LU84902A1 - SYNERGETIC ANTIHYPERTENSIVE PHARMACEUTICAL COMPOSITION - Google Patents

Description

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The present invention relates to a pharmaceutical composition containing a combination of two active substances, indapamide and 8- (3-tert-butylamino-2-hydroxypropoxy) -thiachroman.

5 Indapamide, or N- (3-sulfamoyl 4-chloro benza-mido) 2-methylindoline, or even 4-chloro N- (2-methyl 1r indolinyl) 3-sulfamoyl-benzamide, was described in the example 1 of French patent N ° 69 * 06023 (published under N ° 2.003.311) "as a diuretic usable in particular * ·· * 10 in the treatment of arterial hypertension.

8- (3-tert-butylamino-2-hydroxypropoxy) -thia- - chroman (or its addition salts) is described in example _ __ __ p 2 of French patent N0. · 71.11445 (published under the N ° 2.092.004) which indicates cardio-vascular properties, in particular beta-blockers.

However, we have now found that the two preceding compounds combined have unexpected sy-nergetic properties, making it possible to predict their simultaneous therapeutic use.

The invention therefore relates to a pharmaceutical composition containing a combination of indapamide and 8- (3-tert-butylamino 2-hydroxypropoxy) -thiachroman or one of its pharmaceutically acceptable addition salts, each of these two compounds possibly being find in racemic or optical isomeric form, as well as a therapeutically compatible excipient or vehicle.

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For convenience, 8- (3-tert-butylamino 2-hydro’xypropoxy) -thiachromanne will be referred to as "THPT" below.

Pharmacological study 5 The compounds studied according to the invention were tested on the functional inhibition of adenylate cyclase menbranaire alone and in combination.

• The pigeon red blood cell is known for its great adenylate cyclase activity since the work of 10 SUTHERLAND and coli. (Adenyl cyclase 1. Distribution, preparation and properties, J. Biol. Chem. (1962) 237: 1220-1227). The experiments were carried out on phantoms of erythrocytes (open cells) prepared from the blood of Strasser pigeons according to the technique of 15 SALESSE R. and GARNIER J., "Effects of drugs on pigeon erythrocyte membrane and asymmetry control of adenylate cyclase by the lipid bilayer "(Biochem. Biophys. Acta (1979) 55¾: 102-103).

The suspensions obtained were incubated for 20 minutes in hypotonic buffer with a volume of product tested at the desired final concentration. The assays for adenylate cyclase activity were carried out according to the method of BIRNBAUMER L. et al. (J.

Biol. Chem. (1969) 2¾¾: 2468-3476) and of RAMACHANDRAN J.; 25 LEE ’V. (Biochem. Biophys. Res. Commun. (1970) 41: 358-366).

The final reagent concentrations were ATP 2 m M including Iji Ci of jk- 32Jp ATP, theophylline 4 m M, phosphoereatin 10 m M, creatine kinase 0.5 g / 1, HgCl2 7.5 m M, tromethamine, HCl (" TRIS ", HCl) 10 m M, pH = 7.4 / Λ - 3 - Γ

The total volume was 75 microliters and the

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number of cells per tube approximately 10.

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The production of cyclic AMP (cAMP) was stimulated either by isoproterenol 0.05 m M in the presence of 0.1 m M of GTP, or by sodium fluoride 10 m M. The compound was added to the concentration final required.

The reaction was stopped after -15 minutes at 37 ° C by the addition of 300 microliters of 0.5N hydrochloric acid and a passage of 3 minutes in a boiling water bath. The tubes were then removed from the water bath, neutralizing the content with 3.Q0 microliters of imidazole 1.55 N.

After centrifugation. (10 min, 4000 g) 500 micro-liters of the supernatant were poured onto a column of activated neutral alumina and eluted with 2.6 ml of imidazole "10 m M, pH s 7.5. These columns have a cAMP yield of 95%.

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The radioactivity of the samples was measured in a "Packard 20 Tricarb" scintillation counter, using the Cérenkov effect after addition to the eluate of 10 ml of an aqueous solution at 1 ° / 0o of 7-amino acid. 1,3 - disulfonic naphthalene (counting efficiency 65%.

We have established, on a logarithmic scale, three concentrations above and three concentrations below the concentration for which we observe in vitro, on the rat mesenteric artery, a half inhibition of vasoconstriction 1 Λ Γ - "- induced by norepinephrine.

The results are given as the average of three tests in picomoles of cyclic AMP produced per minute and per milligram of membrane phospholipids.

5 It is found that the indapamide and the THPT subjected to this test inhibit the activation of adenylate cyclase stimulated by isoproterenol or by sodium fluoride: the quantity of cAMP produced decreases. The nature of the effect observed on the production of cAMP depends on the concentration of the compound used. The active concentration of each compound producing an inhibition of 25% (CA2 ^) and 50% (CA ^ q) was determined graphically, then that of their mixtures at these concentrations:

Indapamide (PM = 365.89): ^ 25 = 9 x 10 ”5 mole / 1 __________ 15 · CA50 = 3 x 10_1} mole / 1 THPT, HCl (PM = 331.90) ': CA25 = 1.2 x 10 ~ 5 mole / 1- CA ^ q = 7 x 10-5 mo / i

We have gathered in the table- “according to the inhibition percentages obtained with each of the 1} 2Q mixtures of the two compounds at these concentrations:

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"^ ·"! 1 1 "- '" -Indapamide 25% Indapamide 50% THPT 25 1> 15% 62% THPT 50% 62% 75% ~ \ / -5-.

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It can be seen that the inhibition obtained with the mixture of the two compounds is much higher than that obtained with each separately. For example, with a, -5 mixed with the concentrations of 9 x 10 mole / 1 of indapamide 5 and 1.2 x 10 “** mole / 1 of THPT, 45% inhibition is obtained, which is not achieved only by 2.5 x 10 “** mole / 1 of indapamide or by 5 x 10“ - * mole / 1 of THPT, ie approximately 3 to 4 times the necessary quantity of each of the compounds taken separately . The two effects corresponding to 10 to 25% inhibition therefore multiply in terms of the overall effect and result in a 45% inhibition of the activity of the enzyme.

Discussion ï 1 As THPT is known as beta-blocker, its inhibitory activity on adenyl cyclase associated with an extracellular beta receptor was predictable. On the other hand, it was unexpected that an antihypertensive diuretic such as indapamide has a sami-lar activity and especially that the activity of these two compounds is conjugated: indeed, the dose / activity curve of these compounds and in particular their CA25 and CA ^ q show that they act in multiplying synergy.

The AMP is a second intracellular messenger triggering the initiation of cellular functions. By inhibiting adenyl cyclase, one therefore slows down all the states of cellular hyperactivity such as they exist in hyperthyroidism, hypertension, hypersensitivity, etc. Consequently, the association of the two compounds acting in synergy according to the invention is particularly indicated for the treatment of beta-dependent hyperactivities such as are encountered in hypertension, in particular in cases - 6 - Γ! severe hypertension with kidney failure, or in diseases such as hyperthyroidism.

The pharmaceutical compositions according to the invention 5 are preferably administered orally, and can be in the form of tablets, coated tablets, capsules, suspensions, etc. They can contain in particular from 0.8 to 3 μg of indapamide , and 3 to 12 mg of THPT or one of its pharmaceutically acceptable addition salts, as well as the usual excipients or vehicles for the oral form, and can be administered in one or two daily doses.

EXAMPLE: 90 mg TABLET.

Indapamide ...................................... 1.25 mg ------- ---- 15 8- (3-tert-butylamino 2-hydroxypropoxy) -thiachroman hydrochloride .................... 5.00 mg

Stearic acid ................................. 0.90 mg

Carboxymethyl-sodium starch ........ 3.00 mg

Microcrystalline cellulose ...............; ..... 33.38 mg 20 Lactose ........... 35.75 mg

Calcium hydrogen phosphate .................. 10.00 mg

Colloidal silica ............................... 0.27 mg

Magnesium stearate ........................... 0, ^ 5 mg

This tablet can be coated.

Claims (3)

1. Pharmaceutical composition containing a combination of indapamide and 8- (3-tert-butylamino-2-hydroxy-propoxy) -thiachroman or one of its pharmaceutically acceptable addition salts, each of these two compounds may be found under racemic or optical isomeric form, as well as a therapeutically compatible excipient or vehicle. 2. Pharmaceutical composition according to claim 10 1, characterized in that it contains from 0.8 to 3 mg of indapamide and from 3 to 12 mg of: 8- (3-tert-buty-lamino-2-hydroxypropoxy ) -thiachromanne or one of its addition salts. 3. Pharmaceutical composition intended for oral administration according to claim 2, characterized in that it contains 1.25 mg of indapamide and 5 mg of 8- (3-tert-butylamino hydrochloride). -2-hydroxy-propoxy) -thiachroman. 1 '\ f r r t ·, \ \ \ \ Y:' '- V \\