:

bisaramyl monohydrochloride 50 g colloidal silicon dioxide 1 stearin 2 pyridone 4 thalacium 6 carboxymethyl starch 6

Yactose 60 Starch 71

From the above materials 200 mg tablets are prepared by conventional wet granulation and compression.

. example

225 mg dragees, containing 50 mg of active ingredient:

Tablets prepared in the above manner are coated in a known manner with a layer of sugar and talc. The dragees are polished with a mixture of beeswax and carnauba wax.

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- 10 4. p e 1 d mg injection:

bisaramyl monohydrochloride propi-1-paraoxybenzoate methyl 1-paraoxybenzoate 10% Hydrochloric Acid Sodium chloride d glycine distilled water

25.0 mg 0.25 II 2.5 II 0,015 ml 31.5 mg 35.0 mg 5.0 ml volume t ig.

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Claims

Claims (7)

► 1 ·) 3-Methyl-7-ethyl-9α- (4-chlorobenzoyl-oxy) -4,3,7-diazabicyclo [3,3,1] nonane, which is an active ingredient in the inhibitory effect on root erosion, or a pharmaceutically acceptable salt thereof, and pharmaceutical formulations known per se which contain fillers, diluents, and other excipients which are conventional in the preparation of pharmaceutical compositions. Compositions according to Claim 1, characterized in that the active substance is the monohydrochloride of 3-methyl-7-ethyl-9a- (4-chlorobenzoyloxy) -3,7-diazabicyclo [3.3.1] nonane. They included. 3.) A process known as 3-methyl-7-ethyl-9a- (4-chlorobenzoyl-oxy) -3,7-diazabicyclo [3,3,1] nonane as an active ingredient for inhibiting root production, or a pharmaceutically acceptable salt thereof. or a salt thereof, characterized in that the 3-methyl-7-ethyl-9a- (4-chlorobenzoyloxy) -3,7-diazabicyclo [3.3.1] nonane is prepared in a known manner, or a pharmaceutically acceptable salt thereof, in admixture with excipients, diluents, and other excipients commonly used in the preparation of pharmaceutical compositions, to form a pharmaceutical composition. 4. A process according to claim 3 wherein the active ingredient is the monohydrochloride of 3-methyl-7-ethyl-9a- (4-chlorobenzoyloxy) -3,7-diazabicyclo [3.3.1] nonane. used. 5.) The known 3-methyl-7-ethyl-9a- (4-chlorobenzoyl-oxy) -3,7-diazabicyclo [3,3,1] nonane, or a pharmaceutically acceptable salt thereof, is a pharmaceutical preparation having an anti-root effect. application. 6.) A method of treating disorders based on oxidative stress or depletion or depletion of endogenous antioxidant defenses in the body, wherein the subject is treated with 3-methyl-7-ethyl-9a- (4-chlorobenzoyl-oxy) -3. Or one or more effective doses of a 7-diazabicyclo [3,3,1] nonane, or a pharmaceutically acceptable salt thereof, alone or in the form of a pharmaceutical composition. 7) A process according to claim 6 for the treatment of myocardial infarction, prizes _ft 1 characterized in that the individual to be treated in the three-meti1-7-eti1-9a- (4-chloro-benzoi1-yloxy) -3,7 • »·« • · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·