JP2556893B2 - Murademycin derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Object of the Invention] The present invention relates to a novel compound, a mulaidomycin derivative.

The present inventors previously applied for Murademycin A, B, C, D, E and F (Japanese Patent Application Laid-Open No. 63-211295).
115639), JP-A-63-218685 (Japanese Patent Application No. 61-137567)
No.) and application filed on November 17, 1988 “Muredomycin.
E, F and its derivatives ").

This time, the inventors of the present invention have examined the results of hydrolysis of mureidomycin A, B, C, D, E and F, and their salts, and the results of mureidomycin AP, BP, CP, DP, EP and FP, and these. The present invention has been completed by finding that a salt has an antibacterial action and can be an intermediate for other useful compounds.

[Structure of Invention]

The present invention is a formula The present invention relates to mulaidomycin AP, BP, CP, DP, EP and FP, salts thereof, and processes for producing them.

Here, mureidomycin AP has R ¹ of 2,4-dioxopyrimidin-1-yl. And R ² is α-amino-3-hydroxyphenethyl Mreidomycin BP has R ¹ of 2,4-dioxodihydropyrimidin-1-yl And R ² is α-amino-3-hydroxyphenethyl; mureidomycin CP is R ¹ is 2,4-dioxopyrimidin-1-yl and R ² is α-glycylamino-3-.
Hydroxy phenethyl Murademycin DP is R ¹ is 2,4-dioxodihydropyrimidin-1-yl, R ² is α-glycylamino-3-hydroxyphenethyl; Murademycin EP is R ¹ is 2,4-dioxopyrimidin-1-yl, wherein R ² is 8-hydroxy-1,2,3,4-tetrahydroisoquinolin-3-yl Murademycin FP has R ¹ of 2,4-dioxopyrimidin-1-yl and R ² of 6-hydroxy-1,2,3,
It is 4-tetrahydroisoquinolin-3-yl.

Examples of the salt of the compound having the general formula (I) include inorganic salts such as hydrochloric acid, sulfuric acid and phosphoric acid; acetic acid, citric acid, tartaric acid, malonic acid, maleic acid, malic acid, fumaric acid, itaconic acid, Examples thereof include salts with organic carboxylic acids such as citraconic acid and succinic acid; organic sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid and p-toluenesulfonic acid.

Murademycin AP, BP, C having the above general formula (I)
P, DP, EP and FP and their salts are all novel compounds of the formula And the salts thereof (wherein mureidomycin A, B, C, D, E and F)
R ¹ and R ² are the corresponding mureidomycin AP, BP, CP, D
It has the same meaning as R ¹ and R ² of P, EP and FP. ) Is hydrolyzed.

The reaction is carried out in the presence or absence of a solvent in the above general formula (I
This is accomplished by contacting the compound having I) with an acid, base or proteolytic enzyme.

As the acid or base used, an acid or base used in a usual hydrolysis reaction is used without particular limitation, and examples thereof include mineral acids such as hydrochloric acid, sulfuric acid and hydrobromic acid, or sodium hydroxide, potassium hydroxide, Alkali metal and alkaline earth metal hydroxides such as barium hydroxide; alkali metal and alkaline earth metal carbonates such as sodium carbonate, potassium carbonate and calcium carbonate are preferably used. It is preferable to use a solvent to smoothly carry out the reaction, and the solvent to be used is not particularly limited as long as it does not participate in the reaction, and examples thereof include water or water and alcohols such as methanol and ethanol, or tetrahydrofuran, dioxane and the like. A mixed solvent with the above ethers is preferably used. The reaction temperature is not particularly limited, and the reaction is carried out from room temperature to the reflux temperature of the solvent. However, this reaction is preferably carried out by contacting with a proteolytic enzyme. Examples of the proteolytic enzyme used include pepsin, cathepsin, pronase, chymotrypsin and the like, with pepsin being particularly preferred. For the reaction, water or a mixed solvent of water and an alcohol such as methanol or ethanol or a sulfoxide such as dimethyl sulfoxide is preferably used. The reaction is diluted hydrochloric acid at pH 2.5.
In the reaction solution adjusted to the front and back, the reaction is carried out for 10 hours to 3 days at a temperature at which normal enzyme reaction proceeds, for example, 10 ° C to 60 ° C, especially about 37 ° C.

After completion of the reaction, the target compound is separated, collected and purified by utilizing its physicochemical properties. That is,
Solvent extraction; ion exchange, eg Dowex SBR-P
Anion exchange resins such as (made by Dow Chemical Co.), cation exchange resins such as Dowex 50W (made by Dow Chemical Co.), IRC-50, CG-50; activated carbon as an adsorbent, Amberlite XAD-2, XAD-4 , XAD-7 etc. (made by Rohm and Haas) and Diaion HP10, HP20, CHP20P, HP50 (made by Mitsubishi Kasei Co., Ltd.) and other non-ionic adsorption resins such as resins, silica gel, alumina treatment Is also used as appropriate. In addition, cellulose such as Avicel (manufactured by Asahi Kasei Co., Ltd.), partition column chromatography using Sephadex LH-20 (manufactured by Pharmacia): Sephadex G-10, G-25, G-50, G-100 ( (Falmatia Co., Ltd.) or Toyopearl HW-40 (Seikagaku Corporation) gel permeation method; crystallization; recrystallization and the like are also effective, and these means may be used alone or in any order, Repeatedly used for separation, collection, and purification.

When muredomycin AP, BP, CP, DP, EP and FP are separated in salt form, they can be converted to the free form by conventional methods using, for example, ion exchange resin or adsorption reverse phase chromatography. it can. Similarly, the free compound can be converted into a salt form by a conventional method. For example, by contacting with a salt-forming acid in water or an aqueous organic solvent.

Muradeomycin AP, BP, CP, D thus obtained
P, EP and FP and their salts are used as antibacterial agents for controlling various bacterial infectious diseases. Examples of the administration form include parenteral administration methods such as subcutaneous injection, intravenous injection, intramuscular injection, and suppository, and oral administration methods such as tablets, capsules, powders, and granules. The dose varies depending on the disease to be treated, the route of administration, the number of administrations, and the like, but for example, it is usually preferable to administer 0.1 g to 10 g per day once or in several divided doses to an adult.

Next, the present invention will be described more specifically with reference to Examples and Reference Examples.

Example 1 Murademycin AP 100 mg of Murademycin A was dissolved in 50 ml of distilled water and adjusted to pH 2.5 by adding an aqueous solution of N-hydrochloric acid, 200 mg of pepsin (manufactured by Boehringer Mannheim) was added, and the mixture was stirred overnight at 37 ° C. did. After the reaction was completed, 1N aqueous sodium hydroxide solution was added to the reaction mixture to adjust the pH to 5.0, and then Diaion C
It was adsorbed on 70 ml of HP20P (manufactured by Mitsubishi Kasei Co., Ltd.). The column was washed with 350 ml of distilled water and then eluted with 210 ml of 0.1% trifluoroacetic acid aqueous solution. The eluate was concentrated and subjected to freeze-drying to obtain 34 mg of mureidomycin AP.

1) Property of substance: Amphoteric water-soluble, white powder 2) Molecular formula: C ₂₃ H ₃₀ N ₆ O ₇ 3) Molecular weight: 502 4) Thin layer chromatography: R _f value; 0.13 Adsorbent; Silica gel plate (Merck) Made by Kieselgel 60
F ₂₅₄ ) Developing solvent; n-butanol: acetic acid: water (4: 1: 2) 5) High performance liquid chromatography: Column: ODS H2151 (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent: 5% acetonitrile-0.1% trifluoroacetic acid Aqueous solution Flow rate: 1.0 ml / min Retention time: 8.3 min. Example 2 Murademycin BP Murademycin B 100 mg and pepsin 200 mg were used to carry out a reaction for 24 hours in the same manner as in Example 1 to obtain the target compound 15
mg was obtained.

1) Property of substance: amphoteric water-soluble, white powder 2) Molecular formula: C ₂₃ H ₃₂ N ₆ O ₇ 3) Molecular weight: 504 4) Thin layer chromatography: R _f value; 0.13 Adsorbent; Silica gel plate (Merck) Made by Kieselgel 60
F ₂₅₄ ) Developing solvent; n-butanol: acetic acid: water (4: 1: 2) 5) High-performance liquid chromatography: column; ODS H2151 (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent; 5% acetonitrile-0.1% trifluoroacetic acid Aqueous solution Flow rate: 1.0 ml / min Retention time: 8.1 min Example 3 Murademycin CP 100 mg of mureidomycin C and 200 mg of pepsin were used and the reaction was carried out for 48 hours in the same manner as in Example 1 to obtain the target compound 28.
mg was obtained.

1) Property of substance: Amphoteric water-soluble, white powder 2) Molecular formula: C ₂₅ H ₃₃ N ₇ O ₈ 3) Molecular weight: 559 4) Thin layer chromatography: R _f value; 0.13 Adsorbent; Silica gel plate (Merck) Made by Kieselgel 60
F ₂₅₄ ) Developing solvent; n-butanol: acetic acid: water (4: 1: 2) 5) High-performance liquid chromatography: column; ODS H2151 (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent; 5% acetonitrile-0.1% trifluoroacetic acid Aqueous solution Flow rate: 1.0 ml / min Retention time: 13.6 min Example 4 Mulaidomycin DP Muredomycin D 100 mg and pepsin 200 mg were used and the reaction was carried out for 48 hours in the same manner as in Example 1 to obtain the target compound 11
mg was obtained.

1) Material properties: amphoteric water-soluble, white powder 2) Molecular formula: C ₂₅ H ₃₃ N ₇ O ₈ 3) Molecular weight: 561 4) Thin layer chromatography: R _f value; 0.13 Adsorbent; Silica gel plate (Merck) Made by Kieselgel 60
F ₂₅₄ ) Developing solvent; n-butanol: acetic acid: water (4: 1: 2) 5) High-performance liquid chromatography: column; ODS H2151 (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent; 5% acetonitrile-0.1% trifluoroacetic acid Aqueous solution Flow rate: 1.0 ml / min Retention time: 9.2 min Example 5 Murademycin EP Murademycin E 100 mg and pepsin 200 mg were used to carry out a reaction for 24 hours in the same manner as in Example 1 to obtain the target compound 38.
mg was obtained.

1) Material properties: amphoteric water-soluble, white powder 2) Molecular formula: C ₂₄ H ₃₀ N ₆ O ₇ 3) Molecular weight: 514 4) Thin layer chromatography: R _f value; 0.13 Adsorbent; Silica gel plate (Merck) Made by Kieselgel 60
F ₂₅₄ ) Developing solvent; n-butanol: acetic acid: water (4: 1: 2) 5) High-performance liquid chromatography: column; ODS H2151 (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent; 5% acetonitrile-0.1% trifluoroacetic acid Aqueous solution Flow rate: 1.0 ml / min Retention time: 10.1 min Example 6 Murademycin FP Murademycin F 100 mg and pepsin 200 mg were used for a reaction for 24 hours in the same manner as in Example 1 to obtain the target compound 29.
mg was obtained.

1) Property of substance: Amphoteric water-soluble, white powder 2) Molecular formula: C ₂₄ H ₃₀ N ₆ O ₇ 3) Molecular weight: 514 4) Thin layer chromatography: R _f value; 0.12 Adsorbent; Silica gel plate (Merck) Made by Kieselgel 60
F ₂₅₄ ) Developing solvent; n-butanol: acetic acid: water (4: 1: 2) 5) High-performance liquid chromatography: column; ODS H2151 (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent; 5% acetonitrile-0.1% trifluoroacetic acid Aqueous solution Flow rate: 1.0 ml / min Retention time: 5.3 minutes Reference Example 1 Streptomyces
flavidovirens) SANK60486 strain (Microtechnology Research Institute Article 1347; FE
RM BP-1347) was added to medium A (glucose: 3%, raw yeast:
1% soybean _{flour: 3%, CaCO 3: 0.4} %, MgSO 4 · 7H 2 O: 0.2%,
NISSAN DISFOHM CB-442: 0.01%, pH7 before sterilization.
2) Inoculate one platinum loop into a 500 ml Erlenmeyer flask containing 80 ml,
The cells were cultured at 22 ° C. for 84 hours in a rotary shaker at 220 rpm.
This culture broth (25 ml) was inoculated into four 2-volume Erlenmeyer flasks containing 500 ml of A medium, and the mixture was cultivated at 220 ° C with a rotary shaker at 22 ° C
It was cultured for 24 hours. 750 ml of this culture broth containing 15 A medium 3
Two 0-volume jar fermenters were inoculated and cultured with aeration and stirring at 22 ° C., rotation speed: 150 rpm, aeration rate: 15 / min for 96 hours. When Celite 545 was added to this culture solution 30 as a super-aid, the solution 30 was obtained. 3 this liquid
Adsorbed on Amberlite XAD-2 column, washed with 15 purified water and 12 15% methanol water successively,
Elution was performed with 15 40% methanol water. Methanol was distilled off from the resulting active surface fraction under reduced pressure, followed by freeze-drying to obtain 17.4 g of a coarse powder. 17 g of the obtained coarse powder was dissolved in 3 purified water and passed through a column of Amberlite CG-50 (H ⁺ ), 800 ml to adsorb an effective substance. The active substance was eluted from this column with 0.5 M aqueous ammonia. The active fraction 4.0 was collected and concentrated to 1.0 under reduced pressure. The concentrated liquid 1.0 was passed through 500 ml of DE-52 (manufactured by Wattman Co.) equilibrated with 0.1 M ammonium hydrogen carbonate for adsorption, and eluted with 0.2 M ammonium hydrogen carbonate. Collect 1000 ml of active fraction,
After adsorbing to 200 ml of HP20 (manufactured by Mitsubishi Kasei Co., Ltd.), the active substance was eluted with 500 ml of 50% acetone. By concentrating the active fractions and freeze-drying, the antibiotics muleidomycin A, B,
15 g of a coarse powder containing C, D, E and F was obtained. 14g of this coarse powder
Was dissolved in 200 ml of purified water, adsorbed on 500 ml of DE-52 equilibrated with 0.05 M ammonium hydrogen carbonate, washed with 0.05 M of the same buffer, and then eluted with 0.1 M of the same buffer, every 20 ml. Fractionated into Fractions 80 to 130 were collected as active fractions, desalted by adsorption and elution on an HP column, concentrated under reduced pressure, freeze-dried, and the antibiotics muleidomycin A, C, E were collected.
3.1 g of partially purified powder containing and F was obtained. Partially refined powder
3.0 g was applied to a 100 g silica gel column, developed with a mixed solvent consisting of n-butanol: n-propanol: water (8: 4: 1), and fractionated every 20 ml. Fractions 13 to 70 were collected, concentrated by adding water, and then freeze-dried to obtain 320 mg of crude powder containing the antibiotics muleidomycin A, E and F. Similarly, 66 mg of crude powder containing the antibiotic muleidomycin C was obtained from fractions 56 to 75. Similarly, as DE-52 elution active fraction, fractions 25 to 60
Were collected, desalted by adsorption and elution on a Diaion HP20 column, concentrated under reduced pressure and freeze-dried to obtain 510 mg of partially purified powder containing the antibiotics muleidomycin B and D. 500 mg of the partially purified powder was applied to a 100 g silica gel column, developed with a mixed solvent of n-butanol: n-propanol: water (8: 4: 1), and fractionated every 20 ml. The activity appeared as two peaks. Fractions 37 to 55 were collected, concentrated by adding water, and then freeze-dried to obtain 74 mg of crude powder containing the antibiotic muleidomycin B. In the same manner, 67.5 mg of crude powder containing the antibiotic muleidomycin D was obtained from fractions 76 to 110.

Reference Example 2 300 mg of the crude powder containing the antibiotics mlaidomycins A, E and F obtained in Reference Example 1 was adsorbed on a 1000 ml Toyopearl column HW-40 prepared with 30% methanol and developed with the same solvent, Fractionated into 10 ml each. Fractions 50 to 70 were collected as active fractions, and 10 ml of Amberlite CG-50 (H ⁺
Type) and then eluted with 0.5 M aqueous ammonia. The active fractions were collected, concentrated, and freeze-dried to obtain 24 mg of the antibiotic muleidomycin A. Similarly, fraction 8
0 to 90 were collected to obtain the antibiotic muleidomycin F 32 mg. Similarly, fractions 95 to 105 were collected to obtain 15 mg of the antibiotic muleidomycin E.

Reference Example 3 70 mg of the crude powder containing the antibiotic muleidomycin B obtained in Reference Example 1 was adsorbed on a 1000 ml Toyopearl column HW-40 prepared with 30% methanol and developed with the same solvent to give 10 ml.
Fractionated into each. Fraction 55-75 as the active fraction
Were collected, adsorbed on 10 ml of Amberlite CG-50 (H ⁺ type), and then eluted with 0.5 M aqueous ammonia. The active fractions were collected, concentrated, and lyophilized to obtain the antibiotic muleidomycin B (45 mg).

Reference Example 4 60 mg of the crude powder containing the antibiotic muleidomycin C obtained in Reference Example 1 was adsorbed on a 1000 ml Toyopearl column HW-40 prepared with 30% methanol and developed with the same solvent to give 10 ml.
Fractionated into each. Fraction 65-85 as the active fraction
Were collected, adsorbed on 10 ml of Amberlite CG-50 (H ⁺ type), and then eluted with 0.5 M aqueous ammonia. The active fractions were collected, concentrated, and freeze-dried to obtain 49 mg of the antibiotic muleidomycin C.

Reference Example 5 65 mg of the crude powder containing the antibiotic muleidomycin D obtained in Reference Example 1 was adsorbed on a 1000 ml Toyopearl column HW-40 prepared with 30% methanol and developed with the same solvent to give 10 ml.
Fractionated into each. Fraction 65-85 as the active fraction
Were collected, adsorbed on 10 ml of Amberlite CG-50 (H ⁺ type), and then eluted with 0.5 M aqueous ammonia. The active fractions were collected, concentrated, and freeze-dried to obtain 40 mg of the antibiotic muleidomycin D.

Muradeomycin A, B, C, D, E thus obtained
The physicochemical properties of F and F are as follows.

1. Murademycin A 1) Property of substance: amphoteric water-soluble, white powder 2) Specific rotation: ▲ [α] ²⁵ _D ▼ ＝ + 40.9 ° (c0.69,50)
% Methanol water) 3) Elemental analysis value (%): C, 49.73; H, 5.65; N, 12.08; S, 3.
40 (as hydrate) 4) Molecular weight: 840 (High resolution mass spectrometry FAB-MASS: 841.31)
798 (QM +)) 5) Molecular formula: C ₃₈ H ₄₈ N ₈ O ₁₂ S ₁ 6) Acid hydrolysis: uracil, m-tyrosine, 2-amino-3-N-methylaminobutyric acid 7) Ultraviolet absorption spectrum: Neutral, 260 nm (348); 0.01 NHCl, 258 nm (358); 0.01 N NaOH,
240nm (499), 265nm (330sh) and 295nm (78sh) 8) Solubility: Soluble in water and methanol, sparingly soluble in acetone, insoluble in ethyl acetate, chloroform and benzene.

9) Color reaction: Positive for ninhydrin, sulfuric acid, iodine, ferric chloride, and Bayer reaction.

10) Thin layer chromatography: R _f value; 0.36 adsorbent; silica gel plate (Kieselgel 60 manufactured by Merck)
F ₂₅₄ ) developing solvent; n-butanol: n-propanol: water (4: 2:
1) 11) High Performance Liquid Chromatography: Column; Acacyl SS372-N (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent: Chloroform: Isopropanol: Methanol: Water (200: 100: 100: 40) Flow rate; 1.0 ml / min Retention time; 3.92 min 2. Murademycin B 1) Property: Amphoteric water-soluble, white powder 2) Specific rotation: ▲ [α] ²³ _D ▼ ＝ -7 ° (c0.3,50% methanol water) 3) Element Analytical value (%): C, 50.67; H, 6.36; N, 12.62; S, 3.
13 (as hydrate) 4) Molecular weight: 8.42 (High resolution mass spectrometry FAB-MASS: 843.3
3289 (QM +)) 5) Molecular formula: C ₃₈ H ₅₀ N ₈ O ₁₂ S ₁ 6) Acid hydrolysis: dihydrouracil, m-tyrosine,
2-Amino-3-N-methylaminobutyric acid 7) UV absorption spectrum: Neutral, 255nm (194); 0.1N HCl, 255nm (186); 0.1N NaOH, 24
5nm (325) and 295nm (85sh) 8) Solubility: Soluble in water and methanol, sparingly soluble in acetone, insoluble in ethyl acetate, chloroform and benzene.

9) Color reaction: Positive for ninhydrin, sulfuric acid, iodine, ferric chloride, and Bayer reaction.

10) Thin layer chromatography: R _f value; 0.34 adsorbent; silica gel plate (Kieselgel 60 manufactured by Merck)
F ₂₅₄ ) developing solvent; n-butanol: n-propanol: water (4: 2:
1) 11) High Performance Liquid Chromatography: Column; Acacyl SS372-N (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent: Chloroform: Isopropanol: Methanol: Water (200: 100: 100: 40) Flow rate; 1.0 ml / min Retention time; 3.94 min 3. Murademycin C 1) Material properties: amphoteric water-soluble, white powder 2) Specific rotation: ▲ [α] ²⁵ _D ▼ ＝ + 16.7 ° (c0.57,50)
% Methanol water) 3) Elemental analysis value (%): C, 49.44; H, 5.50; N, 12.53; S, 3.
09 (as hydrate) 4) Molecular weight: 897 (High resolution mass spectrometry FAB-MASS: 898.33
687 (QM +)) 5) Molecular formula: C ₄₀ H ₅₁ N ₉ O ₁₃ S ₁ 6) Acid hydrolysis: uracil, glycine, m-tyrosine, 2-amino-3-N-methylaminobutyric acid 7) Ultraviolet absorption spectrum: Neutral, 258nm (292); 0.01N HCl, 259nm (312); 0.01N NaOH,
240nm (444), 265nm (276sh) and 295nm (72sh) 8) Solubility: Soluble in water and methanol, sparingly soluble in acetone, insoluble in ethyl acetate, chloroform and benzene.

9) Color reaction: Positive for ninhydrin, sulfuric acid, iodine, ferric chloride, and Bayer reaction.

10) Thin layer chromatography: R _f value; 0.29 adsorbent; silica gel plate (Kieselgel 60 manufactured by Merck)
F ₂₅₄ ) developing solvent; n-butanol: n-propanol: water (4: 2:
1) 11) High Performance Liquid Chromatography: Column; Aquasil SS372-N (manufactured by Senshiyu Kagaku Co., Ltd.) (2) Developing solvent: Chloroform: Isopropanol:
Methanol: Water (200: 100: 100: 40) Flow rate: 1.0 ml / min Retention time: 6.29 min 4. Murademycin D 1) Properties of substance: Amphoteric water-soluble, white powder 2) Specific rotation: ▲ [α ] ²³ _D ▼ ＝ −30 ° (c0.52,50%
Methanol water) 3) Elemental analysis value (%): C, 48.79; H, 5.86; N, 12.42; S, 3.
26 (as hydrate) 4) Molecular weight: 899 (High resolution mass spectrometry FAB-MASS: 900.35)
617 (QM +)) 5) Molecular formula: C ₄₀ H ₅₃ N ₉ O ₁₃ S ₁ 6) Acid hydrolysis: dihydrouracil, glycine, m-
Tyrosine, 2-amino-3-N-methylaminobutyric acid 7) UV absorption spectrum: Neutral, 255nm (191); 0.1N HCl, 255nm (184); 0.1N NaOH, 24
5nm (346) and 295nm (90sh) 8) Solubility: Soluble in water and methanol, sparingly soluble in acetone, insoluble in ethyl acetate, chloroform and benzene.

9) Color reaction: Positive for ninhydrin, sulfuric acid, iodine, ferric chloride, and Bayer reaction.

10) Thin layer chromatography: R _f value; 0.26 adsorbent; silica gel plate (Kieselgel 60 manufactured by Merck)
F ₂₅₄ ) developing solvent; n-butanol: n-propanol: water (4: 2:
1) 11) High Performance Liquid Chromatography: Column; Acacyl SS372-N (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent: Chloroform: Isopropanol: Methanol: Water (200: 100: 100: 40) Flow rate; 1.0 ml / min Retention time; 7.24 min 5. Murademycin E 1) Property: Amphoteric water-soluble, white powder 2) Specific rotation: ▲ [α] ²⁵ _D ▼ ＝ -34.2 ° (c1.17,50)
% Methanol water) 3) Elemental analysis value (%): C, 48.57; H, 5.35; N, 11.34; S, 3.
00 (as hydrate) 4) Molecular weight: 852 (high-resolution mass spectrometry FAB-MASS: 853.32)
12 (QM +)) 5) Molecular _{formula: C 39 H 43 N 8 O} 12 S 1 6) acid hydrolysis: uracil, m- tyrosine, 2-amino -3-N-methylamino acid, 8-hydroxy-1,2 , 3,
4-Tetrahydro-3-isoquinolinecarboxylic acid 7) UV absorption spectrum: Neutral, 258 nm (252); 0.01 NHCl, 258 nm (247); 0.01 N NaOH,
240nm (432), 265nm (235sh) and 295nm (80sh) 8) Solubility: Soluble in water and methanol, sparingly soluble in acetone, insoluble in ethyl acetate, chloroform and benzene.

9) Color reaction: Positive for ninhydrin, sulfuric acid, iodine, ferric chloride, and Bayer reaction.

10) Thin layer chromatography: R _f value; 0.39 adsorbent; silica gel plate (Kieselgel 60 manufactured by Merck)
F ₂₅₄ ) developing solvent; n-butanol: n-propanol: water (4: 2:
1) 11) High Performance Liquid Chromatography: Column; Aquasil SS372-N (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent: Chloroform: Isopropanol: Methanol: Water (200: 100: 100: 40) Flow rate: 1.0 ml / min Retention time; 4.7 min 6. Murademycin F 1) Property: Amphoteric water-soluble, white powder 2) Specific rotation: ▲ [α] ²³ _D ▼ ＝ -40.3 ° (c1.05,50)
% Methanol water) 3) Elemental analysis value (%): C, 50.40; H, 5.53; N, 11.20; S, 2.
92 (as hydrate) 4) Molecular weight: 852 (high-resolution mass spectrometry FAB-MASS: 853.31)
80 (QM ⁺ )) 5) Molecular formula: C ₃₉ H ₄₈ N ₈ O ₁₂ S ₁ 6) Acid hydrolysis: uracil, m-tyrosine, 2-amino-3-N-methylaminobutyric acid, 6-hydroxy-1, 2,3,
4-Tetrahydro-3-isoquinolinecarboxylic acid 7) UV absorption spectrum: Neutral, 258nm (232); 0.1N HCl, 258nm (232); 0.1N NaOH, 24
0nm (352), 265nm (200sh) and 295nm (64sh) 8) Solubility: Soluble in water and methanol, sparingly soluble in acetone, insoluble in ethyl acetate, chloroform and benzene.

9) Color reaction: Positive for ninhydrin, sulfuric acid, iodine, ferric chloride, and Bayer reaction.

10) Thin layer chromatography: R _f value; 0.34 adsorbent; silica gel plate (Kieselgel 60 manufactured by Merck)
F ₂₅₄ ) developing solvent; n-butanol: n-propanol: water (4: 2:
1) 11) High Performance Liquid Chromatography: Column; Acacyl SS372-N (manufactured by Senshiyu Scientific Co., Ltd.) Developing solvent: Chloroform: Isopropanol: Methanol: Water (200: 100: 100: 40) Flow rate; 1.0 ml / min Retention time; 5.3 min [Effect of the invention] Murademycin AP, BP, CP, DP, EP and FP were all effective against Gram-negative bacteria, especially bacteria of the genus Chrysanthomonas.

Regarding toxicity, each of the above compounds was administered to mice at 400 mg /
It was administered intravenously by kg, but no toxicity was observed.

 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor Fujio Isono 1-2-58 Hiromachi, Shinagawa-ku, Tokyo Sankyo Co., Ltd. (72) Yoshiyo Sakaida 1-2-58 Hiromachi, Shinagawa-ku, Tokyo Sankyo Co., Ltd. (72) Inventor Takeshi Kinoshita 1-25-2 Hiromachi, Shinagawa-ku, Tokyo Sankyo Co., Ltd.

Claims (2)

(57) [Claims] 1. A formula Murademycin AP, BP, CP, DP, EP and FP and salts thereof. Here, muridomycin AP is R ¹ is 2,4-dioxopyrimidin-1-yl, R ² is α-amino-3-hydroxyphenethyl; mulaidomycin BP is R ¹ is 2 ,Four
-Dioxodihydropyrimidin-1-yl, R ² is α-amino-3-hydroxyphenethyl; mureidomycin CP is R ¹ is 2,4-dioxopyrimidin-1-yl, R ² is α-glycylamino-3-hydroxyphenethyl; mureidomycin DP is R ¹ is 2,4-dioxodihydropyrimidin-1-yl and R ² is α-glycylamino-3-hydroxyphenyl. It is enetyl; Murademycin EP has R ¹ of 2,4-dioxopyrimidine-1-
And R ² is 8-hydroxy-1,2,3,4-tetrahydroisoquinolin-3-yl; mureidomycin
FP has R ¹ of 2,4-dioxopyrimidin-1-yl, R
² is 6-hydroxy-1,2,3,4-tetrahydroisoquinolin-3-yl. 2. A formula And the salts thereof, wherein M ¹ is 2,4-dioxopyrimidin-1-yl and R ² is α. -Amino-3
-Hydroxyphenethyl; mulaidomycin B is
R ¹ is 2,4-dioxodihydropyrimidin-1-yl, R ² is α-amino-3-hydroxyphenethyl; mureidomycin C is R ¹ is 2,4-dioxopyrimidin- 1-yl, R ² is α-glycylamino-3-hydroxyphenethyl; mureidomycin D is R ¹ is
2,4-dioxodihydropyrimidin-1-yl, R
² is α-glycylamino-3-hydroxyphenethyl; mureidomycin E is R ¹ is 2,4-dioxopyrimidin- ¹ -yl and R ² is 8-hydroxy-1,2,3, Four
-Tetrahydroisoquinolin-3-yl; mureidomycin F has R ¹ is 2,4-dioxopyrimidin-1-yl and R ² is 6-hydroxy-1,2,3,4-tetrahydroisoquinoline- 3-yl. ) Is hydrolyzed Murademycin AP, BP, CP, DP, EP and FP and salts thereof. Here, muridomycin AP is R ¹ is 2,4-dioxopyrimidin-1-yl, R ² is α-amino-3-hydroxyphenethyl; mulaidomycin BP is R ¹ is 2 ,Four
-Dioxodihydropyrimidin-1-yl, R ² is α-amino-3-hydroxyphenethyl; mureidomycin CP is R ¹ is 2,4-dioxopyrimidin-1-yl, R ² is α-glycylamino-3-hydroxyphenethyl; mureidomycin DP is R ¹ is 2,4-dioxodihydropyrimidin-1-yl and R ² is α-glycylamino-3-hydroxyphenyl. It is enetyl; Murademycin EP has R ¹ of 2,4-dioxopyrimidine-1-
And R ² is 8-hydroxy-1,2,3,4-tetrahydroisoquinolin-3-yl; mureidomycin
FP has R ¹ of 2,4-dioxopyrimidin-1-yl, R
² is 6-hydroxy-1,2,3,4-tetrahydroisoquinolin-3-yl.