JP2742586B2 - TAN-1030A and its derivatives, their production and use - Google Patents

Description

The present invention relates to a novel compound TAN-1030A (hereinafter referred to as "TAN-103") useful as a therapeutic agent for infectious diseases such as bacteria and fungi and malignant tumors.
0A ”), TAN-1030A and its derivatives, and the use of TAN-1030A.

2. Description of the Related Art As a common antibiotic in the basic skeleton with the antibiotic TAN-1030A of the present invention, Straurospori
ne) are known [Journal of Antibiotics, 30 , 275-282 (1977) and JCS Chemical Communications (JCSChem.C
omm.), 800-801 (1978)], but has a different structure from TAN-1030A, and also differs in the producing bacteria.

In addition, the antibacterial activity of staurosporine against some microorganisms is known, including staurospon, TAN-103.
There is no description of any other physiological activity of the compound having the same basic skeleton as OA.

Problems to be Solved by the Invention Phagocytic cells such as macrophages usually not only have the ability to phagocytose, digest, and detoxify foreign foreign substances and waste products in vivo, but also have an inhibitory effect on tumor cell proliferation, It has extremely diverse capabilities, such as controlling biological functions by producing active substances, establishing and expressing immunity, and plays an important role in biological defense.

In recent years, if the functions of host phagocytes and lymphocytes are reduced by infection with bacteria, fungi, viruses, etc., malignant tumors and their treatment, administration of immunosuppressants in organ transplantation, etc., resistance to infection and other diseases is reduced. The deterioration of the disease, the susceptibility to various diseases, and the seriousness of the disease have become major problems in the medical field. In order to overcome these problems, there is a need in the art for novel compounds having an activating effect on phagocyte functions such as macrophages or intermediate materials for synthesizing them.

Means for Solving the Problems In view of the current situation, the present inventors have repeatedly studied macrophage activating substances from a new viewpoint, and found that among a large number of microorganisms isolated from soil, That the microorganism belongs to the genus Streptomyces, by culturing the microorganism in a suitable medium, to activate a mouse-derived culture system macrophages, a compound having the effect of significantly changing the morphology in the medium Knowing that it could accumulate, the compound was isolated and its physicochemical and biological properties confirmed that the compound was a novel substance, which was named TAN-1030A. The present inventors have further studied based on these findings and completed the present invention.

That is, the present invention relates to the general formula TAN-1030A or a salt thereof represented by, culturing a TAN-1030A-producing bacterium belonging to the genus Streptomyces in a medium, producing and accumulating TAN-1030A in the culture, collecting the TAN-1030A, and optionally obtaining TAN-1030A. To a catalytic reduction reaction and, if desired, to an argylation or (and) acylation reaction. [Wherein Q 'is C = N-OH or CH-NR ¹ ' R ² '
Wherein R ¹ ′ and R ² ′ each represent hydrogen, lower alkyl or lower alkanoyl.] Or a macrophage comprising TAN-1030A or a salt thereof. An activator is provided.

Regarding Q ′ in the compound (I ′), the lower alkyl represented by R ¹ ′ and R ² ′ includes C _1-6 alkyl,
The alkyl may be linear or branched, and examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, hexyl and the like.

As lower alkanoyl represented by R ¹ ′ and R ² ′, C _1-6
Alkanoyl is mentioned, for example, formyl, acetyl, propionyl, isopropionyl, butyryl, pentanoyl, hexanoyl and the like.

When Q ′ is an amino group or a derivative thereof, compound (I ′) may form a physiologically acceptable salt such as an inorganic acid or an organic acid.
Mineral acids such as sulfuric acid, phosphoric acid and the like, and the organic acids include acetic acid, oxalic acid, citric acid, malic acid, maleic acid and the like.

Further, when Q ′ is an amino group or a derivative thereof,
It may be a quaternary salt with a lower alkyl, and its counter anion is halogen (eg, fluorine, chlorine, bromine,
Iodine) ion.

The TAN-1030A-producing bacterium used in the method of the present invention may be any microorganism that belongs to the genus Streptomyces and has the ability to produce TAN-1030A. An example is C isolated from soil collected in Okinawa Prefecture.
-71799 strain, and the mycological properties of this strain are as follows. Unless otherwise specified, the properties on various media were measured at 28 ° C for 14 days and observed according to a conventional method, and the color tone was measured using the Color Harmony Manual (Co
lor Harmony Mannual 4th edition, Container Corporation of America
of America), published in 1958.

1. Morphology The formation of aerial mycelia and spores is abundant on culture media such as yeast extract / malt extract agar, starch inorganic salt agar, glycerol / asparagine agar. The branch of the aerial hyphae is a simple branch, no axle branch is observed, and a chain of spores is observed at the tip of the branch. No special structure such as fungal thread, sclerotium, or spore sac is observed. Observation with an electron microscope shows that the surface of the spores is smooth, the spores are ovoid or cylindrical, and their size is 0.4-0.8 × 1.0-1.5 μm
And usually have 10 or more chains. Also, these spores do not show motility.

2. Properties on various classified media (cultured at 28 ° C for 14 days) 1) Sucrose / nitrate agar medium Growth: good Aerial mycelium: abundant, white to pearl (2ba) Back color: light ivory (2ca) to bamboo (2gc) Soluble pigment: None 2) Yeast extract / malt extract agar medium Growth: Good Aerial mycelia: Abundant, white to pearl gray (13cb) Back color: Light melon yellow (2ea) to camel (3ie) Soluble Pigment: Production 3) Oat-meal agar medium Growth: Good Aerial mycelium: Abundant, white to down blue (15dc) Back color: Light ivory (2ca) to P green (24 1/2 ie) Soluble pigment: None 4) Starch inorganic salt agar medium Growth: Good Aerial mycelium: Abundant, white to down blue (15dc) Back color: Light ivory (2ca) to bamboo (2gc) Soluble pigment: None 5) Glycerol asparagi Agar medium Growth: Good Aerial mycelium: Abundant, white to pearl gray (13dc) Back color: Light ivory (2ca) to Olive gray (2ig) Soluble pigment: None 6) Glucose asparagine agar medium Growth: Good Aerial mycelium: Abundant, white to down blue (15dc) Back color: Light ivory (2ca) to Bamboo (2gc) Soluble pigment: None 7) Peptone, yeast extract, iron agar medium Growth: Poor Aerial mycelium: No growth Back color: bamboo (2gc) to cobalt brown (2nl) Soluble pigment: none 8) Tyrosine agar medium Growth: good Aerial mycelium: Abundant, white to dusk (13fe) Back color: light mustard・ Tan (2ie)-Dark brown (2pn) Soluble pigment: None 9) Calcium maleate agar medium Growth: Medium Aerial hyphae: Abundant, white to natural (3dc) Back color Light ivory (2ca) to pea green (24 1/2 ie) Soluble pigment: none 3. Physiological properties 1) Growth temperature: 13 ° C to 42 ° C Optimum temperature: 22 ° C to 38 ° C 2) Liquefaction of gelatin : Positive 3) Starch hydrolysis: Positive 4) Milk peptone: Positive 5) Milk coagulation: Negative 6) Nitrate reduction: Positive 7) Growth of melanin-like pigment: Negative 8) Use of carbon source: D -Sorbitol + sucrose + i -inositol + lactose + D-mannose + raffinose + D-xylose + trehalose + L-arabinose + mannose + D-galactose + soluble starch + D-glucose + glycerol + D-ramose fructose + Maltose + Cellulose-Ribose-No Addition-Basal Medium: Preed Ham and Gottliep Agar medium 4. Analysis This bacteria diaminopimelic acid and sugars in hydrolysis of whole cell contains a LL-2,6-diaminopimelic acid in the hydrolysis of whole cell, it does not contain arabinose and Majurosu.

To summarize the above taxonomic properties, the C-71799 strain is:
The aerial hyphae is bent and the spore surface is smooth.
Does not form mycelium and does not produce motile spores. The growth color is pale yellow or yellowish brown. The aerial mycelium is white or gray. LL-diaminopimelic acid is detected in the cells, but arabinose This strain belongs to cell wall type I and glycoform belongs to NC type because no majulose is detected. Based on the above properties, edited by R.E. Buffanan and N.E. Gibbons, The Barges Manual of Detergent Bacteriology (Be
rgey's Manual of Determinative Bacteriology)
As a result of searching for bacterial species according to the
Since the strain 1799 was found to belong to the genus Streptomyces, the bacterium was transformed into Streptomyces sp.
sp.) C-71799.

The C-71799 strain was deposited with the Fermentation Research Institute (IFO) on November 11, 1987 under the deposit number IFO-14672. It has been deposited with the Research Institute (FRI) under the deposit number FERM P-9714.

TAN-1030A producing bacteria belonging to the genus Streptomyces,
As with other actinomycetes, their properties tend to change,
For example, even if many mutant strains are obtained by mutation by irradiation with ultraviolet rays, X-rays, radiation, etc., single spores, various mutation treatments, or other means or naturally obtained mutant strains, Any substance which is substantially different from the kind in comparison with the properties and has the property of producing the substance can be used in the method of the present invention.

The medium used for culturing the substance-producing bacterium may be liquid or solid as long as it contains a nutrient source that can be used by the bacterium, but it is more appropriate to use a liquid medium when treating a large amount. The medium is appropriately mixed with a carbon source that can be assimilated by the substance-producing bacterium, a nitrogen source that can be assimilated, inorganic substances, and micronutrients. Examples of the carbon source include glucose, lactose, sucrose, malt, dextrin, starch, glycerin, mannitol, sorbitol, fats and oils (eg, soybean oil, lard oil, chicken oil, etc.), n-paraffin, and the like. Examples include meat extract, yeast extract,
Dried yeast, soy flour, corn steep liquor, peptone, cottonseed flour, molasses, urea, ammonium, salts (eg, ammonium sulfate, ammonium chloride, ammonium nitrate, ammonium acetate, etc.) and the like are used. Further, salts containing sodium, potassium, calcium, magnesium, etc., metal salts such as iron, manganese, zinc, cobalt and nickel, salts such as phosphoric acid and boric acid, and salts of organic acids such as acetic acid and propionic acid are appropriately used. . In addition, amino acids (eg, glutamic acid, aspartic acid, alanine, lysine, methionine, proline, etc.), peptides (eg, dipeptide, tripeptide, etc.), vitamins (eg, B ₁ , B ₂ , nicotinic acid, B ₁₂ , C ), Nucleic acids (eg, purine, pyrimidine, derivatives thereof, etc.). Of course, an inorganic or organic acid, alkali, buffer or the like may be added for the purpose of adjusting the pH of the medium, or an appropriate amount of an oil or fat, a surfactant or the like may be added for the purpose of defoaming. During liquid culture, the pH of the medium is preferably around neutral, particularly preferably about 6 to 8. The culture temperature is preferably about 24 ° C. to 30 ° C., and the culture time is preferably about 48 hours to 120 hours.

The titer of TAN-1030A produced during the course of the culture is quantified according to a liquid dilution method using mouse-derived cultured macrophages Mm1 as test cells. Usually, the production of TAN-1030A reaches the maximum in about 2-3 days of culture.

In order to collect the desired compound TAN-1030A from the culture, TAN-1030A is a neutral and fat-soluble compound and can be collected by a general means utilizing this property.

Since TAN-1030A in the culture is contained in the cells and the filtrate, the culture is adjusted to pH 5 to 9, preferably pH 6 to 8, and then an organic solvent immiscible with water, for example, dichloromethane, ethyl acetate, methyl isobutyl ketone. Alternatively, a method of extracting an active substance by adding isobutanol or the like is used. In addition, an organic solvent miscible with water, such as acetone or methanol, is added to the culture, and the mixture is stirred and extracted. After insolubles are removed by filtration, the organic solvent in the extract is distilled off under reduced pressure. Either by the above extraction method or by using an adsorbent resin such as Amberlite XAD-II (manufactured by Rohm and Haas Co., USA), Diaion HP-20 (manufactured by Mitsubishi Kasei Co., Japan) or Amberlite SP-207 (Mitsubishi Chemical) A method of performing a chromatographic method using, for example, Kasei Co., Japan) is advantageously used. In order to elute the desired active substance from the column using the adsorptive resin, water or a water-containing solvent such as water-containing methanol or water-containing acetone is advantageously used. The extract or eluate thus obtained is concentrated under reduced pressure to obtain a crude substance containing TAN-1030A.

Various chromatographic methods are advantageously used to further purify the crude material to obtain pure TAN-1030A. Silica gel, cellulose, Sephadex LH-
20 (manufactured by Pharmacia, Sweden) and the like, which are usually performed by column chromatography. To elute the active substance from the column, a suitable organic solvent such as n-hexane, ethyl acetate, dichloroethane, chloroform, acetone, methanol or the like, alone or in a mixed solvent is used. When the eluate is concentrated and left in a cool place, or the concentrated residue is dissolved in a suitable crystallization solvent such as diethyl ether, chloroform, ethyl acetate, methanol or a mixture thereof, and left in a cool place,
A crystal of TAN-1030A is obtained.

Next, the catalytic reduction method of TAN-1030A will be described. The catalytic reduction method is carried out by a generally known method, and is advantageously carried out under the following conditions. The catalyst used is platinum black, palladium black or palladium-carbon, and the amount is TA
10 to 70% of N-1030A, preferably 20 to 40%
%. As the reaction solvent, ethanol, acetic acid and their aqueous solvents are advantageously used. The reaction time is 2 to 16 hours, preferably 3 to 10 hours. The reaction temperature is 10 to 50 ° C, preferably 15 to 35 ° C.

The amino derivative of TAN-1030A obtained above (I; R ¹ = R ²
= H) can be subjected to an alkylation or alkanoylation reaction to lead to various derivatives.

Alkylation involves alkyl halide, dimethyl sulfate,
Formic acid and formic ester in an organic solvent (alcohol, acetone, ethyl acetate, chloroform, etc.) or a water-containing solvent thereof, optionally in the presence of a basic catalyst, at room temperature to the boiling point of the solvent (15 to 130 ° C.) at a temperature of 0.5 to 0.5%. The reaction is carried out for up to 10 hours.

Alkanoylation is carried out by subjecting the corresponding halide, acid anhydride or the like to the same solvent as in the above alkylation at room temperature in the presence of a basic catalyst (pyridine, potassium or sodium bicarbonate, potassium or sodium hydroxide, etc.). ~
The reaction is carried out at the boiling point of the solvent (15 to 130 ° C.) for 0.5 to 10 hours.

The compound (I) thus obtained can be separated by ordinary separation and purification means (extraction, chromatography, recrystallization, etc.).

The physicochemical properties of TAN-1030A obtained in Example 2 described below are as follows.

1) Appearance: colorless crystals 2) Melting point: 290-295 ° C (decomposition point) 3) Mass spectrum measurement: m / z 467 (M + H) ⁺ (SI-M)
S method) 4) Elemental analysis value: (%) ^* Calculated as including one H ₂ O molecule Observed value: C, 67.23; H, 5.07; N, 11.70 Calculated value ^* ; C, 66.93; H, 4.99; N, 11.56 5) Molecular formula: C ₂₇ H ₂₂ N ₄ O _4. (H ₂ O) 6) Ultraviolet absorption (UV) spectrum: 233 (630), 244 (600, shoulder), 263 (670 shoulder), 275 (900
Shoulder), 289 (1520), 319 (286 shoulder), 333 (380), 352 (26
0), 369 (286) 7) Infrared absorption (IR) spectrum: in KBr, FIG. 2, ν _max cm ⁻¹ , showing main absorption.

3430,3160,2940,2860,1675,1590,1455,1400,1350,132
0,1285,1255,1230,1120,1020,945,920,840,775,740 8) ¹ H nuclear magnetic resonance (NMR) spectrum: 300 MHz, DMSO-d ₆
Medium, δppmJ (Hz), 10.45 (s), 9.31 (d, J = 7.8), 8.58
(S), 8.01 (d, J = 9.1), 7.98 (d, J = 8.0), 7.70 (d,
J = 8.2), 7.50 (t), 7.44 (t), 7.32 (t × 2), 7.04
(D, J = 5.2), 4.96 (s, like), 4.73 (s), 3.63 (d, J =
14.0), 3.43 (s), 3.01 (dd, J = 5.2,14.0), 2.47
(S) 9) ¹³ C NMR spectrum: 75 MHz, in _{DMSO-d 6, δppm 171.77 (} s), 145.12 (s), 139.82 (s), 136.01
(S), 132.25 (s), 128.03 (s), 125.61 (d), 125.
21 (d), 124.64 (d), 124.60 (s), 123.79 (s), 12
2.85 (s), 120.72 (d), 120.13 (d), 119.52 (d),
119.15 (s), 115.58 (d), 114.98 (s), 114.02
(S), 108.88 (d), 96.16 (s), 83.57 (d), 82.17
(D), 58.29 (q), 45.30 (t), 29.71 (t), 28.59
(Q) (However, s: singlet, d: doublet, t: triplet, q: quartet
Represents).

10) Thin layer chromatography (TLC): carrier; silica gel 60F ₂₅₄ (manufactured by Merck, West Germany) Developing solvent: ethyl acetate: methanol (96: 4) Rf = 0.61 11) High-performance liquid chromatography (HPLC): carrier: ODS Column (YMC-Pack A312, manufactured by Yamamura Chemical Laboratory) Moving bed: 55% acetonitrile / 0.01M phosphate buffer (pH 6.3) Flow rate: 2 ml / min Detection: 254 nm Rt = 4.7 (min) 12) Color reaction: positive; Ehrlich and Barton reaction negative; Dragendorff and ninhydrin reaction 13) various analyzes of the physical over-biological properties and NMR spectrum division described above neutral fat-soluble substance substance ⁽¹ H- ¹ HCOSY method, ¹ H- ¹³ C Correlation method and
According to the COLOC method, the chemical structure of TAN-1030A is estimated to be the following formula.

Action Macrophages (Mm) in mouse culture of compound (I ')
1) The effects on cells are shown in Table 1.

Activating action; ability to induce activated macrophage-like morphology
+, Positive, ±, Positive but weak,-, Negative, in drug-containing Git medium (Nippon Pharmaceutical, Japan) at 37 ° C under 5% CO ₂
After culturing for 3 days, the cell morphology was observed.

Next, a 10% proteose peptone (Difco, USA) solution was intraperitoneally administered to C57BL / 6 (10-week-old, female) mice, and peritoneal cells were collected on the fourth day. Were collected and used as macrophages to examine the effect of TAN-1030A on mouse macrophages. As a result, TAN-1030A was cultured in the presence of 1 ng / ml for 2 days [RPMI 1640 containing 10% (V / V) fetal calf serum (Wittercar M. A. Bio Products, USA).
The phagocytic activity-dependent active oxygen release ability (luminol-dependent chemiluminescence method) of macrophages against zymosan (Sigma, USA) in the medium (Wittercar M. A. Biopdaroctes, USA) It was about 1.3 times stronger.

Further, the growth inhibitory effect of TAN-1030A on mouse tumor cells is as shown in Table 2.

As is clear from the above physicochemical and biological properties, TAN-1030A of the present invention is a novel compound, and compound (I ') activates cultured macrophages,
It is a useful substance as a pharmaceutical or animal drug because it has a strong growth inhibitory effect on tumor cells.

When compound (I ') is used as an antitumor agent, it is mixed with a diluent, excipient, carrier and the like to make a pharmaceutically acceptable preparation, and is orally or parenterally administered.

For example, when used as an injection, compound (I ')
0.1mg / kg to 10mg / day
mg / kg is administered intravenously.

Examples Hereinafter, the present invention will be described more specifically with reference to Examples, but the present invention is not limited thereto. The percentage (%) in the medium represents weight / volume percent unless otherwise specified.

Example 1 A Streptomyces sp. C-71799 strain cultured on a yeast extract / malt extract slant agar medium was used in a 200 ml Erlenmeyer flask containing 2% glucose, 3% soluble starch, 1% raw soybean powder, 1% corn steep. Liquor 1%, Peptone 0.5
%, NaCl 0.3%, CaCO ₃ 0.5%, 40ml seed medium (pH 7.0)
And cultured on a rotary shaker at 28 ° C. for 48 hours to obtain a preculture. 5 ml of the obtained preculture was transferred to a 500 ml seed medium in a 2000 ml Sakaguchi flask, and cultured on a reciprocating shaker at 28 ° C. for 48 hours to obtain a seed culture. Add 500 ml of this seed culture to 50
Stainless steel tank seed medium (transplanted to the same composition 30 as the above seed medium, aeration 30 / min, stirring 280 rpm /
The cells were cultured under the conditions of an internal pressure of 1 kg / cm ² for 1 minute. 5 of the obtained culture solution was added to a 200-volume stainless steel tank with 0.5% glucose, 5% dextrin, 3.5% defatted soy flour, and CaC
o Transfer to 120 main media (pH 7.0) containing ₃ 0.7%
Culturing was carried out for 96 hours at a temperature of 120 ° C., aeration of 120 / min, stirring at 180 rpm, and an internal pressure of 1 kg / cm ² .

Example 2 35 was extracted from the culture solution obtained as in Example 1, the pH of the culture solution was adjusted to 7.9, and Hyflo Supercell was added to the culture solution, followed by filtration to obtain bacterial cells. Acetone 15 was added to the cells, and after stirring for 30 minutes, Hyflo Supercell was added and the mixture was filtered to obtain a filtrate. After the filtrate was concentrated to about 6, 500 g of sodium chloride was added, and the mixture was extracted twice with ethyl acetate (3). The obtained organic layer was washed with water (1.5), dehydrated with sodium nitrate, and concentrated to dryness. The residue was subjected to silica gel (125 g) column chromatography, and ethyl acetate (0.8, Fr. (fraction) 1-4),
Ethyl acetate: methanol = 19: 1 (1.0, Fr. No.5-
9), ethyl acetate: methanol = 9: 1 (1.4, Fr. No. 10)
In -16), elution fractionation was performed in 0.2 steps. Fr.No.9-16
After concentrating to dryness, it was crystallized with chloroform-methanol to obtain 1.13 g of staurosporine crystals (melting point: 270 ° C (decomposition)).
I got

Fr.No.2-7 was collected and concentrated, and then subjected to silica gel (50 g) column chromatography.
After washing with 4: 1 (300 ml), elution was performed with ethyl acetate (200 ml). The eluate was concentrated to dryness, and the crude powder of TAN-1030A (1.07
g) was obtained. The resulting coarse powder was subjected to silica gel (50 g) column chromatography, and chloroform (500 m
l), chloroform: methanol = 49: 1 (600m
The fraction was eluted in l). Collect the effective fractions, concentrate to dryness and TAN
A powder of -1030A (600 mg) was obtained. The powder was crystallized from chloroform-methanol and crystalized as TAN-1030A (110 mg).
was gotten.

Example 3 A suspension of TAN-1030A (35 mg) in 60% aqueous acetic acid (10 ml) was
Platinum black (10 mg) was added, and the mixture was stirred at room temperature under a hydrogen gas atmosphere for 7.5 hours. The reaction solution was filtered and then concentrated. 0.05 after concentration
N hydrochloric acid (30 ml) was added, and the mixture was washed with ethyl acetate (30 ml). After adjusting the pH of the aqueous layer to 10, it was extracted twice with ethyl acetate (30 ml), and the obtained organic layer was washed with saturated saline (15 ml) and dried over anhydrous sodium sulfate. The solution was concentrated to dryness to obtain a powder of a dihydro-deoxy form of TAN-1030A (28 mg). Mass spectral measurements: m / z453 (M + H ) + 1
1 H NMR (300 MHz, CDCl ₃ ): δ 9.44 (1 H, d, J = 8.2 Hz), 7.9
7 (1H, d, J = 8.6Hz), 7.89 (1H, d, J = 7.3Hz), 7.50 (1H,
t), 7.44 (1H, t), 7.38 (1H, t), 7.33 (1H, t), 7.29 (1
H, d, J = 7.9Hz), 6.58 (1H, d, J = 4.0Hz), 6.33 (1H, s),
5.02 (2H, ABq), 3.75 (2H, m), 3.46 (3H, s), 2.60 (2H,
m), 3.33 (3H, s) ppm Example 4 To a dihydro-deoxy form of TAN-1030A (30 mg) were added pyridine (0.6 ml) and acetic anhydride (0.3 ml), and the mixture was allowed to stand at room temperature for 3 hours. After the reaction, ethyl acetate (20 ml) was added, and 0.05N
Washing with hydrochloric acid and brine was successively performed. The obtained organic layer was dried over sodium nitrite, concentrated, and ether was added thereto to add TAN-1030A dihydro-.
A powder (28 mg) of a deoxy-form monoacetyl form was obtained. ¹ H NMR (300 MHz, CDCl ₃ ) δ 9.40 (1 H, d, J = 7.9 Hz), 7.97
(1H, d, J = 9.0Hz), 7.94 (1H, d, J = 8.49Hz), 7.51 (1H,
t), 7.48 (1H, t), 7.39 (2H, t), 7.28 (1H, d, J = 9.4H)
z), 6.71 (1H, br), 6.61 (1H, d, J = 5.2Hz), 5.16 (1H, br
d, J = 6.2Hz), 5.04 (2H, ABq), 4.60 (1H, m), 3.91 (1H,
d, J = 4.4Hz, 3.40 (3H, s), 3.09 (1H, dd, J = 3.1,15.0H
z), 2.53 (1H, m), 2.39 (3H, s), 0.81 (3H, s) Example 5 10% sodium hydroxide (7 ml) and dimethyl sulfate (0.7 ml) were added to staurosporine (146 mg). The mixture was stirred in an ice bath for 30 minutes and then at room temperature for 3 hours. The reaction solution was adjusted to pH 8.7, washed three times with ethyl acetate, and the aqueous layer was adjusted to pH 3 and then concentrated. Methanol / ether was added to the concentrated solution to obtain a precipitate (1.17 g). A small amount of water was added to the obtained precipitate, followed by filtration. The residue was dissolved in 20% aqueous methanol (100 ml), and Amberlite IRA-402 (Cl type, Rohm and
(15 mL, Haas, USA) and washed with water (20 mL). After combining the passing solution and the washing solution, the solvent was distilled off under reduced pressure. Add methanol / ether to the residue and add TAN-1030A
Dihydro-deoxy trimethyl hydrochloride powder (92
mg).

Mass spectrum measurement: m / z 495 ¹ H NMR (300 MHz, DMSO-d ₆ ): δ 9.32 (1 H, d, J = 7.9 H)
z), 8.63 (1H, br), 8.10 (2H, d, J = 8.6Hz), 7.62 (1H, br)
d, J = 8.1Hz), 7.56 (1H, t), 7.52 (1H, t), 7.41 (1H,
t), 7.33 (1H, t), 7.03 (1H, dd, J = 3.0,9.0Hz), 5.00
(3H, m), 4.40 (1H, dd, J = 5.5,12.5Hz), 3.49 (1H, m),
3.20 (9H, s), 2.57 (3H, s), 2.51 (1H, m), 2.17 (3H, s
s) Effects of the present invention The TAN-1030A and the compound (I ') of the present invention have a strong macrophage activating effect and are useful as antitumor agents and the like.

[Brief description of the drawings]

1 and 2 show the UV spectrum and IR spectrum of TAN-1030A.

Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI // (C07D 498/22 209: 00 273: 00 311: 00) (C12P 17/18 C12R 1: 465)

Claims (3)

(57) [Claims] (1) General formula Or a salt thereof. 2. A general formula belonging to the genus Streptomyces A TAN-1030A-producing bacterium represented by is cultured in a medium, TAN-1030A is produced and accumulated in the culture, collected, collected, and optionally subjected to a catalytic reduction reaction, and further optionally subjected to alkylation. General formula characterized by subjecting to an acylation or (and) acylation reaction [Wherein Q 'is C = N-OH or CH-NR ¹ ' R ² '
Wherein R ¹ ′ and R ² ′ each represent hydrogen, lower alkyl or lower alkanoyl.] Or a method for producing a salt thereof. 3. The general formula A macrophage activator comprising a compound represented by the formula: or a salt thereof.