GB2222771A - Synergistically active compositions of polymyxin B and diaveridine - Google Patents

Description

A 41 2277771 SYNERGISTICALLY ACTIVE COMPOSITIONS AND PROCESS FOR PREPARING

THE SAME The invention relates to syner.,gistically active compositions and process for preparing the same. More particularly, the invention relates to new synergistically active pharmaceutical composition and/or feed additive useful primarily in the treatment of diarrhoea

and process for preparing the same. The new composition can be used both in human and veterinary field.

According to the data of World Health Organization (WHO), many hundred million people fall ill to gastro-enteritises of different cause, and the number of deaths caused by these kind of illnesses presumably exceeds ten million. Most illnesses and death cases of this kind occur in the developing countries, but the number of enteriti5es caused by the bacteria belonging e.g. tc the genera Salmonella and Campylobacter in the developed countries is also growing. - Most of the economic losses in animal husbandry is caused by illnesses of the digestive tract and respiratory organs. Just in Hungary, these losses are in the range of hundred million forints.

Most of the human and animal enteritises are caused by pathogen microorganisms, but diarrhoea illnesses of non-infective (food or feed) origin also occur. Most 2 - of the enteritises of infective origin are caused by bacteria, viruses and ptotozoa, the most frequent pathogens are the following in humans and animals: Escherichia coli, Salmonella species of different serotypes, Campylobacter species, Yersinia enterolitica; and besides these, in human the Shigella species. In the tropical countries the enteritises caused by protozoas are very wide-spread.

Since there is no specific protection method against most of the human and animal enteritises, the medicinal treatment is playing central role in the protection, besides the improvement of the hygienic conditions. The necessity of the medical treatment must be therefore strongly emphasized since in most cases there is no diagnosis. But from this follows that as far as possible, such a medicine must be used which is effective against a wide range of potential patogens, and can cure the clinical symptoms.

Since the members of the normal intestinal flora supposedly exert a significant antagonistic effect against the different enteropathogen microorganisms, it is important not to disturb the intestinal flora. Namely, disbacteriosis leads to the protraction of the pathological process. Significant portion of the normal intestinal flora consists of Gram-positive bacteria(Enterococcuses and Lactobacilli).

For the treatment of the enteritises antibiotics of wide spectra are used, supplemented with toxin adsorbents and astringent agents. The most widely used 1 I- - 3 antibiotics are the following: D-(-)-threo-2,2-dichloro-N-/-2-hydroxyl-hydroxy-methyl-2-(4-nitrophenyl)eth Y.L7acetamide (chloramphenicol); tetracyclines; potentiated sulphonamides, where 2,4-diamino-5-(31,41,51trimethoxy- benzyl)-pyrimidine (trimethoprim) is used for potentiation; 3-/-5-nitro- furfurilydene-amino7-2-axazolidinone (furazolidone); and the aminoglycosides, /-"Pharmacological Basis of Large Animal Medicine", Editors: J.A. Bogan, P. Lees and A. T. Yoxall, Blackwell Scientific Publications, Oxford, London, Edinburgh, Boston, Melbourne, 19837.

The application of the chemotherapeutics presently used in the therapy has more or less disadvantages. There is a wide range tetracycline and aminoglycoside resistance among the intestinal bacteria which in most cases are bound to plasmides /-"Veterinary Pharmacology and Therapeutics", Editors: L. Meyer Jones, N. H. Booth and L. E. McDonald Ames, The Iowa State University Press, 4th edition, p. 929-938 pages, 19777. If such individuals or stocks are treated with the antibiotics mentioned, this results in wide spreading of the plasmids harboring the resistance. The application of chloramphenicol is more and more repressed because of its harmful effect to health /-Goodman and Gilman: "The Pharmacological Basis of Therapeutics", Macmillan Publishing Co., New York, 7. edition, p. 1179, 19807. Besides, the antibiotics of wide spectra are effective against both the Gram-positive and Gram-negative bacteria and can-therefore easily cause non-desired disbacteriosis. Hardly forms any resistance against furazolidone but its application is less des-ir able because of its carcinogenic effect. The degree of resistance is constantly growing against sulphonamides potentiated by trimethoprim, especially in cases of bacteria causing enteral infection /-Goddman and Gilman:

"The Pharmacological Basis of Therapeutics", MacMillan Publishing Co., New York; 7. edition, p. 1107, 19807.

The aim of the present invention is to provide an effective composition for the treatment of diarrhoea, from which a low dose is enough to kill the pathogens, and this way the formation of resistance against the composition is less likely.

It has been found that 6,9,16-tris(2- amino-ethyl') -15-methyl-4,7,10-trioxo-3,6,9-triaza-heptadecane-amido7aza-cyclotricosane-2,5,8,11,14,17,20-heptaon (polymyxin B) or one of its pharmacologically acceptable acid-addition salts, 2,4-diamino-5-(31,41-dimethoxybenzyl)-pyrimidine (diaveridine) and, if desired, 2-/-2,6-dimethyl-3 -hydroxy-4-(tertiary-butyl)-benzy.L7-2-imidazolidine (oxy methazoline) or one of its pharmaceutically acceptable acid-addition saltsas a combination have very strong anti bacterial effect against those bacterial strains which are the most frequent pathogens of diarrhoeal diseases, and this way the combination of these compounds can be used in the treatment of diarrhoea.

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Polymyxin B is effective mainly against Gram-negative bacteria, its minimal inhibitory concentration is generally around 1-5 /ug/ml. Diaveridine has an inhibitory effect against bacteria only in high concentration, while oxymethazoline has no therapeutical ly applicable antibacterial effect. That is why it is surprising for the experts of this field that polymyxin B and diaveridine or polymixin B, diaveridine and oxymethazoline together exert inhibitory effect in very small concentrations against different Escherichia coli, Yersinia enterolitica, Campylobacter, Shigella and Salmonella strains. This is supported by the following experimental data.

The determination of inhibitory concentration values was carried out in a broth containing phenolred indicator and glucose. The broths containing the compound(s) to be investigated are inoculated with a drop of the 18-20 hours old nutrient broth culture of the bacteria and the cultivation is carried out for 72 hours with constant control of growth. After 72 hours the cultures are plated to solid media to determine the bactericide concentration. The minimal inhibitory concentrations (MIC) providing bactericidal effect are shown in Table 1. The bacterial strains used during the investiga- tions, and appearing in Table 1, are the following:

A B c 1) E F G H I Escherichia coli 11 Escherichia coli 27 Escherichia coli 113 Escherichia coli 258 Escherichia coli 43 Escherichia cali 53 Escherichia coli 128 Escherichia coli 284 Yersinia enterocolitica tHESZ J Yersinia enterocolitica OKI K Campylobacter jejuni Pen. 19 L Campylobacter coli 234 (enteropatogen strains isolated from sucking pigs and calves) (strain isolated from food sample) (strain isolated from human) (strai isolated from chicken meat) (strain isolated from -eritis) human, sickened in ent human with M Shigella sonnei (strain isolated from dysentery) N Salmonella enteritidis (strains isolated from 0 Salmonella infantis human, Ejickened in enteritis, P Salmonella tiphy-murium and strains isolated from R Salmonella cholerae-suis swine paratyphy occurring epidemically) 4 h k Compound investigated Table 1

MIC values, /ug/ml A c 0 E F G H b a c t e r i a 1 s t r a 1 n s Polymyxin B 5 5 5 5 5 5 5 1 5 Diaveridine 200 200 200 200 200 200 200 200 200 Oxymethazoline >200 >200 >200 >200 ?-200 >200 >200 5200 5200 Combination of /ug/ml diaveridine and polymyxin B: 0.5 0.5 0.5 1 0.5 1 0.5 1 1 Combination of /ug/ml diaveridine, 1 /ug/ml oxymethazoline and polymyxin B: 40.5 <0.5 < 0.5 1 <0.5 1 X-0.5 1 continuation of Table 1 Compound investigated MIC values, /ug/ml j 1 m N 0 p R b a c t e r 1 a 1s t r a 1 n s Polymyxin 0 5 Diaveridine 100 Oxymethazoline '>200 Combination of 20 /ug/ml diaveridine arid polymyxin B: Combination of 20 /ug/ml diaveridine, 1 /tig/ml oxymethazoline and polymyxin B:

25 >200 25 >200 0.5 0.25 50 1000 1000 0.25 50 1000 0.25 50 >1000 3.000 1 OD 1 0.5 0.5 0.5 0.25 0.12 0.25 0.25 0.12 4 0.5 40.5 40.5 0.015 0.06 0.12 0.25 0.06 4! It is obvious from Table I that oxymethazoline in itself has no antibacterial effect, and the diaveridine also has some effect in relatively small doses only against to Campylobacter strains. But at the same time the minimal inhibitory concentration of polymyxin B in the presence of only 20 /ug/ml diaveridine, or in the presence of 1 /ug/ml oxymethazoline plus 20 /ug/ml diaveridine drops to one tenth of the original value against most of the bacterial strains investigated.

Thus, the present invention relates to n.ew synergistic pharmaceutical compositions and/or feed additives activ6 primarily against diarrhoea, and for the preparation of the same, which comprises mixing 1-18 part by weight cf polymvyin 6 or one of its acid addition salts to part by weight of diaveridine and, if desired, to 1-66 part by weight of oxymethazoline or one of its acid addition salts, and optionally to pharmaceutically acceptable carriers in order to prepare a pharmaceutical compcsition, or the compounds mentioned are mixed with carrier(s) and/or adjuvant(s) conventionally used in the preparation of feed additives.

Preferably the acid addition salt of Oolymyxin B is its sulphate, and that of oxymethazoline is its hydrochloride.

The active agents of the invention are known, respectively commercially available.

According to a preferred embodiment of the invention the composition comprises I part by weight of oxymethazcline hydrochloride 1 part by weight, 4-14 parts by weight of polymyxin B sulphate and 40-135 parts by weight of diaveridine.

The new synergistic pharmaceutical composition of the invention can be used both in human and veterinary treatment.

The mixture of the active ingredients can be administered per se, for example filled to capsules, or the active ingredients are mixed to usual pharmacological- ly acceptable carriers, and a pharmaceutical composition is formulated from the mixture. Solid compositions, for example tablets, dragees, boluses etc. and liquid compositions, for example suspensions, can be prepared.

As a carrier glucose, starch, silicon dioxide, silicates, lactose, talc, poly(vinyl-pyrrolidone), polyvinyl- polypyrrolidone, calcium phosphate, magnesium-stearate, microcrystalline cellulose, etc. are used.

The carrier used in the preparation of liquid pharmaceutical compositions is generally water and/or an organic solvent. The mixture of different organic solvents can also be used.

The pharmaceutical compositions can contain one or more usual additives, for example emulgeating agents, disperging agents,suspending agents, wetting agents, antioxidants, stabilizing agents, sweeteners, etc.

The active ingredients used according to the present invention can be formulated to any known pharmaceutical composition, see for example Remington's 1 4 ---11 Pharmaceutical Sciences, 16th edition, Mack Publishing Company,Easton, USA, 1980. For this purpose the general ly used, known methods and carriers, adjuvants and additives in the production of pharmaceuticals can be used as it is mentioned in the literature mentioned above.

By using the pharmaceutical composition of the pres-ent invention both in the human and veterinary medicine the treatment is carried out daily 1-4 times. The usual.

daily dose relative to the total amount of the three active ingredients is 0.1 - 100 mg/kg body weight, preferably 1-5 mg/kg body weight.

If the mixtures of the active ingredients of the present invention are to be used in the veterinary treat- ment, then they can be admixed to the fodder.

In order to assure the uniform admixing, preferably feed-premix is prepared first from the actlve ingredients, and the premix is mixed to the fodder.

In preparing the feed premix the actiVe ingredients are mixed with the carrier(s) and/or additive(s) conventionally used in the preparation of premixes. The carriers can be for example: wheat, low- grade flour, wheat bran, oil-free rice bran, corn flour, soy flour, kaolin, zeolite, calcium carbonate, starch, silicon dioxide, polyvinyl- -polypyrrolidone, etc. The additives can be for example trace elements, vitamins, inorganic salts, etc. The fodder premix contains 1-50 % by weight, preferably 1-10% by weight, of active ingredient.

The fodder premix prepared according to the present invention is mixed to the fodder in such a ratio that the total concentration of the active ingredients be 5-100 ppm, preferably 10-50 ppm.

The pharmaceutical compositions according to the present invention can be successfully used in the treat ment of diarrhoea at human beings and at each domestic mammalian farm animal (for example cattle, pig, sheep, goat, etc.), poultry (for example domestic poultry, and wild fowl and water poultry, like goose, duck, turkey, chicken) and pets (for example dog, cat, bird, etc.). For the treatment of animals the premix of the present invention mixed in the fodder can also be used.

The invention is illustrated in detail by the aid of the following nonlimiting examples.

Example 1

400 9 of diaveridine, 40 9 of polymyxine B, 3 g of oxymethazoline and 245 g of polyvinyl-polypyrrolidine are added into a L6dige 5M type laboratory fast-mixer. The compounds are homogenized for five minutes, then the solution of 16 g of polyvinyl- pyrrolidone in 20 ml ethanol is added to the mixture with continuous stirring, and mixing is continued for five more minutes. The granulate prepared this way is dried in a Retsch laboratory fluid bed drier with 60 OC air inlet temperature. (The time of drying is about 10 minutes.) The dry granulate is regranulated with an Erweka laboratory oscillating grainer 1 1 using a sieve of 1.0 mm thread-distance. To the screened granulate 10 g of talc, 4 g of magnesium stearate and 2 9 of colloidal silicon dioxide are added, and the mixture is further homogenized in an Erweka cube-homo genizer for 20 minutes.

The obtained mixed granulate is transformed to flat flanged tablets of 160 mg weight with a press of 8 mm diameter, or 320 mg weight with a press of 10 mm diameter.

The composition of the tablets is the following:

Diameter'of tablets 8 mm 10 mm mg mg 0.75 mg 61.25 mg Diaveridine Polymyxin 6 Oxymethazoline Polyvinyl-polypyrrolidone Poly(vinyl-pyrrolidone) Talc Magnesium-stearate Colloidal silicon dioxide 2. 5 mg mg mg 1. 5 mg 122. 5 mg 8 M9 mg 1 M9 2 mg 0. 5 mq 1 m _q mg 360 mg Example 2 600 g of diaveridine, 80 g of polymyxin B, 20 g of oxymethazoline, 80 g of microcrystalline cellulose and 60 g of potato starch are added into the material container of a Glatt WSG 1 type fluid bed grainer, and the mixture - 14 is mixed for five minutes with a fluidizing air of 50 OC temperature. Granulation is carried out with a solution of 48 g poly(vinyl- pyrrolidone) in 500 ml of distilled water, using 25 ml/min. feeding speed and 0.5 bar vaporizing pressure. After finishing granulation the granulate is driqd for 10 minutes, then 20 g of talc, 12 g of magnesium stearate and 80 g of polyvinyl-polypyrrolidone are added, and the system is further fluidized for 3 minutes. The dry granulate is screened into an Erweka laboratory oscillating grainer and further homogenised for another 10 minutes in an Erweka cubic-mixer.

The homogenized granulate is tabletted, using flat, flanged press of 8 or 10 mm diameter.

The composition of the tablets is the following:

Diaveridine Polymyxin 8 Oxymethazoline Z 0 Diameter of tablets 8 mm 10 mm 100.00 10.0 2.5 Microcrystalline cellulose 10.0 Potato starch 7.5 Poly(vinyl.;pyrrolidone) 6.0 Polyvinyl-polypyrrolidone 10.0 Magnesium stearate 1.5 Talc mg mg mg mg mg mg mg mg 2.5 mg 150.0 mg 200.0 mg 20.0 mg 5.0 mg 20.0 mg 15.0 mg 12.0 mg 20.0 mg 3.0 mg 5.0 mg 300.0 mg 1 1 1 Example 3 28.0 g of diaveridine, 2.8 g of polymyxin 6, 0.30 g of oxymethazoline, 100 g of polyvinyl-pyrrolidone, 4.9 g of colloidal silicon dioxide and 864 g of potato starch are added into a Londige WS 5 type laboratory mixer, and the mixture is homogenized for 6 minutes.

1 part by weight of the premix obtained this way is mixed with 1000 parts by weight of fodder, and the fodder prepared this way is used for feeding animals suffering from diarrhoea, for example pigs.

Example 4 28.0 g of diaveridine, 2.8 9 of polyrnyxin 8, 0.7 g of oxymethazoline, 300 g of polyvinyl-polypyrrol-idone and 668.5 g of wheat flour are added into a LOOdlige WSG 5 type laboratory mixer, and the mixture is homogenized for 5 minutes.

1 part by weight of the premix obtained this way is mixed with 1000 parts by weight of fodder, and the fodder prepared this way is used for feeding animals suffering from diarrhoea, for example pigs.

Example 5

28.0 g of diaveridine, 2.8 g of polymyxin B, 300 g of polyvinylpolypyrrolidine and 669 g of wheat flour are homogenized.

I part by weight of the premix obtained this way 4 L 1.

is mixed with 1000 parts by weight of fodder, and th fodder prepared this way is used for feeding animals suffering from diarrhoea, for example pigs.

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Claims (12)

1. A synergistically active pharmaceutical composition, useful primarily for the treatment of diarrhoea, which c o m p r i s e s 6,9,18-tris(2-amino -ethyl)-15-benzyl-21-/-2,8-bis(2-amino-ethyl)-5-(l-hydroxy-ethyl)-15-methyl-4,7,10-trioxo-3,6,9-triaza-hept-adecane-amido7-3-(l-hydroxy-ethyl)-12-isobutyl- -1, 4, 7, 10, 13, 16,ig-heptaaza-cyclotricosane-2,5,8,11,14,17,20- heptaon or a pharmaceutically acceptable acid addition salt thereof, 2,4-diamino-5-(31,41-dimethoxy-benzyl)- -pyrimidine and, if desired, 2-/-2,6-dimethyl-3-hydroxy ceutically acceptable acid addition salt thereof.

2. A composition as claimed in claim 1, which comprises 1-18 parts by weight of 6,9,18-tris(2-amino -ethyl)-15-benzyl-21-/-2,8-bis(2-amino-ethyl)-5-(l-hydroxy-ethyl)-15-methyl-4,7,10-trioxo-3,6,9-triaza-heptedecane-amido7-3-(l-hydroxy-ethyl)-12-isobuty1-1,4,7,10,13,16,19-heptaaza-cyclotricosane-2,5,8,11,14,17,20- heptaon or a pharmaceutically acceptable acid addition salt thereof, 1-160 parts by weight of 2,4-diamino-5 -(31,41-dimethoxy-benzyl)-pyrimidine and, if desired, 1-66 parts by weight of 2-/-2,6-dimethyl-3-hydroxy-4 (tertiary-butyl)-benzyl7-2-imidazoline or a pharmaceutical ly acceptable acid addition salt thereof, optionally in admixture with carriers and/or additives commonly used in the pharmaceutical industry.

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3. A composition as claimed in claim 2, which c o m p r i s e s 4-14 parts by weight of 6,9,18-tris(2-amino-ethyl)-15-benzyl-21-/-2,8-bis(2amino-ethyl)-5-(l-hydroxy-ethyl)-15-methyl-4,7,10-tri'Loxo-3,6,9-triazaheptadecane-amido7-3-(l-hydroxy-ethyl)-12-isobutyl-1,4, 10,13,16,19heptaaza-cyclotricasane-2,5,8,11,14,17,20-heptaon sulphate, 1 part by weight of 2-/-2,6-dimethyl-3-hydroxy-4-(tertiary-butyl)-benzyl7-2imidazoline-hydrochloride and 40-135 parts by weight 2,4-diamino-5-(31,41-dimethoxy-benzyl)-pyrimidine as active agents.

4. A composition as claimed in claim 1 or 2, which c o m p r i s e s 1-18 parts by weight of 6,9,16-tris(2-amino-ethyl)-15-benzyl-21-/-2,8-bis(2-amino-ethyl)-5-11-hydroxy-ethyl)15-methyl-4,7,'L.0-4#:riLcwxo-3,6,w9-triaza-heptadecane-amido7-3-(l-hydroxy-ethyl)-12-isobutyl-1,4, 7, 10,13,16,19-heptaaza-cyclotricosane-2,5,8,11,14,17,20- -heptaon or a pharmaceutically acceptable acid addition salt thereof, and 1-160 parts by weight of 2,4-diamino-5 (31,41-dimethoxy-benzyl)-pyrimidine.

5. Feed additive, useful primarily for the treatment of diarrhoea, which c o m p r i s e s 6,9,18-tris(2-amino -ethyl)-15-benzyl--P-I-/-2,8-bis(2-amino-ethyl)-5-(l-hidroxy-ethyl)-15-methyl-4,7,10-trioxo-3,6,9-triaza-heptadecane- -amido7- 3 - (1 -hydrox y -ethyl) - 1 2-i sobutyl-1, 4, 7, 20, 13, 16, 19 -heptaaza-cyclotricosane-2,5,8,11,14,17,20-heptaon or a pharmaceutically acceptable acid addition salt thereof, 2,4-diamino-5-(31,41-dimethoxy-benzyl)-pyrimidine and, if desired, 2-/-2,6-dimethyl-3-hydroxy-4-(tertiary-butyl)- -benzyl7-2-imidazoline or a pharmaceutically acceptable acid addition salt thereof, together with carrier(s) and/or additive(s) commonly used in premix preparation.

6. An additive as claimed in claim 5, which c o m p r i s e s 1-18 parts by weight of 6,9,18-tris(2 -amino-ethyl)-15-benzyl-21-/-2,8-bis(2-amino-ethyl)-5(1-hydroxy-eth-yl)-15-methyl-4,7,10-trioxo-3,6,9-triaza-heptadecane-amido7-3-(l-hydroxy-ethyl)-12-isobutyl-1,4, 10,13,16,19-heptaaza-cyclotricosane-2,5,8,11,14,17,20- heptaon or a pharmaceutically acceptable acid addition salt thereof, 1-160 parts by weight of 2,4-diamino-5 -(31,41-dimethoxy-benzyl)-pyrimidine and, if desireff, 1-66 parts by weight of 2-/_2,6-dimethyl-3-hydroxy-4 -(tertiary-butyl)-benzy-17-2-imidazoline or a pharmaceutical ly acceptable acid addition salt thereof, optionally in admixture with carriers and/or additives commonly used in the pharmaceutical industry.

7. An additive as claimed in claim 6, which c o m p r i s e s 4-14 parts by weight of 6,9,18-tris(2 -amino-ethyl)-15-benzyl-21-/-2,8-bis(2-amino-ethyl)-5-(l-hydroxy-ethyl)-15-methyl-4,7,10-trioxo-3,6,9-triaza-heptadecane-amido7-3-(I-hydroxy-ethyl)-12-isobutyl-1, 4, 710,13,16,19heptaaza-cyclotricosane-2,5,8,11,14,17,20-heptaon sulphate, 1 part by weight of 2-/-2,6-dimethyl-3-hydroxy-4-(tertiary-butyl)-benzyl7-2imidazoline-hydrochloride and 40-135 parts by weight of 2,4-diamino-5-(31, 4'-dimethoxy-benzyl)-pyrimidine as active agents.

B. An additive as claimed in claim 5 or 6, which c o m p r i s e s 1-18 parts by weight of 6,9,18-tris(2-amino-ethyl)-15-benzyl-21-/-2,8-bis(2-amino-ethyl)-5-Cl-hydroxy-ethyl)-15-methyl-4,7,10-trioxo-3,6,9-triaza-heptadecane-amido7-3-(l-hydroxy-ethyl)-12-isobutyl-194, 7 10,13,16,15-heptaaza-cyclotricosane-2,5,8,11,14,17,20- -heptaon or a pharmaceutically acceptable acid addition salt thereof, and 1-160 parts by weight of 2.4-diamino-5 (39,41-dimethoxy-benzyl)-pyrimidine.

9. A process for the preparation of pharmaceutical compositions or feed additives, useful particularly for the treatment of diarrhoea, which comprises mixing as active ingredients 1-18 parts by weight of 6,.01,18-tr-is(2-amino-ethyl)-15-benzyl-21-/-2,8-bis(2-amino-etsl,,yl)-5-(l-hydroxy-ethyl)-15-methyl-4,7,10-trioxo-3,6,9-triaze-heptadecane-amido7-3-(l-hydroxy-ethyl)-12-isobutyl- -1, 4, 7, 10, 13, 16, 19-beptaaza-cyclotricosane-2,5, 6, 11 14,17, 20 -heptaon or a pharmaceutically acceptable acid addition salt thereof, with 1-160 parts by weight of 2,4-diamino -5-(31,41-dimethoxy-benzyl)-pyrimidine and, if desired, 1-66 parts by weight 2-/-2,6-dimethyl-3-hydroxy-4-(tertiary-butyl)-benzyl-2-imidazoline or a pharmaceutically accept able acid addition salt thereof to carriers and/or additives commonly used in the pharmaceutical industry, or the active ingredients mentioned above are mixed with carrier(s) and/or additive(s) commonly used in premix preparation.

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10. A composition substantially as hereinbefore described in any one of Examples 1 to 5.

ll. A composition as claimed in any one of claims 1 to 8 for therapy.

12. Use of a mixture of 6, 9, 18-tris(2-amino-ethyl)-15-benzyl- 21- r,?, 8-bis (2-amino-ethyl)-5- (1-hydroxy- ethyl) -15-methyl-4, 7, 10- -trioxo -3, 6, 9 -triaza-heptadecane-amidJ-3-(I-hydroxy-ethyl)-12isobutyl- 1, 4, 7, 10, 13, 16, 19-heptaaza-cyclotricosane-2, 5, 8, 11, 14, 17, 20- heptaon or a pharmaceutically acceptable acid addition salt thereof, 2, 4-diamino-5-(31, 41-dimethoxy-benzyl)-pyrimidine and, if desired, 2-172, 6-dimethyl-3-hydroxy-4-(tertiary-butyl)-benzyl-J- 2imidazoline or a pharmaceutically acceptable acid addition thereof, in the preparation of a medicament for treating diarrhoea.

Pu9d 1990 at The Patent 0Mce, State House. 8#17 tl'ldhlio)bum. londonWC1R4TP PUTthOr copies maybe ob- nod from The Patent 0Mce.

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