NL8201595A - 3-RIBONUCLEOTIDES OF COMPOUNDS OF THE LINCOMYCIN AND CLINDAMYCIN TYPE. - Google Patents

Description

4 <; - 3-Ribonucleotides of compounds of the lincomycin and clindamycin type.

The properties and preparation of the anti-biotic lincomycin (Formula 1) are described in U.S. Patent 3,086,912. Clindamycin (Formula 2) is described in U.S. Patent 3,1 * 96,163. Lincomycin and clindamycin 3 nucleotides are described in U.S. Pat. No. 3,671.61 * 7. The compounds described therein all contain the propylhygrinic acid residue. When tasting at £ 3. Aureus in vivo has been shown to have about a tenth of that of the parent compound.

The invention relates to 3-ribonucleotides of lincomycin and clindamycin type 15 compounds in which the propylhygrinic acid residue has been replaced by other cyclic amino acids. The nucleotides of the invention surprisingly show in vivo as great antibacterial activity as the parent compounds.

The compounds according to the invention are suitable for the prevention and cure of protozone infections in humans and animals, for example in mice infected with Plasmodium berghei. ducks infected with P. lophurae, chicks infected with P. gallinaceum. monkeys infected with Pi cynomolgl, and humans infected with P. falciparum. P. vivax and P. malariae, all malaria parasites. Infections by protozoans of the Sporazoa class of the order of the Coccidia (a microparasite which causes coccidiosis infection) can also be treated with the compounds according to the invention, for example cattle infected with Eimeria zurnii. E. bovis. E. ellipsoidalis, sheep and goats infected with E. parva, E. faurei, pigs infected E. debliecki, E. scabra and Isospora suis, dogs and cats infected with Isospora bigemina. L · felis. E. canis and E. felina, poultry infected with E. tenella, rabbits infected with 5 * stiedae and E. perforans and weasels infected with E. mustelae.

8201595 - 2 -

The compounds of the lincomycin and clinda-mycine type which can be converted into 3 nucleotides according to the invention have the formula 3 wherein one or more hydrogen atoms, whether or not substituted alkyl groups having in the 5 alkyl group 1-6 carbon atoms, whether or not unsubstituted cycloalkyl groups, substituted oxygen atoms, substituted nitrogen atoms, halogen atoms, substituted or unsubstituted phenyl groups or one or more groups - (CHg ^ -QH or (CHgJ ^ NR ^ R ^) where n is a number from 1-6 and R 1 and R 1 each represent a hydrogen atom or alkyl group having 1 to 8 carbon atoms, and

Rg represents a group of the formula X wherein X is a 7 (R) -hydroxy-methyl-1-thio-α-lincosaminide, T (S) -hydroxymethyl-1-thio-e-linco-saminide, 7 (S) - halomethyl-1-thio-α-lincosaminide, 7 (R) -halomethyl-1-thio-α-lincosaminide, 7 (S) -methoxymethyl-1-thio-α-lincosaminide, 15 7-deoxy-7 (S) - (methylthio) methyl-1-thio-α-lincosaminide, 7-deoxy-7 (S) - (2-hydroxyethylthio) -methyl-1-thio-α-lincosaminide, 7-deoxy— 7 (S) - (3- hydroxypropylthio) -methyl-1-thio-α-lincosamine residue. and the pharmaceutically acceptable acid addition salts thereof, the formula wherein Rj and Rg which may be in the ring in the 2, 3, 5, 6, 7, 8, or 9 position have the meaning indicated above, R 1 has a hydrogen atom methyl group, ethyl group or hydroxyethyl group and η is 1-4, as well as the pharmaceutically acceptable salts thereof, the formula 6 wherein A, B and E each represent a nitrogen atom, oxygen atom, sulfur atom or a group CR ^ g in which R 1 and R 5 above have significance and may be bonded to any ring carbon or nitrogen atom, which may be bonded to more than one ring carbon atom, as well as the pharmaceutically acceptable salts thereof, and the formula 7 wherein A, B, D and E each represent a nitrogen atom, oxygen atom, sulfur atom or a group CR ^ Rg in which R ^ and Rg have the meanings indicated above and may be attached to any ring carbon or nitrogen atom, where R ^ may be attached to more than one ring carbon atom, 35 as well as the pharmaceutically acceptable salts thereof.

820 1 5 95 _____ «** · *. _ f- \ - 3 -

In these compounds, the 3-position hydroxyl group in the lincoeamidin residue is replaced by adenylic acid, guanylic acid, cytidyl acid and uridylic acid. The conversion can be carried out by microbiological means. 3- (51-ribonucleotides) obtainable by conversion of U-5 57930 are represented by formulas 8 to 12. Herein is a: U-57930 b: U-57930 3- (5'-cytidylate) c: U- 57930 3- (5 * adenylate) 10 d: U-57930 3- (5'-uridylate) e; U-57930 3- (5'-guanylate)

The starting compounds of formulas 3, 5, 6 and 7 can be prepared as described in U.S. patent application 148,056. The 3- (5'-ribonucleotides) thereof can be prepared according to methods described in U.S. Pat. No. 3,671,647, as well as the salts of these nucleotides.

The invention further relates to pharmaceutical preparations containing the nucleotides of the invention as an active ingredient. For the preparation or production thereof, reference can be made to the above-mentioned United States patent application 148,056.

Determination of the 3- (5'-ribonucleotides) according to the invention.

Since the 3-ribonucleotides of the invention do not show antibacterial activity in vitro, their formation from the antibacterially active parent compounds can be easily monitored by determining the loss of antibiotic activity. The amounts of antibacterially active strain compound in the culture filtrates or reaction mixtures are measured against standard Sarcina lutea ATCC 9341 by standard assay methods.

To determine the presence of the 3-ribonucleotides in fermentation liquids, extracts and purified materials, the phosphodiester bond is first hydrolyzed with crude basic phosphatase or snake venom phosphodiesterase, as indicated below. The antibacterial active 8201595 r λ- Α - h - compound in the hydrolyzate is determined according to standard methods.

Enzymatic hydrolysis.

Basic phosphatase: stock solutions (0.5 5 mg / ml, 0.5 ^ units / mg) of pigeons intestinal basic phosphatase EC 3.1.3.1 (Sigma) are prepared in Tris (hydroxymethyl) amino-methane hydroxychloride buffer, 0.01 M pS 8.0. Samples to be treated are diluted 1: 2 with the enzyme-buffer mixture and then incubated at 28 ° C for 18 hours.

10 Snake venom phosphodiesterase:

Stock solutions (100 mg / ml, 0.026 units / mg) purified snake venom phosphodiesterase EC 3.1.H.1 (Sigma) are prepared in distilled water. Incubation mixtures contain 0.2 ml of a solution (1 mg / ml) of the sample to be treated in water, 0.6 ml 0.01 M Tris hydrochloride buffer pH 9.0, 0.1 ml 0.3 M magnesium chloride and 0.1 ml of the enzyme stock solution. Incubation is carried out at 37 ° C for 18 hours.

Spleen phosphodiesterase:

Stock solutions of spleen phosphodiesterase 20 EC 3.1Λ. 18 (Sigma) are prepared (1 mg / ml, 19.6 units / mg) in distilled water. Incubation mixes contained 0.5 ml of a solution (0.5 mg / ml) of the sample to be treated in water, 0.5 ml. 0.02 M Tris buffer pH 7.0 and 0.1 ml of the enzyme stock solution. Incubation is carried out at 37 ° C for 18 hours.

25 Thin layer chromatographic analysis of preparations and enzymatic hydrolysates.

The preparation and purification of the 3-ribonucleotides is followed by determination against S. lutea as above and thin layer chromatography on silica gel G and methyl ethyl ketone-acetone-water (186: 52: 20, v / v) or ethyl acetate-acetone -water (8: 5: 1, vol./vol.) as solvent systems. The bioactive stem compounds are demonstrated by bioautography on agar inoculated with S, lutea.

The products of enzymatic or chemical hydrolysis of the 3 nucleotides are separated by thin layer chromatography as follows: A: SiXicagel GF plates (Analthec Inc.) water as solvent system.

B: Silica gel GF plates; n-propyl alcohol-5 concentrated ammonia water (55:10:35, by volume).

C: NM-Polygram Cellulose 300 (Brinkman Instruments * Inc.); 1-butanol-water-formic acid (77:13:10, vol./vol.).

UV absorbing products are detected with short wave ultraviolet light. Bio-inactive, non-UV absorbing products are detected with a permanganate-periodate spray reagent. Bioactive nucleotide products are demonstrated by bioautography on agar inoculated with S. lutea.

In the following examples, the fermentation and purification are described for U-57930 E (formulas 8-12). In the same manner, the 3-ribonucleotides of the other compounds of the formulas 3, 5, 6 and 7 are obtained.

Example I

A; Fermentation

Streptowyces rochei NRRL 3533 is grown in a medium consisting of 10 g / 1 glucose, 4 g / 1 Difco pept on, bg / 1 Difco yeast extract, 0.5 g / 1 MgS0 ^ .7H20, 2.0 g / 1 KHgPO ^ and ^ g / 1 KgHPO2 for 3 days at 28 ° C in a rotary shaker. The mycelium from this culture is used to inoculate a fermentation medium of the same composition. The fermentation is carried out in a rotary shaker for 1 * 8 hours at 28 ° C. Afterwards U-57930 is added to a concentration of 50 mg / l, after which the fermentation is continued at 32 ° C. After 12 hours more U-57930 is added to a concentration of 150 mg / l. After another 12 hours, the concentration of U-57930 is finally brought to 250 mg / l, after which fermentation is continued for 2 hours at 32 ° C. The culture filtrates are harvested afterwards. Of the total amount of U-57930, 2k9 mg / l has been converted to bio-inactive product.

Streptomyces rochei NRRL 3533 is made available on request 35 by Northern Utilization and Research 8201595, *

V

. 6 -

Division (HURL), Agricultural Research Service, Ü.S. Department of Agriculture, Feoria, Illinois, America * B. Isolation and Purification.

Isolation of U-57930 3-trionucleotides from the fermentation broth by adsorption on Amberlite XAD-2.

The fermentation broth (about 12 L) containing 3 g of inactivated U-57930 is filtered at a pH

7.7 using filter aid. The mycelium cake is washed with 1.2 L of water and discarded. The clear filtrate 10 and the wash water are combined and after adjusting to pH 6.0, passed through a 600 ml column of Amberlite XAD-2 (Rohm and Haas Co., Philadelphia, PA) at a rate of 1 * 0 ml / min. The run-out is examined for bioactivity before and after treatment with basic phosphatase and discarded. The column is then washed with 2 1 of water. Also: the washing water is bio-inactive before and after treatment with basic phosphatase and is also discarded. The column is then eluted with a mixture of methanol and water (70:30 v / v). 20 ml fractions are collected at a flow rate of 20 ml / min. Determination of the bioactivity before (-E) and after 20 (+ E) treatment with basic phosphatase gives the following picture:

Zone S. lutea Fraction no. -E + E

5 0 0 10 00 25 11 0 0 13 0 36 20 33 55 25 32 5b 1 * 0 31 50 30 ^ 5 29 1 * 8 50 26 39 55 23 38 60 21 37 65 19 27 35 70 17 26 75 17 26 8201 595 è * - 7 -

Group no. Zone S. lutea

-E + E

80 16 26 85 16 26 5 90 16 26 95 16 26 100 16 26

110 16 2T

120 15 21 10 130 15 21 1U0 15 21 150 15 21 160 15 2.1 170 15 21 15 180 15 21 190 15 21 200 1? 21

Fractions 12-80 are combined, concentrated to an aqueous solution and lyophilized to obtain 12.22 g of preparation ADA-3.

Repeatedly 6 L of fermentation broth containing 2 g of inactivated ü-57930 is obtained. This broth is treated in the same manner as above. The methanolic eluates from the Amberlite XAD-2 column are stored under the code ADA-1k3B. This solution is not concentrated to dryness but purified by Dowex-1 chromatography as described below. Dowex-1 chromatography.

A column is prepared with 300 ml Dowex-1 30 (X-k) in the acetate form. The methanolic solution of ADA-1 ^ 3B (pH 8.2) is passed over the column. The spout is operated at a flow rate of 2.5 ml / min. collected in 20 ml fractions (fractions 1-60). After this, the column is washed with 1.5 l of water (10 ml / min, fractions 66-108). The column is then eluted with 5% acetic acid (10 ml / min, fractions 109-310). The political groups 8201595

V

- 8 - 1 are combined as follows:

Combination 1 fractions 1-80 1000 ml (ADA-1A)

Combination 2 fractions 81-110 600 ml (ADA-2A) Combination 3 fractions 111-130 1 * 50 ml (ADA-3A) 5 Combination 1 * fractions 131-150 1 * 50 ml (ADA-1 * A)

Combination 5 fractions 151-190 900 ml (AM-5A) Combination 6 fractions 191-230 900 ml (ADA-6A) Combination 7 fractions 231-270 900 ml (ADA-7A) Combination 8 fractions 271-310 900 ml (ADA- 8A) 10 Determination, before (-E) and after (+ E) treatment with basic phosphatase gives the following picture:

Zone S_. lutea -E + E

Combination 1 31 52 15 Combination 2 36 1 * 9

Combination 3 3¼ 1 * 5

Combination 1 * 18 1 * 0

Combination 5 25 '30

Combination 6 2T 29 20 Combination 7 27 29

Combination 8 29 31

Combinations 1 and 2 are combined, concentrated to an aqueous solution and lyophilized to give 1.1 * 8 g of the preparation. ADA-2.1 is obtained ,.

The combinations 3 and 1 * are also combined and treated in the same manner to obtain 2.5 g of preparation ADA-2.2.

After treatment of the preparations ADA-2.1 and 2.2 with basic phosphatase, U-57930 is obtained.

The preparations ADA-31 * .1, 2.1 and 2.2 are combined and purified by double counter-current distribution as described below.

Double counter-current distribution.

The collected preparations ADA-31 * .1, 2.1 and 2.2 35 (16.20 g) are dissolved in 25 ml of a mixture of equal 8201595 s-9 parts by volume of butanol-1 and water, the material dissolving in both phases. . The solutions are placed in centering tubes of a glass counter-current distribution device (100 x 25 ml tubes). After 150 transfers, the bioactivity for (-E) and after (+ E) treatment with basic phosphatase is determined. The picture is as follows:

Zone (S. lutea sensitive) Lower collector -E + E

5 29 31 10 10 31 33 15 30 33 20 27 33 25 - 22 33 30 17 3Π 15 35 0 3b-

Ho 0 33.5 k5 0 33 50 0 3h 55 0 3Π 20 60 0 35 65 0 36 70 0 38 75 0 39 8o o Ho 25 85 0 Hl 90 0 Π2 95 0 k3 100 0 Π3.5 2> ne (S.lutea -sensitive)

30 Lower part of device -E + E

50 0 Π5 Π5 trace Π6 HO 15 H7 35 17 H7.5 35 30 17 Π7 8201 595 - 10 -

Zone (£. Lutea sensitive) Lower part of device -E -HE

25 18 UT

20 17.5 U8 5 15 17 U8 10 16 U8 5 15 50 0 track 50

Upper part of device 10 5 track U9 10 track U8.5 15 track U8 20 track U8.5 25 track U9 15 30 30 50 35 track 51 U0 15 52 U5 16 53

50. 21 5U

20 Zone (S. lutea sensitive)

Upper collector -E + E

100 17.5 5U

95 17 53.5 90 19 53.5 25 85 20 53.5 80 21 53.5 75 22 53.5 70 2U 53.5 65 26 53.5 30 60 28 53.5 55 30 52.5 50 32.5 52 U5 33 52

Uo 35 52 35 35 36 52 8201 595 - 11 -

Zone (£. Lutea sensitive) Upper collector -E + B

30 39 49 25 U1 47 5 20 43 48 15 43 1 ^ 6 10 U3 43 5 35 '35

The fractions are combined into combinations I, II and III. Each combination is concentrated to an aqueous solution and lyophilized to yield the corresponding preparations.

Combination I: lower collector 1-50; Combination II; lower collector 51-100; 15 lower part device 50-30;

Combination III: bottom part of device 29-0; upper part of device 1-50; top collector 100—30.

Preparations obtained: from combination I, preparation ADA-4 ?. 1. 9.78 g 20 from combination: II, preparation ADA-47.2, 0.30 g from combination III, preparation ADA-47.3, 5.29 g. The preparations ADA-472 and 47.3 are combined and purified by DEAE-Sephadex chromatography as described below.

DEAE-Sephadex chromatography.

DEAE-Sephadex A-25 (300 g) is stirred for 1 hour with. water and 2 hours with 0.5 natriumί sodium hydroxide. The ion exchanger is washed with water to a pH of about 7.5. The material is then stirred for 2 hours with dilute acetic acid (0.5 N), washed with water to neutral, placed in a column and packed under constant pressure of about 14 kPa. The column is then washed with 4 L water, 8 L 0.1% aqueous tris (hydroxymethyl) aminomethane and 3 L 0.03 M tris (hydroxymethyl) amino methane acetate buffer (pH 8.0) prepared by dissolving 3.64 g of 35 tris (hydroxymethyl) aminomethane in 800 ml of water, adjusting the 8201595 - 12 - pH to 8.0 with glacial acetic acid etc. adding water to 1 l).

The preparations ADA-47.2 and 1 + 7.3 (total about 5 * 50 g) are dissolved in 20 ml of 0.03 M tris (hydroxymethyl) amino methane acetate buffer (pH 8.0). The solution is passed over column 5, after which the column is eluted downstream with 0.3 M tris (hydroxymethyl) aminomethane acetate buffer (pH 8.0). 20 ml fractions (1-190) are collected. After this, the column is eluted upstream. Fractions of 1 1 each (A, B, C, D and E) are collected. * Determination of the bioactivity before (-E) and after 10 (+ E) treatment with basic phosphatase gives the following picture:

Zone (i3. Lutea sensitive) Fraction no, -E + E

3 0 0 6 0 0 15 9 0 0 12 0 0 15 0 0 18 0 0 21 0 0 20 2k 0 0 27 0 0 30 0 0 33 0 0 36 38 39 25 39 1 + 3.5 1 + 1 + 1 + 2 36 36 1 + 5 23.5 23 1 + 8 15 16 51 0 0 30 5l + 0 0 57 0 0 60 0 0 63 0 0 66 oo 35 69 15 16 8201 595 - 13 ~

Zone (S. lutea sensitive) Fraction no. -E + E

72 17 20 75 21 26 ..

5 78 23 27 81 23 30 8U 23 29 87 22 2¾ 90 21 23 10 93 21 2k 96 22 26 99 21 35 102 22 l * k 105 23 51 15 108 22.5 5k 111 22.5 52.5 11k 22 52 117 22 5k.5 120 20.5 56 20 123 20 56

The fractions are combined as follows: Combination I fractions 3 ^ -38 280 ml (ADA-69B)

Combination II fractions 75-90 330 ml (ADA-69C) '

Combination III fractions 101-111 180 ml (ADA-69D) 25 Combination IV fractions 11W150 580 ml (ADA-69E)

Combination V fractions 151-16k 100 ml (ADA-69F)

Combination VI fractions 165-186 125 ml (ADA-69G)

Combination VII fraction C, 1 1 (ADA-69A) Combination I (ADA-69B) contains unchanged U-57930 and is discarded.

Combination II (ADA-69C) contains an unknown product that gives U-57930 by treatment with basic phosphatase. UV: X max 275 nm.

Combination III (ADA-69D) contains U-57930 citidylate 35 and is treated as described below, UV: X max 270 nm.

8201595 -11 * -

Combination IV (ADA-69E) contains U-57930 adenylate and is treated as described below. UV: λ max 260 nm.

Combination V (ADA-69?) Contains a mixture of U-57930 adenylate, U-57930 uridylate and U-57930 guanylate. This solution is treated as described below

Combination VI (ADA-69G ·) contains U-57930 guanylate and is treated as described below. UV: λ max 25U; shoulder at 275.

Combination VII (ADA-69A) contains a mixture of 10 U-57930 guanylate and U-57930 uridylate. This solution is treated as described below.

Isolation of practically pure U-57930 eytidylate U-57930 adenylate and U-57930 guanylate from combinations I, II and III, respectively. Removal of tris-acetate buffer by Amberlite XAD-2 chromatography:

The combinations indicated above III ,. IV and VI are passed over Amberlite XAD-2 containing columns. The spouts are discarded. The columns are washed with water and then eluted. with a mixture of methanol and water (70:30 v / v).

The fractions are analyzed by UV and examined for bioactivity before and after treatment with basic phosphatase. Appropriately selected fractions are combined, concentrated to an aqueous solution and freeze-dried. The amount of Amberlite XAD-2 used for each combination, the amount of wash water, the amount of methanolic eluate and the amount of product obtained are shown below.

Combination Amberlite Wash water Methanolisch Product (mg) nation XAD-2 (ml) (ml) eluate (ml) III 50 200 300 150 30 IV 200 800 600 3510 VI 50 200 300 1 * 70

The product of combination III is kept under the code ADA-73.1, that of the combination IV under the code 35 ADA-7I * · 1 and that of the combination VI under the code ADA-75 · 1 · 8201595 - 15 -

Remove tris-acetate buffer from the combination V (ADA-69F) and the combination VII (ADA-69A) by Amberlite XAD-2 chromatography.

The column is prepared from 300 ml Amberlite XAD-2 as indicated above. The combinations V and VII which are a mixture of. U-57930 adenylate, U-57930 uridylate and U-57930 guanylate are passed over the column. The spout is discarded. The column is washed with 600 ml of water and the spout is discarded. The column is then eluted with a mixture of methanol and water (70:30 v / v). The fractions after treatment with basic phosphatase bioactive material are combined (300 ml), concentrated to an aqueous solution and lyophilized to obtain 670 mg of preparation AM-71-1. This preparation is treated in the manner indicated below.

Separation of U-57930 uridylate from U-57930 adenylate and U-57930 guanylate by DEAE-Sephadex chromatography.

DEAE-Sephadex in the acetate form (600 ml), prepared as indicated above, is washed with 0.03 M 20 tris-acetate buffer (pH 8.0) and packed in a glass column (height 40 cm, diameter 4) under hydrostatic pressure. 5 cm). A solution of the preparation ADA-71.1 in 10 ml of 0.03 M tris-acetate buffer (pH 8.0) is passed over the column. The column is eluted with: 25 1) 0.03 M tris acetate, pH 8.0 (fractions 1-79) 2) 0.12 M tris acetate, pH 8.0 (fractions 80-395) 3) 0 .25 M tris acetate, pH 8.0 (fractions 396-750)

20 ml fractions are collected and analyzed by UV and examined for bioactivity before and after treatment with basic phosphatase. Fractions 51-6o contain U-57930 adenylate; frictions 62-73 (ADA-94.B) contain U-57930 uridylate; fractions 75-110 contain U-57930 guanylate.

Insulation of practically pure U-57930 uridylate. Removal of tris-acetate buffer by Amberlite XAD-2 chromato-35graphy.

8201595 '- 16 -

A column is prepared from 50 ml Amberlite XAD-2. The combination ADA-94B containing U-57930 uridylate is passed over the column at a flow rate of 2 ml / min. The spout is discarded. The column is then washed with 200 ml of water. The wash water is discarded. The column is then eluted with a mixture of methanol and water (70:30 v / v). The U-57930 uridylate containing fractions (shown by UV) are combined (200 ml), concentrated to an aqueous solution, and lyophilized to give 60 mg of ADA-95 · 1.

Characterization of U-57930 3 '(5, -cytidylate) 1. IR absorption peaks (Nujol and KBr):

Frequencies 1> Trans-Type Frequencies% Trans-Type cm “^ mission cm” 1 mission 15 3417.3 24 sch. 121 * 9.0 33 sch.

3341.1 19 br. 1214.3 21 avg.

3211.8 19 br. 1146.8 42 sc.

3108.6 25 br. 1089.9 15 br.

2915.4 2 br M 1070.6 12 avg.

20 2926.3 1 br M 1056.1 14 sch.

2854.9 2 br M 992.4 39 avg.

2729.6 48 br M 972.2 39 avg.

2693.9 51 sch. 955.8 49 sch.

2535.7 65 sch.- 930.7 51 avg.

1649.3 8 avg, 889.2 36 avg.

1610.7 26 avg. 860.3 52 avg.

1575.0 35 avg. 849.7 53 avg.

1528.7 31 avg. 804.4 46 sc.

1489.2 23 avg. 788.9 40 avg.

^ 1462.2 9 avg M 721.4 44 avg M

1404.3 41 br. 705.0 47 br.

1377.3 18 avg M 654.9 45 sch.

1368.6 31 sch M 632.7 38 avg.

1286.6 34 avg.

3 * 5 sc: shoulder; br: broad, avg: medium; M: possible interference with mineral oil.

8201595 - 1T - 25 Highest Peaks% Trans-Frequency% Trans-Frequency Mission cm "^ Mission cm" 1 1 2926.2 2b 31 * 17.2 2 2951.3 25 3108.5 2 285M 2 6 1010.6 8 161 * 9.2 31 1528.6 9 11 * 62.1 31 1368.5 12 1070.5 33 121 * 9.0 1L * 1056.0 31 * 1286.5 15 1089.8 35 1575.0 18 1377.2 36 889.1 19 331 * 1.0 38 632.6 19 3211.7 39 992.3 21 121! *, 2 39 972, .1 23 11 * 89.1

Measurement in mineral oil (Muil)

Max:% Transmission 87 ö 181 * 8.0% Transmission at 3800 (cm ~ ^): 83 Density: 0.961 *.

Frequency% Trans- Frequency% Trans- cm "'mission cm" ^ mission 31 * 08.6 10 br. 1088.9 12 br.

3102.8 26 sch. 1071.5 9 avg.

2963.9 28 avg. 1057.1 11 sch.

2930.2 27 br. 992.1 * 35 avg.

2878.1 37 avg. 972.2 36 avg.

2862.7 39 ch. 956.8 1 * 5 sc.

2768.1 51 sch. 928.8 1 * 8 avg.

2511.6 65 br. 889.2 32 avg.

161 * 9.3 1 * avg. 859.3 1 * 7 avg.

1611 *, 6 20 sc. 851.6 1 * 7 sc.

1576.0 30 avg. 8ol *, l * 39 sch.

1528.7 28 avg. 788.9 3l * avg.

11 * 91.1 21 avg. 71 * 3.6 1 * 7 sc.

8201595

Frequency% Trans- Frequency% Trans at "1 mission cm" 1 mission - 18 - 11 + 62.2 33 avg. 705.0 1 + 0 avg.

11 + 50.6 35 ch. 651 *, 9 39ch.

11 + 01 +, 3 37 avg. 631 *, 6 35 avg.

1381 +, 0 33 avg. 595.1 33 avg.

1360.9 1 + 2 sc. 572.9 33 avg.

1286.6 32 avg. 525.6 32 avg.

1251.9 30 ch. 1 + 1 + 7.5 36 avg.

1215.2 18 avg.

25 Strongest peaks% Trans Frequency% Trans Frequency Mission cm "1 Mission cm" 1 1+ 161 + 9.2 30 1251.8 9 1071.5 32 1286.5 10 31 + 08.5 32 889.1 11 1057 .0 32 525.5 12 1088.8 33 11 + 62.1 18 1215.1 33 1381 +, 0 20 1611 +, 5 33 595.0 21 11 + 91.0 33 572.8 26 31.02.7 3I + 788.8 27 2930.1 35 11 * 50.5 28 2963.8 35 992.3 28 1528.6 35 631 + .5 30 1576.0

Measurement of KBr images.

Max. % transmission: 100? 1 * 03.1% Transmission he 1 + 000 (cm "1): J8 Density: 0.961+ 2. UV Absorption spectrum j_ λ max (a) / _ in Water at: pH 2.0: 279 nm (6, 5) 8201595 - 19 - pH 7.0: 270 nm (9.9) pH 11.0: 271 (9.6) 3. Elemental analysis

Gross Formula: SC1P Molecular Weight 725: Calculated C 1 * 3.61 * f \ H 6.01 *; N 9.79%> 0 26.88% \ S 1 *, 1 * 7% \ Cl U, 89% P ^, 33%.

4. Optical rotation /.a_/D2^: + 107 ° (cone. 0.85 ^ water).

5. Solubility

Freely soluble in water, methanol and ethanol. Sparingly soluble in acetone and other ketones, ethyl acetate and other esters, chloroform and methylene chloride. Insoluble in saturated hydrocarbons.

6. Antibacterial activity.

U-57930 3- (5 * -cytidylate) is not active in vitro. Treatment with basic phosphatase or phosphodiesterase I, however, gives U-57930 which is highly effective against a number of G + organisms, both in vitro and in vivo.

7. Melting point: 205-207 ° C (decomposition).

Characterization of U-57930 3 '(5'adenylate), 1. IE absorption peaks (Nujol and KBr):

Frequency l Trans- Frequency% Transom ”! mission cm “1 mission 3335.3 16 br. 121 * 5.1 2k sc.

3267.8 17 br. 1213.3 1T avg.

3210.8 17 br. 1175.7 1 * 3 sc.

295 ^ 3 3 br M 111 * 6.8 1 * 0 sch.

2921 *, 1 + 2 br M 10.89.9 13 avg.

2868.1 * 6 sh M 1069.6 10 avg.

285l * s9 1 * average M 1055.1 13 sch.

2727.6 1 * 6 br M 991.5 35 avg.

2520.2 61 br. 972.2 36 avg.

168U.0 22 sch. 957.7! * 5 sc.

8201595

Frequency% Trans- Frequency% Trans om-1 mission cm-1 mission - 20 - 161 + 1.6 11+ avg. 930.7 1 + 8 avg.

1600.1 28 avg. 889.2 32 avg.

1576.0 31 avg. 861.3 1 + 7 avg.

1550.9 1 + 3 sc. 81 + 8.7 1 + 9 avg.

1509.1+ 53 ch. 818.8 1 + 5 avg.

11 + 63.1 15 avg M 789.6 1 + 0 avg.

11 + 20.7 35 avg. 722.1+ 36 avg M

1377.3 21+ avg M 708.9 1 + 0 sch.

1367.6 35 sch M 61 + 7.1 33 sch.

1332.0 36 avg. 635.6 31 avg.

1299.2 31+ avg.

25 Highest peaks% Trans-Frequency% Trans-Frequency mission cm-1 mission cm “1 2 2921 +, 3 22 1681 +, 0 3 2951 +, 2 21+ 1377.2! + 2851 +, 8 2l + 121 + 5, 0 6 2868.3 28 1600.0 10 1069.5 31 1576.0 13 1089.8 31 635.5 13 1055.0 32 889.1 11+ 161 + 1.5 33 61 + 7.0 15 11 + 63 .0 3I + 1299.1 16 3335.2 35 11 + 20.6 17 3267.7 35 1267.5 17 3210.7 35 991.5 17 1213.2

Measurement in mineral oil (Muil) max.% Transmission: 85 C 1861 +, 1 +% Transmission at 3800 (cm-1): 81 Density: 0.961+ 8201595

Frequency% Trans- Frequency / ^ Traiis- mission cm “mission - 21 - 3375.8 7 br. 1090.8 8 avg.

3223, 4 10 br. 1009.6 5 avg.

3124.0 17 sch. 1050.3 8 sc., 2963.0 22 avg. 990.5 27 avg.

2929.2 22 br. 972.2 29 avg.

2878.1 31 avg. 956.8 38 seh.

2863.6 33 sch. 929.8 42 avg.

2756.6 45 br. 889.2 24 avg.

2521.2 60 br. 861.3 38 avg.

2188.5 75 br. 851.6 40 sc.

1678.2 15 sc. 818.8 36 avg.

1643.5 7 avg. 807.3 36 br.

1602.0 20 avg. 798.6 30 avg.

1576.0 23 avg. 768.7 41 br.

1553.8 35 sch. 721.4 30 avg.

1511.4 49 seh. 706.9 31 br.

1475.7 27 avg. 648.1 26 avg.

1421.7 29 avg. 636.5 26 avg.

1384.0 32 avg. 584.5 28 sc.

1332.0 31 avg. 571.9 26 avg.

1301.1 28 avg. 533.3 27 sch.

1246.1 19 sch. 522.7 25 avg.

1215.2 12 avg. 503.4 25 avg.

1176.7 36 sch.

25 Strongest Peaks% Trans-Frequency% Trans-Frequency Mission. cm ~ ^ mission cm “^ 5 1069.5 23 1576.0 7 3375.7 24 889.1 7 1643.5 25 522.6 8 1090.7 25 503.3 8201595,% Trans-Frequency% Trans-Frequency Mission cm "1 mission cm" 1 - 22 - 8 1050.2 26 61 * 8.0 10 3223.3 26 636.5 12 1215.1 26 571.8 15 1678.1 27 11 * 75.6 17 312l *, 0 27 990.5 19 121 * 6.0 27 533.2 20 1602.0 28 1301.0 22 2963.0 28 581 *, 5 22 2929.1

Measurement on KBr images Max. % transmission: 95 11 * 05.0% Transmission at 1 * 000 (cm “1): 77 Density: 0.961 * 2. UV Absorption spectrum] _ λ max (a) /

In water at: pH 2.0: 258 (16.0) pH 7.0: 261 (16.5) pH 11.0: 261 (16.0) 3. Elemental analysis

Gross Formula: SC1P, Molecular Fact:

723: Calculated C 1 * 1 *, 81 %% H 5.9l *% \ N 13.55% \ 0 22.13% \ S 1 *, 1 * 2 Cl. 1 * -, 81 * -% \ P 14-, 28!: Found N 12.87% i S 5.39 Cl 1 *, 76.1 \ P

3.83%.

1 *. Optical rotation / _ + 9 ** ° (cone. 0.887, water).

5. Solubility.

Freely soluble in water, methanol and ethanol. Sparingly soluble in acetone and other ketones, ethyl acetate and other esters, chloroform and methylene chloride. Insoluble in saturated hydrocarbons.

6. Antibacterial activity.

U-57930 l_ 3- (5'-adenylate W is not active in vitro. Treatment with basic phosphatase or phosphodiester I gives 8201595 - 23 - U-57930, which is however very active against a number of G organisms, both in vitro and in vivo U-57930 3- (5-adenylate) is active in vivo (subcutaneous, mice) and has a CDQ of 0.62 (0.48-0.79) mg / kg, against S. pyogenes.

7. Melting point: 203.5-205 ° C (decomposition).

Characterization of U-57930 3- (5> uridvlaat) 1. IR absorption peaks (guiol and KBr);

Frequency% Trans- Frequency% Trans- cm “1 mission cm ** 1 mission 3330 19 br. 1332.9 IA br.

3224.4 21 br. 1296.3 40 sc.

2952.4 1 br M 1251.9 28 br.

2924.4 0 br M 1215.2 19 avg.

2867.5 4 sc. M 1089.9 13 avg.

2854.0 3 avg M 1071.5 9 avg.

2733.4 49 sch M 1056.1 13 sch.

2695.8 53 sch. 991.5 39 avg.

2532.8 67 sch. 973.2 39 avg.

1757.3 73 sch. 957.7 48 sc.

1685.9 8 avg. 931.7 49 avg.

16 ^ 7, ^ 21 sc. 890.2 3k avg.

1602.0 41 avg. 858.4 50 avg.

1574.1 42 avg. 813.0 1 * 3 Genr.

1555.7 43 br. 798.6 1 * 7 avg.

1462.2 12 avg M 767.7 50 avg.

1425.5 37 sch. 721.4 42 avg M

1378.3 22 avg M 634.6 35 avg.

1367.6 37 sh M

8201595 - 2k - 4 '25 Strongest Perches% Trans-Frequency% Trans-Frequency Mission cm-1 Mission cm-1 0 292U, 3 22 1378.2 1 2952.3 28 1251.8 3 2851 +, 0 3I + 890 * 1 1+ 2867.5 35 631 +, 5 8 1685.8 37 11 + 25.5 9 1071.5 37 1367.5 12 11 + 62.1 39 991.5 13 1089.8 39 973.1 13 1056.0 1 + 0 1296.2 19 3330.3 1 + 1 1602.0 19 1215.1 1 + 2 1571 +, 0 21 3221 +, 3 1 + 2 721.3 21 161 + T, 3

Measured in mineral oil (Muil).

Max. % Transmission: 86 (3 3761 +, 5% Transmission at 3800 (cm ~ ^): 85 Density: 0.961+

Frequency% Trans- Frequency% Trans- cm ~ 1 mission cm “1 mission 3387.1+ 12 br. 1055.1 10 sc.

3111 +, 1 + 25 ch. 992.1+ 35 avg.

2962.0 26 avg. 973.2 36 avg.

2931.1 26 br. 956.8 1 + 5 sc.

2879.1 36 avg. 929.8 1 + 8 avg.

2863.6 38 sch. 889.2 31 avg.

2833.7 1 + 3 sc. 859.3 1 + 6 avg.

2509.6 61+ br. 813.0 1 + 0 avg.

1685.0 6 br. 811.1 1 + 0 sch.

161 + 7.1 + 17 sc. 789.6 1 + 3 avg.

1605.9 35 sc. 782.2 1 + 8 br.

1576.0 37 avg. 768.7 1 + 8 avg.

8201595

Frequency% Trans- Frequency% Trans- cm "1 mission cm" 1 mission - 25 - 1556.7 39 br. 707.9 43 avg.

11463.1 31 avg. 669.3 43 sch.

11423.6 35 avg. 6149.1 40 sc.

1384.0 32 avg. 634.6 37 avg.

1331.0 43 avg. 585.4 38 sc.

1297.2 38 sch. 567.1 34 avg.

1255.8 24 avg. 523.7 35 avg. 1214.3 17 avg. 447.5 39 avg.

1090.8 10 avg.

1070.6 7 avg.

25 Strongest peaks% Trans-Frequency% Trans-Frequency Mission cm ~ ^ Mission cm “^ 6 1685.0 32 1384.0 7 1070.5 34 567.0 10 1090.7 35 1605.8 10 1055.0 35 1423, 5 12 3387.3 35 992.3 17 1647.3 35 523.6 17 1214.2 36 2879.0 24 1255.7 36 973.1 25 3114.3 37 1576.0 26 2962.0 37 634.5 26 2931.0 38 2863.5 31 1463.0 38 1297.1 31 889.1

Measured on KBr images Max. % transmission: 101 δ 405.0% Transmission he 4000 (cm ^): 76 Density: 0.964 82 0 1 5 9 5 - 26 - 2. UV Absorption spectrum (A max (a))

In water at: pH 2.0: 261 (11.5) pH T, 0: 262 (10.7) pH 11.0: 262 (11.5) 3. Elemental analysis

Gross formula: ^ 26 ^ + 2 ^^ 13 8C1P; molecular weight: 716; Calculated: C 1 + 3.57%, H 5.86%, N 7.82%, 29.05%; S 1 +, 1 + 6 Cl 1 +, 89% i P 1 +, 33%.

1+. Optical rotation.

I "Λ_Τ ^: + 105 ° (cone. 0.91+, water).

5. Solubility.

Good, soluble in water, methanol and ethanol. Sparingly soluble in acetone and other ketones, ethyl acetate and other esters, chloroform and methylene chloride. Insoluble in saturated hydrocarbons.

6 .. Antibacterial activity.

Ut57930 3- (5'-uridylate) is not active in vitro. Treatment with basic phosphatase or phosphodiesterase I gives U-57930 which is, however, very active against a number of G + organisms both in vitro and in vivo.

7. Melting point: 202-203 ° C (decomposition).

Characterization of U-57930 3- (5'-guanylate).

1. IR Absorption peaks (Hu, iol and KBr):

Frequency% Trans— Frequency% Transom * "! Mission cm"! Mission 3335.3 16 br. 1250.0 36 sch.

3227.2 19 br. 1213.3 21 avg.

2953.3 2 avg M 1173.8 39 avg.

2925.3 T br M 111 + 9.7 1 + 1 avg.

2868.1+ 6 sc M 1087.9 11+ sc.

2855.91+ avg M 1071.5 9 avg.

2737.3 51 br M 991.5! +2 avg.

8201595

Frequency% Trans.- Frequency% Trans and "mission cm-Ί mission - 27 - 2521.2 73 br. 972.2 42 avg.

1684.0 6 avg. 956.8 52 sc.

1635.8 11 avg. 929.3 51 avg.

1598.2 21 avg. 890.2 36 avg.

1572.1 26 avg. 860.3 51 avg.

1534.5 34 avg. 800.5 46 avg.

1462.2 18 avg M 783.1 42 sharp

1414.9 44 avg. 720.4 43 avg M

1377.3 24 avg M 707.9 45 br.

1365.7 31 avg. 681.9 40 avg.

1312.7 44 avg. 635.6 34 avg.

25 Strongest peaks% Trans-Frequency% Trans-Frequency Mission cm_1 Mission cm “1 1 2925.2 24 1377.2 2 2953.2 26 1572.0 4 2855.8 31 1365.6 6 2868.3 34 1534.5 6 1684.0 34 635.5 9 1071.5 36 1250.0 11 1635.7 36 890.1 14 1087.8 39 1173.7 16 3335.2 4o 681.8 18 1462.1 41 1149.6 19 3227, 1 42 991.5 21 1598.1 42 972.1 21 1213.2

Measured in mineral oil (Muil)

Max. % transmission: 97 S 3762.6% Transmission he 3800 (cm ^): 97 Density: 0.964.

8201595

Frequency% Trans- Frequency% Trans at "^ mission cm" ^ mission - 28 - 3380.6 9 br. 117b, 7 31 avg.

323b, O 13 br. 11b7.7 33 br.

2963.0 22 avg. 1088.9 9 br.

2929.2 21 br. 1070.6 6 avg.

2878.1 31 avg. 991.5 33 avg.

2862.7 33 sch. 972.2 3b avg.

27bb, 0 bb br. 956.8 b3 sch.

2522.2 61 br. 929.8 bb avg.

1683.0 b br. 889.2 28 avg.

163b, 8 6 avg. 860.3 bl av.

1598.2 1b avg. 800.5 36 avg.

1571.2 19 avg. 783.1 32 avg.

153b, 5 26 avg. 715.6 38 sc.

1b82, b 38 avg. 705.0 38 sc.

Ib6l, 2 36 avg. 679.9 33 avg.

1bb8, T 36 br. 635.6 31 - avg.

Ib3.9 3b avg. 58b, 5 3b sch.

138b, 0 29 avg. 571.9 32 avg.

1359.9 30 avg. 523.7 31 avg.

1312.737 avg. 502.5 30 avg.

1250.9 29 sch. bb7.5 3b avg.

1213.3 16 avg.

25 Strongest Peaks% Trans-Frequency% Trans-Frequency Mission cm ** 1 Mission. cm ^ b 1683.0 29 138b, 0 6 163b, 7 29 1250.8 6 1070.5 30 1359.8 9 3380.5 30 502.5 9 1088.8 31 2878.0 13 323b, 0 31 117b, 6 8201595% Trans-Frequency% Trans-Frequency Mission cm “^ Mission cm-1 - 29 - 14 1598.1 31 635.5 16 1213.2 31 523.6 19 1571.1 32 783.0 21 2929.1 32 571.8 22 2963.0 33 2862.6 26 1534.5 33 114T, 6 \ 28 889.1

Measured by KBr images.

Max. % transmission: 97 8 405.0.

% Trans mission at 4000 (cm “1): 77 Density: 0.964.

2. UV Absorption spectrum / A, max (a) /

In water at: pH 2.0: 256 (13.4); 280 (8.4) sh pH 7.0: 254 (14.5); 273 (9.7) sh pH 11.0: 259 (12.6); 266 (12.4) sh.

3. Basic analysis:

Gross formula: Ο ^ Η ^^ ΗγΟ ^ SC1F; mol weight: 739 Calculated G 43.84% H 5.81 lb; H 13.26 JÉ; 23.2T% \ S 4.33 Cl 4.73% v P 4.19 Found: N 13.32 #; S 4.86% \ Cl 4.49% V P 3.25%.

4. Optical rotation J_ + 97 ° (cone. 0.855, water).

5. Solubility.

Freely soluble in water, methanol and ethanol. Sparingly soluble in acetone and other ketones, ethyl acetate and other esters, chloroform and methylene chloride. Insoluble in saturated hydrocarbons.

6. Antibacterial activity.

U-57930 3- (5'-guanylate) is inactive in vitro. Treatment with basic phosphatase or with phosphodiesterase I gives U-57930 which is active against a number of G + organisms both in vitro and in vivo.

8201595 - 30 - 7. Melting point: 219-220 ° C (decomposition).

Since the compounds of the invention are active against various Gram-positive and Gram-negative micro-organisms, they can be used to combat such micro-organisms by inhibiting their growth. For example, they can be used to cut eating utensils and medical equipment, including dental tools that use S_. aureus is contaminated, to be disinfected. Furthermore, they can be used as a bacteriostatic rinse for washed clothes, for impregnating fabrics and paper products, and for inhibiting the growth of microorganisms on culture plates and in other microbiological media.

The compounds of the invention are also suitable for treating conditions caused by microorganisms of the genus Mycoplasma, the best known representatives of which are pleuropneumonia-like microorganisms such as M. hominis, M. Salivarium. M. mycoides, M. hyopneumonia, M. hyorhinis. M. gallisenticum and M. arthriditis, both in humans and animals including livestock, poultry and laboratory * animals.

The U-57930 3- (5'-ribonucleotides) can be used to treat kidney infections and other infections by the L-forms of Gram-negative and Gram-positive bacteria, for example the L-forms of P. mirabilis.

Since the compounds of the invention are amphoteric, they can form salts with both acids and bases. Examples of acids with which pharmaceutically acceptable salts can be formed are hydrogen chloride, sulfuric acid and phosphoric acid. Pharmaceutically acceptable salts with bases are, for example, the lithium, sodium, potassium and calcium salts. The salts 30 can be prepared in the usual manner.

Examples of pharmaceutical preparations according to the invention are tablets, capsules, pills, powders, granules, sterile parenteral solutions and suspensions, solutions and suspensions intended for oral administration, and oil-water emulsions. The compounds according to the invention may be present therein in the form 8201595-31 - of the free base or of a non-toxic salt.

In addition to one or more compounds according to the invention, tablets contain pharmaceutically customary ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium-5 aluminum silicate, calcium sulfate, starch, lactose, acacia gum and / or methyl cellulose. The tablets may be laminated or otherwise designed to achieve sustained release and prolonged action. For example, the tablet may contain an internal and an external dosage unit separated by an entheric layer. This entheric layer serves to prevent disintegration in the stomach and allows the internal dosage unit to enter the duodenum intact or to be delayed in its release. Examples for forming such entheric layers or coatings include polymeric acids or mixtures of polymeric acids and suitable materials such as shellac, cetyl alcohol, cellulose acetate phthalate and styrene maleic acid copolymers. Two-component tablets containing two or more incompatible active ingredients are also among the possible embodiments. Wafers 20 are manufactured in the same manner as tablets and often contain a sweetener such as sucrose and a flavoring agent.

Hard gelatin capsules can contain a mixture of active compound and inert diluent or polymeric acid coated beads containing an active compound. The hard gelatin capsules are manufactured by filling. Soft gelatin capsules are manufactured by encapsulating a suspension of an active compound in a vegetable oil, light liquid petrolatum or other inert oil.

Liquid preparations for oral administration include suspensions, syrups and elixirs. Syrups are prepared by dissolving the active ingredient in an aqueous carrier together with sugar, an aromatic flavoring and a preservative. Elixirs contain an aqueous-alcoholic (ethanol) carrier, a sweetener such as sucrose and an aromatic flavoring agent. For the preparation of suspensions, a carrier of the type such as for syrups can be used together with a suspending agent such as acacia, tragacanth or methyl cellulose.

Ointments suitable for topical application can be prepared by dispersing the active ingredient in a carrier for ointments such as petrolatum, lanolin, polyethylene glycols or mixtures thereof. A suitable method is to finely distribute the active ingredient using a colloid mill using light liquid petrolatum before dispersing in the carrier. Crimes and lotions are prepared by dispersing the active ingredient in an oil and emulsifying the dispersion in water.

Solutions and suspensions intended for parenteral administration preferably contain water as a sterile carrier. Solutions can be prepared by dissolving the active ingredient in water for injections and sterilizing the solution over a filter and then filling and sealing a vial or ampoule. In addition to an active ingredient, the solutions usually contain a local anesthetic, preservative and buffer. To increase stability, the solution in the vial or ampoule can be frozen and the water removed under vacuum. Before use, the contents of the vial or ampoule are reconstituted with water for injection. Parenteral suspensions are prepared in much the same way except that the active ingredient is suspended rather than being dissolved, and sterilization is not performed by filtration but by exposing the active ingredient to ethylene oxide before suspending in the sterile vehicle . To maintain the action of an intramuscular suspension, the active ingredient is used as an insoluble form such as the trimethylsilyl ether or the pamoate salt. A surfactant or wetting agent is usually included to achieve an even distribution of the active ingredient.

For systemic treatment, the amount of active ingredient in a dosage unit at 8201595 - 33 - preferably 10, 25, 50, 100, 250, or 500 mg, and for parenteral administration preferably 5-65 w / v%.

In the preparation examples below, the active ingredient is the 3- (5 * -ribonucleotide) of 5 U-57930 E or U-60970 E, but it is clear that this is only exemplary. U-60970 E is the i + -cis-n-butyl-L-pipeclinic acid amide of 7-chloromethylthiolincosamidine. The preparation thereof is described in Example VII of the above-mentioned U.S. Patent Application No. 8,056.

10 Where the examples refer to U-57930 E.

or U-60970 E is meant its 3- (5'-ribonucleotide). Preparation Examples Example I Capsules.

1000 Two-part hard gelatin capsules for 15 oral administration, each containing 250 mg of U-57930 E or U-60970 E, are made from the following ingredients: U-57930 E or U-60970 E 250 g

Corn starch 100 g

Talc 75 g 20 Magnesium stearate 25 g

The ingredients are intimately mixed and then encapsulated in the usual manner. The capsules can be used for systemic treatment of infections in adults by administering one capsule orally every 6 hours. Likewise, capsules may be prepared, each of which contains 10, 25, 50, 100 and 500 mg of U-57930 E or U-60970 E instead of 250 mg, respectively.

Example II Capsules.

1000 Two-part hard gelatin capsules for oral administration, each containing 200 mg of U-57930 E or U-60970 E and 250 mg of tetracycline hydrochloride, are manufactured from the following ingredients: U-57930 E or U-60970 E 200 g

Tetracycline hydrochloride 250 g 35 Talc 75 g 8201595

V

3b

Magnesium stearate 25 g

The ingredients are intimately mixed and then encapsulated in the usual manner. The capsules are suitable for systemic treatment of infections in adults by administering one capsule orally every 6 hours. Likewise, capsules may be prepared that contain another antibiotic in place of tetracycline hydrochloride, for example, chloramphenicol, oxytetracycline, chlortetracycline, fumagillin, erythromycin, streptomycin, novobiocin, or dihydronovobiocin.

Also, tetracycline hydrochloride can be replaced with a penicillin, for example potassium penicillin G in an amount of 250-000 units per capsule. Such combinations are suitable for systemic treatment of several types of infections simultaneously in adults by administering one capsule every 6 hours orally.

Example III Tablets 1000 Tablets for oral administration, each containing 500 mg of U-57930 E or U-60970 E, are prepared from the following ingredients: 20 U-57930 E or U-60970 E 500 g

Lactose 125 g

Corn starch 65 g

Magnesium stearate 25 g

Light liquid petrolatum 3 g 25 The ingredients are mixed well and agglomerated. The agglomerates are broken by forcing them through a sieve with 1.19 mm openings. The granules formed are then pressed into tablets. The tablets are suitable for systemic treatment of infections, including malaria infections, in adults by administering one tablet three times a day. Likewise, tablets can be prepared each containing 250 mg of U-57930 E or U-60970 E.

Example IV Tablets 1000 Tablets each containing 250 mg of U-57930 E or 35 U-60970 E and a total of 250 mg of sulfadiazine, sulfamerazine and 8201595-35 - sulfamethazine are made from the following ingredients: Ü-5T930 E or U-6097O E 250 g

Sulfadiazine 83.3 g. Sulfamerazine 83.3 g

Sulfamethazine 83.3 g

Lactose 50 g

Corn starch 50 g

Calcium stearate 25 g 10 Light liquid petrolatum 5 g

The ingredients are mixed well and agglomerated. The agglomerates are broken by forcing them through a sieve with 1.19 mm openings. The granules formed are then pressed into tablets suitable for systemic treatment of infections by first administering k tablets and then 1 tablet every 6 hours.

For the treatment of urinary tract infections, the sulfa compounds are replaced with 250 mg of sulfamethyl thiadiazole or 250 mg of sulfaceetamide per tablet.

20 Example V Syrup.

1000 ml of an aqueous suspension for oral administration, containing per dose of 5 ml 250 mg of U-57930 E or U-60970 E and a total of 500 mg of sulfa compounds, is prepared from the following ingredients: 25 U-57930 E or U- 60970 E 50 g

Sulfadiazine 33.3 g

Sulfamerazine 33.3 g

Sulfamethazine 33.3 g

Citric acid 2 g 30 Benzoic acid 1 g

Sucrose 700 g

Tragacanth 5 g

Lemon oil 2 ml

Deionized water up to 1000 ml of citric acid, benzoic acid, sucrose, tragacanth and 8201595-3-6 lemon oil are dissolved in 850 ml of water. In the solution, U-57930 E or U-60970 E and the finely divided sulfa compounds are evenly distributed with stirring, after which water is added to 1000 ml. The syrup is suitable for systemic treatment of pneumonia in adults by administering 1 tablespoon (10 ml) four times a day.

Example VI Parenteral solution

A sterile aqueous solution for intramuscular administration, which contains 200 mg U-57930 E or U-60970 E 10 per ml, is prepared from the following ingredients: U-57930 E or U-60970 E 200 g

Lidocaine hydrochloride k g

Methylparaben 2.5 g

Propylparaben 0.17 g 15 Water for injections up to 1000 ml

The ingredients are dissolved in water and the solution is sterilized by filtration. The sterilized solution is used to fill vials which are then closed.

Example VII Parenteral Solution.

A sterile aqueous solution for intramuscular administration, containing per ml 200 mg U-57930 E or U-60970 E and ^ 00 mg spectinomycin sulfate, is prepared from the following ingredients: 25 U-57930 E or U-60970 E 200 g

Spectinomycin sulfate h-00 g

Lactose 50 g

Water for injection up to 1000 ml of U-75930 E or U-60970 E, spectinomycin sulfate 30 and lactose are dissolved in water and the solution is sterilized by filtration. Sterilized vials are aseptically filled with the sterilized solution in an amount of 2 ml.

Example VIII Ointment 35 1000 G ointment with a concentration of active 8201595 - 37 - component of 0.25 1 * is prepared from the following ingredients: U-57930 E or U-60970 E 2.5 g

Zinc oxide 50 g

Calamine 50 g 5 Liquid petrolatum (heavy) 250 g

Wool grease 200 g

White petrolatum up to 1000 g

White petrolatum and wool fat are melted after which 100 g of liquid petrolatum are added. U-57930 E or U-60970 E, zinc oxide and calamine are added to the remainder of the liquid petrolatum. This mixture is ground until the powders are finely divided and evenly dispersed. The powder mixture is then stirred in the white petrolatum mixture and stirring is continued until an ointment-like consistency is reached. The ointment can be applied to the skin to treat infections in humans and animals. If desired, the zinc oxide and calamine can be omitted. In the same manner, an ointment is prepared containing 0.5, 1, 2, and 5% U-57930 E or U-60970 E, respectively, instead of 2.5 g and 5, 10, 20, and 50 g, respectively. U-57930 E or U-60970 E. Example IX Crime 1000 G vaginal crime is prepared from the following ingredients: U-57930 E or U-60970 E. 50 g • x) 25 Tegacid Regular 150 g

Spermaceti 100 g

Propylene glyeol 50 g

Polysorbate 80 5 g

Methylparaben 1 g 30 deionized water up to 1000 g x)

Self-emulsifying glyceryl monostearate from Goldschmidt Chemical Corporation, New York.

Tegacid Regular and spermaceti are melted together at a temperature of 70-80 ° C. Methylparaben is dissolved in about 500 g of water after which propylene glyeol, Polysor- 8201595 - 38 - 80 and U-57930 E or U-60970 E are added while maintaining of the temperature at 75-80 ° C. The methylparaben mixture is then added to the melt of Tegacid Regular and spermaceti under constant stirring for at least 30 minutes until the temperature has fallen to 1 * 0-45 ° C. Then the pH adjusted to 3.5 by adding 2.5 g citric acid and 0.2 g potassium dihydrogen phosphate dissolved in about 50 g water. Finally, water is added to a final weight of 1000 g and the preparation is allowed to cool and solidify with stirring to maintain homogeneity. The cream is suitable for treating vaginal infections.

Example X Ophthalmic ointment.

1000 G ophthalmic ointment with a concentration of U-57930 E or U-60970 E of 0.5% is prepared from the following 15 ingredients: U-57930 E or U-60970 E 5 g

Bacitracin 12.2 g

Polymyxin B sulfate (10,000 units / mg) 1 g 20 Light liquid petrolatum 250 g

Wool grease, 200 g

White petrolatum up to 1000 g

The solids are finely divided using a compressed air mill and added to the light liquid petrolatum. The mixture is passed through a collold mill to evenly distribute the micronized particles. Wool grease and white petrolatum are melted together at a temperature of 45-50 ° C. To the melt the suspension in liquid petrolatum is added and stirred until an ointment-like consistency is reached. The ointment is packaged in ophthalmic tubes with a content of approximately 4 g. The ointment is suitable for application to the eye for the treatment of local infections in humans and animals. Bacitracin and polymyxin B sulfate can be omitted, and, if desired, 5 g (0.5%) of methylprednisolone 35 can be included to treat inflammation.

8201595 - 39 -

Example XI Eye-ear drops 1000 Ml sterile aqueous solution for eye or ear containing per ml 10 mg U-57930 E or U-60970 E and 5 mg prednisolone is prepared from the following ingredients: 5 U-57930 E or U- 60970 E 10 g

Methylprednisolone sodium phosphate 5 g

Sodium citrate k, 5 g

Sodium bisulfite 1 g

Polyethylene glycol 4000 120 g 10 Myristyl-Y-picolinium chloride 0.2 g

Polyvinylpyrrolidone 1 g

Deionized water up to 1000 ml

The ingredients are dissolved in water and the solution is sterilized by filtration. Sterilized dropper bottles are aseptically filled with the solution. The preparation is suitable for topical treatment of inflammation and infections in sensitive areas such as the eye and ear.

Example XII Trocheen.

10,000 troches are made from the following 20 components: U-57930 E or U-60970 E 100 g

Neomycin sulfate 50 g

Polymyxin B sulfate (10,000 units / mg) 1 g 25 Ethylaminobenzoate 50 g

Calcium stearate 150 g

Powdered sucrose up to 5000 g

The powdered ingredients are mixed well and then pressed into troches of 0.5 g in the manner usual for making pressed tablets. To treat infections in the mouth and throat, the troches are held in the mouth to allow them to slowly dissolve.

Example XIII Suonositoria.

1000 Suppositories of 2.5 g each and each containing 35 1000 mg U-57930 E or U-60970 E are made from 8201595 - 1 * 0 - the following ingredients: U-57930 E or ü-60970 E 100 g

Polymyxin B sulfate (10,000 units / mg) 1.25 g 5 Methylprednisolone 1 g

Ethylaminobenzoate 75 g

Zinc oxide 62.5 g

Propylene glycol 162.5 g

Polyethylene glycol 1 * 000 to 2500 g of 10 U-57930 E or U-60970 E, polymyxin B sulfate, methyl prednisolone, ethyl aminobenzoate and zinc oxide are added to propylene glycol and the mixture is ground until the powders are finely divided and evenly dispersed. Polyethylene glycol 1000 is melted and the melt dispersion in propylene glycol is added slowly and with stirring. The mixture is then poured into 1 ° 0 ° C molds where it is allowed to cool and solidify. The suppositories are then removed from the molds and wrapped in foil. The suppositories are suitable for treating local inflammation and infections. The steroid can be omitted if desired. Example XIV Mastitis ointment.

1Q00 G ointment for the treatment of mastitis in livestock is prepared from the following ingredients: U-57930 E or U-60970 E 25 g 25 Methylprednisolone acetate 0.5 g

Light liquid petrolatum 300 g

Chlorobutanoly anhydrous 5 g

Polysorbate 80 5 g 2% Aluminum Monostearate Arachis 30 Oil Gel 1 * 00 g

White petrolatum to 1000 g U-57930 E or U-60970 E and methylprednisolone acetate are ground together with light liquid petrolatum until a finely divided and uniform dispersion is obtained.

Chlorobutanol, polysorbate 80, arachisole gel and white petrolatum 82 0 1 5 9 5 - 1 + 1 - are heated at 1 + 9 ° C after which the dispersion in liquid petrolatum is stirred in the melt with cooling (and solidifying) to ambient temperature. Mastitis syringes are filled with the preparation in doses of 10 g.

Example XV Animal feed.

1000 G of a feed mixture is prepared from the following ingredients: U-57930 E or U-60970 E 10 g

Soybean meal 1 + 00 g 10 Fish meal 1 + 00 g

Wheat germ oil 50 g

Sorghummolasse 11 + 0 g

The ingredients are mixed and pressed into granules. The composition can be fed to laboratory test animals such as rats, mice, guinea pigs and hamsters for prophylactic purposes during transport. The composition can also be fed to other animals such as poultry eg chickens, ducks, turkeys and geese, the composition being included in the normal food in an amount such that the desired dose of U-57930 E or U-60970 E is administered. Example XVI.

In the same manner as in the previous examples, preparations are prepared cr.q. manufactured which instead of U-57930 E or U-60970 E contain any of the other antibacterial compounds of the invention. In all cases, the free base or a pharmacologically acceptable salt thereof such as the hydrochloride, sulfate, phosphate, lithium salt, sodium salt, potassium salt or calcium salt can be used.

Example XVII Capsules.

1000 1000 Two-part hard gelatin capsules, each containing 200 mg of U-57930 E or U-60970 E and 200 mg of hydroxychlorquinine sulfate, are made from the following ingredients: U-57930 E or U-60970 E 200 g

Hydroxychloroquinine sulfate 200 g 35 Talc 75 g 8201595 - h2 -

Magnesium stearate 25 g

The ingredients are mixed well and encapsulated in the usual manner. The capsules are suitable to prevent residual attacks of P. vivax in adults by administering 1 capsule orally once a week.

Example XVIII Tablets 1000 Tablets each containing 125 mg U-57930 E or U-60970 E and 325 mg quinine sulfate are prepared from the following ingredients: 10 U-57930 E or U-60970 E 125 g

Quinine sulfate 325 g

Lactose 50 g

Corn starch 50 g

Calcium stearate 25 g 15 Light liquid petrolatum 5 g

The ingredients are mixed well and agglomerated. The agglomerates are broken by forcing them through a sieve with 1.19 mm openings. The granules formed are then pressed into tablets suitable for treating malaria by first administering two tablets every 8 hours for 7 days and then 1 tablet three times a day for 7 days. Example XIX Syrup 1000 Ml aqueous suspension containing per 10 ml 250 mg U-57930 E or U-60970 E, 25 mg pyrimethamine and 500 mg sulfa-25 diazine is prepared from the following ingredients: U-57930 E or U-60970 E 25 g

Pyrimethamine 2.5 g - Sulfadiazine 50 g

Citric acid 2 g 30 Benzoic acid 1 g

Sucrose 700 g

Tragacanth 5 g

Lemon oil 2 ml

Deionized water up to 1000 ml of citric acid, benzoic acid, sucrose, tragacanth and 8201595 - ί * 3 - lemon oil are dissolved in 850 ml of water. U-57930 E or U-60970 E, pyrimethamine and sulfadiazine are added to the solution with stirring to achieve an even distribution. Then water is added to a volume of 1000 ml. The syrup is suitable for prophylactic treatment of malaria in adults by each weds: administer one tablespoon (10 ml) Example XiC Parenteral Solution.

A sterile aqueous solution for intramuscular administration, containing 200 mg TJ-57930 E or U-60970 E 10 per ml, is prepared from the following ingredients: U-57930 E or ü-60970 E 200 g

Lactose 50 g

Water for injection up to 1000 ml of U-57930 E or U-60970 E and lactose are dissolved in water and the solution is sterilized by filtration. Sterilized vials are aseptically filled with the solution in an amount of 2 mL.

Example XXI

In the same manner as in the previous examples, preparations are prepared or manufactured which, instead of U-57930 E or U-60970 E, contain any of the other compounds of the invention which are effective against malaria. In all cases, the free bases or a pharmacologically acceptable salt thereof can be used, for example the. hydrochloride, sulfate, phosphate, lithium salt, sodium salt, potassium salt or calcium salt.

Determination of activity against malaria (P.

mountain heather).

Groups of 10 male CF-1 mice (18-30 g) each were infected intraperitoneally with whole blood from mice infected with P. berghei three days previously. The inoculum used was 0.2 ml of heparinized blood diluted 1:10 with saline. This amount of blood contained about 10 ^ parasites.

Four hours after the infection, each group 8201595 '- Μ - was treated with the test compounds indicated below which were administered subcutaneously (0.2 ml) and orally (0.5 ml in the feed), respectively. Administration was done once a day for four days. The mice were observed for survival and protection for 28 days. As a criterion, mortality was taken within 6 days. The results, based on comparison of the treated groups and the untreated groups or the chloroquinine-treated groups, were run on an IBM 370 digital computer.

The results are summarized in the table below.

10 In vivo activity against P. berghei ♦

Compound Average dose Average protection for survival 1 ^ mg / kg dose ^) mg / kg

Subcutaneous oral Subcutaneous Oral U-57930 E 0.16 1.6 16 (12-22)> 50 15 U-21251 F <20 - 53 (U6-61) U-2 ^ 729 A <1.25 - H, T (3.2-6.9) U-828H chloroquinine <5-10 12.5 11.5 (8.8-15) ft

Chloroquinine (PO ^) g <5 - 20 - 20 1 ^ Dosage in which the mean survival time (0T ^ Q) was significantly increased (p * 0.05) compared to the OT ^ q in the control group 2) 'Mean protection dose in mg / kg (95% confidence limits) Other protozoa against which the compounds of the invention are active are intracellular parasites, for example of the species Plasmodia. Toxoplasma and Leishmania. protozoans that digest the red blood cells of treated patients, for example, Entamoeba histolytica and certain Trypanosoma, and other helminths that digest the red blood cells during disease, for example, Schistosomes.

35 8201595

Claims (19)

A compound according to claim 1, wherein R 1 is in the U position in the pyridine ring and represents an alkyl group of 1 to 8 carbon atoms, and R 1 is in the 2 or 3 position of the pyridine ring, and the pharmaceutically acceptable salts thereof. 1 *. A compound according to claim 2, wherein R 1 is in the U-position of the pyridine ring and represents an alkyl group of 1 to 8 carbon atoms, as well as the pharmaceutically acceptable salts thereof. 5. 3- (5'-Ribonucleotide) of a compound of the formula 13 wherein in the 3, 4, 5 'and / or 6 position is present in the pyridine ring and a hydrogen atom, substituted or unsubstituted alkyl group with in the alkyl group 1-8 carbon atoms, 8201595 '- k6 - substituted or unsubstituted cycloalkyl group, substituted oxygen atom, substituted nitrogen atom, halogen atom, substituted or unsubstituted phenyl group or a group - (CHgJ ^ -OH or - (CHg ^ NK ^ R ^ wherein R 1 and R 1 each represent a hydrogen atom or alkyl group 5 having 1 to 8 carbon atoms and n is a number from 1 to 8, and Y represents a 7 (S) halogen atom or 7 (R) halogen atom, as well as the pharmaceutical , acceptable salts thereof. 6. A compound according to claim 5 * wherein Y represents a 7 (S) halogen atom, as well as the pharmaceutically acceptable salts thereof. 7- 3- (5'-Ribonucleotide) of a compound of the formula wherein Rp which may be in the ring at the 3, H, 5, 7, 8 and / or 9 position is a hydrogen atom, whether or not unsubstituted alkyl group with in the alkyl group 1-8 carbon-15 atoms, substituted or unsubstituted cycloalkyl group. substituted oxygen atom, substituted nitrogen atom, halogen atom, substituted or unsubstituted phenyl group or a group - (CH2) n-0H or - (C ^ n-HR ^ where R ^ and R ^ each represent a hydrogen atom or alkyl group of 1-8 carbon atoms and n is a number from 1 to 8, R 1 represents a hydrogen atom, methyl group, ethyl group or hydroxyethyl group, n is a number from 1-U-, and Y is a 7 (S) halogen atom or 7 (R) - halogen atom "as well as the pharmaceutically acceptable salts thereof". 8. A compound according to claim 7, wherein the 7 (S) halogen atom is a 7 (S) chlorine atom. The compound of claim 7 wherein Y represents a 7 (S) halogen atom, R 1 represents an alkyl group of 1 to 8 carbon atoms, and R 1 represents a hydrogen atom, and the pharmaceutically acceptable salts thereof. A compound according to claim 9 wherein Y represents a 7 (S) halogen atom, R 1 represents an ethyl group and R 1 represents a hydrogen atom. The compound of claim 9 wherein Y represents a 7 (S) halogen atom, R 1 represents a butyl group and R 1 represents a hydrogen atom. 8201595 - ^ 7- The compound of claim 9 wherein the 7 (S) halogen atom is a 7 (S) chlorine atom. The compound of claim 10 wherein the 7 (S) halogen atom is a 7 (S) chlorine atom. A compound according to claim 11, wherein the 7 (S) halogen atom is a 7 (S) chlorine atom. 15. 3- (5 * -Ribonucleotide) of a compound of the formula 15 as well as the pharmaceutically acceptable salts thereof. 16. 3- (5'-Ribonucleotide) of a compound of formula 16, the R2 palmitate and its pharmaceutically acceptable salts. 17- 3- (5'-Ribonucleotide) of a compound of the formula 17 wherein X is a 7 (S) -halomethyl-1-thio- (X-lincosamidin or 7 (R) -halomethyl-1 -thio) <X-lincos amine dinner form 15 and its pharmaceutically acceptable salts 18. 3- (5'-Ribonucleotide) of a compound of the formula 18 wherein X has the meaning as defined in claim 17, as well as the pharmaceutically acceptable salts thereof 3- (5'-Ribonucleotide) of a compound 20 of the formula 19 wherein X has the meaning indicated in claim 17, as well as the pharmaceutically acceptable salts thereof 20. 20- (5 * Ribonucleotide) of a compound of the formula 6 wherein A, B and E each represent a nitrogen atom, oxygen atom, sulfur atom or a group CR-jRg wherein R1 and Rg have the meaning defined in claim 1 and may be attached to any ring carbon or nitrogen atom, where Rc may be attached to more than one ring carbon atom, as well as the pharmaceutically acceptable salts thereof. 21. 3- (5'-Ribonucleotide) of a compound 30 of the formula 20 as well as the pharmaceutically acceptable salts thereof. 22. 3- (5'-Ribonucleotide) of a compound of the formula J wherein A, B, D and E each represent a nitrogen atom, oxygen atom, sulfur atom or a group CR ^ Rg wherein R ^ and R2 have the meaning as defined in claim 1 and can be attached to any ring carbon atom or nitrogen atom, which can be bound to more than one ring carbon atom, as well as the pharmaceutically acceptable salts thereof. 23. A compound according to claim 22 of the formula 21 and the pharmaceutically acceptable salts thereof. 2h, 3- (5'-Ribonucleotide) of a compound of the formula 3 wherein R 1 is a hydrogen atom, unsubstituted or substituted alkyl group, with substituted alkyl group 1-8 carbon atoms, substituted or unsubstituted cycloalkyl group, substituted oxygen-10 atom. nitrogen atom, halogen atom, substituted or unsubstituted phenyl group or a group "(CHg ^ -OH or - (CHg ^ NR ^ Rtj) where R ^ and R ^ each represent a hydrogen atom or alkyl group and n is a number from 1-8, where R ^ may be present in a single or multiple sites in the pyridine ring, if not already occupied by Rg which represents a group of the formula k wherein X is a 7 (S) -halomethyl-1-thio-lincosamidine or 7 represents (R) -halomethyl-1-thio-e (-lincosamidine residue), wherein Rg is present in a single place in the pyridine ring, if not already occupied by R1, and the pharmaceutically acceptable salts thereof. 5 * -Ribonucleotide) of a compound of the formula 5 wherein R 1 and R 8, which are o p may be present in the ring at the 2, 3, 5, 6, 7, 8 or 9 position, have the meaning indicated in claim 1, R 1 represents a hydrogen atom, methyl group, ethyl group or hydroxyethyl group and n is a number of 1-k, as well as the pharmaceutically acceptable salts thereof. 26. A compound of formula 8 wherein R represents a group of formula 9. 27. A compound of the formula 8 wherein R represents a group of the formula 10. 28. A compound of the formula 8 wherein R represents a group of the formula 11. 29. A compound of the formula 8 wherein R represents a group of the formula 12. 30. Process for preparing the 3- (5 * - 8201595 - k9 - ribonucleotides) of a compound of the formula 14, which may be present in the ring at the 3, 5, 7, 8 and / or 9 position and a hydrogen atom, al deux unsubstituted alkyl group with 1-8 carbon atoms in the alkyl group, substituted or unsubstituted cycloalkyl group, substituted oxygen atom, substituted nitrogen atom, halogen atom, substituted or unsubstituted phenyl group, or (CHg ^ -Oi) or - (CHg) n-NR ^ R ^ wherein R ^ and R ^ each represent a hydrogen atom or alkyl group with 1-8 carbon atoms and n is a number from 1-8, R ^ represents a hydrogen-10 atom, methyl group, ethyl group or hydroxyethyl group, n represents a number from 1 to 1, and X represents a 7 (S) halogen atom or 7 (R) halogen atom, as well as the pharmaceutically acceptable salts thereof, culturing Streptomyces rochei NRRL 3533 in an aqueous nutrient medium in the presence of the compound of the formula ih- under conversion to the 3- (5'-ribonucleotide) thereof. 31. Pharmaceutical composition with antimicrobial activity, which contains as active ingredient at least one compound according to any one of the preceding claims. 32. Compounds and pharmaceutical preparations 20 as well as methods of obtaining one as described in the description and examples. 25 8201595