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WO1990009435A1 - Composes d'altromycine - Google Patents

Composes d'altromycine Download PDF

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Publication number
WO1990009435A1
WO1990009435A1 PCT/US1990/000694 US9000694W WO9009435A1 WO 1990009435 A1 WO1990009435 A1 WO 1990009435A1 US 9000694 W US9000694 W US 9000694W WO 9009435 A1 WO9009435 A1 WO 9009435A1
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WIPO (PCT)
Prior art keywords
compounds
altromycin
altromycins
compound
microorganism
Prior art date
Application number
PCT/US1990/000694
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English (en)
Inventor
James B. Mcalpine
Marianna Jackson
Robert J. Theriault
Gregory M. Brill
James P. Karwowski
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Abbott Laboratories
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Publication of WO1990009435A1 publication Critical patent/WO1990009435A1/fr

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    • 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
    • C12P19/44—Preparation of O-glycosides, e.g. glucosides
    • C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
    • C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms

Definitions

  • This invention relates to a mixture of compounds, and in particular to novel altromycin compounds having cytotoxic and antimicrobial activity, as well as to a process for making the new altromycin compounds.
  • the compounds of the present invention are related to, but distinct from, compounds of the pluramycin family which are described in U.S. 3,314,853; U.S. 3,334,016; EP 200,818; EP
  • Pluramycin compounds belong to the anthraquinone derivative class of compounds and have been found as metabolites of
  • the present invention comprises a fermentation isolate containing one or more altromycins which are represented by the following structural formula:
  • R 1 is selected from -NH 2 , -NHCH 3 , and -N(CH 3 ) 2
  • R 2 and R 3 are independently selected from hydrogen and hydroxyl, at least one of R 2 and R 3 being hydroxyl.
  • the present invention also comprises the individual altromycins and their pharmaceutically acceptable salts and ester thereof.
  • the inventive compounds are produced by a novel Nocardia-like Actinomycete microorganism which exhibits the identifying characteristics described in Tables 1-3.
  • Actinomycte sp. AB 1246E-26 produces altromycins upon culturing of said microorganism in a nutrient medium.
  • the compounds of the present invention are different from the pluramycin-type compounds in both the arrangement of the substituents on the chromophore and the nature of the glycosidic substituents.
  • the pluramycin is different from the pluramycin-type compounds in both the arrangement of the substituents on the chromophore and the nature of the glycosidic substituents.
  • the members of the pluramycin class have an amino sugar on carbon 8 and another amino sugar on carbon 10 of the D ring of the anthraquinone-gamma-pyrone nucleus.
  • the altromycins of the present invention have a neutral glycoside, a C-glycoside, attached via carbon 13 to carbon 5 of the B ring. Carbon 8 is unsubstituted in the
  • the altromycins have a disaccharide unit, containing a neutral O-glycoside attached to an amino sugar, on carbon 10 of the D ring. Accordingly, the
  • altromycins of the present invention contain neutral sugars which are not found in the pluramycin-type compounds. Moreover, the altromycins are the metabolites of a Nocardia-like organism.
  • FIGURE 1 shows a UV/visible spectrum of altromycin B recorded in acidic, neutral and basic methanol solutions.
  • the altromycins A, C, D , and G exhibit essentially identical spectra.
  • FIGURE 2 is an Infrared (IR) spectrum of altromycin B taken in deuteriochloroform solution (5%).
  • the altromycins A, C, and D exhibit essentially identical spectra.
  • FIGURE 3 is a 500 MHz 1 H NMR spectrum of altromycin A taken in deuteriochloroform (CDCI 3 ).
  • FIGURE 4 is a 500 MHz 1 H NMR spectrum of altromycin B taken in deuteriochloroform (CDCI 3 ).
  • the compounds and mixture of the compounds of the invention are made by cultivating the microorganism Actinomycete sp. AB 1246E-26 of the order Actinomycetales.
  • the microorganism produces branched vegetative hyphae typical of Actinomycetes.
  • the vegetative hyphae have a tendency to fragment into irregular, smaller, often bacillary, units, typical of Actinomycete species.
  • the strain Actinomycete sp. AB 1246E-26 was isolated from soil collected in South Africa. A subculture of the microorganism was deposited in the permanent collection of the Agricultural Research Service at Northern Regional Research Center, United States Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. The accession number for the
  • Actinomycete sp. strain AB 1246E-26 at the depository is NRRL 18371. Morpholo ⁇ y and Culture Characteristics:
  • the microorganism of the present invention Actinomycete sp. AB 1246E-26
  • Actinomycete sp. AB 1246E-26 can be characterized by morphology and other culture characteristics. Further, whole-cell hydrolysates can be used to predict cell wall composition, and the combination of morphology and chemical composition can be used to classify aerobic actinomycetes into groups according to their cell wall type. Lechevalier and Lechevalier, as reported in Inter. J. Syst. Bacteriol., 20:435-443, (1970), have developed such a
  • the nocardioform morphology, the chemical composition and the resistance to lysozyme of strain AB 1246E-26 are all
  • the meso isomer of diaminopimelic acid was found in whole cell hydrolysates. Chromatography of the sugars in the whole cell hydrolysates showed arabinose and galactose as diagnostic sugars.
  • the microorganism of this invention has a cell wall of Type IV, containing meso-2,6,-diaminopimelic acid and having a type A whole-cell sugar pattern (arabinose and galactose).
  • the chemical composition of the microorganism was also characterized by extraction and analysis of cellular menaquinones.
  • the principal menaquinone had a molecular weight of 720, as determined by mass spectrometry, indicating that the menaquinone is tetrahydrogenated with 8 isoprenoid units.
  • Lysozyme is an enzyme which attacks and cleaves the amino acid cross-linking of the aminohexose chains in the peptidoglycan layer of the cell walls of many but not all Gram-positive bacteria, and resistance to lysozyme is a
  • the color of the aerial growth of Actinomycete sp . AB 1246E-26 is white to light gray.
  • the microorganism makes a diffusible, non-melanoid pigment on several nutrient media.
  • the appearance and cultural characteristics of Actinomycete sp. AB 1246E-26 in various media are described in greater detail in Table 1.
  • the ability of the Actinomycete sp. AB 1246E-26 to grow on various carbon compounds in synthetic medium is shown in Table 2.
  • Physiological characteristics are given in Table 3.
  • ISP 3 AM Sparse, white
  • Peptone-yeast extract G Moderate
  • ISP 7 AM Yellowish white (92)
  • Gause#1 modified G Moderate
  • yeast extract 0.01%, agar 1.5%)
  • G growth
  • AM aerial mycelium
  • R reverse
  • the culture is grown in a culture medium which includes a source of carbon and a source of nitrogen.
  • Media which are useful include an assimilable source of carbon such as starch, sugar, molasses, glycerol, a combination of glucose plus molasses, etc.; an assimilable source of nitrogen such as protein, protein hydrolysate, polypeptides, amino acids, peptone plus yeast extract or whole yeast, etc.; and other organic and inorganic ingredients which can be added to stimulate production of the compounds such as, for example, inorganic anions and cations including potassium, magnesium, calcium, ammonium, sulfate, carbonate, phosphate, chloride, etc. Further, buffers such as calcium carbonate can be added to aid in controlling the pH of the fermentation medium.
  • Aeration can be provided by forcing sterile air through the fermentation medium.
  • Agitation can be provided by shaking the container or by stirring the culture, for example with a
  • the fermentation is generally carried out in a temperature range of from about 24oC to about 35oC.
  • the compounds of the invention are recovered from the whole broth by repeated solvent extraction with a water-immiscible solvent, such as methylene chloride.
  • a water-immiscible solvent such as methylene chloride.
  • This is the fermentation isolate of Actinomycete sp. AB 1246E-26 which shows antibacterial activity and which includes the compounds altromycin A, altromycin B, altromycin C, altromycin D, altromycin E, altromycin F, and altromycin G.
  • the compounds, altromycins A through G can be further purified by sequential counter-current chromatographic techniques as are well known in the art.
  • the isolation of the altromycin compounds can be monitored by TLC analysis of samples with appropriate viewing methods sensitive to the anthraquinone portion of the compounds.
  • TLC may be performed using normal silica EM Reagents HPTLC plates (silica gel 60 F 254 precoated 10 ⁇ 10 cm, E. Merck, Darmstadt, West).
  • the altromycin compounds are visualized in visible light as yellow-orange spots, under short wavelength UV light as dark blue spots on a light blue background and under long
  • UV light as bright orange spots on a moderately dark blue background.
  • the compounds are readily charred on these plates using a 5% cone.
  • Solvents used for development of the TLC plates range from 80-90% chloroform with methanol added to make up the remainder to 100%, depending on the particular sample to be run.
  • the altromycin compounds are consistently found to be in the Rf range of 0.2-0.6.
  • Load samples can be as high as 0.8 g of sample/10 mL silica on preparative runs.
  • a simple step elution gradient is employed, starting with 100% chloroform and incrementally increasing the methanol content until a concentration of 20% MeOH is reached.
  • the column fractions are collected and monitored for antibacterial activity by an agar disc diffusion assay.
  • a 20 microliter sample of each fraction is applied to a paper disc.
  • the discs are then placed on agar plates which were previously seeded with sufficient organism to provide a turbid background after incubation, typically 10 5 to 10 6 colony forming units (CFU) per mL.
  • CFU colony forming units
  • Actinomycete sp. AB 1246E-26 (NRRL 18371) was maintained as a frozen inoculum stock by freezing a portion of the original inoculum and storing it at -75oC.
  • the medium 5B7 (Table 4) was used for seed growth and the medium N2B1 (Table 5) was used for the fermentation.
  • Distilled water was added to achieve a volume of 1 L.
  • the pH was adjusted to pH 7.0.
  • the medium was prepared for seeding as follows : Ten mL of the seed medium (Table 4) were dispensed into 25 X 150 mm glass seed tubes. One hundred mL of the seed medium (Table 4) were dispensed into 500 mL Erlenmeyer flasks. The tubes were covered with stainless steel caps (Morton closures) and the flasks were plugged with rayon pharmaceutical coil. Thereafter, the tubes and flasks were sterilized for 35 minutes at 121oC, 15 psi.
  • Inoculum for the fermentation was prepared in two stages. In the first step, 5% of the frozen inoculum was inoculated into the seed tubes containing 10 mL of seed medium 5B7. The tubes were incubated for 96 hours at 28oC on a rotary shaker, operated at 250 rpm, with a stroke of 5.7 cm. In a second step, 5%
  • vegetative inoculum from the tube growth was used to inoculate the seed flasks containing 100 mL of seed medium 5B7.
  • the seed flasks were incubated for 72 hours at 28oC on a rotary shaker, operated at 250 rpm, with a stroke of 5.7 cm.
  • the fermentation broth (18 L) from Example 1 was adjusted to pH 10.0 with 6N HCl and extracted three times with 4 L portions of methylene chloride.
  • the methylene chloride extracts were combined and concentrated on a vertical evaporator to
  • the concentrate was partitioned in the solvent system MeOH-H 2 O-CCl 4 (5:2:5) using a droplet counter-current device with the lower phase as the stationary phase.
  • This device consists of 100 feet of continuous 3/16" ID (internal diameter) teflon tubing wrapped in a 2 X 14 cm repeating pattern around a 1 inch by 6 inch wooden plank.
  • the coil had 96 turns with a total internal volume of 450 mL, approximately one-half of this volume is retained as stationary phase.
  • Fraction A Fraction A
  • Fraction A concentrate (displaying antibacterial activity) was partitioned on a Coil Planet Centrifuge (CPC) counter-current device (P.C. Inc, Potomac, Maryland) employing the following conditions: MeOH-H 2 O-CCl 4 (5:2:5); lower phase stationary; tail as inlet; 4 mL/min flow rate at 800 rpm; 85-90% stationary phase retention; 10 mL fractions were collected.
  • CPC Coil Planet Centrifuge
  • Fraction A 1 fractions numbered 35-41
  • fraction A 2 fractions numbered 71-105
  • the concentrate from fraction A 2 224.7 mg was rechromatographed on the CPC.
  • the conditions for rechromatography of fraction A 2 were as follows : MeOH-0.01 M aqueous NH 4 OAC-CCI 4 (5:2:5); lower phase stationary; tail as inlet; 4 mL/min flow rate at 800 rpm; 85-90% stationary phase retention; 10 mL fractions were collected.
  • Fraction A 2a fractions numbered 16-21
  • Fraction A 2b fractions humbered 48-80
  • the oil obtained from concentrating the fraction A 2a was essentially identical to the material obtained from the fraction A 1 and was identified as altromycin A.
  • the fraction A 2b was found to contain altromycin B.
  • fractions Bi hexane-EtOAc-MeOH-H 2 O (3:7:6:4 lower phase stationary); tail as inlet, flow rate 4 mL/min at 800 rpm; 85-90% stationary phase retention; 8 mL fractions were collected.
  • fractions Bi One band of activity, fractions numbered 37-45 (hereinafter referred to as fractions Bi) was found which yielded 29.7 mg of orange-red oil after
  • the orange-red oil from fraction B 1 was identified as a mixture of altromycin C and altromycin D.
  • Altromycin C and altromycin D can be further separated on the counter-current device described above, using hexane-EtOAc-MeOH- 0.01 M NH 4 OAc.
  • Actinomycete sp . AB 1246E-26 (NRRL 18371) was maintained as a frozen inoculum stock by freezing a portion of the original inoculum and storing it at -75 C. Seed medium for an 80-liter stirred fermentation was prepared as follows :
  • the tubes were incubated for 96 hours at 28oC on a rotary shaker, operated at 225 rpm, with a stroke of 5.7 cm.
  • 5% vegetative inoculum from the tube growth was used to inoculate the seed flasks containing 600 mL of seed medium 5B7 (Table 4).
  • the seed flasks were incubated for 72 hours at 28oC on a rotary shaker, operated at 225 rpm, with a stroke of 5.7 cm.
  • N2B1 Eighty liters of production medium N2B1 (Table 5) were prepared in a 150-liter New Brunswick stainless steel stirred fermentor. The N2B1 medium was sterilized in the fermentor at 121oC and 15 psi for 1 hour. The glucose was sterilized
  • Antifoam was XFO 371 (made by Ivanhoe Chemical Co., Mundelein, IL). It was added initially at 0.01%, then was available on demand.
  • the fermentor was inoculated with 4 liters of the 2nd step seed growth. The temperature was controlled at 28oC. The agitation rate was 200 rpm and the air flow was 0.7 vol/vol/min. A head pressure of 5 psi was maintained. The fermentation was terminated at 120 hours. The harvest volume was 61 liters.
  • the fermentation broth (61 L) from Example 3 was charged into a 200 liter stainless steel roller barrel (Morse).
  • the broth was extracted four times with 20-24 L portions of methylene chloride or chloroform, rolling for periods of 30-45 minutes and draining off the organic solvent.
  • the chlorinated solvent extracts were combined and concentrated on a vertical evaporator to a volume of 1 L. This volume was reduced to approximately 100 ml of oil on a roto-evaporator and then partitioned several times between methanol and n-heptane (1 :2).
  • the combined heptane layers did not contain altromycin-type compounds when analyzed by TLC.
  • the methanolic layer was concentrated in vacuo and and partitioned several times between pH 10.0 (NH4OH) water and chloroform.
  • the PDCC device with the lower phase as the stationary phase.
  • the PDCC device was made in-house and consisted of 60 meters of continuous teflon tubing. The tubing alternated in diameter and was coiled in a 3 ⁇ 28 cm repeating pattern (the tubing being essentially straight except for sharp bends of the smaller
  • the coil had 100 loops with an approximate stationary phase retention volume of 450 ml on standing. 90-95% of this volume of stationary phase was retained during use.
  • the coil had a mobile phase volume of approximately 70 ml on standing. The coil was run at a rate of 3-4 ml/min, collecting 10-15 ml fractions. Two nearly identical runs were made on a 50% portion of sample. Based on TLC analysis of the fractions, two pools of altromycin-type compounds were obtained from each run.
  • Fraction A4 The first 100 fractions, excluding the initial twenty or so which contained significant impurities and little if any altromycin-types, were pooled (hereinafter referred to as Fraction A4).
  • Fraction B4 The second ⁇ 100 fractions, which included the stationary blow-out, also were pooled (hereinafter referred to here as Fraction B4). Partitioning of Fraction A4
  • Fraction A4 1 Fraction A4 1
  • CPC Centrifuge
  • P.C. Inc. Potomac
  • Fraction B4 was concentrated and run on the PDCC device (2 runs with 1/2 sample load) using the solvent system MeOH-H 2 O-n-heptane-CCl 4 (5:2:2:3). From each of these runs the first ⁇ 100 fractions (excluding the initial ⁇ 6), hereinafter referred to as Fraction B4 1 , were combined on the basis of TLC analysis. These fractions were concentrated for further purification using a CPC as outlined in SCHEME 2, below. This portion of the isolation scheme resulted in the isolation of altromycin F (7.8 mg), altromycin A ( ⁇ 400 mg), altromycin B ( ⁇ 1 g), altromycin C ( ⁇ 50 mg) and altromycin D ( ⁇ 100 mg).
  • the altromycins of this invention can be dried to a glass and are orange-red compounds which are readily soluble in methanol (MeOH), chloroform (CHCl 3 ), carbon tetrachloride (CCI 4 ), dimethyl sulfoxide (DMSO) , ethyl acetate (EtOAc), benzene or dilute acid; moderately soluble in 0.01M phosphate buffer (pH 7); and slightly soluble in n-hexane or water.
  • MeOH methanol
  • CHCl 3 chloroform
  • CCI 4 carbon tetrachloride
  • DMSO dimethyl sulfoxide
  • EtOAc ethyl acetate
  • benzene or dilute acid moderately soluble in 0.01M phosphate buffer (pH 7); and slightly soluble in n-hexane or water.
  • HPLC High Performance Liquid Chromatography
  • altromycin A had a retention time of 10.8 minutes and altromycin B a retention time of 12.8 minutes.
  • altromycin C was 896.3686 (calculated 896.3705), corresponding to a molecular formula of C 46 H 57 NO 17
  • the (M+H) + for altromycin D was 910.3868 (calculated 910.3861), corresponding to a molecular formula C 47 H 59 NO 17
  • the (M+H) + parent mass of altromycin E was
  • UV spectroscopic data were essentially identical for altromycins A, B, C ,D and G and place them in the anthraquinone derivative class of compounds.
  • the UV absorption maxima in basic solution are at approximately 490 nm, 368 nm, 304 nm and 252 nm.
  • the UV absorption maxima in neutral solution are at 424 nm, 242 nm, and 212 nm .
  • the UV absorption maxima in acidic solution are at 424 nm, 276 nm, 243 nm, and 215 nm .
  • the UV spectra of altromycins E and F are es sentially
  • NMR spectroscopic data (Tables 7 and 8 and FIGURES 3 and 4 ) establish that altromycins A, B, C, D, E, F, and G are a new group of compounds in the anthraquinone derivative class of compounds .
  • Altromycins E and F are like altromycins A and B respectively, with the exception that altromycins E and F are each found to have a proton on carbon 13 replacing the hydroxyl functionality .
  • Altromycin G differs from altromycins A and B in that carbon 4" of the amino sugar bears a primary amine as opposed to a mono- or dimethyl amine.
  • 13 C NMR chemical shift data and 1 H NMR chemical shift/coupling data are listed in Tables 7 and 8, respectively.
  • BHI Brain Heart Infusion
  • Streptococcus dilutions of overnight cultures of up to 32 different microorganisms, using a Steers replicator block
  • CFU CFU
  • Ciprofloxacin was used as a reference antibacterial agent. After incubation, each agar plate was observed for the presence or absence of microorganism growth. The MIC was defined as the lowest concentration of test compound yielding no growth. A slight haze caused by the inoculum spot as compared to growth on a control plate containing no test compound was disregarded.
  • Actinetobacter sp. CMX 669 100 >100 25 Enterobacter aerogenes ATCC 13048 >100 >100 >100 Enterococcus faecium ATCC 8043 6.2 3.12 3.1 Escherichia coli DC-2 >100 >100 100 Escherichia coli H560 >100 >100 100 Escherichia coli JUHL >100 >100 100 Escherichia coli KNK 437 25 25 25 25 25 25
  • the compounds of the present invention exhibitied potent in vitro activity against both human and murine cell types.
  • the cytotoxity of altromycins A, B, and C+D is against HeLa cells is illustrated in Table 10 and the cytotoxicity of the altromycins A, B, and D against human and murine tumor cell lines is illustrated in Table 11.
  • the inhibitory concentrations capable of killing 50% of the cells were determined in a colorimetric assay for cytotoxic activity against cultured cells according to the
  • a three day microtiter assay was used to measure the metabolic activity of cultured cells exposed to a range of drug concentrations. Metabolic activity was measured by the cell's ability to reduce the tetrazolium dye, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), to a quantifiable colored formasan derivative.
  • MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
  • Test compounds were dissolved in dimethyl sulfoxide (DMSO) and diluted, first with Earle's Balanced Salt Solution, followed by culture medium, to twice the highest concentration of the compound to be tested. From this concentrated stock, two-fold serial dilutions were prepared in 96-well microtiter trays, each well containing twice the desired final concentration of the compound. Each concentration was tested in triplicate and
  • the cells were grown in the same medium used for diluting the compounds. After harvesting by trypsinization as described in Bird, B.R. & F.T. Forrester, Basic Laboratory Techniques in Cell Culture, U.S Dept. of Health & Human Services, Virology Training Branch, Atlanta Georgia, pp 84-85, 104-105, 111 (1981), viable cell counts were determined and cell density was adjusted to
  • Inoculum (0.1 mL) was then added to each well for a final concentration of 2,500 cells per well. Addition of the inoculum was used to dilute the test compounds to the desired final concentration.
  • Microtiter trays were incubated for three days at 36oC in a humidified atmosphere containing 5% carbon dioxide.
  • MEM Minimum Essential Medium
  • the in vivo antitumor activities of the altromycin and D were determined using the ascitic leukemia tumor line P388 . Prior to testing, P388 was propagated intraperitoneally in female DBA/2 mice. The ascitic tumors were intraperitoneally injected in female B 6 D 2 F1 mice at a concentration of 10 6 cells per mouse. The mice were treated with antitumor agents administered once daily for five consecutive days, with fluorouracil being used as a positive control.
  • All drugs were administered intraperitoneally in 5% aqueous dextrose or other suitable diluent. Drug efficacy was determined by prolonged survival. Survival time was calculated as %T/C, the mean or median survival time of treated mice expressed as a percentage of the mean or median survival time of control mice. Compounds having %T/C values greater than 125 were considered to have significant activity. Treatment groups contained 10 mice and control groups contained 20 mice each. The toxicity of antitumor agents in these tests were evaluated by comparing the weight of mice prior to treatment and 5 days after treatment. LD 50 values for single intraperitoneal administration were 0.3, 0.2 and 0.1 mg/kg for altromycins A, B and D, respectively. The altromycins demonstrated reproducible activity in the P388 systemic leukemia model as illustrated in Table 12.
  • QD1-5 Test compounds administered daily for 5 consecutive days.
  • D1,D5 Test compound administered daily at day 1, day 5 only.
  • Altromycins A, B, and D were also tested against two solid tumors, namely, Lewis lung carcinoma and M5076 Ovarian sarcoma. Prior to testing these tumors were propagated subcutaneously in female C57B1/6 mice. Lewis lung carcinoma or M5076 (solid) tumors were implanted subcutaneously as a 1:10 brei in female B 6 D 2 F 1 mice weighing 16-20 grams. For mice implanted with Lewis lung
  • test compounds were administered daily for 9 consecutive days. Cyclophosphamide was used as the positive control drug. For mice implanted with M5076 (solid) tumors, test compounds were administered 4 times daily for 11 consecutive days . Cisplatinum was used as a positive control. All drugs were administered intraperitoneally in 5% aqueous dextrose, Drug efficacy was determined by reduction in tumor weight. Tumor weight inhibition was expressed as a function of T/C, the ratio of tumor weight of the treated mice to the tumor weight of control mice, and is illustrated in Tables 13 and 14, below. Altromycin D produced nearly 90% tumor weight inhibition as compared to
  • altromycin A was slightly more active than altromycin D (Table 14).
  • Cyclophosphamide was administered once on day 1.
  • Altromycins B and D were tested against a human tumor xenograft in the subrenal capsule assay (SRCA) as des-cribed by Bogden et al., Cancer 48:10-20 (1981).
  • Human colon tumor line, LS174T was maintained by serial propagation in nude, athymic BALB/C mice. Tumors were removed from the nude mice after reaching 0.2-0.3 grams and were minced into 1 mm 3 pieces. The tumor fragments were then implanted into the renal capsule of CDF1 mice. Immediately after implantation the tumor mass was measured with a steroscopic dissecting microscope fitted with an ocular micrometer calibrated in ocular units (OMU). The mean tumor mass in OMU was calculated.
  • the compounds of the present invention may be administered orally, nasally, opthalmically, parenterally, vaginally, rectally, topically or in an aerosol, in dose unit formulations containing conventional nontoxic pharmaceutically acceptable carriers, adjuvants and vehicles, as desired.
  • parenteral includes subcutaneous, intravenous, or infusion techniques.
  • sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic
  • diluent or solvent for example, as a solution in 1,3-butanediol.
  • acceptable vehicles and solvents water, Ringer's solution and isotonic sodium chloride solution.
  • sterile fixed oil may be employed including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid find use in the
  • Suppositories for rectal or vaginal administration of the active ingredient may be prepared by mixing the active ingredient with a suitable, nonirritating excipient such as cocoa butter and polyethylene glycols which are a solid at room temperatures but liquid at the rectal/vaginal temperatures and will therefore melt in the rectum or vagina and release the active ingredient.
  • a suitable, nonirritating excipient such as cocoa butter and polyethylene glycols which are a solid at room temperatures but liquid at the rectal/vaginal temperatures and will therefore melt in the rectum or vagina and release the active ingredient.
  • Solid dosage forms for the oral administration may include capsules, tablets, pills, powders and granules.
  • the active ingredient may be admixed with at least one inert diluent such as sucrose, lactose or starch.
  • Such dosage forms may also comprise, as is normal practice, additional substances other than inert diluents, e.g., lubricating agents such as magnesium stearate.
  • the dosage forms may also comprise buffering agents.
  • Tablets and pills can additionally be prepared with enteric coatings.
  • Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixers containing inert diluents commonly used in the art, such as water. Such compositions may also comprise
  • adjuvants such as wetting agents, emulsifying and suspending agents and sweetening, flavoring and perfuming agents.
  • the compounds of the invention can be used for the following reasons.
  • therapeutically treating a mammalian host affected by a microbial infection or an experimental animal host affected by a malignant tumor are useful for the topical treatment of infectious diseases. Additionally, they can be used in disinfectant and sterilizing solutions or as preservatives in plastics, paints or fabrics.
  • Total daily doses administered to a patient in single or divided doses can be in amounts, for example, 0.01 to 50 mg, and more usually 0.01 to 10.0 mg.
  • Dosage unit compositions may contain submultiples thereof to make up the daily dose.
  • the amount of the active ingredient that can be combined with the carrier materials to produce a single dosage form will vary depending upon the patient treated and the particular mode of administration.
  • the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed; the age, body weight, general health, sex and diet of the patient; the time of administration; the route of administration; the rate of excretion of the compound; drugs used in combination and the severity of the particular disease undergoing therapy.
  • the present invention includes the manufacture of the altromycin compounds it is not limited to the use of

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  • Molecular Biology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

Un mélange de nouveaux composés d'altromycine est produit par un microorganisme appartenant à l'ordre des Actinomycetales. Les composés présentent une activité cytotoxique contre des lignées de cellules tumorales mammifères, ainsi qu'une activité antibactérienne contre un large spectre de bactéries ''Gram positif''.
PCT/US1990/000694 1989-02-07 1990-02-07 Composes d'altromycine WO1990009435A1 (fr)

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US30755589A 1989-02-07 1989-02-07
US307,555 1989-02-07

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WO1990009435A1 true WO1990009435A1 (fr) 1990-08-23

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PCT/US1990/000694 WO1990009435A1 (fr) 1989-02-07 1990-02-07 Composes d'altromycine

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Country Link
EP (1) EP0447494A4 (fr)
GR (1) GR900100078A (fr)
PT (1) PT93070A (fr)
WO (1) WO1990009435A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0452683A3 (en) * 1990-03-16 1991-12-27 Sapporo Breweries, Ltd. Streptomyces antitumor compounds and process for preparing the same
WO1994018189A1 (fr) * 1993-02-08 1994-08-18 Glycomed Incorporated Derives d'anthraquinone et d'anthracene inhibant les molecules d'adhesion cellulaire du systeme immunitaire
EP1125944A1 (fr) * 2000-02-18 2001-08-22 Aventis Pharma Deutschland GmbH Pluraflavine et ses dérivés, procédé de préparation et utilisation

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3314853A (en) * 1964-08-25 1967-04-18 Squibb & Sons Inc Rubiflavin and a process for making same using streptomyces griseus
US3334016A (en) * 1966-10-24 1967-08-01 Bristol Myers Co Hedamycin and process for its production
US4245047A (en) * 1977-03-31 1981-01-13 Takeda Chemical Industries, Ltd. Antibiotics produced from the microorganism nocardia
US4368265A (en) * 1980-01-30 1983-01-11 Hoffmann-La Roche Inc. Culturing a strain of nocardia to produce antibiotic X-14868A
US4389486A (en) * 1977-03-24 1983-06-21 Kowa Company, Ltd. Biologically pure culture
US4605624A (en) * 1982-10-21 1986-08-12 Pfizer Inc. Nocardia species capable of producing nargenicin C1

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS61137882A (ja) * 1984-12-06 1986-06-25 Microbial Chem Res Found 新規抗腫瘍性物質及びその製造方法

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3314853A (en) * 1964-08-25 1967-04-18 Squibb & Sons Inc Rubiflavin and a process for making same using streptomyces griseus
US3334016A (en) * 1966-10-24 1967-08-01 Bristol Myers Co Hedamycin and process for its production
US4389486A (en) * 1977-03-24 1983-06-21 Kowa Company, Ltd. Biologically pure culture
US4245047A (en) * 1977-03-31 1981-01-13 Takeda Chemical Industries, Ltd. Antibiotics produced from the microorganism nocardia
US4315989A (en) * 1977-03-31 1982-02-16 Takeda Chemical Industries, Ltd. Biologically pure culture of the microorganism Nocardia ATCC31280
US4368265A (en) * 1980-01-30 1983-01-11 Hoffmann-La Roche Inc. Culturing a strain of nocardia to produce antibiotic X-14868A
US4605624A (en) * 1982-10-21 1986-08-12 Pfizer Inc. Nocardia species capable of producing nargenicin C1

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP0447494A4 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0452683A3 (en) * 1990-03-16 1991-12-27 Sapporo Breweries, Ltd. Streptomyces antitumor compounds and process for preparing the same
US5168100A (en) * 1990-03-16 1992-12-01 Sapporo Breweries Limited Hp530 compounds and process for preparing the same
WO1994018189A1 (fr) * 1993-02-08 1994-08-18 Glycomed Incorporated Derives d'anthraquinone et d'anthracene inhibant les molecules d'adhesion cellulaire du systeme immunitaire
US5412123A (en) * 1993-02-08 1995-05-02 Glycomed Incorporated Anthraquinone and anthracene derivatives as inhibitors of the cell-adhesion molecules of the immune system
EP1125944A1 (fr) * 2000-02-18 2001-08-22 Aventis Pharma Deutschland GmbH Pluraflavine et ses dérivés, procédé de préparation et utilisation
WO2001060832A3 (fr) * 2000-02-18 2002-01-31 Aventis Pharma Gmbh Pluraflavines et leurs derives, processus de preparation et utilisation correspondante
US6500936B2 (en) 2000-02-18 2002-12-31 Aventis Pharma Deutschland Gmbh Pluraflavins and derivatives thereof, process for their preparation and use thereof
RU2255940C2 (ru) * 2000-02-18 2005-07-10 Авентис Фарма Дойчланд Гмбх Плюрафлавины, способ их получения и композиция на их основе
KR100743380B1 (ko) * 2000-02-18 2007-07-30 사노피-아벤티스 도이칠란트 게엠베하 플루라플라빈, 이의 제조 방법 및 이를 포함하는 약제학적 조성물
HRP20020672B1 (en) * 2000-02-18 2010-12-31 Sanofi-Aventis Deutschland Gmbh Pluraflavins and derivatives thereof, process for their preparation and use thereof

Also Published As

Publication number Publication date
EP0447494A4 (en) 1992-01-22
EP0447494A1 (fr) 1991-09-25
GR900100078A (el) 1991-06-28
PT93070A (pt) 1990-08-31

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