JPH1160589A - New physiologically active substance na00226b and c, its production and use - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain the new subject substance useful as an antiasthmatic agent, a bronchodilator, a therapeutic agent for bronchitis, obtainable from a strain belonging to the genus Streptomyces, having inhibitory actions on phosphodiesterase. SOLUTION: This new substance is a physiologically active substance NA00226B showing the following physical and chemical properties or its pharmaceutically acceptable salt. Appearance: red powder. Molecular weight: 447. Molecular formula: C23 H29 NO8 . Solubility: soluble in a lower alcohol and insoluble in hexane, petroleum ether and water. Rf value by ODS thin film chromatography: 0.42 by a developing solvent of acetonitrile-water-formic acid (50:50:1 v/v). Color reaction: positive in vanillin-sulfuric acid, iron chloride and iodine, etc. The physiologically active substance is obtained by culturing a bacterium belonging to the genus Streptomyces capable of producing a physiologically active NA00226B in a medium, forming and accumulating the substance in the culture product and collecting the substance.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel physiologically active substance NA00226B or C or a pharmaceutically acceptable salt thereof, a method for producing the same, and a use thereof. The compound of the present invention has a phosphodiesterase inhibitory activity, for example, an anti-asthmatic drug, a bronchodilator, a therapeutic agent for bronchitis, an anti-allergic drug, an anti-inflammatory drug, an anti-rheumatic drug, an antihypertensive drug, a therapeutic drug for angina pectoris It is a physiologically active substance used as an antiarrhythmic drug, a cerebral circulation metabolism improving drug, a blood coagulation inhibitor, an antidepressant and the like.

[0002]

2. Description of the Related Art As a physiologically active substance, which has a phosphodiesterase inhibitory action and is used as a bronchodilator, a drug for treating angina, a drug for treating arrhythmia, a drug for improving cerebral circulation and metabolism, an antidepressant, etc. , Theophylline, amrinone and the like are known.

[0003]

However, these compounds are not satisfactory because they have strong side effects. The invention of new compounds suitable for these uses is awaited.

[0004]

Means for Solving the Problems Accordingly, the present inventors have:
As a result of various searches for metabolites of microorganisms, one strain belonging to the genus Streptomyces has a novel physiologically active substance NA00226B having a phosphodiesterase inhibitory action.
And / or C was found to be produced. That is,
The present invention relates to the following (1) to (7).

(1) Physiologically active substance NA00226B or a pharmaceutically acceptable salt thereof having the following physicochemical properties: 1) Appearance; red powder 2) Molecular weight: 447 3) Molecular formula; C ₂₃ H ₂₉ NO ₈ 4) Dissolution Soluble; soluble in lower alcohols, insoluble in hexane, petroleum ether and water. 5) Rf value by ODS thin layer chromatography: 0.42 as a developing solvent of acetonitrile-water-formic acid (50: 50: 1 v / v). 6) Ultraviolet absorption spectrum; shown in FIG. 7) Infrared absorption spectrum; shown in FIG. 8) Hydrogen nuclear magnetic resonance spectrum; FIG. 3 shows a spectrum (300 MHz) measured in deuterated chloroform. 9) Carbon nuclear magnetic resonance spectrum; FIG. 4 shows a spectrum (75 MHz) measured in deuterated chloroform. The chemical shift values are shown below. δ (ppm) 189.1 (s), 181.0 (s), 1
70.7 (s), 158.4 (s), 149.7
(S), 141.4 (s), 134.8 (s), 12
8.3 (s), 119.3 (d), 112.7 (s),
99.5 (d), 75.0 (d), 72.5
(S), 69.9 (t), 69.8 (d), 60.
7 (t), 40.6 (t), 35.5 (t), 3
3.8 (t), 29.2 (q), 28.0 (t),
23.4 (q), 14.2 (q). 10) Color reaction; positive for vanillin-sulfuric acid, iron chloride and iodine. (2) A biologically active substance NA00 having the following physicochemical properties:
226C or a pharmaceutically acceptable salt thereof 1) Appearance; red powder 2) Molecular weight; 433 3) Molecular formula; C ₂₂ H ₂₇ NO ₈ 4) Solubility; soluble in lower alcohol, insoluble in hexane, petroleum ether, water . 5) Rf value by ODS thin-layer chromatography; 0.48 as a developing solvent of acetonitrile-water-formic acid (50: 50: 1 v / v). 6) UV absorption spectrum; shown in FIG. 7) Infrared absorption spectrum; shown in FIG. 8) Hydrogen nuclear magnetic resonance spectrum; FIG. 7 shows a spectrum (300 MHz) measured in deuterated chloroform. 9) Carbon nuclear magnetic resonance spectrum; FIG. 8 shows a spectrum (75 MHz) measured in deuterated chloroform. The chemical shift values are shown below. δ (ppm) 189.1 (s), 181.0 (s), 1
71.2 (s), 158.3 (s), 149.7
(S), 141.4 (s), 134.7 (s), 12
8.3 (s), 119.3 (d), 112.7 (s),
99.5 (d), 75.0 (d), 72.6
(S), 69.9 (t), 69.7 (d), 51.
9 (d), 40.3 (t), 35.4 (t), 3
3.6 (t), 29.3 (q), 27.9 (t),
23.4 (q). 10) Color reaction; positive for vanillin-sulfuric acid, iron chloride and iodine. (3) Streptomyces
A microorganism belonging to the genus and having the ability to produce the physiologically active substance NA00226B and / or C is cultured in a culture medium, and the physiologically active substance NA00226B and / or C is produced and accumulated in the culture, and collected. A method for producing a physiologically active substance NA00226B or C or a pharmaceutically acceptable salt thereof. (4) A medicament comprising a physiologically active substance NA00226B or C or a pharmaceutically acceptable salt thereof as an active ingredient. (5) A phosphodiesterase inhibitor comprising, as an active ingredient, a physiologically active substance NA00226B or C or a pharmaceutically acceptable salt thereof.

(6) antiasthmatics, bronchodilators, bronchitis therapeutics, antiallergics, antiinflammatory, antirheumatics, antihypertensives, angina pectoris, arrhythmias, cerebral circulation metabolism improvers, The medicament according to (4), which is any one of a blood coagulation inhibitor and an antidepressant. (7) Streptomyces
A microorganism belonging to the genus and having the ability to produce the physiologically active substance NA00226B and / or C.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION A physiologically active substance NA00226 having a phosphodiesterase inhibitory activity according to the present invention.
B and / or C producing bacteria are Streptomyces (Stre
ptomyces), for example, Streptomyces sp. NA00226 (S) isolated by the present inventors.
treptomyces sp. NA00226) strain (Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology, Accession number FER
MP-16196) is an example of a strain most effectively used in the present invention.

[0008] The strain belonging to the genus Streptomyces used in the present invention, like other strains of the genus Streptomyces, is liable to change its properties, and is easily mutated by, for example, an artificial mutating means using ultraviolet rays, X-rays and drugs. Any mutant strain having the ability to produce the physiologically active substance NA00226B and / or C of the present invention can be used in the present invention.

In order to produce NA00226B and / or C according to the present invention, the strain is first aerobically cultured in a medium containing nutrients that can be used by the strain. As the nutrient source, known ones conventionally used for culturing bacteria can be used.For example, as a carbon source, glucose, fructose, glycerin, sucrose, dextrin, galactose, organic acids, etc. are used alone or in combination. be able to.

As inorganic and organic nitrogen sources, ammonium chloride, ammonium sulfate, urea, ammonium nitrate,
Sodium nitrate, peptone, meat extract, yeast extract, dried yeast, corn steep liquor, soy flour, cottonseed oil scum, casamino acid, bactosoytone, soluble vegetable protein, oatmeal and the like can be used alone or in combination.

In addition, if necessary, inorganic salts such as salt, calcium carbonate, magnesium sulfate, copper sulfate, iron sulfate, zinc sulfate, manganese chloride, phosphate and the like can be added, and organic substances such as amino acids, vitamins, and nucleic acids can be added. And inorganic substances can be appropriately added. The most suitable culture method is a liquid culture method, particularly a submerged stirring culture method.
It is desirable that the culturing is performed at a culturing temperature of 20 ° C. to 45 ° C. and a pH of slightly acidic to slightly alkaline.

In liquid culture, when the culture is usually performed for 3 to 5 days, NA00226B and / or C substances are produced and accumulated in the culture solution. When the production amount in the culture solution reaches the maximum, the culture is stopped, the cells are separated from the culture solution, and the target substance is purified and isolated from the filtrate. The purification and isolation of the substance from the filtrate generally employs the separation and purification method used to isolate the microbial metabolite from the culture filtrate.

That is, the culture solution is separated into a filtrate and cells by a usual filtration method. The obtained filtrate is used as Diaion HP-20.
(Trade name: Mitsubishi Kasei) The solution is passed through a column to adsorb the target substance, washed with water and eluted with a 50% aqueous acetone solution. The eluted fraction was concentrated, extracted with ethyl acetate under hydrochloric acid, and extracted from the ethyl acetate layer with an aqueous sodium hydrogen carbonate solution. Further, the aqueous sodium hydrogen carbonate solution is extracted with n-butanol under acidic conditions with hydrochloric acid, and concentrated under reduced pressure.

Regarding the vacuum concentrate of the n-butanol layer,
Octadecylsilyl (hereinafter abbreviated as ODS) column chromatography developed with water-acetonitrile-formic acid is performed. About the obtained active fraction, Sephadex LH-
20 (trade name; Pharmacia Biotech) column chromatography (mobile layer: ethanol), centrifugal liquid-liquid distribution chromatography (ascending method: chloroform-methanol-water), and ODS column chromatography (mobile layer: water) -Acetonitrile-formic acid) to obtain NA00226B and C.

The physiologically active substance N obtained as described above
The physicochemical properties of A00226B and C are shown below.

Bioactive substance NA00226B 1) Appearance; red powder 2) Molecular weight: 447 3) Molecular formula; C ₂₃ H ₂₉ NO ₈ 4) Solubility; soluble in lower alcohol, insoluble in hexane, petroleum ether and water. 5) Rf value by ODS thin layer chromatography; 0.42 as a developing solvent of acetonitrile-water-formic acid (50: 50: 1 v / v). 6) Ultraviolet absorption spectrum; shown in FIG. 7) Infrared absorption spectrum; shown in FIG. 8) Hydrogen nuclear magnetic resonance spectrum; FIG. 3 shows a spectrum (300 MHz) measured in deuterated chloroform. 9) Carbon nuclear magnetic resonance spectrum; FIG. 4 shows a spectrum (75 MHz) measured in deuterated chloroform. The chemical shift values are shown below. δ (ppm) 189.1 (s), 181.0 (s), 1
70.7 (s), 158.4 (s), 149.7
(S), 141.4 (s), 134.8 (s), 12
8.3 (s), 119.3 (d), 112.7 (s),
99.5 (d), 75.0 (d), 72.5
(S), 69.9 (t), 69.8 (d), 60.
7 (t), 40.6 (t), 35.5 (t), 3
3.8 (t), 29.2 (q), 28.0 (t),
23.4 (q), 14.2 (q). 10) Color reaction; positive for vanillin-sulfuric acid, iron chloride and iodine.

Bioactive substance NA00226C 1) Appearance; red powder 2) Molecular weight; 433 3) Molecular formula; C ₂₂ H ₂₇ NO ₈ 4) Solubility; soluble in lower alcohols, insoluble in hexane, petroleum ether and water. 5) Rf value by ODS thin layer chromatography; 0.48 in a developing solvent of acetonitrile-water-formic acid (50: 50: 1 v / v). 6) UV absorption spectrum; shown in FIG. 7) Infrared absorption spectrum; shown in FIG. 8) Hydrogen nuclear magnetic resonance spectrum; FIG. 7 shows a spectrum (300 MHz) measured in deuterated chloroform. 9) Carbon nuclear magnetic resonance spectrum; FIG. 8 shows a spectrum (75 MHz) measured in deuterated chloroform. The chemical shift values are shown below. δ (ppm) 189.1 (s), 181.0 (s), 1
71.2 (s), 158.3 (s), 149.7
(S), 141.4 (s), 134.7 (s), 12
8.3 (s), 119.3 (d), 112.7 (s),
99.5 (d), 75.0 (d), 72.6
(S), 69.9 (t), 69.7 (d), 51.
9 (d), 40.3 (t), 35.4 (t), 3
3.6 (t), 29.3 (q), 27.9 (t),
23.4 (q). 10) Color reaction; positive for vanillin-sulfuric acid, iron chloride and iodine. Pharmaceutically acceptable salts of NA00226B and C can be produced by known methods, for example, by treating with a solution containing sodium hydroxide, potassium hydroxide and hydrochloric acid, sulfuric acid and the like.

Various methods known in the art can be applied to the preparation and administration of the compound when used as a pharmaceutical. That is, injection, oral, rectal administration and the like can be used as an administration method. The preparation may take the form of injections, powders, granules, tablets, suppositories and the like.

As long as it does not adversely affect NA00226B or C or a salt thereof during formulation, various adjuvants used for medicine, ie, carriers and other adjuvants such as stabilizers, preservatives, and soothing agents , An emulsifier and the like can be used as needed. In the preparation, the content of NA00226B or C or a salt thereof can be varied widely depending on the form of the preparation and the like. In general, the content of NA00226B or C or a salt thereof is 0.01 to 100% (weight), preferably 0.1 to 100%. 70% by weight, with the balance consisting of carriers and other auxiliaries normally used for pharmaceuticals.

The dose of NA00226B or C or a salt thereof varies depending on the symptoms and the like, but is not more than 0.1% per adult per day.
It is about 01 to 800 mg. When continuous throwing is required, it is preferable to reduce the daily consumption.

[0021]

The effect of NA00226B and C on phosphodiesterase inhibition will be described below with reference to experimental examples. Experimental example

Preparation of bovine phosphodiesterase 50 g of bovine airway smooth muscle after sacrifice was cut into small pieces with scissors and a scalpel, suspended in a 20 mM Tris buffer (pH 7.4) containing 5 mM EDTA 2 mM, and polytron homogenizer. To prepare a crude phosphodiesterase enzyme solution. After centrifugation at 10,000 × g for 20 minutes, the supernatant is used as a soluble phosphodiesterase crude enzyme solution. Furthermore, Q-Sepharose Fast Flow Column (trade name; Pharmacia Biotech) (100 ml)
, A crude enzyme solution of soluble phosphodiesterase is added thereto, washed with a buffer, and eluted with a concentration gradient of 50 mM to 1 M sodium acetate. The phosphodiesterase activity fraction eluted with 0.7 M sodium acetate was used as a phosphodiesterase enzyme solution.

Measurement of Phosphodiesterase Activity Phosphodiesterase activity is determined by using 3 ', 5'-cyclic adenosine monophosphate (Sigma, USA, hereinafter referred to as cAMP) as a substrate and remaining after reaction. The cAMP thus obtained was measured by high performance liquid chromatography. That is, 0.02 mg / ml cAMP, 2.5 mM dithiothreitol, 6 mM magnesium chloride, 50 mM Tris buffer (pH 8.0), 1 μl of a phosphodiesterase enzyme solution and an evaluation sample were added to a 0.3 ml plastic tube. 20 final volume with water
The volume was adjusted to 0 μl to obtain tubes having each evaluation sample concentration shown in the table below. The mixture is agitated and incubated at 37 ° C. for 60 minutes. The reaction is stopped by adding 20 μl of a 200 mM aqueous solution of tetraethylenediamine to each tube. 20 μl of this solution was subjected to high performance liquid chromatography (ODS column: φ3.9 × 150 mm, WATE
(RS, USA) to measure residual cAMP. Table 1 shows the phosphodiesterase inhibitory activity of the compound of the present invention measured by this method.

[0024]

Table 1 Inhibitory activity of phosphodiesterase Table 1 Inhibitory activity (%) Concentration (μg / ml) NA0026B NA0026C 36 85.4 88.1 12 86.4 68.6 4.0 69.0 48.9 48.9 33 37.7 36.2 Thus, NA0026B and C exhibit strong inhibitory activity against phosphodiesterase, and their IC ₅₀ is 5.3 × 10 ⁻⁶ M and 1.0 × 10 ⁻⁵ M, respectively. .

[0025]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be described below, but these are merely examples and do not limit the present invention at all, and various modifications are possible.

Production Method In a 500 ml Erlenmeyer flask for rotary shaker, glycerin 2.0%, glucose 1.0%, soybean powder 0.5
%, Peptone 0.3%, yeast extract 0.5%, calcium carbonate 0.2%, dipotassium phosphate 0.05%, magnesium sulfate 0.05%, 100 ml of a medium (pH 7.0) was dispensed. The solution was autoclaved at 20 ° C. for 20 minutes.
This was inoculated with one platinum loop of NA00226 strain (FERM-16196, National Institute of Bioscience and Biotechnology).
Shaking culture was performed for 2 days at 220 ° C. at 220 ° C. to obtain a seed culture. 500ml for rotary shaker
1.0% glucose, galactose in an Erlenmeyer flask.
0%, dextrin 2.0%, corn steep liquor 1.0%, pharma media 1.0%, magnesium sulfate 0.05%, cobalt chloride 0.0001%, calcium carbonate 0.2% medium (pH 7.0) 1) 100 ml was dispensed, and 1 ml of the above seed culture solution was transplanted into a flask autoclaved at 120 ° C for 20 minutes, and cultured with shaking at 25 ° C and 220 rpm for 5 days.

The obtained culture solution (40 L) was separated into a filtrate and cells by a usual filtration method. The obtained filtrate was adsorbed on a Diaion HP-20 column (4.5 L), and water (9
L) Wash and elute with 50% aqueous acetone (8 L). The acetone is distilled off from the eluted fraction, and then acidified with hydrochloric acid (pH
3) Extract with ethyl acetate. The extract was extracted with an aqueous solution of sodium hydrogen carbonate (2 L) under alkaline conditions (pH 8). Further, extraction was performed from the aqueous sodium hydrogen carbonate solution with n-butanol (2 L) under acidic conditions of hydrochloric acid (pH 3). About the vacuum concentrate (5.544 g) of the n-butanol layer, acetonitrile-water-formic acid (3: 5: 0.01) was used.
ODS column chromatography (φ5 × 25 cm) developed at ６6: 2: 0.01 v / v). The active fraction eluted at an acetonitrile concentration of 70 to 75% was concentrated to obtain a concentrated and dried product (1.637 g). The concentrated and dried product was subjected to Sephadex LH-20 column chromatography (φ3.5 × 45 cm, mobile phase: ethanol) to obtain an active fraction (0.378 g), which was further subjected to centrifugal liquid partition chromatography [elevation method: Chloroform-methanol-water (5: 6: 4 v / v) was carried out and the active fraction B (0.08
6g) and active fraction C (0.018 g). Active fraction B was analyzed by ODS column chromatography [φ2.5 × 4
2 cm, acetonitrile-water-formic acid (3: 7: 0.01
~ 6: 4: 0.01 v / v)] and HPLC
[CAPCELL PAK UG120 (Shiseido);
Acetonitrile-5 mM ammonium acetate aqueous solution (33: 6
7v / v)] and purified by SEP-PAK (WATERS
Desalting using NA00226B (2.0
mg). The active fraction C was analyzed by ODS column chromatography [φ2.5 × 42 cm, acetonitrile-
Water-formic acid (3: 7: 0.01 to 6: 4: 0.01 v /
v)] and HPLC [CAPCELL PA]
K UG120 (Shiseido); acetonitrile-5 mM
Ammonium acetate aqueous solution (33:67 v / v)]
Desalting was performed using SEP-PAK (WATERS) to obtain NA00226C (0.9 mg). Using purified NA00226B and C, appearance, molecular weight, solubility, Rf value by ODS thin layer chromatography, ultraviolet absorption spectrum, infrared absorption spectrum, ¹ H-NMR spectrum, and ¹³ C-NMR spectrum were measured. NA0
The physicochemical properties of 0226B and C were as described above.

[0028]

As is clear from the above, the physiologically active substance NA00226B or C or a pharmaceutically acceptable salt thereof of the present invention has a phosphodiesterase inhibitory action, and is, for example, an anti-asthmatic drug, a bronchodilator, a bronchial drug. Anti-inflammatory drugs, anti-allergic drugs, anti-inflammatory drugs, anti-rheumatic drugs, antihypertensive drugs, angina drugs, arrhythmia drugs, cerebral circulation metabolism drugs,
It can be expected as an active ingredient such as an anticoagulant and an antidepressant.

[Brief description of the drawings]

FIG. 1. Ultraviolet absorption spectrum of NA00226B measured in methanol

FIG. 2 Infrared absorption spectrum of NA00226B

FIG. 3. Hydrogen nuclear magnetic resonance spectrum of NA00226B measured in deuterated chloroform

FIG. 4: Carbon nuclear magnetic resonance spectrum of NA00226B measured in deuterated chloroform

FIG. 5: Ultraviolet absorption spectrum of NA00226C measured in methanol

FIG. 6 is an infrared absorption spectrum of NA00226C.

FIG. 7. Hydrogen nuclear magnetic resonance spectrum of NA00226C measured in deuterated chloroform

FIG. 8: Carbon nuclear magnetic resonance spectrum of NA00226C measured in deuterated chloroform.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 45/00 ABN A61K 45/00 ABN ABQ ABQ ACB ACB ACD ACD ADV ADV C12N 1/20 C12N 1/20 A C12P 1/06 C12P 1/06 Z // (C12N 1/20 C12R 1: 465) (C12P 1/06 C12R 1: 465)

Claims (7)

[Claims] 1. A physiologically active substance N having the following physicochemical properties:
A00226B or a pharmaceutically acceptable salt thereof 1) Appearance; red powder 2) Molecular weight; 447 3) Molecular formula; C ₂₃ H ₂₉ NO ₈ 4) Solubility; soluble in lower alcohol, insoluble in hexane, petroleum ether, water . 5) Rf value by ODS thin-layer chromatography; 0.42 in a developing solvent of acetonitrile-water-formic acid (50: 50: 1 v / v). 6) Ultraviolet absorption spectrum; shown in FIG. 7) Infrared absorption spectrum; shown in FIG. 8) Hydrogen nuclear magnetic resonance spectrum; FIG. 3 shows a spectrum (300 MHz) measured in deuterated chloroform. 9) Carbon nuclear magnetic resonance spectrum; FIG. 4 shows a spectrum (75 MHz) measured in deuterated chloroform. The chemical shift values are shown below. δ (ppm) 189.1 (s), 181.0 (s), 1
70.7 (s), 158.4 (s), 149.7
(S), 141.4 (s), 134.8 (s), 12
8.3 (s), 119.3 (d), 112.7 (s),
99.5 (d), 75.0 (d), 72.5
(S), 69.9 (t), 69.8 (d), 60.
7 (t), 40.6 (t), 35.5 (t), 3
3.8 (t), 29.2 (q), 28.0 (t),
23.4 (q), 14.2 (q). 10) Color reaction; positive for vanillin-sulfuric acid, iron chloride and iodine. 2. A physiologically active substance N having the following physicochemical properties:
A00226C or pharmaceutically acceptable salt thereof 1) Appearance; red powder 2) Molecular weight; 433 3) Molecular formula; C ₂₂ H ₂₇ NO ₈ 4) Solubility; soluble in lower alcohol, insoluble in hexane, petroleum ether, water . 5) Rf value by ODS thin layer chromatography: 0.48 as a developing solvent of acetonitrile-water-formic acid (50: 50: 1 v / v). 6) UV absorption spectrum; shown in FIG. 7) Infrared absorption spectrum; shown in FIG. 8) Hydrogen nuclear magnetic resonance spectrum; FIG. 7 shows a spectrum (300 MHz) measured in deuterated chloroform. 9) Carbon nuclear magnetic resonance spectrum; FIG. 8 shows a spectrum (75 MHz) measured in deuterated chloroform. The chemical shift values are shown below. δ (ppm) 189.1 (s), 181.0 (s), 1
71.2 (s), 158.3 (s), 149.7
(S), 141.4 (s), 134.7 (s), 12
8.3 (s), 119.3 (d), 112.7 (s),
99.5 (d), 75.0 (d), 72.6
(S), 69.9 (t), 69.7 (d), 51.
9 (d), 40.3 (t), 35.4 (t), 3
3.6 (t), 29.3 (q), 27.9 (t),
23.4 (q). 10) Color reaction; positive for vanillin-sulfuric acid, iron chloride and iodine. 3. Streptomyces (Streptomyc)
es) belonging to the genus, the biologically active substance NA00226B and / or
Alternatively, a microorganism having the ability to produce C is cultured in a medium, and the physiologically active substance NA00226B and / or C is produced and accumulated in the culture, and the physiologically active substance NA00226B or C or a pharmaceutical thereof is collected. For the production of chemically acceptable salts. 4. A physiologically active substance NA00226B or C
Or a medicament comprising a pharmaceutically acceptable salt thereof as an active ingredient. 5. A physiologically active substance NA00226B or C
Or a phosphodiesterase inhibitor comprising a pharmaceutically acceptable salt thereof as an active ingredient. 6. An anti-asthmatic drug, a bronchodilator, a therapeutic agent for bronchitis, an anti-allergic drug, an anti-inflammatory drug, an anti-rheumatic drug, a hypotensive drug, a therapeutic drug for angina, a therapeutic drug for arrhythmia, a drug for improving cerebral circulation and metabolism,
5. A blood coagulation inhibitor or an antidepressant drug.
The medicament according to claim. 7. Streptomyces (Streptomyc)
es) belonging to the genus, the biologically active substance NA00226B and / or
Or a microorganism capable of producing C.