[go: up one dir, main page]

CA1163938A - Process for treating a protozoan disease with lincomycin 3-(5'-ribonucleotides) - Google Patents

Process for treating a protozoan disease with lincomycin 3-(5'-ribonucleotides)

Info

Publication number
CA1163938A
CA1163938A CA000398814A CA398814A CA1163938A CA 1163938 A CA1163938 A CA 1163938A CA 000398814 A CA000398814 A CA 000398814A CA 398814 A CA398814 A CA 398814A CA 1163938 A CA1163938 A CA 1163938A
Authority
CA
Canada
Prior art keywords
compound
general formula
ribonucleotide
acceptable salt
prepared
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired
Application number
CA000398814A
Other languages
French (fr)
Inventor
Alexander D. Argoudelis
David W. Stroman
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Pharmacia and Upjohn Co
Original Assignee
Upjohn Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US06/255,542 external-priority patent/US4383109A/en
Priority claimed from US06/255,541 external-priority patent/US4368193A/en
Application filed by Upjohn Co filed Critical Upjohn Co
Application granted granted Critical
Publication of CA1163938A publication Critical patent/CA1163938A/en
Expired legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/06—Pyrimidine radicals
    • C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
    • C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
    • C07H19/16—Purine radicals
    • C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/305—Pyrimidine nucleotides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/32—Nucleotides having a condensed ring system containing a six-membered ring having two N-atoms in the same ring, e.g. purine nucleotides, nicotineamide-adenine dinucleotide
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/38—Nucleosides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/465—Streptomyces

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Saccharide Compounds (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

ABSTRACT
Novel and useful ribonucleotides of analogs of the well known antibiotics lincomycin and clindamycin. These ribonucleotides are unexpectedly highly active against Streptococcus hemolyticus and Staphylococcus aureus in vivo.

Description

1 :1 63938 DESCRIPTION

BACKGROUND OF THE INVENTION
The characteristics and preparation of the antibiotic lincomycin are disclosed in U.S. Patent 3,086,912. Clindamycin is disclosed in U.S. Patent 3,496,163. These antibiotics have been extensively used as medicines in humans and animals. A number of patents world-wide have issued concerning these antibiotics and a variety of derivatives thereof.
The structural formulas for lincomycin ~1) and clindamycin (2 are shown in Chart 1.
Lincomycin and clindamycin 3-nucleotides are disclosed and claimed in U.S. Patent 3,671,647. All of the lincomycin and clinda-mycin compounds disclosed in U.S. 3,671,647 have the propyl hygric acid moiety. These 3-nucleotides were found by test against S. aureus ln vivo to have an activity approximately one-tenth of the parent compound.
BRIEF SUMM~RY OF THE INVENTION
The subject invention concerns the 3-ribonucleotides of linco-mycin- and clindamycin-type compounds in which the propyl hygric acid moiety has been replaced by different cyclic amino acids. Unexpect-edly, these nucleotides demonstrate in vivo antibacterial activity ashigh as the parent compounds. ~ecause of these highly relevant char-acteristics, the nucleotides of the subject invention are considered to be prime candidates for medicinal use. These compounds also are useful for the prophylactic and therapeutic treatment of sub,iects hosting a protozoan parasite. For example, when the protozoan is a malarial parasite, the subject can be animal, e.g., mice infected with Plasmodium berghei; birds, e.g. ducks infected with P. lophurae and chic~s infected with P. gallinaceum, and mammals such as primates, e.g., monkeys infected with P. cynomolgi, and humans infected with P.
falciparunl, P. vivax, and P. malariae.
Mammals hosting a parasitic protozoan of the class Sporazoa, order Coccidia (a microparasite producing the disease coccidiosis) can be treated by administration of the compounds of the present inven-~ 3 63938 tion. For example cattle infected with Eimeria zurnl1, E. bovis~ E.ellipsoidalis; sheep and goats with E. parva, E. faurei; swine with .
debliecki, E. scabra, and Isospora suis; dogs and cats with Isospora bigemina, I. felis, E. canis, E. felina; poultry with ~. tenella;
rabbits with E. stiedae, E~ perforans; and mink with E. mustelae can be treated.
The lincomycin- and clindamycin-type compounds which can be converted to the 3~ribonucleotides are shown in Chart 2. In place of the hydroxyl at the three position of the lincosaminide moiety, there is substituted a nucleotide selected from the group consisting of adenylic acid, guanylic acid, cytidylic acid and uridylic acid.
The 3-ribonucleotides of the subject invention can be prepared by microbiological transformation procedures. The 3-(5'-ribonucleotides) obtained by transformation of U-57930 are shown in Chart 3 DETAILED DESCRIPTION
The parent compounds disclosed in ~hart 2 can be prepared by the procedures disclosed in applicant's U.S. pa~ent No. 4,278,7S9, issued July 14, 1981.
The 3-~(5'-ribonucleotides) of the compounds of Chart 2 can be prepared by following the procedures disclosed in U.S. -Patent 3,671,647. Salts of these nucleotides also can be prepared following the procedures in U.S. 3,671,647.
Formulations of the nucleotides of this invention can be made following the composition examples in U.S. patent 4,278,789.
The ~ormulations are prepared by substitutin~ a nucleotide of the subject invention for the active compound in the examples. The substitution can be on an equimolar basis.
General assay and characterization procedures which can be employed to determine and characterize the nucleotides of the inven-tion are as follows:
Assay of 3-~5'-Ribonucleotides) Since the 3-ribonucleotides of this invention lack in vitro antibacterial activity, their formation from the antibacterially-active parent compounds can be followed easily by measuring the lossof such antibiotic activity. To determine the amoun~s of antibac-terially-act1ve parent compound in culture filtrates or reaction mixtures, a standard assay with Sarcina lutea ATCC 9341 is employed.

1 ~ 63938 ~ 713/3841 To assay for the presence of the 3-ribonucleotides in fermentation beers, extracts, and purified materials, the phosphodiester bond is first hydrolyzed with crude alkaline phosphatase, or snake venom phosphodiesterase, by the procedures described below. The 5 antibacterially-active compound in the hydrolysate is determined by standard assay.
Enzymatic Hydrolyses Alkaline Phosphatase: Stock solutions (0.5 mg/ml, 0.54 Units/mg) of pigeon intestine alkaline phosphatase, EC 3.1.3,1 (Sigma) are 10 prepared in Tris (hydroxymethyl) aminomethane hydrochloride buffer7 0.01 M pH 8Ø Samples to be treated are diluted 1:2 with the enzyme buffer mixture and are incubated at 28C for 18 hours.
Snake Venom Phosphodiesterase: Stock solutions (100 mg/ml, 0.026 Units/mg) of purified snake venom phosphodiesterase LC 3.1.4.1 (Sigma) 15 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 of 0.01 M Tr7s-hydrochloride buffer, pH s.n, o.l ml of 0.3 M MgCl2, and 0.1 ml of the enzyme stock solution. Incubation is carried out at 37C for 18 hours.
Spleen Phosphodiesterase: Stock solutions of spleen phosphodi-esterase EC 3.1.4.18 (Sigma) are prepared (1 mg/ml, 19.6 Units/mg) in distilled water. Incubation mixtures contain 0.4 ml of a solution (0.5 mg/ml) of the sample to be treated in water, 0.5 ml of ().(~2 M
Tris buffer, pH 7.0 and 0.1 ml of the enzyme stock solution. Incu-25 bation is carried out at 37C for 18 hours.
Thin-La~r Chromatographic Analysis of Preparations_and Enzy-matic Hydrolysates.
The production and purification of the 3-ribonucleotides is followed by assay against S. lutea (see above) and by TLC using silica 30 gel G and methyl ethyl ketone-acetone-water (186:52:20, v/v) or ethyl acetate-acetone-water (8:5:1) as the solvent systems. The bioactive parent compounds are detected by bioautography on agar seeded with S.
lutea.
The products of enzymatic or chemical hydrolysis of the 3-nucleo-35 tides are separated by the following TLC systems:
A: Silica gel GF plates (Analtech Inc.); water as the solvent system.
B: Silica gel GF plates; n-propyl alcohol-conc. ammonium i 1 6393~

hydroxide-water (55:10:35, v/v).
C: NM-Polygram Cellulose 300 (Brinkman Instruments Inc.); 1-butanol-water-formic acid (77:13:10, v/v).
UV absorbing materials are detected by a short wavelength UV
lamp. Bioinactive, UV-nonabsorbing materials are detected by a permanganate-periodate spray reagent. Bioactive nucleotide materials are detected by bioautography on agar seeded with S. lutea.
The following example shows the fermentation and purification procedures for preparing the nucleotide of the compound designated as U-57930E. The structural formula of U-57930E is shown in Chart 3. Ry following the procedures of this example, or obvious equivalents thereof, there can be made the 3-l~ibonucleotides of the other com-pounds disclosed in Chart 2.
The following example is illustrative of the process and products of the invention, but are not to be construed as limiting. All per-centages are by weight and all solvent mixture proportions are by volume unless otherwise noted.
Exam~le 1 A. Fermentation Procedure Streptomyces rochei, NRRL 3533, is grown in a medium consisting of glucose, 10 g/liter; Difco peptone, 4 g/liter; Difco yeast extract, 4 g/liter; MgS04.7H20, 0.5 g/liter, KH2P04, 2.0 g/liter, K2HP04, 4 gtliter for three days at 28C on a rotary shaker. The mycelium from this growth is used to inoculate a fermentation medium containing the same ingredients. The fermentation is carried out for 48 hours at 28C on a rotary shaker. At the end of this 48-hour incubation, U-57930 is added to a final concentration of 50 mg/liter and the fermentation continued at 32C. After twelve hours, additional U-57930 is added to make the total concentration 150 mg/liter. After 30 twelve additional hours, the U-57930 concentration is increased to 25n mg/liter. The fermentation is continued at 32 for 24 hours after the last addition of U-57930. At this time the culture filtrates are harvested and Found to contain no more than 1 mg/liter of U-5793 The remaining 249 mg/liter is converted to bioinactive material.
S. rochei, NRRL 3533, is a known microbe which is available to the public upon request from the NRRL repository. The address of this repository is as follows: Northern Utilization and Research Division, ~; Agricultural Research Service, U.S. Department of Agriculture, Peoria, 1 1 ~3938 ~ 713/3841 Ill., U.S.A.
B. Isolation and Purification Procedures Isolation of U-57930 3-Ribonucleotides from Ferm_ntation ~roth Adsor~tion on Amberlite XAD-2: Fermentation broth (ca 12 liter) containing 3 g of nactivated U-57930 is filtered at harvest pH (7.7) by using filter aid. The mycelial cake is washed with 1.2 liter of water and discarded. The clear filt,ate and wash are combined and adjusted to pH 6.0 and passed over a column prepared from 600 ml of Amberlite* XAD-2 (Rohm and Haas Co., Philadelphia, PA), at a flow rate of 40 ml/minute. The spent is tested for bioactivity before and after treatment with alkaline phosphatase and is discarded. The column is washed with 2 liters of water. The aqueous wash is also found bio-inactive before and after treatment with alkaline phosphatase and is discarded. The column is then eluted with methanol-water (70:30 v/v).
Fractions of 20 ml are collected at a rate of 20 ml/minute. Testing for bioactivity before (-E) and after (~E) treatment with alkaline phosphatase shows the following.
Zone (S. lutea) Fraction No. -E +E

11 0 o '` *trade mark ., 1 ~ 63938 Zone (S. lutea~
Fraction No. - +E

100 16 ` 26 Fractions 12-80 are combined, concentrated to an aqueous solution and freeze-dried to give prep ADA-34.1, 12.22 g.
In another series of experiments, 6 liters of fermentation broth containing 2 g of "inactivated" U-57930 is treated as described above~
The methanolic eluates from the Amberlite* XAD-2 column are kept as ADA-143B. This solution is not concentrated to dryness; instead, it is purified by Dowex*1 Ohromatography as described below.
r~ The column is prepared from 300 ml of Dowex*1 (X-4) in the acetate form. The methanolic solution, ADA~143B
pH 8.2 is passed through the column. The spent is collected at a rate of 2.5 ml/m~nute in 20 ml-fractions. (Fractlons 1-60). The column is washed with 1.5 liter of water (10 ml/min9 fractions 66-108). The column is then eluted with 5% acetic acid (rate, 10 ml/minute, ~rac-tions 109-310). The following pools are made:
Pool 1 Fractions 1-80 1000 ml (ADA-lA)
2 Fractions 81-110 600 ml (ADA-2A)
3 Fractions 111-130 450 ml (ADA-3A)
4 Fractions 131-150 450 ml (ADA-4A) Fractions 151-190 900 ml (ADA-5A) 6 Fractions 191-230 900 ml (ADA-6A) 7 Frac~ions 231-270 900 ml (ADA-7A) 8 Fractions 271-310 900 ml (ADA-8A) Testing before (-E) and after (~E) ~reatment with alkaline phosphatase shows the following:

*trade mark : 'j 1 31 ~3~3~

E Zone (S. lutea) F_ _ Pool 1 31 52 8 2g 31 Pools 1 and 2 are combined, concentrated to an aqueous solution and freeze-dried to give prep ADA-2.1, 1.48 9.
Pools 3 and 4 are also combined and treated similarly to give ADA-2.2, 2.5 9.
Preparations ADA-2.1 and -2.2 give U-57930 after treatment with alkaline phosphatase.
Preparations ADA-34.1, -2.1 and -2.2 are combined and purified by the counter double current distribution procedure described below.
Counter Double Current Distribution: The material obtained by combination of preparations ADA-34.1, -2.1 and -2.2, 16.20 9, is dissolved in 25 ml of each phase of the solvent system consisting of equal volumes of 1-butanol-water (1:1), The solutions are added in the center tubes of an all-glass counter double current distribution apparatus (100 tubes, 25 ml/phase). The distribution is analyzed, after 150 transfers, for bioactivity before (-E) and after (~E) treat-ment with alkaline phosphatase. Results follow:

1 1 63~38 Zone (S. lutea-sensitive~
Lower Collector -E ~E

0 33.5 o 3~

100 0 43.5 Zone (S. lutea-sensitive) Lower Machine -E +E

trace 46 17 47.5 17.5 48 O trace 50 Upper Machine trace 49 trace 48.5 trace 48 trace 48.5 trace 49 trace 51 .., ~ . ., Zone (S. lutea-sensitive~
Upper Collector _ +
100 17.5 54 17 53.5 19 53.5 ~'0 53.5 ~'1 53.5 22 53.5 24 53.5 26 53.5 32.5 52 ~3 52 The following pools are made. Each pool is concentrated to an aqueous solution and freeze-dried to give the corresponding prepara tions.
Pool I: Lower collector 1-50;
Pool II: Lower collector 51-100, lower machine 50-30;
Pool III: Lower machine 29-0; Upper machine 1-50j Upper collector 100-30.
25Preparations obtained are:
From pool I, prep. ADA-47.1, 9.78 9 From pool ~I, prep. ADA-47.2, 0.30 9 From pool III, prep. ADA-47.3, 5029 9 Preparations ADA-47.2 and -47.3 are combined and purified by DEAE-Sephadex chromatography as described below~
DEAE-Sephadex*Ohromatography: Three hundred g of DEAE-Sephadex~
~A-25) are stirred for 1 hour with water and for 2 hours with 0.5 N
~queous sodium hydroxide. The ionic exchanger is washed with water until the pH is ca 7.5. The material is then stirred for 2 hours with 0.5 N aqueous acetic acid, washed with water to a neutral pH, and poured into a column and packed under 2 lbs pressure to a constant height. The column is washed with 4 liter of water, 8 liter of 0.1X
aqueous solution of tris-(hydroxymethyl)aminomethane (THAM), and 3 ~3 *trade mark 1 1 63g3~

liter of 0.03 M THAM acetate buffer pH 8.0 (prepared by dissolving 3.64 g of THAM in 800 ml water, adjusting the pH to 8.0 with glacial acetic acid and then adjusting the volume to 1 liter).
Starting material, preparations ADA-47.2 and 47.3 ca 5.50 9, is
5 dissolved in 20 ml of 0.03 M THAM acetate pH 8.0 buffer and added on the top of the column. The column is then eluted downflow with 0.3 M
THAM acetate pH 8.0 buffer. Fractions 1-190 (20 ml) are collected.
At this point elution of the column is continued in an upflow manner.
Fractions A, B, C, D, and E (1 liter each) are collected. Testing for bioactivity before (-E) and after l+E) treatment with alkaline phos-phatase shows the following:

Zone (S~ lutea-sensitive) Fraction No. -E +E
6 0 0 12 0 o 21 oO o 27 0 l 0 39 43. 5 44 23.5 23
7 1 63938 3713/38~1 Zone (S. lutea-sensitive) Fraction No. -E +E

6~ 0 0 66 0 o 1~5 23 51 108 22.5 54 111 22.5 52.5 117 22 54.5 120 20.5 56 The following pools are made:
Pool I Fractions 34-38, 280 ml (ADA-69B) Pool II Fractions 75-90, 330 ml (ADA-69C) Pool III Fractions 101-111, 180 ml (ADA-69D) Pool IV Fractions 114-150, 580 ml (ADA-69E) Pool V Fractions 151-164, 100 ml (ADA-69F) Pool VI Fractions 165-186, 125 ml (ADA-69G) Pool VII Fraction C, 1 liter (ADA-69A) Pool I (ADA-69B) contains unchanged U-57930 and is discarded.
Pool II (ADA-69C) contains an unknown material which yields U-57930 by treatment with alkaline phosphatase. UV: ~ max 275 nm.
Pool IIT (ADA-69D) contains U-57930 cytidylate and is treated as described later. UV: ~ max 270 nm.
Pool IV (ADA-69E) contains U-57930 adenylate and is treated as described later. UV: ~ max 260 nm.
Pool V (ADA-69F) contains a mixture of U-57930 adenylate, U-57930 - uridylate and U-57930 guanylate. This solution is treated as des-I ~ 63938 3/1~ 41 cribed later.
Pool VI (ADA-69G) contains U-57930 guanylate and is treated as described later. UV: ~ max 254; sh at 275.
Pool VII (ADA-69A) contains a mixture of U-57930 guanylate and U-57930 uridylate. This solution is treated as described later.
Isolation of Essentially Pure U-57930-Cytidylate, U-57930-Adenylate and U-57930-Guanylate from Po~ls III, IV and VI, Respec-tively. Removal of THAM Acetate_BuFfer by Amberlite XAD-2 Chroma-tography: Pools III, IV and VI, obtained as described above, are passed over columns containing Amberlite XAD-2. The spents are dis-carded. The columns are washed with water and then eluted with methanol-water (70:30 v/v). Fractions are analyzed by UV and by testing for bioactivity before and after treatment with alkaline phosphatase. Appropriate fractions are combined, concentrated to an aqueous solution and freeze-dried. Details on the amount of Amberlite XAD-2 used for each pool, the amount of water wash, the amount of methanolic eluate and the amount of material obtained are listed in the following table.
Amberlite XAD-2 Water Methanolic Isolated 20 PoolUsed (ml) Wash (ml) Fluate tml)_ Material (m~

The material obtained from pool III is kept as ADA-73.1; from poo~ IV as ADA-74.1; and ~rom pool VI as ADA-75.1.
Removal of THAM Acetate Buffer from Pool V (ADA-69F) and Pool VII
(ADA-69A) by Amberlite*XAD-2 Chromato~raphy: The column is prepared from 300 ml of Amberlite XAD-2. Pools V and VII containing a mixture 30 of U-57930 adenylate; U-57930-uridylate and U-57930 guanylate are passed through the column. The spent is discarded. The column is washed with 600 ml of water. The spent is discarded. The column is eluted with methanol-water (70:30). Fractions yielding bioactive material after treatment with alkaline phosphatase are combined, 300 ml, concentrated to an aqueous solution and freeze-dried to give prep ADA-71.1, 670 mg. Prep -71.1 is treated as described below.
Separation of U-57930 Uridylate from U-5?930-Adenylate and U-57930-Guanylate DEAE-Sephadex Chromatography. Six hundred ml of , ~ ,, *trade mark 1 1 6 3 9 3 8 3/~ L

DEAE-Sephadex in the acetate form, prepared as described earlier, are washed with 0.03 M THAM acetate pH 8.0 buffer and packed into a glass column (ID, 4.5 cm; height, 40 cm) under hydrostatic pressure.
Prep ADA-71.1 (see above) is dissolved in 10 ml of 0O03 M THAM
acetate pH 8.0 buffer and added on the top of the column. The column is eluted with:
1) 0.03 M THAM acetate, pH 8.0 (Fractions 1-79) 2) 0.12 M THAM acetate, pH 8.0 (Fractions 80-395) 3) 0.25 M THAM acetate, pH 8.0 (Fractions 396-750) Fractions of 20 ml are collectecl and analyzed by UV and by test-ing for bioactivity before and after treatment with alkaline phospha-tase. Fractions 51-60 contain U-57930 adenylate; fractions 62-73 (ADA-94.B) contain U-57930 uridylate; fractions 75-110 contain U-57930 guanylate).
Isolation of Essentially Pure U-57930 Uridylate. _Removal of THAM-Acetate Buffer by Amberlite XAD-2 Chromatography. The column is prepared from 50 ml of Amberlite XAD-2. Rool ADA-94~, containing U-57930 uridylate, is passed over the column at a rate of 2 ml/minute.
The spent is discarded. The column is washed with 200 ml of water.
The wash is discarded. The column is eluted with methanol-water (70:30 v/v). Fractions containing (by UV) U-57930 uridylate are combined (200 ml), concentrated to an aqueous solution, and freeze-dried to give ADA-95.1, 60 mg.

~` *trade mark :

1 ~ ~3~3~
* 1 CHARACTERIZATION OF U-57~9~ Lll9 1. IR Tabulation Tables listing the IR absorptions (Nujol and KBr) are as follows:

Band Band Freq. _Inten. Type __ Freq. Inten. 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 1089.9 15 ~RD
2951.4 2 BRD M 1070.6 12 AVG
2326.3 1 BRD M 1056.1 14 SH
2854.9 2 BRD M 992.4 39 AVG
2729.6 48 BRD M 972.2 39 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 AVG M 654.9 45 SH
1368.6 31 SH M 632.7 38 AVG
1286.6 34 AVG
Band Freq.: Band Frequencies in Wavenumbers (cm-1 Inten.: Intensity in percent transmittance (YoT ) Data Type in Local Peak Region: BRD - Broad; AVG - Average; SHP
- Sharp; SH - Shoulder Peak list edited. * Indicates peaks added.
M: Possible interference from mineral oil.

*trade mark 1 3 6.~93~

25 Strongest Peaks %T Freq. %T Freq.
1 2926.2 24 3417.2 2 2951.3 25 3108.5 2 2854.8 26 1610.6
8 1649.2 31 152~.6
9 1462.1 31 1368.5 12 1070.5 33 1249.0 14 1056.0 34 1286.5 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 Prep: Mineral Oil Mull Max %T: 87 @ 1848.0 %T at 3800 (cm-1): 83 Density (cm-1/pt): 0.964 Band Band Freq. Inten. Type Freq. Inten. Type 3408.610 BRD 1088.9 12 BRD
3102.826 SH 1071.5 9 AVG
2963.928 AVG 1057.1 11 SH
2930.227 BRD 992.4 35 AVG
2878.137 AVG 972.2 36 AVG
2862.739 SH 956.8 45 SH
2768.151 SH 928.8 48 AVG
2511.665 BRD 889.2 32 AVG
1649.3 4 AVG 859.3 47 AVG
1614.620 SH 851.6 47 SH
1576.030 AVG 804.4 39 SH
1528.728 AVG 788.9 34 AVG
1491.121 AVG 743.6 47 SH
1462.233 AVG 705O0 40 AVG
1450.635 SH 654.9 39 SH
1404.337 AVG 634.6 35 AVG
1384.033 AVG 595.1 33 AVG
1360.942 SH 57209 33 AVG
1286.632 AVG 525.6 32 AVG
1251.930 SH 447.5 36 AVG
1215.218 AVG
Band Freq.: Band Frequencies in wavenumbers (cm-1 Inten.: Intensity in Percent Transmittance (Y~T) Data Type in Local Peak Region: BRD - Broad; AVG - Average; SHP
- Sharp; SH - Shoulder Peak List Edited. *Indicates peaks added.

25 Strongest Peaks %T Freq. %T Freq.
4 1649.2 30 1251.8 9 1071.5 32 1286.5 3408.5 32 889.1 11 1057.0 32 525.5 12 1088.8 33 1462.1 18 1215.1 33 1384.0 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 1576.0 Prep: KBR Pellet Max %T: 100 @ 403.1 %T at 4000 (cm-1): 78 Density (cm-llpt): 0.964 2. UY Absorption Spectrum [~ max (a~]
In water at:
pH 200, 279 nm (6.5) pH 7.0, 270 nm (9.9) pH 11.0, 271 (9.6) 3. Elemental Composition Mol. formula: C26H43NsO~2 SClP. Molecular Weight, 715.
Calcd: C, 43.64; H, 6.01; N, 9.79; O, 26.88; S, 4.47; Cl, 4.89, P, 4.33.
4. Optical Rotation ~]D2s, +107 (C, 0.854, water) 5. Solubilities Highly soluble in water, methanol and ethanol. Slightly soluble in acetone and other ketones, ethyl acetate and other esters, chloroform, methylene chloride. Insoluble in saturated hydrocarbon solvents.
`' .:

~ 1 6 ~ 9 3 8 6. Antibacterial Activit~
U-57930 3-(5'-cytidylate) is not active in vitro. However, treatment with alkaline phosphatase or phosphodiesterase I
yields U-57930 which is highly active against a variety of G+
organisms, both in vitro and in vivo.
7. Melting point: 205-207 (w~th decomposition).

CHARACT~RIZATION OF U-57930 3'(5'-ADENYLATE) 1. IR Tabulation Tables listing the IR absorptions (Nujol and KBr) are as follows:

Band Band Freq. lnten. Type Freq. Inten. _ T~pe_ 3335.316 BRD 1245.1 24 SH
3267.817 BRD 1213.3 17 AVG
3210.817 BRD 1175.7 43 SH
2954.3 3 BRD M 1146.8 40 SH
2924.4 2 BRD M 1089.9 13 AVG
2868.4 6 SH M 1069.6 10 AVG
2854.9 4 AVG M 1055.1 13 SH
2727.646 BRD M 991.5 35 AVG
2520.261 BRD 972.2 36 AVG
1684.022 SH 957.7 45 SH
1641.614 AVG 930.7 48 AVG
1600.128 AVG 889.2 32 AVG
1576.031 AVG 861.3 47 AVG
1550.943 SH 848.7 49 AVG
1509.453 SH 818.8 45 AVG
1463.115 AVG M 798.6 40 AVG
2 1420.735 AVG 722.4 36 AVG M
1377.324 AVG M 708.9 40 SH
1367.635 SH M 647.1 33 SH
1332.036 AVG 635.6 31 AVG
1299.234 AVG
Band Freq.: Band Frequencies in Wavenumbers (cm-l) Inten. Intensity in percent transmittance (ZT) Data Type in Local Peak Region: BRD Broad; AVG - Average; SHP
- Sharp; SH - Shoulder Peak list edited. *Indicates peaks added.
M: Possible interference from mineral oil.

L~ * trade mark ~ ~ ~3938 25 Strongest Peaks %T _ Freq. %T Freq.
2 2924.3 22 1684.0 3 2954.2 24 1377.2 ~ 2854.8 24 1245.0 6 2868.3 28 1600.0 1069.5 31 1576.0 13 1089.8 31 635.5 13 1055.0 32 889.1 14 1641.5 33 647.0 1463.0 34 1299.1 16 3335.2 35 1420.6 17 3267.7 35 1367.5 17 3210.7 35 991.5 17 1213.2 Prep: Mineral Oil Mull Max %T: 85 @ 1864.4 %T at 3800 (cm-1): 81 Density (cm-1/pt): 0.964 Band Band Freq. Inten. Type Freq.Inten. Type 3375.8 7 BRD 1090.8 8 AVG
3223.410 BRD 1069.6 5 AVG
3124.017 SH 1050.3 8 SH
2963.022 AVG 990.5 27 AVG
2929.222 BRD 972.2 29 AVG
2878.131 AVG 956.8 38 SH
2863.633 SH 929.8 42 AVG
2756.645 BRD 889.2 24 AVG
2521.260 BRD 861.3 38 AVG
2188.575 BRD 851.6 40 SH
1678.215 SH 818.8 36 AVG
1643.5 7 AVG 807.3 36 BRD
1602.020 AVG 798.6 30 AVG
1576.023 AVG 768.7 41 BRD
1553.835 SH 721.4 30 AVG
1511.449 SH 706.9 31 BRD
1475.727 AVG 648.1 26 AVG
1421.729 AVG 636.5 26 AVG
1384.032 AVG 584.5 28 SH
1332.031 AVG 571.9 26 AVG
1301.128 AVG 533.3 27 SH
1246.119 SH 522.7 25 AVG
1215.212 AVG 503.4 25 AVG
1176.736 SH
.

~ .

~ ~ ~393~ 37l3/384l Band Freq.: Band Frequencies in Wavenumbers (cm-1) Inten.: Intensity in percent transmittance (~T) Data Type in Local Peak Region: BRD - Broad; AVG - Average; SHP
- Sharp, SH - Shoulder Peak list edited. *Indicates peaks added.

25 Strongest Peaks %T Freq~ ~oT _Freq.
1069.5 23 1576.0 7 3375.7 24 889.1 7 1643.5 25 522.6 8 1090.7 25 503.3 8 1050.2 26 648.0 3223.3 26 636.5 12 1215.1 26 571.8 1678.1 27 1475.6 17 3124.0 27 990.5 19 1246.0 27 533.2 1602.0 28 1301.0 22 2963.0 28 584.5 22 2929.1 20Prep: KBR Pellet Max %T: 95 @ 405.0 %T at 4000 (cm-1): 77 Density (cm-1/pt): 0.964 2. UV Absorption Spectrum [~ 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 Composition Molecular formula: C27H~3N701o SClP. Molecular Weight, 723.
Calcd 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 [~]D2s~ +94 (C, 0.887, water).
5~ Solubilities Highly soluble in water, methanol and ethanol. Slightly ~ ~ ~3938 soluble in acetone and other ketones, ethyl acetate and other esters, chloroform and methylene chloride. Insoluble in saturated hydrocarbon solvents.
6. Antibacterial Activit~
U-57930 [3-(5'-adenylate)~ is not active in vitro. However, treatment with alkaline phosphatase or phosphodiester I
yields U-57930, which is highly active against a variety of G+ organisms b~th in vitro and in vivo. U-57930 3-(5'-aden-ylate) was found active in vivo (subcutaneously, mice) ~ith a CDso of 0.62 (0.48-0.79) mg/kg. against S. pyogenes.
7. Melting Point: 203.5-205 (with decomposition) CHARACTERIZATION OF U-57930 3-(5'-URIDYLATE) 1. IR Tabulation Tables listing the IR absorptions (Nujol and K~r) are as follows:

Band Band Freq. Inten. Type _ Freq. Inten. 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 RRD
2924.4 0 BRD M 1215.2 19 AVG
2867.5 4 SH M 1089.9 13 AVG
2854.0 3 AVG M 1071.5 9 AVG
2733.4 49 SH M 1056.1 13 SH
2695.B 53 SH 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
146202 12 AVG M 767.7 SO AVG
1425.5 37 SH 721.4 42 AVG M
1378.3 22 AVG M 634.6 35 AVG
1367.6 37 SH M
Band Freq.: Band Frequencies in Wavenumbers (cm-1) Inten.: Intensity in percent ~ransmittance (%T) Data Type in Local Peak Region: BRD - Broad, AVG - Average; SHP
- Sharp, SH - Shoulder This peak list is unedited.
F~
i!~i,,, ~ j~
il~ * trade mark - 1 ~ 6~938 M: Possible interference from mineral oil.

25 Stron~est Peaks %T Fre~1~ %T Freq.
0 2924.3 22 1378.2 1 295203 28 1251.8 3 2854.0 34 890.1 4 2~67.5 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 322403 42 721.3 21 1647.3 Prep: Mineral Oil Mull Max %T: 86 ~ 3764.5 %T at 3800 (cm-1): 85 Density (cm-1/pt): 0.964 Band Band Freq. Inten. Type Freg.Inten. Ty~e 3387.412 BRD 1055.1 10 SH
3114.425 SH 992.4 35 AVG
2962.026 AVG 973.2 36 AVG
2931.126 BRD 956.8 45 SH
2879.136 AVG 929.8 48 AVG
2863.638 SH 889.2 31 AVG
2833.743 SH 859.3 46 AVG
2509.664 BRD 813.0 40 AVG
1685.0 6 BRD 811.1 40 SH
1647.417 SH 798.6 43 AVG
1605.935 SH 782.2 48 RRD
1576.037 AVG 768.7 48 AVG
1556.739 BRD 707.9 43 AVG
1463.131 AVG 669.3 43 SH
1423.635 AVG 649.1 40 SH
1384.032 AVG 634.6 37 AVG
1331.043 AVG 585.4 38 SH
1297.238 SH 567.1 34 AVG
1255.824 AVG 523.7 35 AVG
1214.317 AVG 447.5 39 AVG
1090.810 AVG
1070.6 7 AVG
., Band Freq.: Band Frequencies in Wavenumbers (cm-1) ~ 1 B~938 Inten.: Intensity in percent transmittance (q~T) Data Type in Local Peak Region: BRD - Broad; AVG - Average; SHP
- Sharp; SH - Shoulder Peak List Edited. *Indicates peaks added.

25 Stronge.st Peaks %T _ _ Freq. %T Freq.
6 1685.0 32 1384.0 7 1070.5 34 567.0 1090.7 35 1605.8 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 3114~3 37 1576.0 26 2962.0 37 634.5 26 2931.0 38 2863.5 31 1463.0 38 1297.1 31 &89.1 Prep: K~R Pellet Max %T: 101 @ 405.0 %T at 4000 (cm-1): 76 Density (cm-1/pt): 0.964 2. UV Absorption Spectrum [~max (a~
In water at:
pH 2.0, 261 (11.5) pH 7.0, 262 (10.7) pH 11.0, 262 (11.5) 3. Elemental Composition Molecular formula: C26H42N4013 SClP. Molecular Weight, 716.
Calcd C, 43.57; H, 5.86; N, 7.82; O, 29.05; S, 4.46; Cl, 4.89; P, 4.33.
4. Op~ical Rotation ~]D25 +105~ (C, 0.94, water) 5. Solubilities Highly soluble in water, methanol and ethanol. Slightly soluble in acetone and other ketones, ethyl acetate and other --~ esters, chloroform and methylene chloride. Insoluble in ~ ~ 6~938 3713/3841 saturated hydrocarbon solvents.
6. Antibacterial Activity U-57930 3-(5'-uridylate) is not active in vitro. However, treatment with alkaline phosphatase or phosphodiesterase I
yields U-57930 which is highly active against a variety of G
organisms both in vitro and in vivo.
7. Melting Point: 202-203 (with decomposition) CHARACTERIZATION OF U-57930 3-(5'-GUANYLATE) 1. Tables l1sting the IR absorptions (Nujol and KBr) are as follows:

Band Band Freq. Inten. Type Freq. Inten. Type 3335.316 BRD 1250.0 36 SH
3227.219 BRD 1213.3 21 AVG
2953.3 2 AVG M 1173.8 39 AVG
2925.3 1 BRD M 1149.7 41 AVG
2 2868.4 6 SH M 1087.9 14 S~
2855.9 4 AVG M 1071.5 9 AVG
2737.351 BRD M 991.5 42 AVG
2521.273 BRD 972.2 42 AVG
1684.0 6 AVG 956.8 52 SH
1635.811 AVG 929.8 51 AVG
- 1598.221 AVG 890.2 36 AVG
1572.126 AVG 860.3 51 AVG
1534.534 AVG 800.5 46 AVG
1462.218 AVG M 783.1 42 SHP
1414.944 AVG 720.4 43 AVG M
1377.324 AVG M 707.9 45 BRD
1365.731 AVG 681.9 4n AVG
1312.744 AVG 635.6 34 AVG
Band Freq.: Band Frequencies in Wavenumbers (cm-1) Inten.: Intensity in Percent Transmittance (%T~
Data Type in Local Peak Region: BRD - Broad; AVG - Average; SHP
- Sharp; SH - Shoulder Peak List Edited. *Indicates Peaks Added.
M: Possible interference from Mineral Oil *trade mark ::: . .J

1 ~ 6 3 9 3 ~ 3713/3841 25 Strongest Peaks %T Freq. %T _req.
1 2925.2 24 1377.~
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 21 1213.2 Prep: Mineral Oil Mull Max %T: 97 @ 3762.6 %T at 3800~cm-1): 97 Density (cm-1/pt): 0.964 Band Band Freq. Inten. Type Freq. Inten. 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
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 860.3 41 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
~, Band Freq.: Band Frequencies in Wavenumbers (cm-1) ..~

1 ~ ~393~

Inten.: Intensity in Percent Transmittance (%T) Data Type in Local Peak Region: BRD - Broad; AVG - Average; SHP
- Sharp, SH - Shoulder Peak List Edited. *Indicates Peaks Added.

25 Strongest Peaks %T Freq. %T Freq.
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 lQ88.8 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 1534.5 33 1147.6 28 889.1 Prep: KBR Pellet Max %T: 97 @ 405.0 %T at 4000(cm-1): 77 Density (cm-1/pt): 0.964 2. UV Absorption Spectrum ~Amax (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. Elemental Composition Molecular formula: C27H43N701l SClP. Molecular Weight 739.
Calcd C, 43.84; H, 5.81; N, 13.26; O, 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 [~D25~ +97 (C, 0.855, water) 5. Solubilities Highly soluble in water, methanol and ethanol. Slightly soluble in acetone and other ketones, ethyl acetate and other esters, chloroform and methylene chloride. Insoluble in ~ 3 ~3938 saturated hydrocarbon solvents.
6. Antibacterial Activit~
U-57930 3-(5'-guanylate) is not active ln vitro. However, treatment with alkaline phosphatase or phosphodiesterase I
yields U-57930 which is highly active against a variety of G
organisms both in vitro and in vivo.
7. Melting Point: 219-220 (with decomposition) Since the compounds of the subject invention are active against various Gram-positive and Gram-negative microbes, they can be used in various environments to inh;bit such microbes. For example, they can be used as disinfectants to inhibit S. aureus on washed and stacked food utensils contaminated with this bacterium. They also can be used as disinfectants on various dental and medical equipment contaminated with S. aureus. Further, the compounds of the invention can be used as bacteriostatic rinses for laundered clothes, and for impregnating papers and fabrics; and, they are also useful for suppressing the growth of sensitive organisms in plate assays and other microbio-logical media.
The compounds of the present invention are also useful in the treatment of diseases caused by members of the genus Mycoplasma, the most commonly known forms are PPL0 (pleuropneumonia-like organisms) such as M. hominis, M. salivarium, M. mycoides, M. hyopneumonia, M.
hyorhinis, M. gallisepticum, M. arthriditis and other species in man and animals, including domestic animals such as sheep, dogs, cattle, swine, and poultry (e.g., chickens, turkeys9 ducks, and geese) and laboratory animals (e.g., rats and mice).
The U-57930 3-(5'-ribonucleotides) can be used in the treatment of kidney and other infections when L forms of gram-negative and gram-positive bacteria are present, for example, L forms of P. mirabilis.
Since the compounds of the subject invention are amphotericsubstances, they can form salts with both acids and bases by using standard procedures. Examples of inorganic acids which can be used to form salts are hydrochloric, sulfuric, phosphoric, and the like.
Examples of inorganic bases are sodium, potassium, calcium, lithium, and the like. Salts of the compounds can be used for the same pur-poses as the parent compounds.
The compounds of the subject invention are useful as antibac-1 ~ 6393~3 terial agents in suitable compositions. These compositions arepreferably presented for administration to humans and animals in unit dosage forms, such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil-water emulsions containing suitable quantities of the active compound in the form of the free base, or its pharmacolog-ically acceptable salts.
For oral administration, either solid or fluid unit dosage forms can be prepared. For preparing solid compositions such as tablets, the principal active ingredient is mixed with conventional ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, acacia, methyl-cellulose, and functionally similar materials as pharmaceutical dilu-ents or carriers. The tablets can be laminated or otherwise com-pounded to provide a dosage form affording the advantage of prolongedor delayed action or predetermined successive action of the enclosed medication. For example, the tablet can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids or mixture of polymeric acids with such materials as shellac, cetyl alcohol, cellulose acetate phthalate, styrene maleic acid co-polymer and the like. Alternatively, the two component system can be utilized for preparing tablets containing two or more incompatible active ingredients. Wafers are prepared in the same manner as tab-lets, differing only in shape and the inclusion of sucrose or other sweetener and flavor. In their simplest embodiment, capsules, like tablets, are prepared by mixing the compound of the formulation with an inert pharmaceutical diluent and filling the mixture into a hard gelatin capsule of appropriate size. In another embodiment, capsules are prepared by filling hard gelatin capsules with polymeric acid coated beads containing a compound of the formulas. Soft gelatin capsules are prepared by machine encapsulation of a slurry of a com-pound of the formulas with an acceptable vegetable oil, light liquid petrolatum or other inert oil.

l ~ ~ 3 g 3 8 3713/3841 Fluid unit dosage forms for oral administration such as syrups,elixirs, and suspensions can be prepared. The water-soluble forms of a compound of the formulas can be dissolved in an~aqueous vehicle together with sugar, aromatic flavoring agents and preservatives to form a syrup. An elixir is prepared by using a hydroalcoholic (ethan-ol) vehicle with suitable sweeteners such as sucrose together with an aromatic flavoring agent. Suspensions can be prepared of the insol-uble forms with a syrup vehicle with the aid of a suspending agent such as acacia, tragacanth, methylcellulose, and the like.
Topical ointments can be prepared by dispersing the active com-pound in a suitable ointment base such as petrolatum, lanolin, poly-ethylene glycols, mixtures thereof, and the like. Advantageously, the compound is finely divided by means of a colloid mill utilizing light liquid petrolatum as a levigating agent prior to dispersing in the ointment base. Topical creams and lotions are prepared by dispersing the compound in the oil phase prior to the emulsification of the oil phase in water.
For parenteral administration, fluid unit dosage forms are pre-pared utilizing a compound of the formulas and a sterile vehicle, water being preferred. The compound, depending on the form and con-centration used, can be either suspended or dissolved in the vehicle.
In preparing solutions, a water-soluble form of a compound of the formulas can be dissolved in water for injection and filter sterilized before filling into a suitable vial or ampul and sealing. Advantage-ously, adjuvants such as a local anesthetic, preservative and buffer-ing agents can be dissolved in the vehicle. To enhance the stability, the composition can be frozen after filling into the vial and the water removed under vacuum. The dry lyophilized powder is then sealed in the vial and an accompanying vial of water for injection is sup-plied to reconstitute the powder prior to use. Parenteral suspensionsare prepared in substantially the same manner except that the compound is suspended in the vehicle instead of being dissolved and steriliza-tion cannot be accomplished by filtration. The compound can be ster-ilized by exposure to ethylene oxide before suspending in the sterile vehicle. For sustained action, an intramuscular suspension is pre-pared with an insoluble form such as the trimethylsilyl ether or the pamoate salt. Advantageously, a surfactant or wetting agent is in-cluded in the composition to facilitate uniform distribution of the 1 1 63938 3713/3~41 compound.
The term unit dosage form as used in the specification and claimsrefers to physically discrete units suitable as unitary dosages for human subjects and animals~ each unit containing a predetermined quantity of active material calculated to produce the desired thera-peutic effect in association w;th the required pharmaceutical diluent, carrier or vehicle. The specifications for the novel unit dosage forms o~ this invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding such an active material for therapeutic use in humans and animals, as disclosed in detail in this specification, these being features of the present invention. Examples of suitable unit dosage forms in accord with this invention are tablets, capsules, pills, troches, suppositories, powder packets, granules, wafers, cachets, teaspoonfuls, tablespoonfuls, dropperfuls, ampuls, vials, segregated multiples of any of the foregoing, and other forms as herein described.
An active compound is compounded with a suitable pharmaceutical carrier in unit dosage form for convenient and effective administra-tion. In the preferred embodiments of this invention, the dosage units contain 10, 25, 50, 100, 250, and 500 mg amounts of a compound of the formulas for systemic treatment~ 5 to 65 percent w/v for parenteral treatment. The dosage of compositions containing an active compound and one or more other active ingredients is to be determined with reference to the usual dosage of each such ingredient.
The following examples are illustrative of the best mode contem-plated by the inventor for carrying out his invention and are not to be construed as limiting.
The examples use the 3-(5'-ribonucleotide) of u-57~93nE or U-60,970E as the active compound, but it should be understood that this is only exemplary of the other active compounds of the subject invention. U-60,970E is the 4-cis-n-butyl-L-pipecolic acid a~ide of 7-Cl-methylthiolincosaminide. Its preparation 1s shown in Example 7 of U.S. patent 4,27~,789, Reference hereinafter to U-57,930E or U-60~970E means the 3-~5'-ribonucleotide) of these compounds. The 3-ribonucleotides are those as disclosed herein.

~ 1 ~393~

Composition Example 1 - Capsules One thousand two-piece hard gelatin capsules for oral use, each containing 250 mg of U-57,930E or U-60,970E, are prepared from the following types and amounts of materials:
U-57,930E or U-60,970E 250 gm Corn starch 100 gm Talc 75 gm Magnesium stearate 25 gm The materials are thoroughly mixed and then encapsulated in the usual manner.
The foregoing capsules are useful for the systemic treatment of infection in adult humans by oral administration of one capsule every 6 hours.
Using the procedure above, capsules are similarly prepared con-taining U-57,930E or U-60,970E in 10, 25, 50, 100, and 500 mg amounts by substituting 10, 25, 50, 100 and 500 gm of U-57,930E or U-6n,97nE
for the 250 gm used above.
Composition Example 2 - Capsules One thousand two-piece hard gelatin capsules for oral use, each containing 200 mg of U-57,930E or U-60,970E and 250 mg of tetracy-cline hydrochloride, are prepared from the following types and amounts of ingredients:
U-57,930E or U-60,970E 200 gm Tetracycline hydrochloride250 gm Talc 75 gm Magnesium stearate 25 gm The ingredients are thoroughly mixed and then encapsulated in the usual manner.
The foregoing capsules are useful for the systemic treatment of infection in adult humans by the oral administration of one capsule every 6 hours.
Using the procedure above, capsules are similarly prepared con-taining U-57,930E or U-60,970E and each of the following antibiotics in place of tetracycline by substituting 250 gm of such other anti-biotic for tetracycline: chloramphenicol, oxytetracy~line, chlor-tetracycline, fumagillin, erythromycin, streptomycin, dihydronovo-biocin and novobiocin. When a penicillin, such as potassium peni-cillin G, is to be used in place of tetracycline, 25n,0no units per i 1 63~38 capsule is employed.
Such combination products are useful for the systemic treatmentof mixed infections in adult humans by the oral administration of one capsule every 6 hours.
Composition Example 3 - Tablets One thousand tablets for oral use, each containing 500 mg of U-57,930E or U-60,970E, are prepared from the following types and amounts of materials:
U-57,930E or U-60,970E 500 gm Lactose 125 gm Corn starch 65 gm Magnesium stearate 25 gm Light liquid petrolatum 3 gm The ingredients are thoroughly mixed and slugged. The slugs are broken down by forcing through a number 16 screen. The resulting granules are then compressed into tablets, each tablet containing 50n mg of U-57,930E or U-60,970E.
The foregoing tablets are useful for systemic treatment of infec-tions, including malarial infections, in adult humans by oral admini-stration of one tablet three times a day.
Using the above procedure, except for reducing the amount of U-57,930E or U-60,970E to 250 gm, tablets containing 250 mg of U-57,930E or U-603970E are prepared.
Composition Example 4 - Tablets One thousand oral tablets, each containing 25n mg of U-57,930E or U-60,970E and total of 250 mg (83.3 mg each) of sulfadiazine, sulfa-merazine, and sulfamethazine, are prepared from the following types and amounts of materials:
U-57,930E or U-60,970E 250 gm Sulfadiazine 83.3 gm Sulfamerazine 83.3 gm Sulfamethazine 83.3 gm Lactose 50 gm Corn starch 50 gm Calcium stearate 25 gm Light liquid petrolatum 5 gm The ingredients are thoroughly mixed and slugged. The slugs are broken down by forcing through a number sixteen screen. The resulting ~ :3 ~393~

granules are then compressed into tablets, each containing 250 mg ofU-57,930E or U-60,970E and a total of 250 mg (83.3 mg each) of sulfa-diazine, sulfamerazine, and sulfamethazine.
The foregoing tablets are useful for systemic treatment of infec-tions by the oral administration of four tablets first and then oneevery six hours.
For the treatment of urinary infections, the triple sulfas in the above formulation is advantageously replaced by 250 gm of sulfamethyl-thiadiazole or 250 gm of sulfacetamide.
Composition Example 5 - Oral Syrup One thousand cc of an aqueous suspension for oral use, containing in each 5 cc dose 250 mg of U-57,930E or U-60,970E and 500 mg of total sulfas is prepared from the following types and amounts of ingredi-ents:
U-57,930E or U-60,970E 50 gm Sulfadiazine 33.3 gm Sulfamerazine 33.3 gm Sulfamethazine 33.3 gm Citric acid 2 gm Benzoic acid 1 gm Sucrose 700 gm Tragacanth 5 gm Lemon oil 2 cc Deionized water, q.s. 1000 cc The citric aid, benzoic acid, sucrose, tragacanthf and lemon oil are dispersed in sufficient water to make 850 cc of solution. The U-57,930E or U-60,970E and finely divided sulfas are stirred into the syrup until uniformly distributed. Sufficient water is added to make 1000 cc.
The composition so prepared is useful in the systemic treatment of pneumonia in adult humans at a dose of 1 tablespoonful (10 cc) 4 times a day.
Composition Example 6 - Parenteral Solution A sterile aqueous solution for intramuscular use, containing 20n 35 mg of U-57,930E or U-60,970E in 1 cc is prepared from the following types and amounts of materials:
U-57~930E or U-60,970E 200 gm - Lidocaine hydrochloride 4 gm 3 ~

Methylparaben 2.5 gm Propylparaben 0.17 gm Water for injection, q.s.19000 cc The ingredients are dissolved in the water and the solution sterilized by filtration. The sterile solution is filled into vials and the vials sealed.
Composition Example 7 - Parenteral Preparation A sterile aqueous composition for intramuscular use, containing in 1 cc 200 mg of U-57,930E or U-60,970E and ~00 mg of spectinomycin sulfate is prepared from the following types and amounts of ingre-dients:
U-57,930E or U-60,970E 200 gm Spectinomycin sulfate 400 gm Lactose 50 gm Water for injection, q.s.1,000 cc The U-57,930E or U-60,970E, spectinomycin sulfate, and lactose are dispersed in the water and sterilized. The sterile composition, in the amount of 2 cc, is filled aseptically into sterile vials.
Composition Example 8 - Top cal Ointment One thousand gm of 0.25% ointment is prepared from the following types and amounts of ingredients:
U-57,930E or U-60,970E 2.5 gm Zinc oxide 50 gm Calamine 50 gm Liquid petrolatum (heavy)250 gm Wool fat 200 gm White petrolatum, q.s. 1,000 gm The white petrolatum and wool fat are melted and lOn gm of liquid petrolatum added thereto. The U-57,93nE or U-60,970E, zinc oxide and calamine are added to the remaining liquid petrolatum and the mixture milled until the powders are finely divided and uniformly dispersed.
The powder mixture is stirred into the white petrolatum mixture and stirring continued until the ointment congeals.
The foregoing ointment is usefully applied topically to the skin of mammals for the treatment of infection.
The foregoing composition can be prepared by omitting the zinc oxide and calamine.
Following the procedure above, ointments are similarly prepared .

containing U-57,930E or U-60,970E in 0.5, l, 2, and 57~ amounts by substituting 5, 10, 20 and 50 gm of U-57,930E or U-60,970E for the 2.5 gm used above.
Composition Example 9 - Cream One thousand gm of a vaginal cream are prepared from the follow-ing types and amounts of ingredients:
U-57,930E or U-60,970E 50 gm Tegacid Regularl 150 gm Spermaceti 100 gm Propylene glycol 50 gm Polysorbate 80 5 gm Methylparaben 1 gm Deionized water, q.s. 1,000 gm 1Self-emulsifying glyceryl monostearate from Goldschmidt Chemical Corporation, New York, N.Y.
The Tegacid and spermaceti are melted together at a temperature of 70-80C. The methylparaben is dissolved in about 500 gm of water and the propylene glycol, Polysorbate ~30, and U-57,930E or U-60,970E
are added in turn, maintaining a temperature of 75-80C. The methyl-paraben mixture is added slowly to the -Tegacid and spermaceti melt, with constant stirring. The addition is continued for at least 30 minutes with continued stirring until the temperature has dropped to 40-45C. The pH of the final cream is adjusted to 3.5 by incorpor-ating 2.5 gm of citric acid and 0.2 g of dibasic sodium phosphate dissolved in about 50 gm of water. Finally, sufficient water is added to bring the final weight to 1,000 gm and the preparation stirred to maintain homogeneity until cooled and congealed.
The foregoing composition is useful for the treatment of vaginal infections in humans.
Composition Example 10 - Ointment, Ophthalmic One thousand gm of an ophthalmic ointment containing 0.5~ U-57,930E or U-60,970E are prepared from the following types and amounts of ingredients:
U-57,930E or U-60,97nE 5 gm Bacitracin 12~2 gm Polymyxin B sulfate (10,000 units/mg 1 gm Light liquid petrolatum250 gm ~ ~ ~3g3~

Wool fat 200 gm White petrolatum, q.s. 1,000 gm The solid ingredients are finely divided by means of an airmicronizer and added to the ligh liquid petrolatum. The mixture is passed through a colloid mill to uniformly distribute the micronized particles. The wool fat and white petrolatum are melted together, strained, and the temperature adjusted to 45-50C. The liquid petro-latum slurry is added and the ointment stirred until congealed.
Suitably the ointment is packaged in one dram ophthalmic tubes.
The foregoing ointment is usefully applied to the eye for treat-ment of localized infection in humans and other animals.
Advantageously the foregoing composition can contain 5 gm (0.5~0) of methylprednisolone for the treatment of inflammation, and, alter-natively, the bacitracin and polymyxin B sulfate can be omitted.
Compos tion Example 11 - Eye-Ear Drops One thousand cc of a sterile aqueous solution for eye or ear use containing 10 mg of U-57,930E or U-60,970E and 5 mg of methylpred-nisolone in each cc is prepared from the following types and amounts of ingredients:
U-57,930E or U-60,970E 10 gm Methylprednisolone phosphate sodium 5 gm Sodium citrate 4.5 gm Sodium bisulfite 1 gm Polyethylene glycol 400n120 gm Myristyl-y-picolinium chloride0.2 gm Polyvinylpyrrolidone 1 gm Deionized water, q.s. ad 1000 cc The ingredients are dissolved in the water and the resulting solution is sterilized by filtration. The solution is aseptically filled into sterile dropper containers.
The composition so prepared is useful in the topical treatment of inflammation and infection of the eye and ear as well as other sensi-tive tissues of the animal body.
Composition Example 12 - Troches Ten thousand troches are prepared from the following types and amounts of ingredients:
U-57~930E or U-60,970E 100 gm Neomycin sulfate 50 gm 1 ~ 63938 Polymyxin ~ sulfate (10,000 units/mg) 1 gm Ethyl aminobenzoate 50 gm Calcium stearate 150 gm Powdered sucrose, q.s. 5,000 gm The powdered materials are mixed thoroughly and then compressed into half gram troches following the usual techniques for the prep-aration of compressed tablets.
The troches are held in the mouth and allowed to dissolve slowly to provide treatment for the mouth and throat of humans.
Composition Example 13 - Suppository, Rectal One thousand suppositories, each weighing 2.5 gm and containing 100 mg of U-57,930E or U-60,970E are prepared from the following types and amounts of ingredients:
U-57,930E or U-60,970E lOO gm Polymyxin B sulfate (10,000 units/mg) 1.25 gm Methylprednisolone 1 gm Ethyl aminobenzoate 75 gm Zinc oxide 62.5 gm Propylene glycol 162.5 gm Polyethylene glycol 4,000 q.s. 2,500 gm The U-57,930E or U-60,970E, polymyxin B sulfate, methylpred-nisolone, ethyl aminobenzoate, and zinc oxide are added to the propyl-ene glycol and the mixture milled until the powders are finely dividedand uniformly dispersed. The polyethylene glycol 4000 is melted and the propylene glycol dispersion added slowly with stirring. The suspension is poured into unchilled molds at 40C.
The composition is allowed to cool and solidify and then removed from the mold and each suppository foil wrapped.
The foregoing suppositories are inserted rectally for local treatment of inflammation and infection.
Alternatively, the foregoing composition can be prepared omitting the steroid.
Composition Example 14 - Mastitis Ointment One thousand gm of an ointment for the treatment of mastitis in dairy cattle is prepared from the following types and amounts o, - ingredients:

~ 1 6393~

U-57,930E or U-60,970E25 gm Methylprednisolone acetate 0.5 gm Light liquid petrolatum 300 gm Chlorobutanol, anhydrous 5 gm Polysorbate 80 5 gm 2% Aluminum monostearate-peanut oil gel 400 gm White petrolatum, q.s. 1000 gm The U-57,930E or U-60,970E and methylprednisolone acetate are milled with the light liquid petrolatum until finely divided and uniformly dispersed. The chlorobutanol, polysorbate 80, peanut oil gel and white petrolatum are heated to 12nF to form a melt and the liquid petrolatum dispersion stirred in. With continued stirring, the dispersion is allowed to cool (and congeal) to room temperature and is filled into disposable mastitis syringes in 10 gm doses.
Composition Example 15 - Animal Feed One thousand gm of a feed mix is prepared from the following types and amounts of ingredients:
U-57,930E or U-60,970E10 gm Soybean meal 400 gm Fish meal 400 gm Wheat germ oil 50 gm Sorghum molasses140 gm The ingredients are mixed together and pressed into pellets~ The composition can be fed to laboratory animals, i.e., rats, mice, guinea pigs, and hamsters for prophylaxis during shipping.
For other animals such as poultry, e.g., chickens, ducks, tur-keys, and geese, the composition can be added to the animal's regular feed in an amount calculated to give the desired dose of U-57,93nE or U-60,970E.
Composition Example 16 Following the procedure of each of the preceding Composition Examples 1-15, inclusive, each antibacterially-active compound of the subject invention is substituted in an equivalent amount for the U-57,930E or U-60,970E shown in the example to provide therapeutic properties.
Similarly, each of the above free base compounds can be used in the form of a pharmaceutically (or pharmacologically) acceptable salt, ~ g 3 8 3713/3841 e.g., hydrochloride, sulfate, phosphoric, sodium, potassium, calcium,and lithium.
Composition Example 17 - Capsules One thousand two-piece hard gelatin capsules for oral use, each containing 200 mg of U-57,930E or U-60,970E and 200mg of hydroxy-chloroquine sulfate, are prepared from the following types and amounts of ingredients:
U-57,930E or U-60,970E 200 gm Hydroxychloroquine sulfate200 gm Talc 75 gm Magnesium stearate 25 gm The ingredients are thoroughly mixed and then encapsulated in the usual manner.
The foregoing capsules are useful to prevent recurrent attacks of P. vivax in adult humans by the oral administration of 1 capsule weekly.
Composition Example 18 - Tablets One thousand oral tablets, each containing 125 mg of U-579930E or U-60,970E and 325 mg of quinine sulfate, are prepared from the follow-ing types and amounts of materials:
U-579930E or U-60,970E 125 gm Quinine sulfate 325 gm Lactose 50 gm Corn starch 50 gm Calcium stearate 25 gm Light liquid petrolatum 5 gm The ingredients are thoroughly mixed and slugged. The slugs are broken down by forcing through a number 16 screen. The resulting granules are then compressed into tablets, each containing 125 mg of U-57,930E or U-60,970E and 325 mg of quinine sulfate.
The foregoing tablets are useful for treatment of malaria by the oral administration of two tablets every 8 hours for 7 days, then one tablet three times a day for 7 days.
Composition Example 19 - Oral Syrup One thousand cc of an aqueous suspension for oral use, containing in each 10 cc dose 25 mg of pyrimethamine, 250 mg of U-57,93nE or U-60,970E and 500 mg of sulfadiazine is prepared from the following types and amounts of ingredients:

'~ 1 63g38 , 3713/3841 U-57,930E or U-60,970E 25 gm Pyrimethamine 2.5 gm Sulfadiazine 50 gm Citric acid 2 gm ~enzoic acid 1 gm Sucrose 700 gm Tragacanth 5 gm Lemon oil 2 cc Deionized water, q~s. 1000 cc The citric acid, benzoic acid, sucrose, tragacanth, and lemon oil are dispersed in sufficient water to make 850 cc of solution. The U-57,930E or U-60,970E pyrimethamine and sulfadiazine are stirred into the syrup until uniformly distributed. Sufficient water is added to make 1000 cc.
The composition so prepared is useful in the prophylactic treat-ment of malaria in adult humans at a dose of 1 tablespoonful (10 cc) weekly.
Composition Example 20 - Parenteral Preparation A sterile aqueous composition for intramuscular use, containing in 1 cc 200 mg of U-57,930E or U-60,970E is prepared from the follow-ing types and amounts of ingredients:
U-57,930E or U-60,970E 200 gm Lactose 50 gm Water for injection, q.s. 1000 cc The U-57,930E or U-60,970E and lactose are dispersed in the water and sterilized. The sterile composition, in the amount of 2 cc, is filled aseptically into sterile vials.
Composition Example 21 Following the procedure of each of the preceding examples, each antimalarially-active compound of the subject invention is substituted in an equivalent amount for the U-57,930E or U-60,97nE shown in the example to provide therapeutic properties. Similarly, each of the above compounds can be used in the form of a pharmaceutically (or pharmacologically) acceptable salt, e.g., hydrochloride, sulfate, phosphoric, sodium, potassium, calcium, and lithium.

~ 3 63~38 In Vivo Results vs. P. ber~hei MEDI(mg/kg) ~so2(mg/k9) COMPOUND Sub~ Q3 OI Sub n _ OI
U-57,g30E 0.16 1.6 16 (12-22) >50 U-21,251F <20 __ 53 (46-61) --U-24,729A < 1.25 -- 4.7 (3.2-6.9) --U-8,284 Chloroquine < 5-10 12.5 11.5 (808-15) 14
10 Chloroquine (PO4)2 < 5 ~~ <20 --MED = Dosage at which median survival time (STso) was increased signiFicantly (p=0.05) over ST50 of untreated controls.
2 CDso = Median protective dose in mg/kg 95~0 limits.
3 Route of administration Anti-Malarial_Test (P. berghei~
Test method. Male, CF-1 mice (18-20 g) were housed in groups of and were infected intraperitoneally with whole blood from mice infected with P. berghei 3 days prior to bleeding. A n.2-ml amount of heparinized blood, diluted 1:10 with saline, served as the inoculum.
This volume contained approximately 106 parasites.
At 4 hr postinfection, each group of 10 mice was treated, either subcutaneously with 0.2 ml or orally by gavage, with n.5 ml of the desired drug concentration. Treatment was continued once each day for 4 days. The animals were observed for 28 days and deaths were recorded. Deaths prior to the 6th day were considered traumatic.
Evaluation for efficacy of the various analogues and drug con-centrations of individual analogues was based on the median survival time of animals at each treatment level and the median protective dose of the individual analogue. Calculations were computed on an IBM 370 digital computer. Results based on the treated groups were compared with those of untreated groups or groups treated with chloroquine.
Other protozoans within the concept of the subject invention process are intracellular parasites, for example~ the species of Plasmodia, Toxoplasma, and Leishmania; protozoa that digest the red blood cells (RBC's) of treated patients, for example, Entamoeba histolytica and certain Tryp nosoma; and other helminths which ingest ~ ~ 63938 RBC's during the disease processes, for example the Schistosomes.

~ ~ ~3938 ÇH3 LH,N H Ho_lCH3H

H~--N--- C-H
o Hl~ O\H

H~SCH3 H OH

Cl H3 CH3 HV N \IH H-C-Cl H~--N--C- H ( 2 ) H H O HIOk9\lH
H~SCH3 H OH

1 ~ 63938 3713/3841 C~ART 2 ~1 .
l ~---R2 N

wherein Rl, which can be singly or multiply substituted in any posi-tion of the pyridine ring not alreaqy substituted by R2, is selected from the group consisting of hydrogen, alkyl and substltuted alkyl wherein the alkyl portion is from 1 to 8 carbon atoms, inclus1ve, and isomeric forms thereof, cycloalkyl and substituted cycloalkyl, sub-stituted oxygen, substituted nitrogen, halogen, phenyl, and substi-tuted phenyl, -(CH2)m-OH, -(CH2~ -NR4R5, and isomeric forms l:hereof, wherein m is an integer of from 1 to 8, inclusive, R4 and R5 are H or alkyl of from 1 to 8 carbon atoms, inclusive, and isomeric forms thereof, wherein R2, which can be singly substituted in any position of the pyridine ring not already substituted by Rl, is . -C-X
and X is the amino function of a compound selected from the group 7(R)-hydroxy-methyl l-thio--lincosaminide, 7(S)-hydroxy-methyl 1-thio-a-lincosaminide~ 7(S)-halo-methyl 1-th1o-a-lincosaminide~ 7(R)-halo-methyl l-thiO-a-lincosaminide~ 7(S)-methoxy-methyl l-thio-a-lincosaminide, 7-deoxy-7(S)-(methylthio)-methyl 1-thio-a-lincos-aminide, 7-deoxy-7(S)-(2-hydroxyethylthio)-methyl 1-thio-a-lincos-aminide and 7-deoxy-7(S)-(3-hydroxypropylthio)-methyl l-thio-a-lincos-aminide, and the pharmaceutically acceptable salts thereof.

~CHa) n R1 ~R2 N ~

1 ,, ,~, i ~ 6~938 CHART 2 (cont.) wherein Rl and R2, which can be in the 2, 3, 4, 5, 6, 7, 8, or 9 position of the ring, are as defined above; wherein R3 is selected from the group consisting of H, CH3, C2Hs, and -CH2-CH2-OH; wherein n is an integer of from 1 to 4, inclusive; and the pharmaceutically acceptable salts thereof.

R ~ ~ R
E / A
B

wherein A, B and E are selected from the group consisting of nitrogen, oxygen, sulfur and CRlRl; Rl and R2 are as defined above, and can be attached to any ring carbon or nitrogen atom; Rl can be multiply attached to any ring carbon atom, and the pharmaceutically acceptable salts thereof.

E ~ B

wherein A, B, D and E are selected ~rom the group consisting of nitro-gen, oxygen, sulfur and CRlRl; Rl and R2 are as defined above and can be attached to any ring carbon or nitrogen atom; Rl can be multiply attached to any ring carbon atom, and the pharmaceutically acceptable salts thereof.

i ~ S3g3~ 3713/3g41 c. ~

5 ~3l H-C-Cl 10 1: R= H
OH

15 ~3 6 ~N'~ 8 1l 51 o 5~
20OH ~ 1 OH ~ 2 OH OH OH OH
OH ~H
N ~ NH 1 ~ ~ N

4: R= -P-O- CH~ 5: R= -P-O-CH2 30OH ~ 2l OH

OH OH OH OH
1: U-57930 2: U-57930 3-(5'-cytidylate) 3: U-57930 3-(5'-adenylate) 4: U-57930 3-(5'-uridylate) _: U-57930 3-(5'-guanylate)

Claims (56)

    THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
    PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

    1. A process for preparing the 3-(5'-ribonucleo-tide) of a compound having a general formula selected from the group consisting of:

    (I), (II), (III) and (IV) wherein for the compound of general formula I:
    R1, which can be singly or multiply substituted in the 2, 3, 4, 5 or 6 position of the pyridine ring not already substi-tuted by R2, represents a group selected from H, (C1-C8)-alkyl, substituted alkyl and isomeric forms thereof, cycloalkyl and substituted cycloalkyl, substituted oxygen, substituted nitrogen, halogen, phenyl and substituted phenyl, -(CH2)m-OH, -(CH2)m-NR4R5 and isomeric forms there-of, wherein R4 and R5 represent a group selected from H
    and (C1-C8)alkyl and isomeric forms thereof, and m is an integer of 1 to 8, inclusive; and R2, which can be singly substituted in any position of the pyridine ring not already substituted by R1, represents:
    -?-X

    Claim 1.....cont'd. (2) wherein X represents the amino function of a group selected from 7(S)-halo-methyl 1-thio-.alpha.-lincosaminide and 7(R)-halo-methyl 1-thio-.alpha.-lincosaminide;
    wherein for the compound of general formula II:
    R1, which can be singly or multiply substituted in the 2, 3, 4, 5, 6, 7, 8 or 9 position of the ring not already substituted by R2, is as defined above for the compound of general formula I;
    R2, which can be singly substituted in any position of the ring not already substituted by R1, represents:

    -?-X' wherein X' represents the amino function of a group of general formula:

    wherein R represents a group selected from H, Claim 1.....cont'd. (3) ;

    and R3 represents a group selected from H, -CH3, -C2H5 and -CH2-CH2-OH; and n is an integer of 1 to 4, inclusive;
    wherein for the compound of general formula III:
    R1, which can be singly substituted at any ring nitrogen atom, or singly or multiply substituted at any ring carbon atom not already substituted by R2, is as defined above for the compound of general formula I;
  1. Claim 1.....cont'd. (4) R2, which can be singly substituted at any ring nitrogen or carbon atom not already substituted by R1, is as defined above for the compound of general formula I; and A, B and E, independently, represent a group selected from nitrogen, N-CH3, oxygen, sulfur and CR1R1, wherein R1 is as defined above; and for the compound of general formula IV:
    R1, which can be singly substituted at any ring nitrogen atom, or singly or multiply substituted at any ring carbon atom not already substituted by R2, is as defined above for the compound of general formula I;
    R2, which can be singly substituted at any ring nitrogen or carbon atom not already substituted by R1 is as defined above for the compound of general formula I, and A, B, D and E, independently, represent a group selected from nitrogen, , oxygen, sulfur and CR1R1, wherein R1 is as defined above;
    said process comprising:
    cultivating streptomyces rochei, NRRL 3533, in an aqueous medium containing suitable nutrients, in the presence of a compound with a general formula I, II, III or IV, and recovering the desired 3-(5'-ribonucleotide) from the culti-vated medium; and, when required, producing a pharmaceutically acceptable salt of the 3-(5'-ribonucleotide).
  2. 2. The 3-(5'-ribonucleotide) of a compound having a general formula selected from the group consisting of:

    (I), (II), (III), and (IV) wherein R1, R2, R3, n, A, B, D and E are as defined in claim 1, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 1 or an obvious chemical equivalent thereof.
  3. 3. A process for preparing the 3-(5'ribonucleo-tide) of a compound of general formula:

    (I) wherein R1 and R2 are as defined in claim 1; said process comprising: cultivating streptomyces rochei, NRRL 3533, in an aqueous medium containing suitable nutrients, in the presence of the compound of general formula I, and recovering the desired 3-(5'-ribonucleotide) from the cultivated medium; and, when required, producing a pharmaceutically acceptable salt of the 3-(5'-ribonucleotide).
  4. 4. The process defined in claim 3, wherein for the compound of general formula I, R1 is in the 4-position and represents (C1-C8)alkyl and isomeric forms thereof, and R2 is in the 2 or 3- position.
  5. 5. The process defined in claim 3, wherein the compound of general formula I has the general formula:

    wherein R1, which can be singly or multiply substituted in the 3, 4, 5 or 6 position of the pyridine ring, is as defined in claim 3 and Y represents a group selected from 7(S)-halo and 7(R)-halo.
  6. 6. The process defined in claim 5, wherein Y
    represents 7(S)-halo.
  7. 7. The 3-(5'-ribonucleotide) of a compound of general formula:

    (I) wherein R1 and R2 are as defined in claim 3, and a pharma-ceutically acceptable salt thereof, when prepared by the process defined in claim 3 or an obvious chemical equivalent thereof.
  8. 8. The 3 (5'-ribonucleotide) defined in claim 7, wherein R1 is in the 4-position and represents (C1-C8)alkyl and isomeric forms thereof, and R2 is in the 2- or 3-position, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 4 or an obvious chemical equivalent thereof.
  9. 9. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein R1 and Y are as defined in claim 5, and a pharmaceut-ically acceptable salt thereof, when prepared by the process defined in claim 5 or an obvious chemical equivalent thereof.
  10. 10. The 3-(5'-ribonucleotide) defined in claim 9, wherein Y represents 7(S)-halo, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 6 or an obvious chemical equivalent thereof.
  11. 11. A process for preparing the 3-(5'-ribonucleo-tide) of a compound of general formula:

    (II) wherein R1, R2, R3 and n are as defined in claim 1, said pro-cess comprising: cultivating sreptomyces rochei, NRRL 3533, in an aqueous medium containing suitable nutrients, in the presence of the compound of general formula II, and recovering the desired 3-(5'-ribonucleotide) from the cultivated medium;
    and, when required, producing a pharmaceutically acceptable salt of the 3-(5'-ribonucleotide).
  12. 12. The process defined in claim 11, wherein for the compound of general formula II, R1 is in the 4-position and represents (C1-C4)alkyl and isomeric forms thereof.
  13. 13. The process defined in claim 11, wherein the compound of general formula II has the general formula:

    wherein R1, which can be singly or multiply substituted in the 3, 4, 5, 7, 8 or 9 position of the ring, is as defined in claim 11; R3 and n are as defined in claim 11; and Y represents a group selected from 7(S)-halo and 7(R)-halo.
  14. 14. The process defined in claim 13, wherein Y
    represents 7(S)-chloro.
  15. 15. The process defined in claim 13, wherein R1 represents (C1-C8)alkyl and isomeric forms thereof; R3 represents H; and Y represents 7(S)-halo.
  16. 16. The process defined in claim 15, wherein R1 represents -C2H5.
  17. 17. The process defined in claim 15, wherein R1 represents -C4H9.
  18. 18. The process defined in claim 15, wherein Y
    represents 7(S)-chloro.
  19. 19. The process defined in claim 16, wherein Y
    represents 7(S)-chloro.
  20. 20. The process defined in claim 17, wherein Y
    represents 7(S)-chloro.
  21. 21. The process defined in claim 11, wherein the compound of general formula II has the formula:

  22. 22. The process defined in claim 11, wherein the compound of general formula II has the formula:

  23. 23. The process defined in claim 11, wherein the compound of general formula II has the formula:

    wherein X is as defined in claim 11.
  24. 24. The process defined in claim 11, wherein the compound of general formula II has the fonmula.

    wherein X is as defined in claim 11.
  25. 25. The process defined in claim 11, wherein the compound of general formula II has the formula:

    wherein X is as defined in claim 11.
  26. 26. The process defined in claim 11, wherein the compound of general formula II has the general formula:

    wherein R represents:

  27. 27. The process defined in claim 11, wherein the compound of general formula II has the general formula:

    wherein R represents:

  28. 28. The process defined in claim 11, wherein the compound of general formula II has the general formula:

    wherein R represents:

  29. 29. The process defined in claim 11, wherein the compound of general formula II has the general formula:

    wherein R represents:

  30. 30. The 3-(5'-ribonucleotide) of a compound of general formula:

    (II) wherein R1, R2, R3 and n are as defined in claim 11, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 11 or an obvious chemical equivalent thereof.
  31. 31. The 3-(5'-ribonucleotide) defined in claim 30, wherein R1 is in the 4-position and represents (C1-C8)alkyl and isomeric forms thereof, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 12 or an obvious chemical equivalent thereof.
  32. 32. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein R1, R3, n and Y are as defined in claim 13, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 13 or an obvious chemical equivalent thereof.
  33. 33. The 3-(5'-ribonucleotide) defined in claim 32, wherein Y represents 7(S)-chloro, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 14 or an obvious chemical equivalent thereof.
  34. 34. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein R1, R3 and Y are as defined in claim 15, and n is an integer of 1 to 4, inclusive, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 15 or an obvious chemical equivalent thereof.
  35. 35. The 3-(5'-ribonucleotide) defined in claim 34, wherein R1 represents -C2H5, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 16 or an obvious chemical equivalent thereof.
  36. 36. The 3-(5'-ribonucleotide) defined in claim 34, wherein R1 represents -C4H9, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 17 or an obvious chemical equivalent thereof.
  37. 37. The 3-(5'-ribonucleotide) defined in claim 34, wherein Y represents 7(S)-chloro, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 18 or an obvious chemical equivalent thereof.
  38. 38. The 3-(5'-ribonucleotide) defined in claim 34, wherein R1 represents -C2H5 and Y represents 7(S)-chloro, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 19 or an obvious chemical equivalent thereof.
  39. 39. The 3-(5'-ribonucleotide) defined in claim 34, wherein R1 represents -C4H9 and Y represents 7(S)-chloro, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 20 or an obvious chemical equivalent thereof.
  40. 40. The 3-(5'-ribonucleotide) of a compound of formula:

    and a pharmaceutically acceptable salt thereof, when pre-pared by the process defined in claim 21 or an obvious chemi-cal equivalent thereof.
  41. 41. The 3-(5'-ribonucleotide) of a compound of formula:

    and a pharmaceutically acceptable salt thereof, when pre-pared by the process defined in claim 22 or an obvious chem-ical equivalent thereof.
  42. 42, The 3-(5'-ribonucleotide) of a compound of general formula:
    wherein X is as defined in claim 23, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 23 or an obyious chemical equivalent thereof.
  43. 43. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein X is as defined in claim 24, and pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 24 or an obvious chemical equivalent thereof.
  44. 44. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein X is as defined in claim 25, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 25 or an obvious chemical equivalent thereof.
  45. 45. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein R is as defined in claim 26, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 26 or an obvious chemical equivalent thereof.
  46. 46. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein R is as defined in claim 27, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 27 or an obvious chemical equivalent thereof.
  47. 47. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein R is as defined in claim 28, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 28 or an obvious chemical equivalent thereof.
  48. 48. The 3-(5'-ribonucleotide) of a compound of general formula:

    wherein R is as defined in claim 29, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 29 or an obvious chemical equivalent thereof.
  49. 49. A process for preparing the 3-(5'-ribonucleo-tide) of a compound of general formula:

    (III) wherein R1, R2, A, B and E are as defined in claim 1, said process comprising: cultivating streptomyces rochei, NRRL
    3533, in an aqueous medium containing suitable nutrients, in the presence of the compound of general formula III, and recovering the desired 3-(5'-ribonucleotide) from the culti-vated medium; and, when required, producing a pharmaceutically acceptable salt of the 3-(5'-ribonucleotide).
  50. 50. The process defined in claim 49, wherein the compound of general formula III has the formula:

  51. 51. The 3-(5'-ribonucleotide) of a compound of general formula:

    (III) wherein R1, R2, A, B and E are as defined in claim 49, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 49 or an obvious chemical equivalent thereof.
  52. 52. The 3-(5'-ribonucleotide) of a compound of formula:

    and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 50 or an obvious chemical equivalent thereof.
  53. 53. A process for preparing the 3-(5'-ribonucleo-tide) of a compound of general formula:

    (IV) wherein R1, R2, A, B, D and E are as defined in claim 1;
    said process comprising: cultivating streptomyces rochei, NRRL 3533, in an aqueous medium containing suitable nutrients, in the presence of the compound of general formula IV, and recovering the desired 3-(5'-ribonucleotide) from the cul-tivated medium; and, when required, producing a pharmaceu-tically acceptable salt of the 3-(5'-ribonucleotide).
  54. 54. The process defined in claim 53, wherein the compound of general formula IV has the formula:

  55. 55. The 3-(5'-ribonucleotide) of a compound of general formula:

    (IV) wherein R1, R2, A, B, D and E are as defined in claim 53, and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 53 or an obvious chemical equivalent thereof.
  56. 56. The 3-(5'-ribonucleotide) of a compound of formula:

    and a pharmaceutically acceptable salt thereof, when prepared by the process defined in claim 54 or an obvious chemical equivalent thereof.
CA000398814A 1981-04-20 1982-03-19 Process for treating a protozoan disease with lincomycin 3-(5'-ribonucleotides) Expired CA1163938A (en)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US255,542 1981-04-20
US06/255,542 US4383109A (en) 1981-04-20 1981-04-20 Lincomycin nucleotides
US06/255,541 US4368193A (en) 1981-04-20 1981-04-20 Process for treating malaria
US255,541 1988-10-07

Publications (1)

Publication Number Publication Date
CA1163938A true CA1163938A (en) 1984-03-20

Family

ID=26944757

Family Applications (1)

Application Number Title Priority Date Filing Date
CA000398814A Expired CA1163938A (en) 1981-04-20 1982-03-19 Process for treating a protozoan disease with lincomycin 3-(5'-ribonucleotides)

Country Status (15)

Country Link
KR (1) KR880002416B1 (en)
AU (1) AU545748B2 (en)
CA (1) CA1163938A (en)
CH (1) CH653037A5 (en)
DE (1) DE3213921A1 (en)
ES (1) ES511499A0 (en)
FR (1) FR2504142B1 (en)
GB (1) GB2097002B (en)
HU (1) HU191085B (en)
IL (1) IL65385A (en)
IT (1) IT1151719B (en)
NL (1) NL8201595A (en)
PL (1) PL133485B1 (en)
SE (1) SE459861B (en)
SU (1) SU1303036A3 (en)

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3671647A (en) * 1971-03-24 1972-06-20 Upjohn Co Lincomycin 3-nucleotides and the salts thereof

Also Published As

Publication number Publication date
KR830010195A (en) 1983-12-26
ES8308927A1 (en) 1983-10-01
GB2097002A (en) 1982-10-27
SE459861B (en) 1989-08-14
IL65385A0 (en) 1982-05-31
SU1303036A3 (en) 1987-04-07
NL8201595A (en) 1982-11-16
PL236043A1 (en) 1982-11-08
HU191085B (en) 1987-01-28
PL133485B1 (en) 1985-06-29
AU8198882A (en) 1982-10-28
AU545748B2 (en) 1985-08-01
IT1151719B (en) 1986-12-24
CH653037A5 (en) 1985-12-13
ES511499A0 (en) 1983-10-01
FR2504142B1 (en) 1985-07-26
IL65385A (en) 1985-10-31
DE3213921A1 (en) 1983-01-05
IT8220810A0 (en) 1982-04-19
FR2504142A1 (en) 1982-10-22
KR880002416B1 (en) 1988-11-08
GB2097002B (en) 1985-06-19
SE8202433L (en) 1982-10-21

Similar Documents

Publication Publication Date Title
EP0182315B1 (en) Novel antibiotic nk84-0218 pharmaceutical compositions containing it and process for the production of the same
PL96314B1 (en) METHOD OF MANUFACTURING THE ANTIBIOTIC THIENAMYCIN
US3857948A (en) Salinomycin
US4464466A (en) Process of producing lincomycin nucleotides
US4022889A (en) Therapeutic compositions of antibiotic U-44,590 and methods for using the same
US4383109A (en) Lincomycin nucleotides
HU179463B (en) Process for preparing deoxy-narasin antibiotic complex
EP0001567B1 (en) Thienamycin related antibiotics ps-6 and ps-7, process for their production and compositions containing them
US3751459A (en) L-trans-2-amino-4-(2-aminoethoxy)-3-butenoic acid
CA1163938A (en) Process for treating a protozoan disease with lincomycin 3-(5&#39;-ribonucleotides)
US4368193A (en) Process for treating malaria
US4069316A (en) Method for producing antibiotic T-42082 and antibiotic T-42082
PL80273B1 (en)
GB2111495A (en) Aminoglycoside antibiotics
US3207750A (en) Derivatives of decoyinine
EP0156193B1 (en) Compounds and compositions for treating protozoal infections with a novel antibiotic
US5286649A (en) Antiviral antibiotic BU-3889V
US5098708A (en) Antiviral antibiotic BU-3889V
CA1046965A (en) Naphthyridinomycin antibiotics from streptomyces
US4690926A (en) Boxazomycin A and B, new antibiotics containing benzoxazole nucleus
US5064855A (en) Acid polycyclic ether antibiotic
CS238399B2 (en) 19-epi-dianemycine production method
US5089478A (en) Method for the control of pneumocystis carinii
US4728612A (en) Boxazomycin A and B, new antibiotics containing benzoxazole nucleus
US5155097A (en) Polycyclic ether antibiotic having anthelmintic, anticoccidial and growth promotant activity

Legal Events

Date Code Title Description
MKEX Expiry