NL194535C - Pharmaceutical preparation for the treatment of leprosy containing diaminodiphenyl sulfone. - Google Patents

Description

. 1 194535

Pharmaceutical preparation for the treatment of leprosy containing diaminodiphenyl sulfone 4

The invention relates to a pharmaceutical preparation containing diaminodiphenyl sulfone for the treatment of leprosy and / or other mycobacterioses.

A pharmaceutical preparation for the treatment of leprosy containing diaminodiphenyl sulfone (DDS) is known from US patent 4,005,207. This document describes compositions containing isonicotinic acid hydrazide, 2-propylthioisonicotinic acid amide and diaminodiphenylsulfone. These compositions exhibit synergistic activity against, inter alia, Mycobacterium marinum, a suitable model system for Mycobacterium leprae.

In view of the continuing need for better and more efficient drugs, the object of the invention is to provide a pharmaceutical preparation for the treatment of leprosy containing diaminodiphenyl sulfone with improved performance.

Surprisingly, it was found that a mixture of DDS and 2,4-diamino-5- (4-bromo-3,5-dimethoxybenzyl) pyrimidine (Brodimoprim) described below with respect to leprosy agents compared with the action of the individual components develops synergistically enhanced activity. It is known from British patent application 2,082,063 that a mixture of a sulfone and a pyrimidine derivative can exhibit a synergistic antibacterial action. To this end, a combination of DDS and Trimethoprim is compared with DDS only, with a combination of Trimethoprim and Sulfamethazine, and with only Sulfamethazine. Although it is known to use DDS for the treatment of leprosy, this British patent application does not in any way discuss the possible efficacy of a mixture of DDS and Trimethoprim against Mycobacterium strains. This literature therefore gives no reason for an expert to investigate whether Brodimoprim - a per se known analogue of Trimethoprim - in combination with DDS has a synergistic effect from the point of view of activity with respect to Mycobacterium strains.

The invention therefore relates to a pharmaceutical preparation as described in the preamble, characterized in that the preparation contains diaminodiphenyl sulfone and 2,4-diamino-5- (4-bromo-3,5-dimethoxybenzylpyrimidine in a molar ratio of 20 : 1 to 1: 220.

The pronounced synergistic effect of the combination of DDS and Brodimoprim according to the invention with respect to leprosy agents is apparent from the tests described below: A Dubos-modified albumin (fraction V) modified with 0.25% (weight / volume) of bovine serum Davis nutrient solution was contaminated with Mycobacterium lufu from a slanted Gottsacker culture.

The Dubos-Davis nutritional solution consists of:

Substance g / l 35 ------- 1 KH2 PO4 1 2 Na2 HPO4.12H20 6.5 3 iron ammonium citrate 0.05 4 Na citrate.5.5 H2 O 1.5 40 5 MgSO4.7H20 0.005 6 casein hydrolyzate 1 7 asparagine 1 8 Tween 80; pH = 6.5-6.8 0.5 ml 45 bovine serum albumin (fraction V) 2.5

The Gottsacker culture is prepared as follows: 1 part Sauton solution and 1 part egg yolk (without protein) are mixed gently. A 50% solution of malachite green is added to this in a ratio of 1: 8500. Tubes are filled with this and coagulated for 15 minutes at 100 ° C and set obliquely in the coagulator.

1 · 194535 2

Sauton solution: 5 asparagine 4 g 20 g citric acid 2 g 10 g K2 HPO4 0.5 g 2.5 g

MgSO 4 .7H 2 O 0.5 g 2.5 g iron ammonium citrate 0.05 g 0.25 g glycerine bidest. 20 ml 100 ml water dest. 100 ml 5000 ml

Dissolve components separately, adjust pH to a value between 6.8-7.2 with 25% ammonia solution or citric acid solution, and autoclave for 45 minutes at 115 ° C.

To obtain a uniform suspension, the bacteria were homogenized in 5 ml of medium. The suspension was then diluted with 15 ml of medium and centrifuged at 150 g for 4 minutes.

Part of the supernatant was used for electronic counting and the rest was used to start the cultures. The suspension was diluted to about 10 5 cells / ml and 50 ml portions were placed in 300 ml Erlenmeyer flasks as culture flasks containing a magnetic stirrer.

The inhibitory substance was added and the cultures were kept at 31 ° C. Before taking samples for counting, the cultures were vigorously magnetically stirred for 1 minute. The counting was performed with a Coulter Counter. For the determination of the total germ count, culture samples were diluted with particle-free table salt (0.85% by weight) - formaldehyde (0.2% by weight) - solution, so that a number of 500-20,000 organisms was obtained. For the determination of the living bacterial count, samples of 0.5 ml were taken from time to time, placed in 50 ml of fresh nutrient solution and incubated at 31 ° C. The cultures were counted after 8 days - as described - with the Coulter Counter.

The change in the growth rates of the cultures in the presence of the individual inhibiting substances was determined. To that end, the rate constant kapp [sec.'1] for the observed first-order multiplication rate, in the absence and presence of the inhibiting substances, was determined from the rise in the growth curves according to the equation log N = t / 2.303 + log N0, where N is the number of microorganisms in a defined culture volume, t is the time in seconds and N0 is the number of germs at the time t = 0.

The results are summarized in Table A for DDS-sensitive and in Table B for DDS-resistant (> 100 μΜ DDS) mycobacteria (M. lufu). The constants for the observed sustained multiplication rate (steady state generation rate) for M. lufu at 31 ° C in the presence of different inhibitors are indicated at different concentrations.

40 TABLE A

Inhibitor Prof 1 Test 2 k [sec1.10-®] k [sec'1.10-6] 45 Control value 7.69 8.98 DDS (0.2 μΜ / ml 5.76 6.22

Brodimoprim (10 μΜ / ml) 7.70 8.90 DDS (0.2 μΜ / ml + 0 -0.50

Brodimoprim (10 μΜ / ml (total germ count) living germ count) 50 - 3 194535

TABLE

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Inhibitor k [sec'MO "6] 5 Control value 7.75 DDS 50 μΜ / ml 7.22 100 μΜ / ml 7.61 200 μΜ / ml 4.88 10 Brodimoprim 5 μΜ / ml 7.77 10 μΜ / ml 7 83 DDS + Brodimoprim 10+ 10 μΜ / ml 2.55 15 50 + 5 μΜ / ml 1.27 100+ 5 μΜ / ml 0.7 100 + 10 μΜ / ml 0.24 20 The values in Table A show that that in the presence of the active substance combination according to the invention, no growth of the microorganisms and even a reduction of the living bacterial count in the culture was observed, while the growth of the culture - albeit in a delayed manner - progresses when the active substances were present separately Also with respect to DDS-resistant cultures the combination of DDS and Brodimoprim were completely inhibited in suitable concentrations.

The pharmaceutical compositions according to the invention contain the said active ingredient combination together with a compatible pharmaceutical carrier. This carrier can be an organic or inorganic carrier material suitable for enteral, percutaneous or parenteral administration, such as, for example, water, gelatin, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols or vaseline. In addition, the pharmaceutical compositions may contain other pharmaceutically valuable substances, such as fever-lowering agents, analgesic agents or anti-inflammatory agents. The pharmaceutical preparations can be administered orally, for example in the form of tablets, capsules, pills, powders, granules, solutions, syrups, suspensions or elixirs. However, administration can also take place parenterally, for example in the form of sterile solutions, suspensions or emulsions, or locally in the form of solutions, suspensions, ointments, powders or aerosols. The pharmaceutical compositions may be sterilized and / or contain components such as preservatives, stabilizers, wetting agents, emulsifiers, salts to vary the osmotic pressure, and buffer substances.

The compositions according to the invention can be used for the treatment of leprosy, also with respect to DDS-resistant leprosy. The compositions are particularly suitable for non-physician-controlled leprosy therapy and for avoiding the development of resistance in 40 DDS monotherapy.

For therapeutic use, the compositions according to the invention can be administered in daily doses of 500-100 mg DDS and an appropriate amount of Brodimoprim in adults.

The dose can be administered as a single dose or in several daily doses. Preferably one tablet is administered per day.

45 An example of a typical dosage form is the following:

Tablet: DDS 100 mg 50 Brodimoprim 300 mg PRIMOJEL (starch derivative) 6 mg

Povidone K 30 (polyvinylpyrrolidone) 8 mg magnesium sterate 6 mg

Total weight 420 mg 55 -

Claims (3)

194535 4 The beneficial efficacy of the pharmaceutical preparation according to the invention against Mycobacterium strains is further illustrated by the following comparative tests:, * A Comparative test in vitro The efficacy of a preparation of diaminodiphenyl sulfone (DDS) and Brodimoprim (BDP) according to the invention was compared with a preparation of DDS and Trimethoprim (TMP) according to the prior art. In the tests, the constants for the observed steady state generation rate of DDS-sensitive microbacteria (M.lufu) were observed. The results are shown in Table C below: TABLE C Preparation Rate constant Inhibition 15 k (hour1) Blank 4.43.10'2 DDS (0.2 pM / ml) + TMP (5 pM / ml) 0.81.10'2 69, 55% DDS (0.1 pM / ml) + BDP (5 pM / ml) 0.51.10'2 88.5-100% 20 - B. Comparative test in vivo The efficacy of a combination of DDS and BDP (according to the invention) was compared in a mouse paw test (see Lepr.Rev. 49.1978, 7) to the efficacy of a combination of DDS and TMP (prior art). In this test, a combination of 0.0001% DDS and 0.1% TMP (prior art) showed no inhibition. A combination of 0.0001% DDS and 0.03% DBP, on the other hand, resulted in a 120-day bacterial propagation delay (M. Leprae). The above comparative tests show that the combination of DDS and BDP according to the invention is considerably more effective than the combination of DDS and TMP according to the prior art, despite the fact that in the above compositions according to the invention a considerably lower concentration of DDS than a considerably lower concentration of BDP (compared to TMP) was applied. Pharmaceutical composition containing diaminodiphenyl sulfone for the treatment of leprosy and / or other mycobacterioses, characterized in that the composition contains diaminodiphenyl sulfone and 2,4-diamino-5- (4-bromo-3,5-40 dimethoxybenzylpyrimidine in a molar ratio of 20: 1 to 1: 220.