HU191085B - Process for producing 3-/5-comma above-ribonucleotide/derivatives - Google Patents

Abstract

The 3-(5'-ribonucleotide) of a compound as claimed in any claims 1 to 35 of British Patent Specification No. 2,063,252A, in which X is 7(S)- halomethyl 1-thio-alpha- lincosaminide or 7(R)-halomethyl 1- thio-alpha-lincosaminide; or a pharmaceutically acceptable salt thereof.

Description

(57) EXTRAS

The present invention relates to a process for the preparation of S-S'-ribonucleotide derivatives of compounds of formula (I). According to the method, the strain Streptomyces rochei is cultivated in the presence of a compound of the formula (R = H) in an aqueous medium containing carbon, nitrogen, oxygen and phosphorus, and the resulting 3- (5'-ribonucleotides) are isolated, υ The resulting ribonucleotides have strong in vivo activity against various microorganisms.

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The present invention relates to novel 3-ribonucleotides of lincomycin and clindamycin type compounds in which the cyclic amino acids are substituted for the propyl ligric acid unit.

The properties and preparation of antibiotic lincomycin are disclosed in U.S. Patent No. 3,086,912. Clindamycin is disclosed in U.S. Patent 3,486,163. These antibiotics have widespread therapeutic use inipd in human and veterinary medicine. Numerous patents have been granted worldwide for these antibiotics and their various derivatives.

U.S. Patent 3,671,647 discloses 3-nucleotides of linkamycin and clindamycin. All lincomycin derivatives of clindainicin disclosed in this patent have a common structural unit: propylhygric acid. In vivo experiments on S. aureus have shown that the efficacy of these 3-nucleotides is approximately one tenth of that of the parent compound.

The compounds of the present invention are 3- (5'-ribonucleotide) compounds of formula (1). In formula (1), R is a group of formula (a), (b), (c) or (d).

In accordance with the present invention, compounds of formula (1) are prepared by culturing Streptomyces rochei NRRL 3533 in an aqueous medium containing appropriate nutrients in the presence of a compound of formula (I) wherein R is hydrogen and then from the culture medium. isolating the desired 3- (5'-ribonucleotide) derivative.

Surprisingly, it has been found that the in vivo activity of the nucleotides of the present invention is the same as that of the? Because these properties are very important, the nucleotides of the invention are considered to be particularly useful for therapeutic applications. These compounds are useful in the prophylactic and therapeutic treatment of individuals carrying a protozoan parasite. For example, if the protozoan is a malaria parasite, it can also infect animals: for example, mice may be infected with Plasinodium bcrglici-vcl; birds such as ducks can be infected by P. ophurae, chickens by P. gallinacium; from mammals, monkeys may be attacked by P. cynomolgi and humans may be infected by P. falciparum, P. vivas and P. malariae.

Mammals carrying protozoan parasites of the order Coccidia of the Sporozoa class (this micro-parasite produces coccidiosis) can be treated by administration of the compounds of the invention. These compounds are useful in veterinary medicine for example in the following cases: bovine animals infected with Ejmeria zurnii, E. bovis and E. ellipsoidalis; Sheep and goats infected with E. parva and E. faurei, pigs infected with E. debliecki, E. scabra or Isospora suis, Isospora bigemina, 1. dogs and cats infected with felis, E. canis, or E. felina; Poultry infected with E. tenella; Rabbits infected with E. stiedae- or E. perforans; and Weasel infected with E. mustelae.

Those lincomycin and clindamycin derivatives. which can be converted to 3-ribouucleotides are included in Annex 1. Instead of the 3-hydroxyl group of the lysosaminamine moiety, the molecule contains adenylic, guanylic, cytidylic or uridylic acids as nucleotides. 2. ______ - - · 1

The parent compounds defined in Annex 1 may be prepared by the methods described in U.S. Patent 4,278,789.

Compounds disclosed in Annex 1. 3- ^(5-f -ribonukleotidj derivatives by the methods disclosed in U.S. 3,671,647 discloses prepared. C nucleotides salts can be prepared by following methods for the same patent.

The nucleotides of the present invention can be formulated into pharmaceutical compositions as described in the composition examples of U.S. Patent No. 4,278,789. These compositions may be formulated by substituting a nucleotide of the invention for the active ingredient in these examples. This substitution is made on the basis of the nickel cell counts.

The following general and specific methods can be used to determine and characterize the nucleotides of the invention.

Measurement of 3- (5'-ribonucleotide) derivatives

Since the 3-ribonucleotides of the present invention have no antibacterial activity in vitro, their formation from the parent bacterial parent compound can be easily monitored by measuring the reduction in antibiotic efficacy. The standard determination of the parent compound having an antibacterial activity in culture filtrates or reaction mixtures is performed using Sarcina lutea strain ATCC 9341. In the presence of 3-ribonucleotides in fermentation broths, extracts and purified materials, the assay is carried out by first hydrolyzing the phosphodiester linkage with crude alkaline phosphatase or serum phosphodiesterase as described below. The antimicrobial compound in the hydrolyzate is determined by a standard method.

Enzymatic hydrolysis

Alkaline phosphatase: Stock solution (0.5 mg / l, 0.54 cfu / mg) was prepared from pigeon with alkaline phosphatase / EC 3.1.3.1 (Sigma) / Tris (hydroxymethyl) aminomethane hydrochloride (hereinafter Tris) with a buffer of 0, 01M, pH 8.0. Samples to be assayed were diluted 1: 2 with cnzim buffer mixtures and incubated at 28 ° C for 18 hours.

Snake venom phosphodiesterase: Stock solution (100 mg / ml, 0.026 units / mg) is prepared from purified snake venom phosphodiesterase / EC 2.1.4.1 (Sigma) / distilled water. Incubation mixture: 0.2 ml of a 1 mg / ml aqueous solution of the test sample, 0.6 ml of 0.01 M Tris buffer, pH 9.0, and 0.1 ml of 0.3 M magnesium chloride solution and 0.1 ml enzyme stock solution. Incubation was carried out at 37 ° C for 18 hours.

Spleen phosphodiesterase: Stock solution (1 mg / ml, 19.6 units / mg) was prepared from spleen phosphodiesterase (EC 3.1.4.18 (Sigma)) in distilled water. The incubation mixture contains 0.4 ml of a 0.5 mg / ml aqueous solution of the test sample, and 0.5 ml of 0.02 M Tris buffer pH 7.0 and 0.1 ml of enzyme stock solution. Incubation was performed at 37 ° C for 18 hours.

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Thin-layer chromatography (TLC) of preparations and enzymatic hydrolysates

Production and purification of the 3-ribonucleotides are monitored by measuring against S. lutea (see above) and by TLC assay. Silica gel G was used as the adsorbent for TLC and mixtures of butanone-acetone-water (186: 52:20, v / v) or butanone-acetone-water (8: 5: 1, v / v) were used as eluent. Biologically active starting materials are detected by bioautography on agar inoculated with S. lutea.

The enzymatic or chemical hydrolysis products of the 3-nucleotides are separated by the following TLC systems:

A: Silica gel GF plate (Aualtech Inc.), pellet water;

B: silica gel FG plate, eluent n-propanol, concentrated ammonium hydroxide / water 55:10:35 (v / v);

C: NM-Polygram Cellulose 300 (Brinkman Instruments Inc.) plate, eluent 1-butanol / water / formic acid 77: 13: 10 (v / v).

Substances absorbed in the UV spectral region are detected using a short wavelength UV lamp. Biologically inactive substances that do not absorb UV are detected with permanganate periodate reagent. Biologically active nucleotides are detected by bioautography on agar inoculated with S. lutea.

The following example illustrates the fermentation and purification procedures used to prepare the nucleotide derivatives of U-57 930 and U-57 930E respectively. The 3-ribonucleotides of the other compounds as defined in Annex 1 may be prepared by the methods described in this Example or equivalent methods. (U-57 930E is the hydrochloride salt of U-57 930.)

The process of the invention will be described in detail in the following embodiment. Unless otherwise noted, percentages are always by weight and solvents are always by volume.

Example 1

A) The fermentation process

Streptomyces rocke strain NRRL 3533 was cultured in a medium containing 10 g / l glucose, 4 g / l Difco-peptone, 4 g / l Difco yeast extract, 0.5 g / l MgSO ₄ · 7H ₂ O, 2, 0 g / liter KH ₂ PO _4l 4 g / liter K ₂ HPO ₄ . The culture was maintained on a rotary shaker at 28 ° C for 3 days. Mycelial strains from this culture are used to inoculate the fermentation medium containing the same components. The fermentation was carried out on a rotary shaker for 48 hours at 28 ° C. At the end of the 48 hour incubation, enough U-57 930 (R) is added to 50 mg / L, and the fermentation is continued at 32 ° C, and after 12 hours additional U-57 930 is added. to a concentration of 150 mg / liter. After another 12 hours, .......... - ...... 1, the concentration of U-57 930 was increased to 250 mg / liter. The fermentation is carried out at 32 ° C for 24 hours after the addition of the last portion of U-57 930. The measurement at this time shows that the culture filtrate does not contain more than 1 mg / liter of U-57 930, ie the remainder of this compound, i.e. 249 mg / liter, converted to a biologically inactive compound.

S. rochei NRRL 3533 is a known microbe that is commonly available on request in the NRR1 bank. Address of this bank: Northern Utilization and Research Division, Agricultural Research Service, U.S.A. Department of Agriculture, Peoria, 111. USA.

B) Isolation and purification procedures

Isolation of the 3-ribonucleotides of U-57 930 from the fermentation broth Adsorption on Amberlite XAD-2: About 12 L of fermentation broth containing 3 g of inactivated U-57 930 was filtered at pH 7.7 using a filter aid. The mycelial mass was washed with 1.2 liters of water and discarded. The clear filtrate was combined with the wash liquid, adjusted to pH 6.0, and passed through a column of 600 ml Amberlite XAD-2 (Rohm and Haas Co., Philadelphia, PA) at a flow rate of 40 ml / min. The biological activity of the effluent is assayed before and after treatment with alkaline phosphatase and discarded. The column was washed with 2 liters of water. The aqueous niosalt liquid before and after treatment with alkaline phosphatase is biologically ineffective and is therefore discarded. The column was then eluted with methanol / water (70:30 v / v). Elution was carried out at 20 ml / min and 20 ml fractions were collected. The results of the efficacy study before (-E) and after (+ E) treatment with alkaline phosphatase are as follows:

Serial number of the fraction Zone (S. lutea) (mm) -E E + 5 0 0 10 0 0 11 0 0 13 0 36 20 33 55 . 25 32 54 40 31 50 45 29 48 50 26 39 55 23 38 60 21 37 65 19 27 70 17 26 75 17 26 80 16 26 85 16 26 90 16 26 95 15 26 100 16 26 110 16 21 120 16 21 130 15 21 140 15 21

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Zone (S. lutea) (mm) Fraction number —Ε + E

150 15 21 160 15 21 170 15 21 180 15 21 190 15 21 200 15 21

Fractions of Λ 12 80 were combined, concentrated on aqueous side and lyophilized. This gives 12.22 g of the preparation), which is ADA-34.1.

In another set of experiments, 6 liters of fcrmcntl containing 2 g of "inactive" U-57 930 was treated as described above. The methanol eluate from the Amberliter XAD-2 column was retained as ADA-143B. This solution is not concentrated to dryness but purified by Dowex-1 chromatography as described below.

Dowex-1 chromatography: The column is prepared from 300 ml of Dowex-1 (X-4) acetate form. The ADA-143B methanolic solution, pH 8.2, was passed through the column and the effluent was collected in fractions of 20 ml (fractions 1-60) at a flow rate of 2.5 ml / min. The column was then washed with 1.5 L of water (flow rate 10 mL / min, fractions 66-108). The column was then eluted with 5% acetic acid (10 mL / min, fractions 109-310). The following fractions are combined:

1st collection

2nd collection

3rd collection

4. Collection

5. Collection

6. Collection

7th collection

8. collection

Fractions 1-80 Fractions 81-110 Fractions 111-130 Fractions 131-150 Fractions 151-190 Fractions 191-230 Fractions 231-270 Fractions 271-310 Fractions

1000 ml (ADA-1A)

600 mL (ADA-2A) 450 mL (ADA-3A) 450 mL (ADA-4A) 900 mL (ADA-5A) 900 mL (ADA-6A) 900 mL (ΛΗΛ-7Λ) 900 mL (ADA-8A)

The results of the efficacy study before (-E) and after (+ E) treatment with alkaline phosphatase are as follows:

Zone (S. lutea) (mm) -E + E 1. Collection 31 52 2. Collection 36 49 3. Collection 34 45 4. Collection 18 40 5. Collection 25 30 6. Collection 27 29 7. Collection 27 29 8. Collection 29 31

Collections 1 and 2 were combined, concentrated to an aqueous solution and lyophilized. This yields 1.48 g of material, designated ADA-2.1 Collection 3 and 4, and is treated similarly to give 2.5 g of material, designated ADA-2.2.

When treated with alkaline phosphatase, the ADA-2.1 and ADA-2.2 formulations yield U-57 930.

The ADA-34.1, ADA-2.1, and ADA-2.2 formulations were combined and purified by the following dual countercurrent distribution procedure.

Dual countercurrent distribution: 16.20 g of ADA-34.1, ADA5 2.1 and ADA-2.2 are combined in 25 ml of each of the two solvent systems consisting of 1-butanol / water. The solutions are poured into the middle tubes of a glass double counterflow dispenser (100 tubes / 25 ml / phase). The distribution is analyzed after 150 transmissions for biological efficacy, after treatment with alkaline phosphatase is banned (E) and after (+ E). The results are as follows:

Zone (S. lutea Lower Collector Sensitive) (mm)

-Ε + E

5 29 31 10 31 33 15 30 33 20 27 33 25 22 33 30 17 34 35 0 34 40 0 33.5 45 0 33 50 0 34 55 0 34 60 0 35 65 0 36 70 0 38 75 0 39 80 0 40 85 0 41 90 0 42 95 0 43 100 0 43.5

Zone (S. lutea-

Lower machine sensitive) (nini) -E E + 50 0 45 45 trace 46 40 15 47 35 17 47.5 30 17 47 25 18 47 20 17.5 48 15 17 48 10 16 48 5 15 50 0 trace 50 Top Machine ' 5 trace 49 10 trace 48.5 15 trace .48 20 trace 48.5 25 trace 49

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Lower collector Zone (sensitive to S. lutea) (mm) -E + E 30 30 50 35 trace 51 40 15 52 45 16 53 50 21 54

Upper collector Zone (sensitive to S. lutea) (mm) -E + E 100 17.5 54 95 17 53.5 90 19 53.5 85 20 53.5 80 21 53.5 75 22 53.5 70 24 53.5 65 26 53.5 60 28 53.5 55 30 52.5 50 32.5 52 45 33 52 40 35 52 35 36 52 30 39 49 25 41 47 20 43 48 15 43 46 10 43 43 5 35 35

The following collections are made (see below). Each collection was concentrated to an aqueous solution and lyophilized to the appropriate formulation.

I. Collection: Lower Collector 1-50;

II. Collection; Lower collector 51-100; lower machine 50-30;

III. Collection: Lower Machine 29-0; upper machine 1-50;

upper collector 100-30. The following preparations are obtained:

From Collection I: ADA-47.1, 9.78 g

II. From Collection: ADA-47.2, 0.30 g

Alii. From Collection: ADA-47.3, 5.29g

ADA-47.2 and ADA-47.3 were combined and purified by DEAE-Sephadex chromatography as described below.

DEAE-Sephadex chromatography: 300 g of DEAE-Sephadex (A-25) resin was stirred for 1 hour with water and then for 2 hours with 0.5N aqueous sodium hydroxide solution. Wash the ion exchanger with water until the pH is about 7.5. The material was then stirred for 2 hours with 0.5N aqueous acetic acid, washed with water to an inert pH, then poured into a column and compacted to a constant height of 1 kg. The column was washed with 4 L of water, 8 L of 0.1% aqueous tris (hydroxymethyl) aninomethane (hereinafter THAM) solution, and 3 L of 0.03M THAM acetate buffer pH 8.0 (prepared to dissolve 3.64 g of THAM in 800 ml of water and adjust the pH to 8.0 with acetic acid and make up to 1 liter).

By combining ADA-47.2 and ADA-47.3, approx. Dissolve 5.50 g in 20 ml of 0.03M THAM acetate pH 8.0 and pour onto the top of the column. The column is then eluted with 0.3M THAMacetate buffer pH 8.0. Fractions 1 to 186 (about 20 mL) were collected and the column eluted with upward flow. Fractions Λ, B, C, D, and E are collected (each volume is 1 liter). The test for biological activity before (-E) and after (+ E) treatment with alkaline phosphatase shows the following results.

Serial number of fraction Zone (sensitive to S. lutea) (mm) -E + E 3 0 0 6 0 0 9 0 0 12 0 0 15 0 0 18 0 0 21 0 0 24 0 0 27 0 0 30 0 0 33 0 0 36 38 39 39 43.5 44 42 36 36 45 23.5 23 48 15 16 51 0 0 54 0 0 57 0 0 60 0 0 63 0 0 66 0 0 69 15 16 72 17 20 75 21 26 78 23 27 81 23 30 84 23 29 87 22 24 90 21 23 93 21 24 96 22 26 99 21 35 102 22 44 105 23 51 108 22.5 54 111 22.5 52.5 114 22 52 117 22 54.5 120 20.5 56 123 20 56

191,035

Serial number of the fraction Zone (sensitive to S. lutea) (nmi) -E. + E 126 20.5 56 129 20 55 132 20.5 55 135 21 55 138 21 54 141 20 54 • 144 20.5 54 147 21 54 150 23 54 153 21.5 52

Serial number of the fraction Zone (sensitive to S. lutea) (mm) -E E + 156 20 53 159 20 54 162 20 54.5 165 20 54.5 168 21 55 171 22 55 174 22.5 54 177 23 54 180 24 54 183 24 54 186 0 46.5 THE 24 23 B 15 28 C 20 44 D 22 23 E 22 24

The following collections were made:

I. Collection 34-48 Fractions, 280 mL (ADA-69B)

II. Collection 75-90 fractions, 330 ml (ADA-69C)

III. Collection 101-111 Fractions, 180 ml (ADA-69D)

ARC. Collection 114-150 Fractions, 580 mL (ADA-69E)

V. Collection 151-164 Fractions, 100 ml (ADA-69F)

VI. Collection 165-186 Fractions, 125 mL (ADA-69G)

VII. Collection C fraction, 1 liter (ADA-69A)

Collection I (ADA-69B) contains unchanged U-57 930 and is therefore discarded.

Of fractions A-E, only fraction C is retained, the rest being discarded.

II. Collection (ADA-69C) contains an unknown substance which, when treated with alkaline phosphatase, yields U-57 930. EJV:? _Max = 275 nm.

Á III. (ADA-69D) 3- (cytidine-5'-phosphate) U-57 930 is turlulase and treated as described below. UV: λ _max 270 nm.

The IV. (ADA-69E) contains 3- (adenosine-5'-phosphate) U-57 930 and is treated as described below. UV: λ _max 260 nm.

Collection V (ADA-69F) S-fadenosine S'-phosphate U-57 390, 3- (uridine-5'-phosphate) - U-57 930 and 3- (guanosine S'-phosphate) - U-57 930 contains a mixture. This solution was treated as described below.

VI. (ADA-69G) contains 3- (guanosine-5'-phosphate) U-57 930 and is treated as described below. UV:? _Max = 254 nm; sh at 275.

VII. Collection (ADA-69A) is a mixture of 3- (guanosine-5'-phosphate) U-57 930, 3- (uridine-5'-phosphate) and U-57 930. This solution was treated as described below.

Substantially pure isolation of 3- (cytidine-5'-phosphate) - U-57 930, 3- (adenosine-5'-phosphate) - U-57 930 and S-fguanosine-S'-phosphate) - III, IV and VI respectively collections: Removal of THAM acetate buffer by Amberlite XAD-2 chromatography.

A III., IV. and VI. collections obtained as described above are passed through columns containing Amberlite XAD-2. The dripping liquid was discarded. The columns were washed with water followed by methanol and water. Elute with a 70:30 mixture. The fractions were analyzed by UV, and before and after treatment with alkaline phosphatase for biological efficacy. The appropriate fractions were combined, concentrated to an aqueous solution and lyophilized. The following table summarizes the amount of Amberlite XAD-2 used for each collection, the amount of aqueous wash liquid, the methanol eluate, and the amount of material obtained.

Collection Used Amber Lite XAD-2, ml Aqueous washer- liquid, ml methanol effluent, ml Separated substance, ml III. 50 200 300 150 ARC. 200 800 600 3510 VI. 50 200 300 470

In III. collection material ADA-73.1; and ADA-74.1; collection material is stored under the ADA-75.1 mark.

Removal of THAM acetate buffer from Collection V (ADA-69F) and VII. (ADA-69A) by Amberlite XAD-2 chromatography.

The column is prepared from 300 ml Amberlite XAD-2 resin. Transfer columns V and VII through the column. collections comprising a mixture of 3- (adenosine-5'-phosphate) -U-57,930, 3- (uridine-5'-phosphate) -U-57,930 and 3- (guanosine-5'-phosphate) -U-57,930. The dripping liquid is discarded. The column was washed with 600 mL of water and the dripping liquid was also discarded. The column was then eluted with 70-30 methanol / water. The fractions which, after treatment with alkaline phosphatase, combine to form a biologically active substance are 300 ml which is concentrated to an aqueous solution and lyophilized. This gives ADA-71.1 670 shirt weight. ADA71.1 is treated as described below.

Separation of 3- (uridine-5'-phosphate) - U-57 930 from 3- (adenosine-5'-phosphate) - U-57 930 and 3- (guanosine-5'-phosphate) - U-57 930 DEAE- Sephadex chromatography: 600 ml

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--------- J

The DEAE-Sephadex acetate resin prepared as described above was washed with 0.03M THAM acetate buffer pH 8.0 and compacted in a glass column (4.5 cm internal diameter, 40 cm high) under hydrostatic pressure.

ADA-71.1 (see above) was dissolved in 10 mL of 0.03M THAM acetate buffer pH 8.0 and poured onto the top of the column. The column is eluted as follows.

1. 0.03M THAM acetate, pH 8.0 (fractions 1-79);

2. 0.12M THAM acetate, pH 8.0 (fractions 80-395);

3. 0.25M THAM steel, pH 8.0 (fractions 396-750);

fractions were collected and analyzed on the one hand by UV and on the other hand before and after treatment with alkaline phosphatase. ADA-94B) 3- (uridine-5'-phosphate) -U-57 930 and fractions 75-110 contain 3- (guanosine-5'-phosphate) -U-57 930.

Isolation of substantially pure 3- (uridine-5'-phosphate) -U-57 930; removing the THAM acetate buffer by Amberlite XAD-2 chromatography; A column is prepared from 50 ml of Amberlite XAD-2 resin. The ADA-94B collection containing 3- (uridine-5'-phosphate) -U-57 930 was passed through the column at 2 µm / min. The dripping liquid was discarded. The column was washed with 200 mL of water and this wash was discarded. The column was then eluted with 70-30 methanol / water. Fractions containing 3- (uridine-5-phosphate) -U-57930 (as determined by UV) were combined (200 mL), concentrated to an aqueous solution and lyophilized. This gives 60 mg of ADA-95.1.

Characterization of 3- (uridine-5'-phosphate) -U-5,7930

1. IR spectral data

The IR spectral absorption band (Nujol and Kbr) is as follows:

a) Nujol

Acid- frequency Strong- ness Type Acid- frequency Strong- ness Type 3417.3 24 SH 1249.0 33 SH 3341.1 19 BRD 1214.3 21 AVG 3211.8 19 BRD 1146.8 42 SH 3108.6 25 BRD 1080.9 15 BRD 2951.4 2 BRDM 1070.6 12 AVG 2926.3 1 BRDM 1056.1 14 SH 2854.9 2 BRDM 992.4 39 AVG 2729.6 48 BRDM 972.2 29 AVG 2693.9 51 SH 955.8 49 SH 2535.7 65 SH 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 SH 1489.2 23 AVG 788.9 40 AVG 1462.2 9 AVG M 721.4 44 AVG M 1404.3 41 BRD 705.0 47 BRD 1377.3 18 AVGM 654.9 45 SH 1368.6 31 SHM 632.7 38 Avg 1286.6 34 Avg

Explanation to Table:

Band frequency: at wavelength (cm ^-1 )

Strength: as a percentage of transmittance (% T).

Peak environment data: BRD has a broad meaning;

AVG means average; SHP stands for sharp; SH stands for shoulder. M stands for mineral oil disturbance.

The 25 strongest peaks

T% Frequency T% Frequency 1 2926.2 24 3417.2 2 2951.3 25 3108.5 2 2854.8 26 1610.6 8 1649.2 31 1528.6 9 1462.1 31 1368.5 12 1070.5 33 1249.0 14 1056.0 34 1286.5 15 1089.8 35 1575.0 18 1377.2 36 889.1 19 3341.0 38 632.6 19 3211.7 39 992.3 21 1214.2 39 972.1 23 1489.1

Preparation: rubbing with mineral oil. Max% T: 87 @ 1848.0% T at 3800 cm ^-1 : 83 Density (cm ^-1 / pt): 0.964

b) KBr

Acid- frequency Strong- ness Type Acid- frequency Strong- ness Type 3408.6 10 BRD 1088.9 12 BRD 3102.8 26 SH 1071.5 9 Avg 2963.9 28 Avg 1057.1 11 SH 2930.2 27 BRD 992.4 35 Avg 2878.1 37 Avg 972.2 36 Avg 2862.7 39 SH 956.8 45 SH 2768.1 51 SH 928.8 48 Avg 2511.6 . 65 BRD 889.2 32 Avg 1649.3 4 Avg 859.3 47 Avg 1614.6 20 SH 851.6 47 SH 1576.0 30 Avg 804.4 39 SH 1528.7 28 Avg 788.9 34 Avg 1491.1 21 Avg 743.6 47 SH 1462.2 33 Avg 705.0 40 Avg 1450.6 35 SH 654.9 39 SH 1404.3 37 Avg 634.6 35 Avg 1384.0 33 Avg 595.1 33 Avg 1360.9 42 SH 572.9 33 Avg 1286.6 32 Avg 525.6 32 Avg 1251.9 30 SH 447.5 36 Avg 1215.2 18 Avg

Explanation to Table:

Band frequency: waveguide (cm ^-1 ) Intensity: Percentage of transmittance (% T)

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Peak environment data: BRD has a broad meaning; AVG means average; SHP stands for sharp; SH stands for shoulder.

The 25 strongest peaks

T% F requisition T% Frequency 4 1649.2 30 1251.8 9 1071.5 32 1286.5 10 3408.5 32 889.1 11 1057.0 32 525.5 12 1088.8 33 1462.1 18 1215.1 33 1384.0 20 1614.5 ' 33 595.0 21 1491.0 33 572.8 26 3102.7 34 788.8 27 2930.1 35 1450.5 28 2963.8 35 992.3 28 1528.6 35 634.5 30 1576.0

Preparation: KBr Pastilles Max% Tr: 100 @ 403.1% T: 4000 cm ^-1 : 78 Density (cm ^-1 / pt): 0.964

2. UV absorption spectrum / X _max (a) /

In water, the breasts have the following values: pH 2.0. 279 nm (6.5) pH 7.0, 270 nm (9.9) pH 11.0, 271 nm (9.6)

3. Elemental composition

Molecular formula; C ₂₆ H43ClN ₅ 0nPS. Molecular Weight: 715. Calculated; C 43.64, H 6.01, N 9.79, 0 26.88, S 4.47,

Cl 4.89, P 4.33%.

4. Optical rotation;

(α) θ-107 ° (c = 0.854, water).

5. Solubility:

Very soluble in water, methanol and ethanol. Slightly soluble in acetone and other ketones, ethyl acetate and other esters, chloroform, dichloromethane. Insoluble in saturated hydrocarbon type solvents.

6. Anti-bacterial effect:

3- (Cytidine-5'-phosphate) - U-57 930 is ineffective in vitro. It is converted to U-57 930 by alkaline phosphatase or phosphodiesterase I, which is highly potent in vitro and in vivo against many Gram positive organisms.

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

Characterization of 3- (adenosine-5'-phosphate) -U-57 930

1. IR Spectrum Data

IR spectral absorption bands (Nujol and KBr) are as follows:

Acid- frequency Strong- ness Type Acid- frequency Strong- ness Type 3335.3 16 BRD 1245.1 24 SH 3267.8 17 BRD 1213.3 17 Avg 3210.8 17 BRD 1175.7 43 SH 2954.3 3 BRDM 1146.8 40 SH 2924.4 2 BRDM 1089.9 13 Avg 2868.4 6 SH M 1069.6 10 Avg 2854.9 4 AVG M 1055.1 13 SH 2727.6 46 BRD M 991.5 35 Avg 2520.2 61 BRD 972.2 36 Avg 1684.0 22 SH 957.7 45 SH 1641.6 14 Avg 930.7 48 Avg 1600.1 28 Avg 889.2 32 Avg 1576.0 31 Avg 861.3 47 Avg 1550.9 43 SH 848.7 49 Avg 1509.4 53 SH 818.8 45 Avg 1463.1 15 AVGM 798.6 40 Avg 1420.7 35 Avg 722.4 36 AVGM 1377.3 24 AVGM 708.9 40 SH 1367.6 35 SH M 647.1 33 SH 1332.0 36 Avg 635.6 31 Avg 1299.2 34 Avg

Explanation to Table:

Band frequency: in waves (cm ^-1 )

Strength: as a percentage of transmittance (% T)

Peak environment data: BRD has a broad meaning; AVG means average; SHP stands for sharp; SH is shoulder; M stands for mineral oil disturbance.

The 25 strongest peaks

T% Frequency T% Frequency 2 2924.3 22 1684.0 3 2954.2 24 1377.2 4 2854.8 24 1245.0 6 2868.3 28 1600.0 10 1069.5 31 1576.0 13 1089.8 31 635.5 13 1055.0 32 889.1 14 1641.5 33 647.0 15 1463.0 34 1299.1 16 3335.2 35 1420.6 17 3267.7 35 1367.5 17 3210.7 35 991,5-

1213.2

Preparation: rubbing with mineral oil. Max% T: 85 @ 1864.4% T: 3800 cm ^-1 : 81 Density (cm ^{- 1} / pl): 0.964

b) KBr Acid- frequency Strong- ness Type Acid- frequency Strong. ness Type 3375.8 7 BRD 1090.8 8 Avg 3223.4 10 BRD 1069.6 5 Avg 3124.0 17 SH 1050.3 8 SH

-815

191 085

Acid- frequency Strong- ness Type Acid- frequency Strong- ness Type 2963.0 22 Avg 990.5 27 Avg 2929.2 22 BRD 972.2 29 Avg 2878.1 31 Avg 956.8 38 SH 2863.6 33 SH 929.8 42 Avg 2756.6 45 BRD 889.2 24 Avg 2521.2 60. BRD 861.3 38 Avg 2188.5 75 BRD 851.6 40 SH 1678.2 15 SH 818.8 36 Avg 1643.5 7 AVG ' 807.3 36 BRD 1602.0 20 Avg 798.6 30 Avg 1576.0 23 Avg 768.7 41 BRD 1553.8 35 SH 721.4 30 Avg 1511.4 49 SH 706.9 31 BRD 1475.7 27 Avg 648.1 26 Avg 1421.7 29 Avg 636.5 26 Avg 1384.0 32 Avg 584.5 28 SH 1332.0 31 Avg 571.9 26 Avg 1301.1 28 Avg 533.3 27 sll 1246.1 19 SII 522.7 25 Avg 1215.2 12 Avg 503.4 25 Avg 1176.7 36 SH

Explanation to Table:

Band frequency: in waves (cm ^-1 )

Strength: as a percentage of transmittance (% T)

Peak environment data: BRD has a broad meaning; AVG means average; SHP stands for sharp; 'SH stands for shoulder.

3. Elementary analysis Calcd: C _I7 I1 ₄₃ C1N ₇ 0 | OPS. Molecular Weight: 723. Calculated: C, 44.81; H, 5.94; N, 13.55; O, 22.13; S, 4.42.

Cl 4.84, P 4.28%.

Found: N, 12.87; S, 5.39; Cl, 4.76; P, 3.83.

4. Optical rotation:

(α) θ + 94 ° (c = 0.887, water).

5. Solubility

Very soluble in water, methanol and ethanol. Slightly soluble on steel and other ketones, ethyl acetate and other esters, chloroform and dichloromethane. Insoluble in saturated hydrocarbon solvents.

6. Anti-bacterial writing

3- (adenosine-5'-phosphate) - U-57 930 is ineffective in vitro. When treated with alkaline phosphatase or phosphodiesterase I, U-57,930 is produced, which is highly effective against many Gram-positive organisms both in vitro and in vivo. 3- (adenosine-5'-phosphate) - U-57 930 was found to be active in vivo (subcutaneously in mice) with a CK _S0 of 0.62 (0.48-0.79) mg / kg in S. Pyogenes against the pathogen.

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

Characterization of 3- (uridine-5'-phosphate) - U-57 930

1. IR Spectrum Data

IR spectral absorption bands (Nujol and Kbr) are as follows:

The 25 strongest peaks

T% Frequency T% Frequency 5 1069.5 23 1576.0 77 3375.7 24 889.1 '7 1643.5 25 522.6 8 1090.7 25 503.3 8 1050.2 26 648.0 10 3223.3 26 636.5 12 1215.1 26 571.8 15 1678.1 27 1475.6 17 3124.0 27 990.5 19 1246.0 27 533.2 20 1602.0 28 1301.0 22 2963.0 28 584.5 22 2929.1

Preparation: KBr Pastilles Max% T: 95 @ 405.0% T: 4000 cm ^-1 : 77 Density (cm ^-1 / pt): 0.964

2. UV Absorption _Spectrum / X _mnx (a) / In water, at pH 2.0, 258 nm (16.0) pH 7.0, 261 nm (16.5) pH 11.0, 261 nm (16.0)

a) Nujol

Acid- rekvencia Strong- ness Type Acid- frequency Strong- ness Type 3330.4 19 BRD 1332.9 44 BRD 3224.4 21 BRD 1296.3 40 SH 2952.4 1 BRD M 1251.9 28 BRD 2924.4 0 BRDM 1215.2 19 Avg 2867.5 4 SHM 1089.9 13 Avg 2854.0 3 AVGM 1071.5 9 Avg 2733.4 49 Siim 1056.1 13 SH 2695.8 53 sII 991.5 39 Avg 2532.8 67 SH 973.2 39 Avg 1757.3 73 SH 957.7 48 SH 1685.9 8 Avg 931.7 49 Avg 1647.4 21 SH 890.2 34 Avg 1602.0 * 41 Avg 858.4 50 Avg 1574.1 42 Avg 813.0 43 Avg 1555.7 43 BRD 798.6 47 Avg 1462.2 12 AVGM 767.7 50 Avg 1425.5 37 SH 721.4 42 AVGM 1378.3 22 AVGM 634.6 35 Avg 1367.6 37 SHM

Explanation to Table:

Band frequency: in waves (cm ^-1 )

Strength: as a percentage of transmittance (% T)

Peak environment data: BRD present9

-917

191 085 wide; AVG means average; SHP stands for sharp; SH is shoulder; M stands for mineral oil disturbance.

The 25 strongest peaks

T% Frequency T% Frequency 0 2924.3 22 1378.2 1 2952.3 28 1251.8 3 2854.0 34 890.1 4 2867.5 35 634.5 8 1685.8 37 1425.5 9 1071.5 37 1367.5 12 1462.1 39 991.5 13 1089.8 39 973.1 13 1056.0 40 1296.2 19 3330.3 41 1602.0 19 1215.1 42 1574.0 21 3224.3 42 721.3 21 J, 647.3

Preparation: rubbing with mineral oil. Max% T: 86 @ 3764.5% T: 3800cm ^-1 : 85 Density (cm ^- * / pt): 0.964

b) KBr

Acid- frequency Strong- ness Type Acid- frequency Strong- ness Type 3387.4 12 BRD 1055.1 10 Sll 3114.4 25 SH 992.4 35 Avg 2962.0 26 Avg 973.2 36 Avg 2931.1 26 BRD 956.8 45 SH 2879.1 36 Avg 929.8 48 Avg 2863.6 38 Sll 889.2 31 Avg 2833.7 43 SH 852.3 46 Avg 2509.6 64 BRD 813.0 40 Avg 1685.0 6 BRD 811.1 40 SH 1647.4 17 SH 798.6 43 Avg 1605.9 35 SH 782.2 48 BRD 1576.0 37 Avg 768.7 48 Avg 1556.7 39 BRD 707.9 43 Avg 1463.1 31 Avg 669.3 43 SH 1423.6 35 Avg 649.1 40 SH 1384.0 32 Avg 634.6 37 Avg 1331.0 43 Avg 585.4 38 SH 1297.2 38 SH 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

Explanation to Table:

Band frequency: in wave number (cm ^-1 )

Strength: as a percentage of transmittance (% T)

Λ peak area data: BRI) meaning broad; AVG means average; SHP stands for sharp; SH stands for shoulder.

The 25 strongest peaks

T% Frequency T% Frequency 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 1464.0 38 1297.1 31 889.1

Preparation: KBr Pastilles Max% T: 101 @ 405.0% T: 4000 cm ^-1 : 76 Density (cm ^-, / pt): 0.964

2. UV absorption spectrum | „, _ax (a) /

In water, with the following values. pH 2.0, 261 nm (11.5) pH 7.0, 262 nm (10.7) pH 11.0, 262 nm (11.5)

3. Elemental formula: C ₂₆ H _{4 2} N ₄ C 10 ₁₃ PS. Molecular Weight: 716. Calculated: C, 43.57. II 5.86. N, 7.82, 0, 29.05, S, 4.46,

Cl 4.89, P 4.33%.

4. Optical rotation (α) + 105 ° (c = 0.94, water).

5. Solubility

Very soluble in water, methanol and ethanol. Slightly soluble in acetone, other ketones, ethyl acetate and other esters, chloroform and dichloromethane. Insoluble in saturated hydrocarbon solvents.

6. Anti-bacterial effect

3- (uridine-5'-phosphate) - U-57 930 is ineffective in vitro. It is converted to U-57 930 by alkaline phosphatase or phosphodiesterase I, which is very potent against many Gram-positive organisms both in vitro and in vivo.

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

Characterization of 3- (guanosine 5'-phosphate) -U-57 930

1. IR Spectrum Data

IR spectral absorption bands (Nujol and KBr) are as follows:

-1019 '91 085

a) Nujol

Acid- frequency Strong- ness Type Acid- frequency Strong- ness Type 3335.3 16 BRD 1250.0 36 SH 3227.2 19 BRD 1213.3 21 Avg 2953.3 2 AVGM 1173.8 39 Avg 2925.3 1 BRDM 1149.7 41 Avg 2868.4 · 6 SHM 1087.9 14 SH 2855.9 4 AVGM 1071.5 9 Avg 2737.3 51 BRD.M 991.5 42 Avg 2521.2 73 BRD 972.2 42 Avg 1684.0 6 Avg 956.8 52 SH 1635.8 11 Avg 929.8 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 AVGM 783.1 42 SHP 1414.9 44 Avg 720.4 43 AVGM 1377.3 24 AVGM 707.9 45 BRD 1365.7 31 Avg 681.9 40 Avg 1312.7 44 Avg 635.6 34 Avg

Table Explanation: Band Frequency: Hulam (cin ^-1 )

Strength: as a percentage of transmittance (% T) Peak environment data: BRD has a broad meaning; AVG means average; SHP stands for sharp; SH is shoulder; M stands for mineral oil disturbance. The 25 strongest peaks T% Frequency T% Frequency 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 40 681.8 18 1462.1 41 1149.6 19 3227.1 42 991.5 . 21 1598.1 42 972.1

1213.2

Preparation: rubbing with mineral oil.

T max% : 97 @ 3,762.6 . % T: 3800 cm ^-1 : 97 Density / (cm ^-1 / pt): 0.964 b) KBr Acid- Strong- Acid- Strong- frequency ^Tl P ^us frequency ness Type 3380.6 9 BRD 1174.7 31 Avg 3234.0 13 BRD 1147.7 33 BRD 2963.0 22 AVG 1088.9 9 BRD

b) KBr

Acid- frequency Strong- ness Type Acid- frequency Strong- ness Type 2929.2 21 BRD 1070.6 6 Avg 2878.1 31 Avg 991.5 33 Avg 2862.7 33 SH 972.2 34 Avg 2744.0 44 BRD 956.8 43 SH 2522.2 61 BRD 929.8 44 Avg 1683.0 4 BRD 889.2 28 Avg 1634.8 6 Avg 8 (.0,3 4J Avg 1598.2 14 Avg 800.5 36 Avg 1571.2 19 Avg 783.1 32 Avg 1534.5 26 Avg 715.6 38 SH 1482.4 38 Avg 705.0 38 SH 1461.2 36 Avg 679.9 33 Avg 1448.7 36 BRD 635.6 31 Avg 1413.9 34 Avg 584.5 34 SH 1384.0 29 Avg 571.9 32 Avg 1359.9 30 Avg 523.7 31 Avg 1312.7 37 Avg 502.5 30 Avg 1250.9 29 SH 447.5 34 Avg 1213.3 16 Avg

Explanation to Table:

Band frequency: in waves (cm ^-1 )

Strength: Percentage of Tiansmittance (% T)

Peak environment data: BRD broad, AVG average; SHP stands for sharp; SH stands for shoulder.

Λ 25 strongest peaks

T% Frequency T% Frequency 4 1683.0 29 1384.0 6 1634.7 29 1250.8 6 1070.5 30 1359.8 9 3380.5 30 502.5 9 1088.8 31 2878.0 13 3234.0 31 1174.6 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 28 1534.5 889.1 33 1147.6

Preparation: KBr Pastilles Max% T: 97 @ 405,0% T 4000 cm ^- Hymen: 77 Density (cm ^-1 / pt): 0.964

2. UV absorption spectrum / A, _nax (a) /

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

Ί F.lpmi n ^? 7Pfptpl 'Formula: C ^ IUjClN.jOnPS. Molecular Weight: 739.

-1119 J C85. 21

Calculated: C 43.84, H 5.81, N 13.26, 0 23.27, S 4.33, Cl 4.73, P 4.19%.

Found: N 13.32, S 4.86, Cl 4.49, P 3.25%

4. Optical rotation:

(-) + 97 ° (c = 0.855, water).

5. Solubility

Very soluble in water, methanol and ethanol. Slightly soluble in aceion, other ketones, ethyl acetate, and other esters, chloroform and dichloromethane. Insoluble in saturated hydrocarbon type solvents.

6. Anti-bacterial effect

3- (guanosine-5'-phosphate) - U-57 930 is ineffective in vitro. It is converted to U-57 930 by alkaline phosphatase or phosphodiesterase I, which is effective against many Gram-positive organisms both in vitro and in vivo.

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

Because the compounds of the present invention are active against various Gram-positive and Gram-negative microbes, they can be used in various environments to inhibit these microbes; Thus, for example, disinfectants can be used in a qualitative manner to inhibit S. aureus in washed cookware. They can also be used as disinfectants on dental and medical devices infected with various S. aureus microorganisms. The compounds of the invention may also be used for bacteriostatic rinse of underwear, impregnation of papers and various webs; in addition, they can be used to negatively inhibit the growth of sensitive microorganisms in plate assays and other microbiological media.

The compounds of the invention may also be used to treat diseases caused by (non) members of the Mycoplasma genus. The most well-known of these pathogens are PPLO (pneumonia-like and similar) microorganisms such as M. honünnis, M. salivarium, M. mycoides, M. Hypopneutnodia, M. hyorhinis, M. gallisepticum, M. arthritis and other bacterial species that are infectious to humans and animals such as domestic animals such as sheep, dogs, cattle, pigs and poultry (such as chickens, turkeys, ducks and geese) and laboratory animals (such as rats and mice) .

U-57 930-3- (5'-ribonucleotides) are useful in the treatment of kidney infections and other infections when L-forms of Gram-negative or Gram-positive bacteria are present, for example, L-forms of P. mirabilis.

Because the compounds of the invention are amphoteric, they can form salts with both bases and acids. These salts may be prepared by conventional procedures. Organic acids which may be used include hydrochloric, sulfuric, phosphoric and the like. Examples of inorganic bases include all basic inorganic hydroxides and salts with which the sodium, potassium, calcium and lithium salts and the like can be formed. Salts of the compounds of the invention may be used for the same purposes as the parent compounds.

The compounds of the present invention are antimicrobial agents suitable for the preparation of suitable compositions. These compositions may advantageously be formulated for administration to humans and animals in unit dosage forms such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, orally (oral) solutions or suspensions, and oil / water emulsions. containing an appropriate amount of the free base form or pharmaceutically acceptable salts of the active ingredient.

For oral administration, the unit dosage form can be prepared from lyophilisate. even in liquid form. For the preparation of solid compositions such as tablets, the main active ingredient is admixed with the usual tablet ingredients such as talc, magnesium stearate, dicalcium phosphate, inagnesium aluminum silicate, calcium sulfate, starch, lactose, acacia, diluents and carriers used by pharmacists. The tablets may be layered or otherwise formulated so as to provide a dosage form which preferably provides a sustained or delayed action or the effect of the active ingredient incorporated in the tablet at a predetermined time. Thus, for example, the tablet may comprise an inner and an outer part, the outer of which may roughly cover the inner. The two components may be separated by an enteric coating, which is intended to prevent the tablet from disintegrating in the stomach, thereby allowing the internal component to enter the duodcnum intact or to be released in a delayed manner. A number of substances may be used for such enteric coatings or camelids; by way of example, several polymeric acids or mixtures of polymeric acids with substances such as shellac, cetyl alcohol, cellulose acetate phthalate, styrene-maleic acid copolymer, and the like. Alternatively, this dual component system may be used to make tablets containing two or more incompatible active ingredients. Pills can be made in the same way as tablets; the only difference is that they have a different shape and contain sucrose or other sweetener and flavoring. In their simplest embodiments, capsules, like tablets, may be prepared by mixing the active ingredient with an inert, pharmaceutically acceptable diluent, and filling the resulting mixture into a hard gelatin capsule of appropriate size. In another embodiment, the capsules are prepared by filling the hard gelatin capsules with polymeric acid coated beads containing the active ingredient of the present invention. Soft gelatine capsules are prepared by mechanically encapsulating a pasty mixture of the active ingredient with a pharmaceutically acceptable vegetable oil or other inert oil.

Liquid dosage forms such as syrups, elutes, and suspensions may be prepared for oral (oral) administration. An active ingredient of the present invention may be dissolved in an aqueous carrier with sugar, aromatic flavors and preservatives to form a syrup. Elixir is prepared by using aqueous ethanol as a carrier, with suitable flavoring agents such as cane sugar and aromatic flavoring.

Including 191,085 times. Suspensions may be prepared from the insoluble derivatives of the active compounds of the present invention in a syrup vehicle and suspending agent. Suitable suspending agents are, for example, acacia gum, tragacanth, methylcellulose and the like. Lubricating tubes for topical (topical) use may be prepared by incorporating the active ingredient in a suitable ointment base such as petroleum jelly, lanolin, polyethylene glycols, mixtures thereof, or other. dispersed in similar materials. Preferably, the active ingredient is finely ground in a colloidal mill using liquid petrolatum as a grinding agent and then dispersed in the ointment base. Topical cream and lotions are formulated by first dispersing the active ingredient in an oily phase and then emulsifying the oily phase in vk.

For parenteral administration, liquid dosage unit forms are prepared using an active ingredient of the invention and a sterile vehicle. A preferred vehicle for this purpose is water. The active ingredient, depending on the dosage form and concentration, may be suspended or dissolved in the vehicle. For the preparation of solutions, the water-soluble form of an active compound of the invention may be dissolved in water for injection and sterilized by filtration and filled into a suitable vial or ampoule and then sealed. It is preferred to dissolve the adjuvants in the vehicle, for this purpose a topical anesthetic, preservative and buffering agents may be used. To increase stability, the composition may be lyophilized by freezing it after filling into an ampoule and removing the water under vacuum in a frozen state. The dry, lyophilized powder is then sealed in an ampoule and an ampoule of water for injection is provided to reconstitute the lyophilized powder before use. For parenteral purposes, suspensions may be formulated in substantially the same manner except that the active ingredient is suspended in the vehicle rather than dissolved. In this case, sterilization cannot be performed by filtration. The active ingredient may be sterilized by exposure to ethylene oxide prior to suspension in the sterile vehicle. For prolonged action, an intramuscular suspension of an insoluble derivative of the active ingredient, such as trimethylsilyl ether or its pamoic acid salt, is prepared. It is preferable to include a surfactant or wetting agent in the composition to facilitate uniform distribution of the active ingredient.

The term unit dosage form as used herein and in the claims means physically discrete units that can be administered in units to humans or animals. Each unit contains a predetermined amount of the active ingredient which is determined to produce the desired therapeutic effect. This amount of active ingredient is admixed with the required pharmaceutically acceptable diluent and / or excipient. The new dosage unit forms have been established based on (a) the individual characteristics of the active ingredient and the therapeutic effect sought, and (b) the limitations that must always be considered when administering an active ingredient for therapeutic use in humans or animals. for the purpose of compositions. This will be explained in detail herein and are also typical features of the present application. Examples of suitable dosage unit forms according to the invention include tablets, capsules, pills, suppositories, lozenges, granules, teaspoonfuls and tablespoonfuls, ampoules, vials, and any of the foregoing multiple forms.

The active ingredient is mixed in a unit dosage form with a suitable pharmaceutical carrier for convenient and effective administration. A preferred embodiment is that the unit dosage forms for systemic administration comprise 10, 25, 50, 100, 250 or 500 mg of active ingredient; for parenteral administration, the preferred concentration is 5 to 65 ppm / volume. Dosages of compositions containing one or more other active ingredients in addition to an active ingredient of the invention will be determined with reference to the appropriate dosage of each of the active ingredients present.

The following examples illustrate the process of the invention in detail, but are not limiting.

In the examples, the 3- (5'-ribolucleotide) derivative of U-57 930E is used as the active ingredient, but it is to be understood that these are only examples and that the other active compounds of the present invention can be used.

Example 1 Preparation of Compositions: Capsules

From the indicated amounts of the following substances, 1000 two-part capsules of cumin gelatine are prepared for oral use. Each capsule contains 250 mg of U 57 930E 3- (5'-ribonucleotide).

U-57 930E-3- (5'-ribonucleotide) 250g wheat starch 100 g talc 75g Magnesium stearate 25g These materials thoroughly mixed

encapsulate as usual.

These capsules are suitable for systemic treatment of infections in adult individuals by administering 1 capsule every 6 hours.

Using the above procedure, capsules containing the S-fS'-ribonucleotide of U-57 930E can be prepared in the same manner by replacing 250 g of the active ingredient with 10, 25, 50, 100 and 503 g of U-57 in the above formula. 930E is substituted for 3- (5'-ribonucleotide).

Example 2: Compositions: Capsules

From the indicated amounts of the following substances, 1000 pieces of two-piece capsule gelatin capsules are prepared for oral administration. Each capsule contains 200 mg of U-57 930E 3- (5'-ribonucleotide) and 250 mg of tetracycline hydrochloride.

U-57 930E- 3- (5'-ribonucleotides) 200 g Tetracycline hydrochloride 250g talc 75g Magnesium stearate 25g

-1 325

191 085

The components are thoroughly mixed and then encapsulated as usual.

These capsules are suitable for systemic treatment of infections in adult individuals by administering 1 capsule every 6 hours.

- Using the above procedure, capsules containing, in addition to U-57 930E-3- (5'-ribonucleotide), any of the following antibiotics may be prepared instead of tetracycline hydrochloride. In this case, 250 g of the following antibiotics are replaced by the above formula. Such antibiotics include chloramphenicol, oxytetracycline, chlorotetracycline, fumagillin, errotrotnicin, streptomycin, dilidro-novobiocin and novobiocin. When using a penicillin such as potassium penicillin G instead of tctracycline, ¹⁵ 25 0 000 units are used per capsule.

These combinations are suitable for systemic treatment of mixed infections in adult individuals by administering 1 capsule every 6 hours. 20

Example 3 for the preparation of compositions: tablets

From the indicated amounts of the following substances, 1000 tablets are prepared for oral administration. Each 25 tablet contains 500 mg of U-57 930E-3- (5'-ribonucleotide).

U-57 930E-3- (5'-ribonucleotide) 500 g Lactose 125 g wheat starch 65g Magnesium stearate 25g Liquid light petroleum jelly 3g The materials are thoroughly mixed and fine

grind and pass through sieve # 16. The resulting granules were compressed into tablets each containing 500 mg of U-57 930E-3- (5'-ribonucleotide).

These tablets are suitable for systemic treatment of infections of adults, including infections of malaria, by oral administration of 1 tablet three times daily.

According to the above procedure, with the exception of reducing 500 g of active ingredient to 250 g, tablets containing 250 mg of U-57 930E3 (5'-ribonucleotide) can be prepared.

Example 4 Preparation of Compositions: Tablets

From the indicated amounts of the following substances, 1000 tablets are prepared for oral administration. Each tablet contains 250 mg of U-57 930E 3- (5'-ribonucleotide) plus a total of 250 mg (83.3 mg each) of sulfadiazine, sulfamerazine and sulfamethazine 55.

U-57 930E-3- (5'-ribonucleotide) 250g sulfadiazine 83.3 g sulfamerazine 83.3 g sulfamethazine 83.3 g Lactose 50g BÚ2akeményítő 50g Calcium stearate 25g Light fluid petroleum jelly 5g

The components are thoroughly mixed and passed through a 1.66 mesh screen. The granules thus obtained are compressed into tablets each containing 250 mg of U-57 930E-S-S'-ribonucleotide and 6 total 250 mg (83.3 mg each) of sulfadiazine, sulfamerazine, and sulfamethazine.

These tablets can be used for the systemic treatment of infections by oral administration of 4 tablets, followed by 1 tablet every 6 hours.

For the treatment of urinary tract infections, the above-mentioned three sulfolamides are preferably replaced by 250 g of sulfamethylthiadiazole or 250 g of sulfacetamide.

Example 5: Compositions: Syrup for oral administration

For oral use, 100 ml of an aqueous suspension are prepared containing 250 mg of U-57 930E-3- (5'-ribomyclotide) in each 5 ml, or a total of 500 mg of sulfonainide, from the following amounts:

U-57 930E-3- (5'-ribonucleotide) 50 g

Sulfadiazine 33.3 g

Sulfamerazine 33.3 g

Sulfamethazine 33.3 g

Citric acid 2 g

Benzoic acid 1 g

Sucrose 700 g

Tragacanth 5 g

Lemon oil 2 ml

Add deionised water as needed to 1000 ml.

Citric acid, benzoic acid, sucrose, tragacanth and lemon oil are dispersed in water to give 850 ml of the mixture. U-57 930E-3- (5'-ribonucleotide) and finely divided sulfonamides were mixed with the syrup until uniform in distribution. The suspension was then made up to 1000 ml with water.

The composition thus prepared can be used to systemically treat adult pneumonia by administering it four times a day with 1 tablespoon (10 ml).

Example 6 Preparation of compositions: Solution for parenteral administration

A sterile aqueous solution for intramuscular administration containing 200 mg of U-57 930E3- (5'-ribonucleotide) in 1 ml was prepared. The solution is prepared from the following amounts of the following substances:

U-57 930E- 3- (5'-ribonucleotides) 200 g

Lidocain hydrochloride 4 g

Methyl paraben 2.5 g

Propyl Paraben 0.17 g

Water for injections (1000 ml h).

The components are dissolved in water and the solution is sterilized by filtration. The sterile solution is filled into ampoules and the ampoules are sealed.

-1 427

191 085

Example 7 Preparation of Compositions: A composition suitable for parenteral administration

From the indicated amounts of the following substances for intramuscular use, a sterile aqueous composition is prepared containing 1 ml of 200 mg of U-57 930E-3- (5'ribonucleotide) and 400 mg of spectinomycin sulfate.

U-57 930E-3- (5'-ribonucleotide) 200 g

Spectinomycin sulfate 400 g

Lactose 50 g

For injection, v (z) is required for 10000mi.

U-57 930E-S-S'-ribonucleotide, spectinomycin sulfate and lactose are dispersed in water and sterilized. The sterile composition is aseptically filled into sterile ampoules such that each ampoule contains 2 ml.

Example 8 Preparation of compositions: ointment for topical (topical) treatment

1000 g of an ointment containing 0.25% of the active ingredient are prepared from the indicated amounts of the following substances:

U-57 930E-3- (5'-ribonucleotide) 2.5 g

Zinc oxide 50 g

Kalatnin 50 g

Heavy Fluid Vaseline 250 g

Wool fat 200 g

White petrolatum needed for 1000 g.

The white petrolatum and wool grease are melted and 100 g of liquid petrolatum are added. U-57 930E-3- (5'-ribonucleotide), zinc oxide and calamine are added followed by the remaining liquid petrolatum and the mixture is kneaded until the powder is finely divided and dispersed evenly. The powder mixture is mixed with the white petrolatum mixture and the mixing is continued until the ointment solidifies.

This ointment may be applied topically to the skin of mammals for the treatment of infections.

The above composition can also be prepared by omitting zinc oxide and calamine.

Be prepared by the above process was obtained as ointments containing the U-930E- 57 3- (5'-ribonucleotide) was in an amount of 0.5, 1, 2, and 5% ^- 2.5 in this case in the above fornrulázásban g of the active ingredient are added with 5, 10, 20 and 50 g of U-57 930E-3- (5'r.ibonucleotide) respectively.

Example 9 for the preparation of a composition: cream

From the indicated amounts of the following substances, 1000 g of inert cream are prepared.

U-57 930E-3- (5'-ribonucleotide) 50g Tegacid Regular ¹ 150 g spermaceti 100 g propylene 50g Polysorbate 80 5g Methyl paraben 1g lonmen demineralized water as needed 1000 g.

Note: ^J Tegacid Regular is a self-emulsifying glyceryl monostearate (Goldschmidt Chemical Corpoartion, New York, Y.Y).

Tegacid and Spermacet were fused at 70-80 ° C. Methyl paraben is dissolved in 500 µl of water and propylene glycol, polysorbate 80, and U-57 930E-3- (5 ribonucleotides) are added, maintaining the temperature at 75-80 ° C. The methyl paraben-containing mixture is added slowly to the melt containing Tegacid and Spermacet, with continuous stirring, for at least 30 minutes, while allowing the temperature to fall to 40-45 ° C. The cream is adjusted to pH 3.5 with a solution of 2.5 g of citric acid and 0.2 g of twice basic sodium phosphate in 50 g of water. Finally, sufficient water is added to the mixture to achieve a weight of 1000 g, and the co-position is stirred until the mixture cools and solidifies to maintain homogeneity.

This composition is useful for the treatment of vaginal infections in women.

Example 10 Preparation of compositions: Eye ointment

1000 g of the indicated amounts of the following substances

A zinc quill is prepared containing 0.5% (5'-libonucleotide). U-57 930E- U-57 93OE-3- (5'-ribonucleotide) 5g bacitracin 12.2 g Polymixin B sulfate (10,000 units / shirt) 1g Light fluid petroleum jelly 250g wool grease 200 g White Vaseline as needed 1000 g. Solid active ingredients with the help of air micronizer

finely disperse and add to light, flowing petroleum jelly. The mixture is passed through a colloidal mill to uniformly distribute the micronized particles. The wool fat and white petrolatum were melted and the temperature was adjusted to 45-50 ° C. Flowing petroleum jelly is added and the cream is stirred until it solidifies. The ointment is conveniently packaged in eye ointment tubes containing 1 dram (i.e., 3,888 g).

The ointment is suitable for treating eye infections in humans and animals.

A preferred form of the above composition may comprise 5 g (0.5%) of methyl prednisolone for the treatment of inflammation; in another embodiment, bacitracin and polymyxin B sulfate may be omitted.

Example 11 Preparation of compositions: eye and ear drops

From the indicated amounts of the following substances, 1000 ml of sterile aqueous solution is prepared for ophthalmic and otic use. 1 nd of the solution contains 10 mg of U-57 93OE-3- (5'-ribonucleotide) and 5 mg of inctilis-proline diol.

U-57 930E-3- (5'-ribonucleotide) 10g Sodium-methyl-prednisolone phosphate 5g Sodium citrate 4.5 g Sodium bisulfite Year 1 Polyethylene glycol 4000 120g T myristyl picolinium chloride 0.2 g Polyvinylpyrrolidone 1g water as needed For 1000 mi.

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085

The components were dissolved in brine and the resulting solvent was sterilized by filtration. The filtrate is placed in a sterile aseptic drip tray.

The composition thus prepared can be used for the local (topical) treatment of inflammation and infections of the eye and ear, and for the topical treatment of other sensitive tissues of the human body.

Example 12 Preparation of the composition: lozenges

000 pastilles are prepared from the specified quantities of the following materials:

U-57 93 0E-3- (5'-ribonucleotide) 100 g

Neomycin sulfate 50 g

Polymyxin B sulphate (10,000 units / mg) 1 g of p-Ainino-benzoic acid ethyl ester 50 g

Calcium starch 150g

Powdered sucrose as needed for 5000 g.

The powdered ingredients are thoroughly mixed and then compressed to a tablet weight of 0.5 g using standard tableting techniques.

During use, the pellet should be kept in the mouth and allowed to dissolve slowly to treat infections of the mouth and throat.

Example 13 Preparation of Compositions: Suppository

From the indicated amounts of the following substances, 1000 suppositories are prepared each weighing 2.5 g; one suppository contains 100 mg of U-57 930E 3- (5'-ribonucleotide).

U-57 930E-3- (5'-ribonucleotide) 100 g

Polymixin B sulfate (10,000 units / mg) 1.25 g Methyl prednisolone 1 g p-Aminobenzoic acid ethyl ester 75 g

Zinc oxide 62.5 g

Propylene glycol 162.5 g

Polyethylene glycol 4000 as needed for 2500 g.

U-57 930E-3- (5'-ribonucleotide), polymyxin B sulfate, methylprednisolone, ethyl p-aminobenzoic acid, and zinc oxide were added to propylene glycol and ground until the the powders are finely divided and evenly dispersed. The polyethylene glycol 4000 is melted and the propylene glycol dispersion is slowly added with stirring. The suspension is poured into non-refrigerated molds at 40 ° C.

The composition is allowed to cool and solidify, then removed from the mold and wrapped with a foil on each suppository.

Suppositories are administered into the rectum for the local treatment of inflammation and infection.

The above composition can also be prepared without steroid.

Example 14 Preparation of compositions: ointment for the treatment of mastitis (inflammation of the breast)

From the indicated amounts of the following substances, 1000 g of ointment is prepared which is suitable for the treatment of mastitis in a dairy cow.

U-57 930E-3- (5'-ribonucleotide) Methyl Prednisolone Acetate Light Fluid Vaseline Anhydrous chlorobutanol

Polysorbate 80

2% Aluminum Monostearate Peanut Oil Gel

White petrolatum as necessary g

0.5 g 300 g g 5 g

400 g to 1000 g.

U-57 930E-3- (5'-ribonucleotide) and methylprodnisolone acetate were ground with light, flowing petroleum jelly until a fine distribution and uniform dispersion were obtained. Chlorobutanol, polysorbate 80, peanut gel and white petroleum jelly are heated to 50 ° C to melt and mixed with the dispersed petroleum jelly. While stirring, the dispersion is allowed to cool (and solidify) at room temperature and then filled into disposable mastitis syringes containing 10 g.

2C

Example 75 Preparation of Compositions: Animal Feed

1000 g of the compound feed are prepared from the indicated amounts of the following substances.

U-57 930E-3- (5'ribonucleotide) 10g soybean meal 400g fish meal 400g Wheat germ oil 50g Ciro Kine Halasz 140g The parts are mixed together and put into grains

provide. The composition can be administered to laboratory animals, such as rats, mice, guinea pigs, and hamsters, as laboratory food for prophylactic use prior to shipping on board.

For other decoy, such as poultry such as chicken, duck, turkey and goose, the composition may be added to the daily diet of the decoy in an amount that provides the desired dose of U-57 930E-3- (5'-ribonucleotide).

Example 76 Preparation of Compositions: Capsules

From the indicated amounts of the following substances, 1000 two-part hard gelatin capsules were prepared for oral administration, each containing 200 mg of U-57 930E-3- (5'-ribonucleotide) and 200 mg of hydroxychloroquine sulfate.

U-57 93OE-3- (5'-ribonucleotide) 200 g HRIroxi klorocliin-sulfate 200 g talc 75g Magnesium stearate 25g The components are thoroughly mixed and then a

encapsulation in a conventional manner.

These capsules are suitable for oral treatment of adult subjects against the recurrent attack of P vivax by administering 1 capsule per week.

Example 7 Preparation of Compositions: Tablets

From the indicated amounts of the following substances, 1000 to 65 tablets each of 125 mg are prepared

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191 085

It contains U-57 930E-3- (5'-ribonucleotide) and 325 mg of kin.sulfate.

U-57 930E-3- (5'-ribonucleotide) 125 g

Quinine sulfate 325 g

Lactose 50 g

Wheat starch 50 g

Calcium starch 25 g

Light fluid petroleum jelly 5 g

The parts are mixed thoroughly, the pieces thus formed are broken and crushed by passing it through a No. 16 (fine) sieve. The resulting granules are compressed into tablets each containing 125 mg of U-57 930E-3- (5'-ribonucleotide) and 325 mg of quinine sulfate. These tablets are suitable for oral treatment of malaria by administering 2 tablets every 8 hours for 7 days, then 1 tablet 3 times daily for 7 days.

Example 18 Preparation of Compositions: Syrup for oral administration

From the indicated amounts of the following substances, 1000 ml of an aqueous suspension for oral use are prepared containing 25 ng of pyrimethamine, 250 mg of U-57 930E-3- (5'-ribonucleotide) and 500 mg of sulfadiazine per 10 ml.

U-57 930E-3- (5'-ribonucleotide) 25 g

Pyrimethamin 2.5 g

Sulfadiazine 50 g

Citric acid 2 g

Benzoic acid 1 g

Sucrose 700 g

- Tragacanth, 5 g

Lemon oil 2 ml deionized water needed for 1000 ml.

Citric acid, benzoic acid, sucrose, tragacanth and lemon oil are dispersed in water to give a volume of 850 ml. U-57 93 OE-S-S '-S ribonucleotide), pyrimethamine and sulfadiazine are added to the syrup until uniform distribution is achieved. The volume was then made up to 1000 ml. This composition is useful for the prophylactic treatment of adult malaria by administering 1 tablespoon (10 ml) per week.

Example 19 for the preparation of compositions: parenteral composition

Sterile aqueous compositions containing 200 mg of U-57 930E3- (5'-ribolucleotide) per ml are prepared for intramuscular administration from the indicated amounts of the following substances.

U-57 930E-3- (5'-ribonucleotide) 200 g

Lactose 50 g

Water for Injection, needed in 10,000 ml-hc.

U-57 930E-S5S'-ribonucleotide and lactose are dispersed in water and sterilized. The sterile composition is aseptically filled into sterile ampoules such that each ampoule contains 2 ml.

Effects shown in vivo against the oilmodium berghei

agent MED ¹ (mg / kg) CDjo ² (ing / kg) Sub Q ³ 01 Sub Q OI U-57 930E 0.16 1.6 16 (12-22) > 50 U-21 251F <20 - 53 (46-61) - U-24 729 A <1.25 - 4.7 (3.2 to 6.9) - U-82.84 (klorochin) <5,10 12.5 11.5 (8.8 to 15) 14 Chlorochin- (PO ₄ ) 2 <5 - <20 -

Explanation to Table:

* MED: The dose at which the median survival time (ST ₅₀ ) was significantly increased (p = 0.05) compared to the ST ₅₀ value in untreated controls.

² CD ₅₀ : Medium protective dose in mg / kg with 95% confidence limits,

3: Method of administration.

U-57 930E: 3- (adenosine-5 ^, phosphate) U-57 930 in the form of its hydrochloride salt.

U-21 251F: Clindamycin Hydrochloride Hydrate.

U-24 729A: 1'-Dimethyl-4 ^, -depropyl-4 ^, - (R, S) -n-pentylclindamycin hydrochloride.

Examination of the anti-macarean effect (against P. berghei)

Test method

Male CF-1 mice weighing 18-20 g were placed in cages in groups of 10 and intraperitoneally infected with whole blood of mice infected with P. berghei 3 days prior to blood collection. 0.2 ml of heparinized blood diluted 1:10 in physiological saline was used for inoculation. This volume contained approximately 10 ⁶ parasites.

At 4 hours post-infection, all groups of 10 mice were treated either 0.2 ml subcutaneously or 0.5 ml orally (by gavage) with the desired concentration of drug. This treatment was performed once daily for 4 days. The animals were then observed for 28 days. cs recorded the death. Death before day 6 was considered thiomatic.

The different drug concentrations of the various analogs and the individual analogs were evaluated for efficacy on the basis of the average survival time of the animals at each treatment level, and the average protective dose of the individual analogue was also taken as a basis for evaluation. Calculations were performed on an IBM 370 digital computer. Λ The results of the treated groups were compared with those of the untreated groups and those of the chloroquine treated groups.

The agents of the invention are also active against other intracellular parasites such as Plasmolia, Toxoplasma and Leislimania; against protozoa that digest red blood cells such as Entamoeba histolytica, 17

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191 085 and certain Trypanosoma species; against some worms that bleed red blood cells during the course of the disease, such as Schistosomes.

ANNEX 1 ι.

A compound of formula (II) wherein R 1 is. may be mono- or polysubstituted at any position on the pyridine ring which is not yet substituted with R ₂ and represents hydrogen, C 1-8 alkyl or substituted alkyl and its isomers, and cycloalkyl and substituted cycloalkyl, substituted with oxygen or substituted nitrogen alkyl, halogen, phenyl or substituted phenyl group, and - _(CH2) "-, -OII, - (C1I _2)", NR ₄ R _s composition groups and its isomers, where - n is 1 to 8 to integer;

R _{4 and} R ₅ are hydrogen or C 1-8 alkyl and isomers thereof;

R 2 may be singly or multiply substituted at any position on the pyridine ring that does not yet have Rt

Substituted with port II and represents -C-X where -

X α 7 (R) -Hydroxy-methyl-1-mercapto-α-lincosuloid, 7 (S) -hydroxymethyl-1-m -capto-α-lincosaminide, 7 (S) -Halo-methyl-1-mercapto-lincosaminide, 7 (R) -Halo-methyl-1-mercapto-a-lincosaminide, 7 (S) -methoxymethyl-1-mercapto-a-lincosaminide, 7-deoxy-7 (S) -methylthiomethyl-1-mercapto- α-lincosaminide, 7-deoxy-7 (S) - (2-hydroxyethyl-lio) methyl-1-mercapto-α-lithicosaininide or 7-deoxy-7 (S) (3-hydroxypropylthio) and the pharmaceutically acceptable salts thereof.

The compound of formula (III) wherein -

R, and R ₂ the ring may be a 2-, 3-, 4-, 5-, 6-, 7-, 8- or

9- and have the same meaning as above;

R ₃ is hydrogen, methyl, ethyl and 2-hydroxyethyl;

n is 1, 2, 3 or 4 and pharmaceutically acceptable salts thereof.

A compound of Formula IV wherein - θ A, B and E are nitrogen, oxygen, sulfur or -CR 1 R 1 -;

Ri and _R2 are as defined above, and capitalization 'csolódhatnak any carbon or nitrogen atom on the ring;

R, the ring may be multiple bonded to any one

· Its carbon atom;

and pharmaceutically acceptable salts thereof.

Compound of Formula V wherein - θ A, B, D and E are nitrogen, oxygen, sulfur or

-CR, R 1 -;

Ri and _R2 are significantly the same as above and attached to any ring carbon or nitrogen atom;

R j may be multiple-linked to any carbon atom in the ring;

and pharmaceutically acceptable salts thereof.

PATENT CLAIMS

Claims (2)

A process for the preparation of a compound of formula (I) wherein R is (a), (b), (c) or (d) characterized in that Streptomyces rochci, strain NRRL 3533 is an aqueous, 5-carbon strain. culture medium containing oxygen, nilogenene and phosphorus sources in the presence of a compound of formula (I) wherein R is hydrogen and the resulting S-S 'ribonucleotides). A process for the preparation of a medicament having antimicrobial activity, which comprises mixing a compound of the formula (I) obtained by the process of claim 1 with a diluent, carrier and other auxiliaries customary in the preparation of a medicament. It is converted into 5 pharmaceutical compositions. 2 sheets drawing -18 to 085 NSZ0 ₄ : C 12 P 19/64; C 12 P 19/30; A 61 K 31/71 Hl) (IV) l * 3 ARC) i9 -1991-085 NGO ₄ : 3 12 P 19/64; C 12 P 19/30; A 61 K 31/71