JP2002255969A - New physiologically active substance fo-7711cd4 and/or cd6 and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a physiologically active substance FO-7711 CD4 and/or CD6 having antimicrobial activity against Staphylococcus aureus having resistance against existing antibiotics and a method for producing the above substance. SOLUTION: A microorganism having ability to produce a physiologically active substance FO-7711 CD4 and/or CD6 belonging to the genus Penicillium is cultured in a culture medium and the physiologically active substance FO-7711 CD4 and/or CD6 is accumulated in the culture products and the physiologically active substance FO-7711 CD4 and/or CD6 is collected from the culture products. The above substance is expected to have effects on various diseases in which resistant mechanism is concerned, e.g. food poisoning caused by antibiotic- resisting bacterium, particularly hospital acquired infection, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel bioactive substance FO-7711 CD4 and / or CD6 (hereinafter sometimes referred to as FO-7711 CD4 and / or CD6) which suppresses the growth of multidrug-resistant Staphylococcus aureus (KB324). FO-7711)
The present invention relates to a novel method for producing a physiologically active substance FO-7711 CD4 and / or CD6 in which a CD4 and / or CD6 substance is produced by culturing a microorganism-producing bacterium.

[0002]

2. Description of the Related Art Infectious diseases are mainly treated by chemotherapy, and so far imipenem (Imipene) has been used.
nem), kanamycin, and various other therapeutic agents for infectious diseases have been developed. Although it seems that humans have overcome infectious diseases with these infectious disease drugs, the emergence of resistant bacteria against these infectious disease drugs is increasing. Particularly in recent years, nosocomial infection, which is difficult to treat among infectious diseases, has become a major social problem.

[0003]

In recent years, patients with infectious diseases have become multidrug resistant in many cases, making treatment more difficult. Originally, these nosocomial infections are mainly caused by patients with reduced resistance to infection, that is, patients with serious underlying illness or after major surgery, and the infection is fatal It is possible. Thus, there has been a strong demand for the development of a new therapeutic agent having antibacterial activity against bacteria that have become resistant to conventional infectious disease therapeutic agents.

[0004] The present invention provides a novel bioactive substance FO-7711CD4 and / or CD which can satisfy such a demand.
6 is provided. Further, the present invention provides a bioactive substance FO-7711 CD4 belonging to the genus Penicillium and / or
Alternatively, a microorganism having the ability to produce CD6 is cultured in a medium, and the physiologically active substance FO-7711CD4 is contained in the culture.
And / or accumulating CD6, and collecting a physiologically active substance FO-7711 CD4 and / or CD6 from the culture.
The present invention provides a method for producing CD4 and / or CD6.

[0005]

DISCLOSURE OF THE INVENTION The present inventors have isolated bacterial strains from various soils for the purpose of searching for new physiologically active substances, and have continued to study metabolites produced by the same. Strain FO-77 isolated from soils
It has been found that a substance that suppresses the growth of multidrug-resistant Staphylococcus aureus is produced in 11 cultures. Next, as a result of separating and purifying the active substance from the culture, no substance having such a chemical structure and its physicochemical properties has been known so far.
O-7711 CD4 and / or CD6 material. The present invention has been completed based on such findings, and has the following formula [I] and / or formula [II]

[0006]

Embedded image A novel physiologically active substance FO-7711CD4 and / or CD6 characterized by being a compound represented by the formula:

The present invention further provides a novel physiologically active substance FO-7711 CD4 having the following physicochemical properties:
Or regarding CD6. [1] Bioactive substance FO-7711 CD4 (1) Property: white powder (2) Molecular weight: 503 [High-resolution fast atom bombardment (FA
B) Mass spectrum] (3) Molecular formula: C ₃₂ H ₄₁ O ₄ N ₁

(4) Ultraviolet / visible absorption spectrum: The ultraviolet absorption spectrum measured in methanol is as shown in FIG. 1 and is 266 (log ε = 3.44), 235
(Log ε = 3.56), 212 (log ε = 3.
74) Characteristic absorption maximum near nm. (5) Infrared absorption spectrum: The infrared absorption spectrum measured by the potassium bromide method is as shown in FIG.
19, 2949, 2924, 1687, 1614, 15
06, with a characteristic absorption band at 1226 cm ^-1

(6) Specific rotation [α] _D ²⁷ + 50.0 °
(C = 0.1, CHCl ₃ ) (7) Solubility in solvent: acetone, chloroform,
Soluble in ethyl acetate, methanol and ethanol, poorly soluble in water (8) Color reaction: positive for sulfuric acid and iodine

[2] Bioactive substance FO-7711 CD
6 (1) Property: white powder (2) Molecular weight: 503 [High-resolution high-speed atom bombardment (FA
B) Mass spectrum] (3) Molecular formula: C ₃₂ H ₄₁ O ₄ N ₁

(4) Ultraviolet / visible absorption spectrum: The ultraviolet absorption spectrum measured in methanol is as shown in FIG. 5 and is 271 (shoulder, log ε = 3.5)
2) shows a characteristic absorption maximum near 224 (log ε = 4.19) nm. (5) Infrared absorption spectrum: The infrared absorption spectrum measured by the potassium bromide method is as shown in FIG. 34
23, 2949, 2924, 1689, 1612, 15
06, having a characteristic absorption band at 1227 cm ^-1

(6) Specific rotation [α] _D ²⁷ + 12.8 °
(C = 0.1, CHCl ₃ ) (7) Solubility in solvent: acetone, chloroform,
Soluble in ethyl acetate, methanol and ethanol, poorly soluble in water (8) Color reaction: positive for sulfuric acid and iodine

Further, the present invention further comprises culturing a microorganism having the ability to produce the physiologically active substance FO-7711 CD4 and / or CD6 belonging to the genus Penicillium in a medium, and adding the physiologically active substance FO-7711 CD4 and And / or allow CD6 to accumulate and allow the bioactive substance FO
-7711 CD4, a novel bioactive substance characterized by collecting CD4 and / or CD6
And / or a method for producing CD6.

The present invention further relates to a microorganism belonging to the genus Penicillium and capable of producing the physiologically active substance FO-7711 CD4 and / or CD6, wherein the microorganism is Penicillium sp. FO-7711.
The present invention also relates to a strain belonging to the genus Penicillium and having the ability to produce the physiologically active substance FO-7711 CD4 and / or CD6, wherein the microorganism is Penicillium sp.
FO-7711 (FERM P-18211).

The physiologically active substance FO-7711 C of the present invention
The D4 and / or CD6 substance producing bacterium is not particularly limited, as long as it is a genus Penicillium and has an ability to produce FO-7711 CD4 and / or CD6 substance. A preferred example of the strain used for producing the FO-7711 CD4 and / or CD6 substance of the present invention is Penicillium sp. Newly isolated from the soil of Nagasaki Prefecture by the present inventors.
ium sp. ) FO-7711 strain. The bacteriological properties of this strain are as follows.

1. Morphological properties This strain grows well on Tsakku yeast agar medium, 25% glycerin / nitrate agar medium, corn / meal agar medium, etc., and has good conidium formation. corn·
When the colonies grown on the meal agar medium were observed with a microscope, the hyphae were transparent and had partition walls, and the conidiophores (12.5
７５75 × 2.5-3.0 μm) stand upright from the basal hyphae or extend from the branched aerial hyphae (1.8-3.0 μm).

Penicillus is a two-wheeler and metre (10-20
× 1.8-3.0 μm), phialide (8.8-15)
× 1.0 to 3.0 μm). Metre is 1
It branches into three, and the phialides are ampoule-shaped and grow in groups of three to four. At first, one phialo-conidium attaches to the apex of phialide and becomes chain-like as the culture time elapses. The conidium is spherical and has a size of 1.8 to 3.0 μm, and the surface is rough.

2. Various properties on various media Table 1 shows the results of macroscopic observation when cultured at 25 ° C for 2 weeks on various agar media.

[0019]

[Table 1]

3. Physiological and ecological properties (1) Optimal growth conditions The optimal growth conditions for this fungus are pH 4-7, and the growth temperature is 13-
32 ° C. (2) Growth range The growth range of this fungus is pH 3 to 10, and the growth temperature is 7.0 to 7.0.
36 ° C. (3) Distinguishing between aerobic and anaerobic conditions This bacterium is aerobic.

Based on the morphological characteristics, culture characteristics and physiological characteristics of the FO-7711 strain described above, the strain was compared with known bacterial strains. As a result, the strain was identified as Penicillium (Peni).
and identified as Penicillium sp. FO-7711. This strain is Penicillium sp. FO-7711 (Penic
illium sp. FO-7711) is deposited at the Institute of Biotechnology and Industrial Technology, located at 1-3 1-3 Higashi, Tsukuba, Ibaraki Prefecture. Deposit date is 2001
On February 19, the accession number is FERM P-18211.

As a preferred strain of the present invention, the present FO-7
711 CD4 and / or CD6 substance-producing bacteria have been described, but as a general property of filamentous fungi, the mycological properties are extremely variable and not constant. It is a well-known fact that mutation is performed by artificial mutation means using irradiation or a mutant derivative agent. Such artificial mutants, as well as natural mutants, belong to the genus Penicillium and
Any strain capable of producing -7711 CD4 and / or CD6 material can be used in the present invention. In addition, strains mutated by cell engineering such as cell fusion and genetic manipulation are also used in the present FO-7711 CD4 and / or C
Included as D6 substance producing bacteria.

In the present invention, first, the present FO-7711 CD4 and / or CD6 substance producing bacterium belonging to the genus Penicillium is cultured in an appropriate medium. In culturing this strain, a general method for culturing filamentous fungi is generally applied.
As the medium, a nutrient medium containing a carbon source that can be assimilated by microorganisms, a nitrogen source that can be digested, and, if necessary, inorganic salts and the like is used.

The nutrient source suitable for producing the above-mentioned FO-7711 CD4 and / or CD6 substance may be any nutrient source that can be used as a nutrient source for filamentous fungi. For example, commercially available peptone, meat extract, corn steep liquor, cottonseed flour, peanut flour, soybean flour, yeast extract, NZ-amine, casein hydrate, sodium nitrate, ammonium nitrate, nitrogen source such as ammonium sulfate, glycerin ,starch,
Glucose, galactose, carbohydrates such as mannose,
Alternatively, a carbon source such as fat, and an inorganic salt such as salt, phosphate, calcium carbonate, and magnesium sulfate can be used alone or in combination.

In addition, if necessary, a trace amount of a metal salt or the like can be added. If foaming occurs in the liquid culture, an antifoaming agent such as silicone oil, vegetable oil, or a surfactant may be appropriately used. . Any of these can be used as long as they utilize the producing bacteria and can contribute to the production of the FO-7711 CD4 and / or CD6 substance. All known culture materials of filamentous fungi can be used. For large-scale culture of the present FO-7711 CD4 and / or CD6 substance, liquid culture is preferred.
Applicable to the extent that D4 and / or CD6 substances can be produced. When culturing, the conditions described above were applied to the FO-77.
11 It can be appropriately selected according to the properties of the bacteria producing CD4 and / or CD6 substance.

The FO-7711 CD4 and / or CD6 substance accumulated in the culture solution thus obtained is
Since it is contained in the culture filtrate or cultured cells, the culture filtrate may be filtered by adding a filter aid, for example, celite, high flow supercell, or the like, or may be centrifuged to separate the culture filtrate. The FO-7 is separated from the mixture of the filtrate obtained by concentrating the organic solvent extract of the culture filtrate and the cells.
It is advantageous to collect 711 CD4 and / or CD6 material.

Further, the culture filtrate and the cells are extracted with a hydrophilic organic solvent such as ethanol or acetone without separation, and the obtained extract is concentrated under reduced pressure, and the concentrate is non-hydrophilic such as ethyl acetate or chloroform. It can be extracted with a neutral solvent. Further, when the content of the FO-7711 CD4 and / or CD6 substance is extremely large in either the culture filtrate or the bacterial cells, the FO-711 may be extracted from the larger one. In order to separate and purify the FO-7711 CD4 and / or CD6 substance from the cultured cells, usually, a mixture of the organic solvent extract of the culture filtrate and the cells is concentrated or a mixture of the non-hydrophilic solvent such as acetic acid is used. The FO-711 CD4 and / or CD is extracted by extracting with ethyl and chloroform.
Six substances are dissolved in the organic solvent.

The organic solvent layer thus obtained is dehydrated by adding various dehydrating agents, for example, anhydrous sodium sulfate, bead gel or the like, if necessary, and then the organic solvent is distilled off under reduced pressure. In this concentration operation, the FO-
7711 CD4 and / or CD6 substance is stable, but it is usually preferable to carry out the heating at a temperature of 50 ° C. or lower, particularly preferably 30 ° C. to 40 ° C.

In order to further purify the obtained extract, the difference in solubility between the FO-7711 CD4 and / or CD6 substance and a mixture thereof, the difference in partition between two immiscible liquid layers, various Many means utilizing the difference in adsorptive power to the adsorbent are possible. In particular, chromatography is performed using the FO-7711 CD4 and / or CD6.
This is an effective method for purifying substances.

The FO-7711 CD4 and / or CD
Chromatography effective for the purification of the six substances includes adsorption chromatography using an adsorption resin such as silica gel, alumina, activated carbon cellulose, hydroxyapatite, and HP-20, and reversed-phase partition chromatography using silanized silica gel, octadecyl silanized silica gel, and the like. , Sephadex LH-20, Toyopearl (trade name,
HW-40) and the like, and ion-exchange chromatography using gel filtration chromatography based on a molecular sieve.

The present FO-7711 CD4 and / or CD
The six substances can be separated and purified by using these means such as chromatography, electrophoresis, countercurrent distribution, and ultrafiltration singly or in combination or repetition in any order. For example, the above crude product is dissolved in a small amount of chloroform, adsorbed on silica gel, and subjected to column chromatography using a chloroform-methanol mixed solvent, and the active fraction is concentrated under reduced pressure, and then dissolved in a small amount of methanol. Can be separated and purified by reverse-phase high performance liquid chromatography.

Next, the physicochemical properties of the FO-7711 CD4 substance of the present invention will be described. (1) Property: white powder (2) Molecular weight: 503 [High-resolution high-speed atom bombardment (FA
B) by mass spectrum] (3) Molecular formula: C ₃₂ H ₄₁ O ₄ N ₁ (4) Specific rotation: [α] _D ²⁷ + 50.0 ° (c = 0.
1, CHCl ₃ ) (5) High resolution spectrum: FAB-MS 502 (M
-H) ^-

(6) Ultraviolet / visible absorption spectrum: The ultraviolet absorption spectrum measured in methanol is as shown in FIG. 1 and is 266 (log ε = 3.44), 235
(Log ε = 3.56), 212 (log ε = 3.
74) shows a characteristic absorption maximum near nm. (7) Infrared absorption spectrum: The infrared absorption spectrum measured by the potassium bromide method is as shown in FIG.
19, 2949, 2924, 1687, 1614, 15
06, having a characteristic absorption band at 1226 cm ^-1 (8) Solubility in solvents: acetone, chloroform,
Soluble in ethyl acetate, methanol and ethanol, poorly soluble in water

(9) Color reaction: positive for sulfuric acid and iodine (10) Proton nuclear magnetic resonance spectrum: Varian
XL-400, manufactured by Varian Japan
Was measured using a type NMR spectrometer. ¹H-nuclear magnet
Gas resonance spectrum (measured in deuterated dimethyl sulfoxide solution)
400 MHz) is as shown in FIG.
And chemical shifts (ppm) of carbon are shown in Table 2 below.
(11)¹³C-nuclear magnetic resonance spectrum (deuterated dimethyl sulf
(Measured in a hydroxide solution, 100.58 MHz) is shown in FIG.
As shown, the chemical shifts of hydrogen and carbon (pp
m) is as shown in Table 2 below.

[0035]

[Table 2] In the table, s is a singlet, d is a doublet, t is a triplet,
q indicates a quadruple, m indicates a multiplet, H indicates the number of protons, and J indicates a spin coupling constant (Hz). (12) Chemical structure: as in the following formula [I]

[0036]

Embedded image

Next, the physicochemical properties of the FO-7711 CD6 substance of the present invention will be described. (1) Property: white powder (2) Molecular weight: 503 [High-resolution high-speed atom bombardment (FA
B) Mass spectrum] (3) Molecular formula: C ₃₂ H ₄₁ O ₄ N ₁ (4) Specific rotation [α] _D ²⁷ + 12.8 ° (c = 0.
1, CHCl ₃ ) (5) High resolution spectrum: FAB-MS 502 (M
-H) ^-

(6) Ultraviolet / visible absorption spectrum: The ultraviolet absorption spectrum measured in methanol is as shown in FIG. 5, and is 271 (shoulder, log ε = 3.5)
2) shows a characteristic absorption maximum near 224 (log ε = 4.19) nm. (7) Infrared absorption spectrum: The infrared absorption spectrum measured by the potassium bromide method is as shown in FIG. 34
23, 2949, 2924, 1689, 1612, 15
06, having a characteristic absorption band at 1227 cm ^-1 (8) Solubility in solvents: acetone, chloroform,
Soluble in ethyl acetate, methanol and ethanol, poorly soluble in water

(9) Color reaction: positive for sulfuric acid and iodine (10) Proton nuclear magnetic resonance spectrum: UNITY manufactured by Varian Japan
Measured using an INOVA type NMR spectrometer
^{The 1} H-nuclear magnetic resonance spectrum (in deuterated dimethyl sulfoxide solution, 600 MHz) is as shown in FIG. 7, and the chemical shifts (ppm) of hydrogen and carbon are as shown in Table 3 below. (11) ¹³ C -The nuclear magnetic resonance spectrum (150.84 MHz in deuterated dimethyl sulfoxide solution) is as shown in FIG. 8, and the chemical shifts (ppm) of hydrogen and carbon are as follows:
It is as shown in Table 3 below.

[0040]

[Table 3] In the table, s is a singlet, d is a doublet, t is a triplet,
q indicates a quadruple, m indicates a multiplet, H indicates the number of protons, and J indicates a spin coupling constant (Hz). (12) Chemical structure: as in the following formula [II]

[0041]

Embedded image

As described above, the present FO-7711 CD4
As a result of examining various physical properties and spectrum data of the FO-7711 CD6 substance in detail, the physiologically active substance FO-7711 CD4 and / or FO-77 was obtained.
11 CD6 was determined to have a chemical structure represented by the following formulas [I] and / or [II].

[0043]

Embedded image

As described above, the present FO-7711 CD
4 and / or various physical-chemical properties of the FO-7711 CD6 substance have been described in detail, but no compound corresponding to such properties has been reported so far, and the compound of the formula [I]
And / or the biologically active substance FO-77 represented by [II]
11 CD4 and / or FO-7711 CD6 were determined to be novel substances.

Next, the biological properties of the FO-7711 CD4 substance of the present invention will be described below. Book FO-77
11 Regarding the antimicrobial activity of the CD4 substance, the standard method of the Japanese Society of Chemotherapy (Chemotherapy 29,76)
(1981), the minimum inhibitory concentration (MIC) was measured. Muller-Hilon Broth (D
ifco), 1.5% agar was added to dissolve the medium, and when the temperature reached 60 to 50 ° C, 100% was added.
The antibacterial agent solution diluted 2-fold from 0 μg / mL was added to 1/9
The resulting mixture was mixed well, mixed well, and dispensed into petri dishes to form flat plates. After culturing in a culture medium for enrichment, the number of bacteria was adjusted to about 10 ⁶ / mL and used as an inoculum, and one spot was inoculated. 18-2
After culturing at 37 ° C. for 0 hour, the MIC value was defined as the lowest concentration at which culturing was completely inhibited. This FO-7711 C
The results for substance D4 are shown in Table 4.

[0046]

[Table 4]

Next, the FO-7711 CD of the present invention
The biological properties of the six substances are described below. This FO-
7711 The antimicrobial activity of the CD6 substance is determined by the standard method of the Japanese Chemotherapy Society (Chemotherapy 29,7).
6 (1981), minimum inhibitory concentration (MIC)
Was measured. Muller-Hilon Broth
(Manufactured by Difco), 1.5% agar was added to dissolve the medium, and when the temperature reached 60 to 50 ° C, 10%
The antibacterial agent solution diluted 2-fold from 00 μg / mL was added to 1 /
Nine amounts were added, mixed well, and dispensed into a petri dish to form a plate. After culturing in a culture medium for enrichment, the number of bacteria was adjusted to about 10 ⁶ / mL and used as an inoculum, and one spot was inoculated. 18 ~
After culturing at 37 ° C. for 20 hours, the MIC value was defined as the lowest concentration at which culturing was completely inhibited. This FO-7711
The results for the CD6 material are shown in Table 5.

[0048]

[Table 5]

Next, the present invention will be described specifically with reference to examples, but the present invention is not limited to these examples.

[Example] Penicillium sp. FO-7711
(Penicillium sp. FO-7711)
(FERM P-18211) spore suspension (10 ⁷ /
mL) of glucose 2.0%, polypeptone (Nippon Pharmaceutical Co., Ltd.) 0.5%, agar 0.1%, yeast extract (Oriental Yeast Co., Ltd.) 0.2%, magnesium sulfate heptahydrate 0 0.05%, potassium dihydrogen phosphate 0.
A 1% liquid medium (pH 6.0) was inoculated into one 500 ml Erlenmeyer flask dispensed 100 ml at a time.
C. for 3 days with shaking to obtain a seed culture solution.

Maltose 2.0%, dried yeast 3.0%,
Potassium bromide 1.0%, sodium dihydrogen phosphate 0.0
After adding 20 liters of a liquid medium (pH 6.0) in which 5% and 0.05% of manganese sulfate heptahydrate were dissolved in a 30-liter jar fermenter, sterilized, and then 200 mL of a seed culture solution obtained by the above method was used. (1%) was transplanted to a sterile solution, and cultured with aeration and agitation at 27 ° C. for 6 days to obtain about 20 liters of the culture solution.

The culture solution was centrifuged, and the obtained cells were extracted with 5 L of acetone, and 3 L of distilled water was added.
After concentration under reduced pressure, acetone was distilled off and the solution was dissolved in water. An equal amount of ethyl acetate was added thereto, and the mixture was sufficiently stirred, and then an ethyl acetate extract was separated by centrifugation. Further, the culture solution was added with an equal volume of ethyl acetate, stirred sufficiently, and then an ethyl acetate extract was separated by centrifugation. The ethyl acetate extracts obtained from each were mixed and concentrated under reduced pressure using a rotary evaporator to obtain 5.2 g of a crude extract.

The crude extract was adsorbed on a silica gel (70-230 mesh, Merck) column previously filled with chloroform, and the chloroform-
Elution with a mixed solvent of methanol and FO-7711 CD
An active fraction containing 4 and / or CD6 material was obtained. Further, this active fraction was adsorbed onto a silica gel (70-230 mesh, Merck) column previously filled with chloroform, eluted with a chloroform-methanol mixed solvent system, and FO-7711 CD4 and / or CD6 was eluted.
An active fraction containing the substance was obtained.

Further, this active fraction was dissolved in a small amount of methanol and subjected to high performance liquid chromatography (Sensepak Pegasil B, inner diameter 20 mm, length 250 mm), and an 80% acetonitrile-H ₂ O solution was used as a mobile phase. By fractionation, 36.4 mg of a pure product of the present FO-7711 CD4 substance and 68.8 mg of a pure product of the FO-7711 CD6 substance were obtained.

[0054]

As described above, the physiologically active substance FO-7711 belonging to the genus Penicillium CD4 and / or CD6
A microorganism having the ability to produce E. coli is cultured in a culture medium, and the physiologically active substance FO-7711 CD4 and / or
Alternatively, CD6 is accumulated and the physiologically active substance F
The substance from which O-7711 CD4 and / or CD6 was collected was found to inhibit the growth of multidrug-resistant Staphylococcus aureus. Therefore, it is expected to be effective against diseases considered to involve multidrug-resistant bacteria, for example, infectious diseases such as hospital-acquired infections.

[Brief description of the drawings]

FIG. 1 shows a physiologically active substance FO-7711 C according to the present invention.
It is an ultraviolet absorption spectrum (in methanol) of D4.

FIG. 2 shows a physiologically active substance FO-7711 C according to the present invention.
It is an infrared absorption spectrum (potassium bromide) of D4.

FIG. 3 is a physiologically active substance FO-7711 C according to the present invention.
^{1 is} a ¹ H-nuclear magnetic resonance spectrum of D4 (400 MHz in a heavy dimethyl sulfoxide solution).

FIG. 4 is a physiologically active substance FO-7711 C according to the present invention.
It is a ¹³ C-nuclear magnetic resonance spectrum (100.58 MHz in deuterated dimethyl sulfoxide solution) of D4.

FIG. 5 is a physiologically active substance FO-7711 C according to the present invention.
It is an ultraviolet absorption spectrum (in methanol) of D6.

FIG. 6 shows a physiologically active substance FO-7711 C according to the present invention.
It is an infrared absorption spectrum (potassium bromide) of D6.

FIG. 7 shows a physiologically active substance FO-7711 C according to the present invention.
^{1 is} a ¹ H-nuclear magnetic resonance spectrum of D6 (in a heavy dimethyl sulfoxide solution, 600 MHz).

FIG. 8 is a physiologically active substance FO-7711 C according to the present invention.
It is a ¹³ C-nuclear magnetic resonance spectrum of D6 (150.84 MHz in deuterated dimethyl sulfoxide solution).

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) (C12P 17/18 (C12P 17/18 C C12R 1:80) C12R 1:80) (72) Inventor Hayashi Masahiko 5-9-1, Shirogane, Minato-ku, Tokyo Inside Kitasato Institute (72) Inventor Rokuro Masuma 5-9-1, Shirogane, Minato-ku, Tokyo Inside Kitasato Institute (72) Inventor Akiko Fukami Tokyo 5-9-1 Shirogane, Minato-ku, Tokyo F-term in Kitasato Institute (reference) 4B064 AE58 BA06 BE07 BE09 BE10 BE14 BG01 BH01 BH02 BH06 BH10 CA05 CE08 CE09 CE10 DA01 4B065 AA67X AC14 BA22 BD16 CA18 CA34 CA44 4C050 AA01EE0104 CC GG03 HH01

Claims (6)

[Claims] 1. The following formula [I] and / or formula [II] A novel physiologically active substance FO-7711CD4 and / or CD6, which is a compound represented by the formula: 2. A novel physiologically active substance FO-7711 CD4 and / or CD6 having the following physicochemical properties. [1] Bioactive substance FO-7711 CD4 (1) Property: white powder (2) Molecular weight: 503 [High-resolution fast atom bombardment (FA
B) Mass spectrum] (3) Molecular formula: C ₃₂ H ₄₁ O ₄ N ₁ (4) Ultraviolet / visible absorption spectrum: The ultraviolet absorption spectrum measured in methanol is as shown in FIG.
266 (log ε = 3.44), 235 (log ε
= 3.56) and a characteristic absorption maximum near 212 (log ε = 3.74) nm. (5) Infrared absorption spectrum: The infrared absorption spectrum measured by the potassium bromide method is as shown in FIG. And 34
19, 2949, 2924, 1687, 1614, 15
06, having a characteristic absorption band at 1226 cm ^-1 (6) Specific rotation [α] _D ²⁷ + 50.0 ° (c = 0.1,
CHCl ₃ ) (7) Solubility in solvent: acetone, chloroform,
Soluble in ethyl acetate, methanol and ethanol, poorly soluble in water (8) Color reaction: positive for sulfuric acid and iodine [2] Bioactive substance FO-7711 CD6 (1) Property: white powder (2) Molecular weight: 503 [ High resolution fast atom bombardment (FA
B) Mass spectrum] (3) Molecular formula: C ₃₂ H ₄₁ O ₄ N ₁ (4) Ultraviolet / visible absorption spectrum: The ultraviolet absorption spectrum measured in methanol is as shown in FIG.
271 (shoulder, log ε = 3.52), 224
(Log ε = 4.19) shows a characteristic absorption maximum near nm. (5) Infrared absorption spectrum: The infrared absorption spectrum measured by the potassium bromide method is as shown in FIG.
23, 2949, 2924, 1689, 1612, 15
06, having a characteristic absorption band at 1227 cm ^-1 (6) Specific rotation [α] _D ²⁷ + 12.8 ° (c = 0.
1, CHCl ₃ ) (7) Solubility in solvent: acetone, chloroform,
Soluble in ethyl acetate, methanol and ethanol, poorly soluble in water (8) Color reaction: positive for sulfuric acid and iodine 3. A physiologically active substance F belonging to the genus Penicillium
A microorganism having the ability to produce O-7711 CD4 and / or CD6 is cultured in a medium to accumulate the physiologically active substance FO-7711 CD4 and / or CD6 in the culture. −7711
A novel bioactive substance FO-7711 CD4 and / or C characterized by collecting CD4 and / or CD6
Method for producing D6. 4. A bioactive substance F belonging to the genus Penicillium
A microorganism capable of producing O-7711 CD4 and / or CD6 is Penicillium sp.
C. sp. 4. The method according to claim 3, which is FO-7711 strain. 5. A bioactive substance F belonging to the genus Penicillium
O-7711 A microorganism capable of producing CD4 and / or CD6. 6. The method according to claim 6, wherein the microorganism is penicillium sp.
enicilium sp. ) FO-7711 (FE
The microorganism according to claim 5, which is RMP-18211).