NO802858L - INGREDIENTS AND MANUFACTURING THEREOF - Google Patents

Description

The present invention relates to combinations of active ingredients

.with antibiotic properties, production and use of these as well as pharmaceutical preparations based on these. .Compounds with the general formula

<;>wherein R1 is hydrogen, methyl, hydroxymethyl, a mono-,

di - or trihalomethyl group;

1 'R 2 is a lower alkyl, hydroxyl lower alkyl or guanidino lower alkyl residue, which is characteristic of an α-amino acid which'

not normally occur in proteins,” and

the configuration of the c atoms (a) and (b) is R (when. R<1>f h)

respectively L, and their physiologically tolerable salts are known. These compounds have anti-bacterial activity.

It has now been found that the active substance combination containing a

compound with' formula I. or one of its physiologically tolerable salts together with a nocardicin antibiotic, . i.e. combinations that have not been described so far are special

interesting with regard to the potential detection of a nocardicin antibiotic.

In the present application and the associated claims, the term "lower alkyl" means straight or branched alkyl residues with preferably up to .8 carbon atoms, such as e.g. ethyl, propyl, butyl. tart. butyl, pentyl and hexyl. Examples of hydroxyl lower alkyl groups are hydroxyethyl, 3-hydroxypropyl and 4-hydroxybutyl. The term "halogen" includes fluorine, chlorine, bromine and iodine; examples of halomethyl groups are chloromethyl, dichloromethyl and trifluoromethyl.

Preferred compounds of formula I are those in which R"<*>"

is hydrogen or methyl. Likewise, preferred compounds with (formula I> in which R 2. is lower alkyl, especially ethyl, n-propyl or n-butyl. Examples of the aforementioned stated compounds., with formula I are

(1R)-1-[L(α-aminobutyryl)-amino]-ethylphosphonic acid

(IR)-1-(L-norleucylamino)-ethylphosphonic acid

i (IR)-1-(L-norvalylamino)-ethylphosphonic acid (L-α-aminobutyrylamino)-methylphosphonic acid

(L-norleucylamirio)-methylphosphonic acid

(L-norvalylamino)-methylphosphonic acid

(IR)-1-(L-homoarginylamino)-ethylphosphonic acid (L-homoarginylamino)-methylphosphonic acid.

.The front forty compounds are

(IR)-1-(L-norvalylamino)-ethylphosphonic acid).

Physiologically tolerable salts of compounds of formula I are formed with physiologically tolerable strong acids (e.g. HCl, HBr, I^SO^, methanesulfonic acid and p-toluenesulfonic acid) and bases (e.g. NaOH and KOH).

in

The nocardicin antibiotic in the present combinations can be one of those which are, for example, mentioned in the German Offen-, legungsschrift Nr. 25 29 941, but is preferably nocardicin

A.

The weight ratio of the compound I or one of its salts in relation to the nocardicin antibiotic can vary within wide limits in the combinations according to the invention. Generally, it is 1:100 to 100:1, preferably on

1:64 to 64:1. Particularly preferred is a ratio from 1:16 to 16:1.

The active substance combination in the present application is prepared

"by mixing a compound of formula I or a"

.a physiologically tolerable salt thereof, and a nocardicin antibiotic in the above-mentioned weight ratios with each other.. The active ingredient combination according to the invention has anti-bacterial activity, as the activities against pseudomonas, .proteus and serratia species (e.g. pseudomonas aeruginosa , proteus mirabilis, p-roteus morganii, proteus vulgaris and 'serratia marcescens) are of particular interest. They can therefore be used for therapy and prophylaxis of bacterial infections. Active ingredient combinations can be given orally or parenterally.

The in vitro activity of the active substance combination according to

.the invention was demonstrated in the following way:

Concentrated solutions of mixtures of the compounds of formula I and nocardicin A were prepared in the desired weight ratios, which were then diluted as desired. Aliquots of the diluted solutions were mixed with a suitable nutrient agar in petri dishes. For comparison, similar agar dishes were prepared, which, in addition to the nutrient medium, contained the compound I or nocardicin A alone. After surface inoculation with the microorganisms, the dishes were kept for 24 hours at 37°C, then the minimum inhibitory concentration (M.I.C.) was determined and the F.I.C. indices were calculated. The results using representative' compounds of formula ■ I, namely (IR)-1-(L-norvalylamino).-ethylphosphonic acid and 1-(L-norvalylamino).-methylphosphonic acid are summarized in Tables I - IV.

In ..vitro the activity of the combination according to the invention can also be shown by the microtiter checkerboard method..In this method, filter-sterilized base solutions of compounds I and a nocardicin antibiotic are prepared, which are 4.8 times as concentrated as the solutions to be examined. Aliquot portions of the culture medium (50 ul) were placed in all 12 (1-12) x 8 (A-H) wells of the microtiter plate with the exception of the holes 12 H. 50 ul of the peptide solution was then placed

.• in each hole in 12 rows and 100 ul L in hole 12 H. The peptide-containing solutions were then diluted within the rows

A-H by transferring 50 ul from row 12 to row 11 each time, mixing well and again transferring 50 ul to row 10, and this procedure was repeated up to row 2, so that row 1 was omitted. Then 50 µl of a solution of a nocardicin antibiotic was dosed into each hole in row H and similarly diluted in the previously described manner during rows 1-12, during which the last hole in each row was again omitted. After the volumes were all brought to 100 µl by the addition of 50 µl curing fluid, 20 µl of a 1/1000 dilution of an overnight culture of a test organism was added. By shaking, an even distribution was ensured, after which the plates were closed with adhesive strips. The minimum inhibitory concentration was determined as the minimum concentration after 18 hours of incubation

.| of the cultures prevented a visible cloudiness. The results with 1 (IR)-1-(L-norvalylamino)-ethylphosphonic acid and nbcardicin D

is composed.in table V.

The in vivo activity of the combinations according to the present invention was investigated by pseu.domonas septicemia in mice, during which the administration occurred 1 hour, 3 hours and 5 hours after intraperitoneal infection. The results obtained with (IR)-1-(L-norvalylamino)-ethylphosphonic acid and nocardicin A are summarized in the following Table VI. The active substance combinations according to the invention can be given in the form of pharmaceutical preparations which are likewise the subject of the present invention. For the preparation of preparations, the usual pharmaceutical carriers that are compatible with the compounds of formula I, respectively. their "salts and nocardicin antibiotics on. speech. Carrier materials can

be liquid or solid, of an organic or inorganic nature and e.g. include water, gelatin, mannitol, mineral oils, vegetable oils, gum arabic, propylene glycols or polyalkylene glycols.

The production of the pharmaceutical preparations can take place in a manner known per se, e.g. by mixing the individual components with the suitable carrier materials and bringing them into a suitable galenic form.

The pharmaceutical preparations can be in solid form (e.g. as lyophilisates) or in liquid form (e.g.

as solutions, suspensions or emulsions}. They are possibly sterilized and resp. or contains further auxiliaries such as preservatives, stabilising, wetting or emulsifying agents, agents for flavor improvement, salts for changing the osmotic pressure or puffer substances. When using buffers, the preparations' pH values may vary within the usual limits.

The content of the active substance combination according to the invention in pharmaceutical preparations can also vary within wide limits. The optimal daily dosage depends on the practically used components, the route of application, the type of infection, etc. Thus, e.g. the daily dose by parenteral application of approx. 200 - 2000 mg of mixtures of the active components. This dose can be given as a single dose or in partial doses - and in special situations can be increased or decreased according to the doctor's prescriptions. individual needs.

EXAMPLES

; Example' L

Mixture, for the preparation of injection solutions which "contains the following components:

The ground individual components were thoroughly mixed with each other and filled into suitable containers under sterile conditions. For the preparation of an injection solution, the obtained mixture can be dissolved in 2 ml of water for injection purposes.

Example 2

Lyofilisat for the preparation of injection solutions containing the following ingredients:

An aqueous solution of these components was freeze-dried. To prepare an injection solution, the lyophilisate obtained was dissolved in 2 ml of water for injection purposes.

Example 3

■ An injection solution is prepared by dissolving 100 ml of nocardicin A sodium salt in 1 ml of a solution containing 100 mg (IR).-1-(L-norvalylamino)-ethylphosphonic acid per

•ml of a suitable buffer. A suitable puffer can have the following composition:

Example 4

A powder mixture for the production of injection solutions can have the following composition:

This mixture is prepared in an analogous manner as in example 1. For the preparation of an injection solution, the mixture is dissolved in 2 ml of water for injection purposes.

Example 5

Powder mixture for the production of injection solutions with the following composition:

The mixture is prepared in an analogous way as in example 1.

To prepare an injection solution, the mixture is dissolved in 2 ml of water for injection purposes.

Claims (6)

1. Process for the production of active ingredient combinations with antibiotic properties containing a compound of the general formula wherein R"*" is hydrogen, methyl, hydroxymethyl, a mono-, di- or trihalomethyl group; 2 R is a lower alkyl, hydroxyl lower alkyl- or guanidione lower alkyl residue that is characteristic of an α-amino acid that does not normally occur in proteins and the configuration of the C atoms (a) and (b) is R- (whenR<1> f K) or L, or a physiologically tolerable salt of such a compound and a nocardicin antibiotic,. characterized by mixing a compound of formula I or a physiologically tolerable salt thereof and a nocardicin antibiotic with each other. 2. Method according to claim 1, characterized in that nocardicin A is used as the nocardicin antibiotic. 3. Method according to claim 1 or 2, characterized by . that R"'" in connection with formula I is hydrogen or methyl. 4. Method according to one of claims 1-3, characterized in that a compound of formula I is used in which R 2 is a lower alkyl residue. 5. Method according to one of claims 1-4, characterized in that a compound of formula I is used which is .(IR)-1-(L-norvalylamino)-ethylphosphonic acid. 6. Process for the production of pharmaceutical preparations based on active ingredient combinations according to claims 1-5, characterized by mixing a peptide of formula I or a physiologically tolerable salt thereof and a nocardicin antibiotic with a non-toxic, inert, therapeutic .acceptable solid or liquid carrier and brings the resulting mixture into the desired galenic form.