WO2005061461A1 - INHIBITORS AGAINST MEMBERS OF THE HEAT SHOCK PROTEIN 90 (hsp90) FAMILY - Google Patents

Abstract

Inhibitors against members of the heat shock protein 90 (hsp90) family, containing as the active ingredient benzenoid ansamycin derivatives represented by the general formula (I) which are useful as antitumor drug or other drugs or pharmacologically acceptable salts thereof: (I) wherein R2 is hydrogen or methyl; R11 and R18 may be the same or different from each other and are each hydroxy, substituted or unsubstituted lower alkoxy, or substituted or unsubstituted lower alkanoyloxy; and R17 is hydrogen, hydroxy, substituted or unsubstituted lower alkoxy, or substituted or unsubstituted lower alkanoyloxy.

Description

 Heat shock protein 90 (hsp90) family one protein inhibitor

Technical field

 The present invention relates to a heat shock protein 90 (hsp90) family one protein inhibitor useful for pharmaceuticals such as an antitumor agent and an angiogenesis inhibitor, and a benzenoid ansamycin derivative. Background art

 As the hsp90 family one protein, hsp90 o: protein, hsp90 ^ protein, grp94, hsp75 / TRAPl, etc. are known [for example, "Pharmacology & Therapeutics", 1998, No. Vol. 79, p. 129-168 and "Molecular Endocrinology", 1999, Vol. 13, p.1435-1448].

Conventionally, compounds that bind to hsp90 family proteins include geldanamycin (Gelda recitation ycin) and nodibimycin (Herbimycin).

Antibiotics and Radicicol are known (eg, "Cell's

Stress & Chape rones) ⁵⁵ , 1998, vol. 3, p. 100-108 and "Journal of Medicinal Chemistry", 1999, vol. 42, p. 260- 266). It has been reported that all of these compounds bind to the hsp90 family 1 protein and exhibit pharmacological activities such as antitumor activity by inhibiting the function of the hsp90 family 1 protein. Therefore, compounds that bind to the hsp90 family protein are considered to be useful as therapeutic agents for diseases involving the hsp90 family protein or the protein to which the sp90 family protein binds (hsp90 client protein).

Geldanamycin derivatives (for example, see "Investigational New Drugs", 1999, Vol. 17, p. 361-373, etc.) and radicicol derivatives (for example, "Cancer , Research (Cancer Research) ", 1999, Vol. 59, p.2931-2938," Blood ", 2000, Vol. 96, p.2284-2291, "Cancer Chemotherapy and Pharmacology", 2001, Vol. 48, p.435-445, etc.) have been reported to exhibit antitumor effects.

 Novobiocin and PU3 have also been reported as compounds that bind to the] isp90 family protein (eg, "Journal of the National Cancer Institute"). , 2000, Vol. 92, pp. 242-248 and "Chemistry & Biology", 2001, Vol. 8, pp. 289-299, etc.).

On the other hand, as benzenoidoansamycin derivatives, Reblastatin, Autolytimycin, and 17-0-Demethylreblastatin are known. Reblastatin has been reported to arrest the cell cycle in the G1 phase by inhibiting the phosphorylation of retinoblastoma (Rb) protein (see, for example, JP-A-9-286779 and "Journal of Ob'an"). 'See Journal of Antibiotics ", 2000, 53, .1310-1312. Autolytimycin and 17-0-Deraethylreblastatin are anti-HSV-1 agents (see, for example, "Chinese Chemistry Letters", 2001, Vol. 12, p. 903-906). ) And Oncostatin M inhibitors (see, for example, "Journal of Antibiotics", 2000, Vol. 53, p. 657-663). .

Reblastatin Autolytimycin

 17-O-Demethylreblastatin

Disclosure of the invention

 An object of the present invention is to provide an hsp90 family protein inhibitor useful as a pharmaceutical. More specifically, it is an object of the present invention to provide a medicament useful for prevention and / or treatment of cancer and the like. Still another object of the present invention is to provide a therapeutic agent for various diseases associated with hsp90 family protein and a protein to which hsp90 family protein binds, such as an antitumor agent, an angiogenesis inhibitor, etc. An object of the present invention is to provide a novel benzenoid ansamycin derivative useful as a component. The present invention relates to the following (i) to (29).

(1 set

(Wherein, R ² represents hydrogen or methyl, R ¹¹ and R ¹⁸ are the same or different and each represents a hydroxy, a substituted or unsubstituted lower alkoxy or a substituted or unsubstituted lower alkanoyloxy, and R ¹⁷ represents A benzenoid doansamycin derivative represented by hydrogen, hydroxy, substituted or unsubstituted lower alkoxy or substituted or unsubstituted lower alkanoyloxy [hereinafter, a compound represented by the formula (I) I). The same applies to compounds of other formula numbers] or a heat shock protein 90 (hsp90) family protein inhibitor which comprises a pharmacologically acceptable salt thereof as an active ingredient.

(2) The hsp90 family protein inhibitor according to (1), wherein R ¹⁷ is hydrogen.

(3) The hsp90 family protein inhibitor according to the above (1) or (2), wherein R ² is hydrogen.

(4) The hsp90 family protein according to (1) or (2), wherein R ² is methyl and R ¹¹ is substituted or unsubstituted lower alkoxy or substituted or unsubstituted lower alkanoyloxy. Inhibitors.

(5) The hsp90 family protein inhibitor according to any of (1) to (3), wherein R ¹¹ is hydroxy.

(6) The hsp90 family protein inhibitor according to (1), wherein R ² is methyl, R ¹¹ is hydroxy, R ¹⁷ is hydrogen or hydroxy, and R ¹⁸ is hydroxy.

(7) The hsp90 family protein inhibitor according to any one of the above (1) to (5), wherein R ¹⁸ is a substituted or unsubstituted lower alkoxy or a substituted or unsubstituted lower alkanoyloxy.

(8) The above-mentioned (1) to (5), wherein R ¹⁸ is a substituted or unsubstituted lower alkanoyloxy. An hsp90 family one protein inhibitor according to any of the preceding claims.

(9) Equation (la)

(In the formula, R ^2a , R ^Ua , R ^17a and R ^18a have the same ^meanings as R ^h , R ¹⁷ and R ¹⁸ , respectively, provided that R ^2a is methyl, and both R ^11a and R ^18a are hydroxy. when it is, ^{17 a} are hydrogen, hydroxy and Penzenoi door Nsamaishin derivative or a pharmacologically acceptable salt thereof represented by any neither) methoxy.

(10) The benzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to the above (9), wherein R ^2a is hydrogen.

 (11) The benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to the above (9) or (10), wherein R is hydrogen.

(12). R one or both of ^lla and R ^18a is, according to the above (9) to (11) or Re noise is lower alkanoyloxy Noi Ruo alkoxy lower alkoxy, or substituted or unsubstituted substituted or unsubstituted Benzenoy doansamycin derivative or its pharmacologically acceptable

(13) A medicament comprising the benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to any of (9) to (12) as an active ingredient.

 (14) An antitumor agent comprising, as an active ingredient, the benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to any of (9) to (12).

(15) An hsp90 family protein or an hsp90 family protein containing, as an active ingredient, the benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to any of (9) to (12) above. A therapeutic agent for diseases involving proteins. (16) An hsp90 family protein inhibitor comprising, as an active ingredient, the benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to any of (9) to (12).

 (17) A method for inhibiting hsp90 family protein, which comprises administering an effective amount of compound (I) or a pharmacologically acceptable salt thereof.

 (18) A method for treating a malignant tumor, which comprises administering an effective amount of the penzenoid ansamycin derivative or the pharmacologically acceptable salt thereof according to any of (9) to (12). .

 (19) An hsp90 family protein, comprising administering an effective amount of the benzenoidoansamycin derivative or a pharmaceutically acceptable salt thereof according to any of (9) to (12). A method for treating a disease involving a protein to which the hsp90 family protein binds.

 (20) Inhibiting hsp90 family protein, which comprises administering an effective amount of the benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to any of (9) to (12). how to.

 (21) Use of the compound (I) or a pharmacologically acceptable salt thereof for the production of an hsp90 family one protein inhibitor.

 (22) Use of the benzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to any of (9) to (12) for the manufacture of an antitumor agent.

 (23) The benzenoid ansamycin derivative or the benzenoid ansamycin derivative according to any of (9) to (12) for the manufacture of a therapeutic agent for a disease associated with hsp90 family protein or a protein to which hsp90 family protein binds. Use of its pharmacologically acceptable salts.

 (24) Use of the penzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to any of (9) to (12) for the production of an hsp90 family protein inhibitor.

Equation (25) (lb)

A microorganism belonging to the genus Streptomyces, which produces a benzenoid ansamycin derivative represented by the formula:

 (26) Streptomyces sp.EH21 (Streptomyces sp. EH21, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary (1-1 Tsukuba, Higashi, Ibaraki Pref., Japan 1) Chuo No. 6 Postcode 305-8566) FERM BP-08574] strain.

 (27) A method for producing a compound (lb), comprising culturing the microorganism according to (25) and isolating a compound produced from the obtained culture solution.

 (28) Streptomyces sp.EH21 (Streptomyces sp.EH21, Accession No.FERM BP-08574, Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology), and the compound produced from the resulting culture solution is isolated. The method for producing a benzenoidoansamycin derivative according to any one of the above (9) to (12), comprising a separating step.

(29) Streptomyces sp.EH21 '(Streptomyces sp.EH21, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary No. FERM BP-08574) A method for producing a compound (lb), comprising a step of isolating the compound. In the definition of each group of compound (I) and compound (la), the lower alkyl moiety of lower alkoxy is, for example, linear or branched alkyl having 1 to 8 carbons or cycloalkyl having 3 to 8 carbons. Alkyl is mentioned. Examples of the linear or branched alkyl having 1 to 8 carbon atoms include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, neopentyl, Examples of tert-pentyl, hexyl, heptyl, octyl and the like, and examples of cycloalkyl having 3 to 8 carbon atoms include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.

 Examples of the lower alkynyl moiety of lower alkynyloxy include linear or branched alkanols having 1 to 7 carbon atoms, specifically, formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, Vivaloyl, hexanoyl, heptanoyl and the like can be mentioned.

 Substituents in the substituted lower alkoxy and the substituted lower alkanoyloxy are the same or different and include, for example, 1 to 3 substituents, specifically, hydroxy, cyano, carboxy, substituted or unsubstituted lower alkoxy and the like. can give. The substitution position is not particularly limited. Here, the lower alkoxy has the same meaning as the lower alkoxy, and the substituents in the substituted lower alkoxy are the same or different and include, for example, hydroxy having 1 to 3 substituents.

 Some of the compounds (I) and (la) may have stereoisomers such as geometric isomers and optical isomers, and all possible isomers, including these, and mixtures thereof Can be used as the hp90 family one protein inhibitors of the present invention. Similarly, possible isomers of compound (la) and mixtures thereof are included in the compounds of the present invention.

The pharmacologically acceptable salts of compound (I) and compound (la) include, for example, pharmacologically acceptable metal salts, ammonium salts, acid addition salts, organic amine addition salts, amino acid addition salts and the like. I do. Pharmacologically acceptable metal salts include, for example, alkali metal salts such as lithium salt, sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt, zinc salt and the like. Examples of pharmacologically acceptable ammonium salts include salts of ammonium, tetramethylammonium and the like. Examples of pharmacologically acceptable acid addition salts include hydrochlorides and sulfates , Nitrates, phosphates and other inorganic salts, acetates, maleates, fumarates, citrates and other organic acid salts. Examples of pharmacologically acceptable organic amine addition salts include: Examples thereof include addition salts of morpholine and piperidine. Examples of the pharmacologically acceptable addition salts of amino acids include glycine, phenylalanine, aspartic acid, and Evening Min acid addition salts such as lysine and the like. Compound (I) and compound (la) and their pharmacologically acceptable salts may exist in the form of adducts with water or various solvents, and these adducts are also disclosed in the present invention. Hsp90 family one protein inhibitor. Similarly, adducts of compound (la) with water or various solvents are included in the compounds of the present invention.

 Compound (I) or compound (la) or a pharmacologically acceptable salt thereof has hsp90 family protein binding activity, and is used to prevent various diseases in which hsp90 family protein is involved. And / or can be used as a medicament for treatment. Compound (I) or compound (la) or a pharmacologically acceptable salt thereof can be administered alone as it is, but it is usually desirable to provide it as various pharmaceutical preparations. The pharmaceutical preparations are used for animals and humans.

hsp90 family protein inhibition means inhibiting the binding of the hsp90 family protein to the protein to which the hsp90 family protein binds (hsp90 client protein). Examples of the hsp90 family protein include hsp90 sphing protein, hsp90-protein, grp94 hsp75 / TRAPl, etc. [eg, “Pharmacology & Therapeutics”, 1998, Vol. 79 , P.129-168 and "Molecular Endocrinology", 1999, Vol. 13, p.1435-1448]. The hsp90 client protein, hsp90 Fuamiri although protein may be any protein which binds, for example EGFR, ErbB2, Bcr- AbU src, Raf-l s AKT, Fit - 3ヽPLK, Weel, FAK, cMET, hTERT , HIF, mutant p53, estrogen receptor, etc. [For example, "Expert Opinion on Biological Therapy", 2002, Vol. 2, p.3-24 reference].

As described above, for example, benzoquinone ansamycin antibiotics such as geldanamycin and herbimycin, and Radidicol are known (for example, "cell 'stress"). Cell Stress & Chaperones ", 1998, Vol. 3, p.100-108 and" Journal of Medicinal Chemistry ", 1999, No. 42 Vol., Ρ.260-266 etc.). All of these compounds bind to the hsp90 family protein and inhibit the function of the hsp90 family protein, thereby providing pharmacological activities such as antitumor activity. It is reported to show activity.

 Geldanamycin derivatives (for example, see "Investigational New Drugs", 1999, Vol. 17, p. 361-373, etc.) and radicicol derivatives (for example, "Cancer "Research (Cancer Research)", 1999, Vol. 59, p. 2931-2938, "Blood", 2000, Vol. 96, p. 2284-2291, "Cancer-Chemotherapy & Pharmacology ( Cancer Chemotherapy and Pharmacology) ", 2001, Vol. 48, p. 435-445, etc.) have been reported to exhibit antitumor effects.

 Therefore, compound (I) and compound (la) and their pharmacologically acceptable salts can be used as therapeutic agents for diseases involving the hsp90 family protein or the protein to which the hsp90 family protein binds (hsp90 client protein). (For example, it is considered to be useful as an antitumor agent and the like, specifically, a therapeutic agent for solid cancer such as breast cancer and a therapeutic agent for blood cancer such as myeloid leukemia).

 The pharmaceutical preparation according to the present invention may contain Compound (I) or Compound (Ia) or a pharmacologically acceptable salt thereof alone or as an active ingredient in combination with any other active ingredient for treatment. It can be contained as a mixture. In addition, these pharmaceutical preparations are produced by mixing the active ingredient with one or more pharmacologically acceptable additives and by any method well known in the technical field of pharmaceutical preparation.

 It is desirable to use the most effective route for treatment, and it can be oral or parenteral, for example, intravenous.

 Examples of the administration form include tablets, powders, granules, syrups, injections and the like.

 Tablets and the like suitable for oral administration can be produced using excipients such as lactose, disintegrants such as corn starch, lubricants such as magnesium stearate, binders such as hydroxypropylcell mouth, and the like. .

 Suitable for parenteral administration, for example, in the case of an injection, can be produced using a salt solution, a glucose solution or a mixture of a salt solution and a pudose solution, or the like.

The dosage and frequency of compound (I) or compound (la) or a pharmaceutically acceptable salt thereof will depend on the mode of administration, the age and weight of the patient, the nature or seriousness of the condition to be treated. Orally, 0.01 mg to lg, preferably 0.05 to 50 mg per adult is administered once or several times a day, depending on the degree. In the case of parenteral administration such as intravenous administration, 0.001 to L00 mg, preferably 0.01 to 10 mg, per adult is administered once or several times a day. However, the dose and the number of administrations vary depending on the various conditions described above.

 The method for producing the compound of the present invention is not particularly limited. For example, the compound can be produced according to the steps described below.

Manufacturing method 1

Among the compounds (I), the compound (lb) in which R ² is hydrogen, R ¹¹ and R ¹⁸ are hydroxy, and R ¹⁷ is hydrogen belongs to the genus Streptomyces and is represented by the formula (lb). A microorganism having the ability to produce a penzenoid doansamycin derivative is cultured in a medium, a compound (lb) is produced and accumulated in the culture, and the compound (lb) is collected from the culture. The microorganism having the ability to produce the benzenoid ansamycin derivative represented by the formula (lb) is a strain belonging to the genus Streptomyces and having the ability to produce the benzenoid ansamycin derivative represented by the formula (lb). Any strain can be used. In addition, even if these strains are mutated by an artificial mutation method, for example, ultraviolet irradiation, X-ray irradiation, treatment with a mutagenic agent or the like, or naturally mutated, the benzenoid represented by the formula (lb) Any substance having an ansamycin derivative-producing ability can be used in the present invention. A specific preferred example is Streptomyces sp. EH21 strain.

The present inventors have found that strain EH21 belonging to the genus Streptomyces newly isolated from soil produces a compound (lb). In addition, the present inventors have proposed that the above EH21 strain produces a compound (II) in which R ² is methyl, R ¹¹ and R ¹⁸ are hydroxy, and is hydrogen, hydroxy or methoxy in the compound (I). I also found something to do. The representative strain EH21 producing compound (lb) was isolated from soil and has the following bacteriological properties.

 This paper describes the morphology, culture characteristics, and physiological characteristics of the actinomycete EH21 belonging to the genus Streptomyces newly isolated from soil.

1.morphological quality 1) mycelium

 Aerial mycelium formation: Yes

 Aerial hyphal fragmentation and motility: None

 Disruption and motility of the underlying mycelium: None

 2) spores

 Presence or absence of spore formation and location: formation on aerial hyphae

 Presence or absence of sporangia and location: No

 Spores on spores «Number of spores: 10 or more

 Shape when many spores are linked: Coiled

 Spore characteristics

 Surface structure: Circular

 Shape and size: rod, about 1.0-1.3 X 1.9〃m

 Motility and presence of flagella: none

 3) Other

 Chlamydospore: nothing

 Focusing hypha: None

 Pseudospore: None

 Hyphal division mode: simple branching

2.Cultural properties

 The EH21 strain shows normal or vigorous growth on synthetic and natural media used in the United States, and the underlying mycelium shows a pale yellow to deep reddish yellow. The medium may produce a yellow colored soluble dye.

 The characteristics of growth and color when cultured on various media at 28 ° C for 14 days are shown below. The color display was in accordance with the classification of colors by the JIS Color Name Book (Japan Standards Association).

 1) Sucrose-salt salt agar medium

 Growing condition: poor

 Color of the underlying hypha: very light yellow to yellowish white (7.5Y 9/3 to 5Y 9/1)

Epiphytic state of aerial mycelium and its color tone: vigorous, yellowish dark gray to white (5Υ48 to 9.59.5) Soluble pigment: None ) GlucoseAsparagine agar

 Growth ^: poor

 Color of the underlying mycelium: dull yellow to pale yellow (5Y 7.5 / 7-5Y 9/6) Aerial mycelial growth and its color: normal, white (Ν 9.5)

 Soluble dye: None

) Glycerin'asparagine agar

 Growth ϋ: Normal

 Color of the underlying mycelium: dull reddish yellow to pale yellow (10YR 7/7-5Υ 9/6) Aerial mycelial epiphytes and their color: poor, white (Ν 9.5)

 Soluble pigment: Production (yellow)

) Suyuichi's inorganic salt agar medium

 Growth status ϋ: Active

 Color of the underlying mycelium: very light yellow to light yellow (10YR 9/3 to 5Υ 9/6) Epiphytic state of the aerial mycelium and its color: vigorous, reddish gray to white (5R 5.5 / 1 to 9.5 9.5) Soluble pigment :Nothing

) Tyrosine agar medium

 Growth ϋ: Normal

 Color of the underlying mycelium: dark reddish yellow to pale yellow (10YR 5.5 / 10 to 5Υ 9/6) Aerial mycelial growth and its color: poor, light grayish yellow (7.5Υ 8/3) Soluble pigment: slight Produced in (yellow)

) Nutrient agar medium

 Growth status ϋ: Active

 Color of the underlying mycelium: pale reddish yellow (10YR 8.5 / 6)

 Epiphytic state of aerial mycelium and its color: poor, very pale yellow (10YR 9/3) Soluble pigment: none

) List 'malt agar medium

 Growth status ϋ: Active

Color of the underlying mycelium: dull yellow-red to gray-yellow (5YR 6/7 to 5YR 5/3) Aerial mycelial growth and its color: vigorous, gray to white (N 5.5 to N 9.5) Soluble dye: None

 8) Oatmeal agar

 Growing condition: strong

 Color of the underlying mycelium: Light reddish yellow to very light yellow (10YR 8.5 / 6 to lQYR 9/3) Aerial mycelial growth and its color: vigorous, yellowish dark gray to white (5Y 4/1 ~ Ν 9.5) Soluble dye: None

Physiological properties

 The physiological properties of the EH21 strain are shown below. The growth temperature range is the result after culturing for U days, and the others are the results after culturing for 2 weeks.

 1) Growth temperature range: 15.0 ° C ~ 43, 5 ° C, optimum temperature around 39 ° C.

 2) Liquefaction of gelatin: None

 3) Starch water splitting angle: Yes

 4) Coagulation and peptization of skim milk powder: with paptonization

 5) Melanin-like pigment formation

 a) Peptone 'East' iron agar: None

 b) Tyrosine agar: none

 6) Utilization of carbon source

 As a basal medium, pre-ham's Gottlieb agar medium was used.

 In the following, + indicates use, and one indicates not use.

 L-arabinose: +

 D-Kishi mouth: +

 D-glucose: +

 Sucrose: +

 Raffinose: +

 D-Fructose: +

 Rhamnose: +

 Inositol: +

 D-mannitol: +

Chemical taxonomic properties 1) Optical isomer of diaminopimelic acid in bacterial cells: LL type

 As described above, morphologically, spore chains are formed in aerial hyphae, sporangia is not formed, and basal hyphae are not disrupted.Chemically, the cell wall is type I (LL-diaminopimelic acid). ), This strain is classified as Streptomyces among actinomycetes.

 Therefore, this strain was named Streptomyces sp. EH21 (Streptomyces sp. EH21) and dated December 16, 2003, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary (Japan). Deposit No. FERM BP-08574 at Tsukuba East 1-chome, 1-chome, Chuo No. 6 Postcode 305-8566, Ibaraki Pref.

 In culturing the benzenoid ansamycin derivative-producing bacterium of the present invention, a usual method of culturing actinomycetes is applied. As the medium, any of a synthetic medium and a natural medium can be used as long as the medium appropriately contains one selected from a carbon source, a nitrogen source, an inorganic ion, an organic nutrient source, and the like, which can be used by microorganisms.

 As the carbon source, glucose, starch, dextrin, mannose, fructose, sucrose, lactose, xylose, arabinose, mannitol, molasses, etc. are used alone or in combination. In addition, hydrocarbons, alcohols, organic acids, etc. are used depending on the assimilation ability of the bacteria.

 Nitrogen sources include ammonium chloride, ammonium nitrate, ammonium sulfate, sodium nitrate, urea and water. Ton, meat extract, yeast extract, dried yeast, corn-steep rice, soybean flour, pupa flour, casamino acid, etc. are used alone or in combination.

 In addition, as necessary, sodium chloride, potassium chloride, magnesium sulfate, calcium carbonate, potassium dihydrogen phosphate, magnesium phosphate octahydrate, ferrous sulfate, calcium chloride, manganese sulfate, zinc sulfate, copper sulfate, etc. Of inorganic salts. Furthermore, trace components that promote the growth of the bacterium used and the production of the benzenoid ansamycin derivative can be appropriately added.

As a culture method, a liquid culture method, particularly a submerged stirring culture method, is suitable. The cultivation is performed at a temperature of 16 to 37 ° C, preferably 25 to 32 ° C, at a pH of 4 to 10, preferably at a pH of 6 to 8, and the pH of the culture medium is adjusted with ammonia water or an ammonium carbonate solution. . Culture Usually complete in 1 to 10 days, but stop culturing when compound (lb) or compound (II) is produced and accumulated in the culture solution and cells, and the amount of production in the culture reaches the maximum. Is preferred.

 In isolating and purifying the compound (lb) or compound (11) accumulated in the culture, the conventional method for isolating and purifying normal microbial metabolites from the culture can be performed according to the method commonly used for isolating and purifying the compound from the culture. Done. For example, add methanol, ethanol, or 2-propanol directly to the culture and extract the active ingredient, or perform two-layer partition extraction with 2-butanol, tert-butanol, and n-butanol. The active ingredient is thereby extracted. The culture is separated into a culture filtrate and cells by filtration, and cell components are extracted from the cells with a solvent such as black-mouthed form, acetone, and methanol. Then, the extract or the culture filtrate is passed through a polystyrene adsorbent, for example, Diaion HP-20, HP-20ss (manufactured by Mitsubishi Chemical Corporation) or the like, to adsorb the active ingredient, and then eluted with methanol, acetone, or the like. Next, the compound (C) is subjected to gel filtration using, for example, Sephadex LH-20, Toyopearl HW40 or the like, column chromatography using octadecyl-bonded silica gel (0DS), high performance liquid chromatography, column chromatography using silica gel, etc. lb) or compound (II). The detection of compound (lb) or compound (II) during the culturing, isolation and purification procedures is carried out by thin layer chromatography, high performance liquid chromatography, or the like.

Manufacturing method 2

Process 1

In the compound (la), R ^Ua and R ^18a are hydroxy or substituted or unsubstituted lower alkoxy, R is hydrogen, hydroxy or substituted or unsubstituted lower alkoxy, R ^lla , R ^17a and R Compound (III) in which at least one of ^18a is substituted or unsubstituted lower alkoxy is a compound (IV) {commonly used in organic synthetic chemistry from compound (Ib), compound (II) or compound (11) Known methods [for example, by R. C. Larock, "Comprehensive Organic T-ransformations", John, United States-Sands, Inc., USA John Wiley & Sons Inc.), 1989].

(Wherein, R ^2a are as defined above, ³ and R ¹⁸ - ³ represents hydroxy or substituted or unsubstituted lower alkoxy, R ¹ represents hydrogen, hydroxy or substituted or non-replacement of the lower alkoxy, R ¹¹ - ^{3, 3} and R ¹⁸ - at least one of ³ substituted or represents unsubstituted lower alkoxy, R ¹⁷ - when ³ is hydrogen, R ¹⁷ - ⁴ will display the hydrogen, R ¹⁷ When R ³ — is methoxy, R ¹⁷ — ⁴ represents hydroxy or methoxy; when R ¹⁷ — ³ is substituted or unsubstituted lower alkoxy or hydroxy except for methoxy, R ¹⁷ — ⁴ represents hydroxy)

 Compound (III) can be synthesized by alkylating hydroxy of compound (IV) with an appropriate alkylating agent in an inert solvent, if necessary, in the presence of 1 to 100 equivalents of a base. Examples of the inert solvent include methanol, acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide, chloroform and dichloromethane. As the base, for example, sodium hydride, oxide, silver, carbonated rim, diisopropylethylamine, pyridine, triethylamine, Ν, Ν-dimethylaniline or 1,8-bis (dimethylamino) naphthalene, etc. can give. Any suitable alkylating agent may be used as long as it is generally used in the method for alkylating hydroxy, such as trimethylsilyldiazomethane, alkyl halide, alkoxyalkyl halide, alkyl alkane sulfonate or trialkoxy tetra. Fluoroborate and the like. The reaction temperature is preferably from -30 ° C to 150 ° C, and the reaction time is usually from 5 minutes to 150 hours.

In the compound (III), the compound in which at least one of R ^{1 i} , R ¹⁷ — ³ and R ¹⁸ — ³ is hydroxy can form the hydroxy by adjusting the reaction conditions and the equivalent of the alkylating agent. As a mixture with a substituted or unsubstituted lower alkoxy compound It can be obtained and can be isolated and purified by a separation and purification method commonly used in organic synthetic chemistry, for example, filtration, extraction, washing, drying, 'concentration, recrystallization, various types of chromatography, and the like. Also, protection and deprotection of hydroxy groups as required [eg, by TW Greene, "Protective Groups in Organic Synthesis", John, "Ili-ichi", USA And 'Sands' Incorporated (John Wiley Sons Inc.), 1981] to selectively produce the target compound.

Process 2

In the compound (la), R ^11la and R ^18a are hydroxy, substituted or unsubstituted lower alkoxy or substituted or unsubstituted lower alkanoyloxy, and R ^17a is hydrogen, hydroxy, substituted or unsubstituted lower. Compound (V) which is alkoxy or substituted or unsubstituted lower alkanoyloxy and wherein at least one of R ^11a , R ^17a and R ^18a is substituted or unsubstituted lower alkanoyloxy is a compound (VI) ) [The compound (III) or compound (IV)] can be synthesized.

 (VI) (V)

(Wherein, 'R ^2a are as defined above, R ¹¹ - ⁵ and R ¹⁸ - ⁵ is hydroxy, a lower alkanoyloxy Noi Ruo alkoxy lower alkoxy, or substituted or unsubstituted substituted or unsubstituted replacement and table, R ^1M represents hydrogen, hydroxy, lower alkanoyloxy Noi Ruo alkoxy lower alkoxy or substituted or unsubstituted or substituted ^{unsubstituted, R "- 5, R 17} - 5 and R ¹⁸ - one Kutomo 1 less out of ⁵ represents a lower alkanoyloxy Noi Ruo carboxymethyl substituted or unsubstituted, R ¹¹ - for ⁵ properly be substituted unsubstituted lower alkanoyloxy Noi Ruo alkoxy or hydroxy, R ^{11 '6} are hydroxy Shi, R ¹¹ - ⁵ If it is substituted or unsubstituted lower alkoxy, ⁶ represent the same Ku substituted or unsubstituted lower alkoxy and ^{R "_ 5, R 17 -} 5 are lower substituted or unsubstituted For Arca Noi Ruo alkoxy or hydroxy, R ¹⁷ - ⁶ represents hydroxy, R ¹⁷ - case ⁵ is hydrogen or a substituted or unsubstituted lower alkoxy, the R ^1M R ¹⁷ - ⁵ and was likewise hydrogen or substituted or represents lower § ^ Kokishi unsubstituted, R ¹⁸ - case ⁵ is of a low grade Arca Noi Ruo alkoxy or hydroxy substituted or unsubstituted, R ¹ represents a hydroxy, R ¹⁸ - ⁵ is lower substituted or unsubstituted If alkoxy, R ¹⁸ - ⁶ represents a lower alkoxy same substituted or unsubstituted and R ¹⁸ _ ^5).

 Compound (V) is prepared by subjecting compound (VI) to alkanoylation of hydroxy using 1 to 100 equivalents of an appropriate alkanoinolelating agent in the presence of 1 equivalent to a solvent in a solvent without solvent or in a solvent. be able to. Examples of the solvent include ,, Ν-dimethylformamide, chloroform, or dichloromethane. Examples of the base include pyridine, triethylamine, Ν, ミ ン -dimethylaniline, ethyldiisopropylamine, 4-dimethylaminopyridine and the like. The alkanoylating agent to be used may be any one usually used in a method for alkanoylating hydroxy, such as an acid anhydride or an acid halide. The reaction temperature is preferably from -30 ° C to 150 ° C, and the reaction time is usually from 5 to 150 hours. Compound (V) can also be obtained by reacting compound (VI) with 1 to 100 equivalents of carboxylic acid in a solvent in the presence of 1 to 100 equivalents of a base and 1 to 100 equivalents of a condensing agent. Can be synthesized. Examples of the solvent include N, N-dimethylformamide, chloroform, dichloromethane and the like. Examples of the base include pyridine, triethylamine, Ν, Ν-dimethylaniline, ethyldiisopropylamine, 4-dimethylaminoviridine and the like. Examples of the condensing agent include 1,3-dicyclohexylcarbodiimide, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide and carbonyldiimidazole. The reaction temperature is preferably from -30 ° C to 150 ° C, and the reaction time is usually from 5 to 150 hours.

If necessary, continue in a solvent such as water, methanol, acetonitrile, tetrahydrofuran, dimethylsulfoxide, dimethylformamide, chloroform, or dichloromethane, if necessary, in an amount of 0.1 to 1000 equivalents of potassium carbonate or carbonate. In a compound in which hydroxy which is not intended for alkanoylation is reacted with a base such as sodium hydrogen, sodium methylate, etc., the alkanol is The group can be removed. The reaction temperature is preferably from -30 ° C to 150 ° C, and the reaction time is usually from 5 minutes to 150 hours.

Further, among the compounds ^{(V), R "- 5} , R 17 - 5 and R ¹⁸ - by compound at least one of the ^five is a hydro alkoxy is, to adjust the equivalent reaction conditions and Arukanoiru agent, the In some cases, hydroxy can be obtained as a mixture with a substituted or unsubstituted lower alkanoyloxy compound, and can be isolated and purified or selectively produced in the same manner as in Step 2.

 The intermediates and target compounds in the above Steps 1 and 2 are subjected to separation and purification methods commonly used in organic synthetic chemistry, for example, filtration, extraction, washing, drying, concentration, recrystallization, various chromatographies, etc. It can be isolated and purified. Further, the intermediate can be subjected to the next reaction without purification.

In addition, known methods commonly used in organic synthetic chemistry [eg, R. Larock, "Comprehensive Organic Transformations ⁵⁵ , John Wiley and Sands, USA"'Incorporated (John Wiley & Sons Inc.), 1989] can convert various functional groups to produce various compounds (I) and compounds (la). Protection and deprotection of each functional group [eg, TW Greene, "Protective Groups in Organic Synthesis", John, "Ili-ichi", USA And 'Sands' Incorporated (see John Wiley & Sons Inc., 1981) can be combined to produce the desired compound with various functional groups.

 When it is desired to obtain a salt of the compound (I) and the compound (la), when the compound (I) and the compound (la) are obtained in the form of a salt, they may be purified as they are, and in the free form. When used, compound (I) and compound (la) may be dissolved or suspended in an appropriate solvent, and then isolated and purified by adding an acid or a base.

Representative compounds of the compound (I) suitably used as the active ingredient of the present invention are shown below, and their production methods are described in Reference Examples described later. However, the active ingredient of the present invention is not limited to these compounds. Table 1 Specific examples of compounds ([)

Compound R ² R ¹¹ R ¹⁷ R ¹⁸

1 CH ₃ OH OCH3 OH

2 CH ₃ OH H OH

3 CH ₃ OH OH OH

 4 H OH H OH

5 CH ₃ OH OCH ₃ OCH3

6 CH ₃ OH OCH3 OCOCH3

7 CH ₃ OCOCH3 OCH3 OH

8 CH ₃ OCOCH3 OCH3 OCOCH3

Next, the pharmacological action of the representative compound (I) will be specifically described with reference to test examples. Test example 1 hsp90 protein binding activity test

Purified from cereal brain according to the method described in the literature [see "Archives of Biochemistry and Biophysics", 1990, Vol. 282, pp. 290-296]. The diluted hsp90 protein (58% purity) was diluted to 1 g / mL with Tris-buffered saline (TBS, pH 7.4) and added to a 96-well ELISA assay plate manufactured by Sumitomo Belite Co., Ltd. After dispensing at a volume of 〃L / well, the mixture was allowed to stand at 4 ° C for 1 化 for immobilization. The supernatant was removed, and TBS containing 1% serum albumin (BSA) was dispensed at a volume of 300 zL / ml for blocking. After removing the blocking solution, add Tris-buffered saline (TBST) containing 0.05% Tween 20 at a volume of 500 / L / ゥ ヱ. The operation of washing the solid phase was repeated 3 'times. Each solution was prepared by diluting the test compound with TBST in eight steps at a concentration of 10 square root from the highest concentration of 0.5 ol / L. This test compound solution was used as a protease inhibitor confe. Reed EDTA-free (Roche) and Perfloc SC (Pefabloc SC, Roche), 1 tablet / 40 mL and 0.4 TB ol / L TBST containing 80 〃L / ゥ L, respectively. Add 20 zL / well to the poured assay plate! And left at room temperature for 1 hour. Here, dimethyl sulfoxide (DMS0) was used as a positive control at a final concentration of 1 μL / well, and radicicol was used as a negative control at a final concentration of 0.25 μmol / L on the same plate as the test compound. The operation was performed. The biotinylated radicicol represented by formula (A) ["Bioorganic & Medicinal Chemistry", 200 years, 10 volumes, so that the final concentration is 0.1 mol / L. , P.3445- 3454], and left at room temperature for 1 hour to conduct a competitive reaction of the binding of the test compound to the immobilized hsp90 protein.

 (A)

After removing the reaction solution, the procedure of adding TBST to the wells and washing the solid phase was repeated three times. Europium-labeled streptavidin [Perak Oy (Wallac Oy)] was diluted with Atsushi's buffer [Wallac Oy] to a final concentration of 0.1 _g / mL. After dispensing at a volume of 100 L / well, the mixture was allowed to stand at room temperature for 1 hour to carry out a biotin-avidin binding reaction. After removing the reaction solution, the operation of washing the solid phase by adding TBST in an amount of 500 µL / well was repeated three times. A fluorescence enhancing solution [Wallac Oy] was added in an amount of 100 µL / µl, and a color reaction was performed at room temperature for 5 minutes. A multilabel counter [ARV0 ^™ , Wallac Oy's] was used. The time-resolved fluorescence was measured at an excitation wavelength of 340 nm and a measurement wavelength of 615 nm. Positive control Binding rate of 100% a measurement of time-resolved fluorescence, as binding 0% measurement value of the negative control, the concentration IC ₅ test compound combine 50% inhibition of the Piochin of Radeishikoru and hsp90. (nmol / L) was calculated.

The results are shown in Table 2.

Table 2 hs _P 90 protein binding activity

 Piotinylated Radicicol

 Inhibition of binding to hsp90

 Compound

 1 2.1

 2 2.6

 3 6.0

 4 12

 5 35

 6 26

Test Example 2 Growth inhibition test on human breast cancer-derived KPL-4 cells

In a 96-well microplate (manufactured by Nunc), 2,000 KPL-4 cells derived from human breast cancer are plated per well, and in Dulbecco's modified Eagle's medium containing 10% fetal calf serum (FCS) (DMEM) Preincubation was performed at 37 ° for 24 hours in a 5% carbon dioxide gas incubator overnight. Thereafter, a DMS0 solution of each test compound prepared at 10 ol / L was serially diluted with a culture medium and added to make a total of 100 L per well. Thereafter, the cells were cultured at 37 ° C. for another 72 hours in a 5% CO 2 incubator. After completion of the culture, WST-1 {4- [3- (4-iodophenyl) -2- (4-nitrophenyl) -2H-5-tetrazolio] -1,3 diluted 2-fold with the culture medium -Benzenedisulfonic acid (4- [3- (4-Iodophenyl) -2-(^ nitrophenyD-SH-S-tetrazol iol ^ -benzene disul.fonate) labeled mixture (Roche Diagnostics) }, Add 20 L per liter, and incubate for 1 hour at 37 ° C in a 5% CO 2 incubator, and use a microplate spectrophotometer (Bio-Rad, Model 550). The absorbance was measured at 450 nm and 655 ° C. The cell growth inhibitory activity was shown as a 50% growth inhibitory concentration GI _5G GI _{5. The} calculation method is described below: From the absorbance at 450 nm of each well at 655 ° C. The value (difference in absorbance) obtained by subtracting the absorbance of the test compound was calculated. The difference in absorbance obtained from the control cells was 100 ° /. And then, by comparing the difference absorbance obtained in cells treated with the compounds of the test concentration, and calculate the concentration of the compound in inhibiting the growth of cells by 50%, which the GI _5. Indicated by. Table 3 shows the results. According to Table 3, the test compound shows a cell growth inhibitory activity against human breast cancer-derived KPL-4 cells, and is useful as an antitumor agent.

 Table 3 Inhibition of human breast cancer-derived KPL-4 cell growth

 Growth inhibition

 Compound GISD (μηιο1 / Ρ

 1 0.43

 2 0.71

 4 3.6

 6 0.2 Test Example 3 Elimination of intracellular hsp90 client proteins Raf-1 and ErbB2

In a 24-well microplate (manufactured by Nunc), 50,000 KPL-4 cells derived from human breast cancer per 1 μl are seeded, and the medium is added to a Dulbecco's modified single medium containing 10% fetal calf serum (FCS) ( (DMEM) and 5% CO 2 incubator at 37 ° C for 24 hours. Thereafter, a DMS0 solution of each test compound adjusted to 10 mol / L was diluted stepwise with a culture medium and added to each well to the test concentration. Then, the cells were further cultured at 37 ° C for 40 hours in a 5% CO 2 incubator. Cooled lysis buffer [HEPES 50 NaOH, pH 7.4, 250 mmol / L NaCl, lmmol / L Ethylenediaminetetraacetic acid (EDTA), 1% Nonidet P-40 (NP -40), one thigh ol / L dithiothreitol (DTT), one thigh ol / L futsudani phenylmethylsulfonyl (PMSF), 5 jg / mL leptin (leupeptin)] ' After lysing the cells for 30 minutes at ° C, they were centrifuged at 20000G for 10 minutes. The protein concentration of the obtained supernatant was measured, and samples were prepared to have the same protein content in each lane.The protein was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Done. The separated protein sample was transferred to a polyvinylidene difluoride (PVDF) membrane (Millipo) and then used as a primary antibody as an anti-ErbB2 antibody [anti-c-Ne, Ab-3, Oncogene). Anti-Erk-2 antibody [anti-Erk2, manufactured by Upstate Biotechnolgy], Anti-Raf-1 antibody [anti-Raf-1 (C-12), manufactured by Santa Cruz Biotechnology, Inc.] was added and reacted with the protein on the membrane. Then, as a secondary antibody, a horseradish peroxidase (Horseradish Peroxidase) -labeled secondary antibody that reacts with each primary antibody [anti-Egret Ig antibody or anti-mouse Ig antibody, Amersham] is reacted. Was. Detection was performed using the ECL reagent [PIERCE], and the band obtained on the X-ray film was examined. ErbB2 and Raf-1 disappeared using the non-hsp90 client protein Erk-2 as a control. The concentration at which was first detected was defined as the minimum effective concentration (Fig. 1). The results are shown in Table 4. According to Table 4, the test compound is considered to be useful as a therapeutic agent for human breast cancer because it has a selective elimination effect on Raf-l and ErbB2 involved in the growth and malignancy of human breast cancer .

Table 4 disappearance act minimal effective concentration of intracellular hs _P 90 client proteins RalM and ErbB2

 Compound mol / _

 0.33

 twenty one

 4 3.3

 6 0.33 Test Example 4 Measurement of differentiation-inducing activity on human chronic myeloid leukemia cell line K562 using expression of surface marker as an index

Bcr-Abl ', a translocation gene product, is known as a causative gene of human chronic myeloid leukemia (CML), but this protein itself is a hsp90 client protein, and Bcr-Abl-expressing human chronic bone marrow Hsp90 inhibitors such as radicicol and its derivatives, or benzoquinon ansamycin conjugates (geldanamycin, herbimycin A), etc., indirectly inhibit the function of Bcr-Abl on myeloid leukemia cell line K562 It has been reported that it has anticellular activity and also has the activity of differentiating leukemic cells into erythroid cells [see Blood, 2000, vol. 96, p. 3312-3318]. Therefore, in order to clarify the hsp90 inhibitory action of the benzenoidoansamycin derivative and its usefulness as a therapeutic agent for leukemia, the following method was used to measure the differentiation inducing activity on 1 (562 cells). K562 cells (1 × 10 ⁵ cel ls / mL) were seeded in 6-well plates (Falcon) at 5 mL per well, and 0.1% of a test compound diluted to each concentration with DMSO was added. After 40 hours the cells were added to the drug at 37 ° C, 5% C0 ₂ under cultivation, suspended in 10mL cell pack (Toa Medical Electrical) by sampling the fine alveolar fluid of 0.2mL for anti-cell activity measurement The cells became turbid, and the number of cells was measured using an automatic blood cell counter F-300 (Toa Medical Electric). The remaining cell solution was transferred to a 15πΛ tube (Falcon), centrifuged at 100 rpm for 10 minutes, the medium was removed by suction, and the cells were washed with a phosphate buffer (PBS) by the same operation. After washing, the cells collected by centrifugation are suspended in PBS containing 0.1% serum albumin (BSA), and glycophorin A, a cell surface protein that is specifically expressed in erythroid cells, is recognized. A primary antibody against [Glycophorin A (GPA)] {mouse antr GPA antibody (Pharmindin)] was added to a final concentration of 2.5 / g / mL. After reacting on ice for 30 minutes, wash the cells again with PBS, and then again at 0.1 ° /. Suspend in PBS containing BSA and add a secondary antibody, eg, rabbit anti mouse IgG-FIC conjugate (Bec Yuichisha) to a final concentration of 2 ク / 111. did. After reacting on ice for 30 minutes, the cells obtained by washing with PBS were fixed with a PBS solution containing 2.5% glutenaldehyde. The expression level of GPA surface antigen (FITC fluorescence wavelength: 515-545 nm) was measured for 10,000 cells of the obtained cell sample by FACScan (Becton Dickinson) as an index of erythroid differentiation.

 FIG. 2 shows the results of an experiment using compound 6 (the acetyl compound of compound 1). Formula of the control compound (B)

 OH

 (B)

Radicicol derivative [See "Bioorganic & Medicinal Chemistry", 2002, Vol. 10, p. 3445-3454] In addition to showing the inhibition of proliferation of K562 cells as well as the increase in erythroid markers, it was confirmed that it has the activity of inducing the hsp90 inhibitor, which is one of the activities of the hsp90 inhibitor. . As shown in Table 5, the same activity was confirmed for other evaluated benzenoid ansamycin derivatives.

 In FIG. 2, the anticellular activity was expressed as a relative value when the number of cells of the test compound untreated cells after 40 hours of culture was 100%. Differentiation-inducing activity is expressed as the ratio of cells with increased fluorescence intensity out of 10,000 cells based on the GPA expression level (fluorescence intensity) of cells in which test compound-untreated cells were labeled only with the secondary antibody after 40 hours of culture. expressed.

 Table 5 Differentiation I Excellent activity Differentiation inducing activity

 (Concentration that increases to GPA force 0% or more)

 Compd. (JAmol / L) ■

 1 1.1

 2 3.3

 4 30

 6 30 Brief description of drawings

FIG. 1A is a view showing the action of compound 1 for eliminating intracellular Hsp90 client proteins Raf-1 and Raf-2. Similarly, B represents compound 2, C represents compound 4 and D represents compound 6. FIG. 2 is a diagram showing the anti-cell activity and differentiation-inducing activity of compound 6 and the Radicicol derivative on K562 cells.

Explanation of symbols

 ▲: Anti-cell activity of compound 6

 Δ: differentiation-inducing activity of compound 6

 Hata: Anticellular activity of Radicicol derivatives

〇: Differentiation-inducing activity of Radicicol derivatives Best form for carrying out the invention J

 Hereinafter, embodiments of the present invention are described with reference to Examples and Reference Examples.

Example 1 Production of Compound 4 by Streptomyces sp. EH21

 Type 1 medium and type 2 medium include glucose 10 g / L, soluble starch 10 g / L, meat extract 3 g / L, yeast extract 5 g / L, tryptone 5 g / L, and lithium dihydrogen phosphate lg / A medium having a composition of L (pH 7.0) was used. Glycerol-preserved inoculum was inoculated into a total of eight tubes of 0.5 mL each in 10 mL of a first-class medium in a 70 mL test tube, and cultured with shaking at 28 ° C for 192 hours. This first-class culture solution was inoculated into a total of 31 tubes of 2.5 mL each in 50 mL of a second-type medium in a 300 mL Erlenmeyer flask, and cultured with shaking at 28 ° C. for 96 hours. The second-type culture obtained was inoculated into 12 liters (36 L in total) of 75 mL each in 3 L of the main fermentation medium placed in a 5-L jar amen, and aerated and agitated at 28 ° C for 144 hours (rotation speed). 200 rpm). The main fermentation medium is glucose 10 g / L, soluble starch 10 g / L, pupal flour 10 g / L, roasted tea 5 g / L, yeast extract / L, nitrohumic acid 0.25 g / L, M0PS2 g / L, zinc sulfate-7. Water salt 10 mg / L, manganese sulfate · pentahydrate 10 mg / L, iron (II) sulfate · 7 hydrate 10 mg / L, cobalt chloride · 6 hydrate 10 mg / L, magnesium sulfate · 7 hydrate 0.5 g / A medium having a composition of L (pH 6.4) was used.

To 29 L of the obtained fermentation culture broth, a filter aid (Radiolite 600, manufactured by Showa Chemical Industry Co., Ltd.) was added at a ratio of 10%, followed by suction filtration to separate the culture filtrate and the cells. To the separated cells, 16 L of methanol was added, and the mixture was sufficiently stirred and extracted. The mixture was filtered again with a suction filter, and the obtained methanol extract was diluted 3-fold with water to obtain a 48 L cell extract. The culture filtrate and the cell extract were separately passed through a column packed with 2 L of Diaion HP-20 to adsorb the active ingredients. After washing each column with a 30% aqueous methanol solution, the active components were eluted with 50% and 75% aqueous methanol solutions. Next, the active fractions were passed through a column packed with 500 mL of Diaion HP-20ss to adsorb the active ingredients. After that, each column was washed with a 50% aqueous solution of methanol. , 60% and 70 ° /. The active ingredient was eluted with an aqueous solution of methanol. After evaporating the organic solvent under reduced pressure from the active fraction derived from the culture filtrate, the residue was dissolved in methanol, 35 mL of silica gel was added thereto, and methanol was distilled off to adsorb the active ingredient onto the silica gel. . Next, the silica gel on which the active ingredient was adsorbed was placed on a column filled with 300 mL of silica gel, When the mixture was developed with a mixed solution of formaldehyde and methanol, active ingredients eluted in 10% and 20% methanol / chloroform fractions, respectively. The active fraction derived from the bacterial cell extract was similarly adsorbed on 75 mL of silica gel, placed on 300 mL of silica gel that had already been filled, and developed with a methanol / chloroform mixture, resulting in 10% methanol / chloroform. The active component eluted in the lume fraction. The active fraction derived from the supernatant and the cell extract are mixed, and purified on reversed-phase silica gel YMC-GEL ODS-AQ 120-S5O 400 mL MM column (flow rate 2.5 mL / min, mobile phase 55% aqueous methanol solution). The active fraction eluted at a retention time of 260 to 325 minutes was further fractionated. High performance liquid chromatography (column Inertsil 0DS, 020 x 250 mm, column temperature 35 ° C; flow rate 8 mL / min, mobile phase 25% aqueous acetonitrile solution) The active fraction with a retention time of 25 to 28 minutes was collected in the above). Further, purification was carried out by preparative thin-layer chromatography [methanol / form-form mixture (1: 9)] to obtain 7.2 mg of compound 4.

 The physicochemical properties of Compound 4 are as follows.

 The physicochemical properties were measured with the following instruments.

 FAB mass spectrum and high resolution g FAB mass spectrum: JEOL JMS-HX / HX110A mass spectrometer

 Nuclear magnetic resonance spectrum: Bruker AM-50O (500 MHz)

Physicochemical data for compound 4

 Property: White powdery substance

Molecular formula: C ₂₇ H _4. N ₂ 0 ₇

 . FAB mass spectrum (m / z): 505 [M + H] +, 503 [M-H] "

 High resolution FAB mass spectrum (m / z):

 Measured value 503.2750

Theory 503.2742 (as _{C 27 H 39 N 2 0 7} )

¾ - nuclear magnetic resonance scan Bae spectrum: 6 (CD ₃ 0D)

0.84 (3H, d, J-6.8 Hz), 1.00 (3H, d, J = 6.4 Hz), 1.02 (1H, m), 1.32 (1H, m), 1.34 (3H, s), 1.44 (1H, m ), 1.64 (1H, m), 1.97 (1H, m), 2.30 (2H, m), 2.32 (1H, m) „2.35 (1H, m), 2.82 (1H, dd, J2 3.7Hz, 12, 9Hz), 3.08 (1H, ddd, J = 2.7Hz, 2.9Hz, 10.0Hz), 3.28 (1H, m), 3.32 (3H, s), 3.40 (3H, s), 3.46 (1H, in), 4.87 (1H, d, J = 8.3Hz), 5.23 (1H, d, J = 11.2Hz), 5.96 (1H, ddd, J2 1.7Hz, 2.0Hz, 15.6Hz), 6.34 (1H, m), 6.43 (1H, dd, J = 2.0Hz, 2.2Hz), 6.52 (1H, dd, J2 1,5Hz, 2.2Hz) , 6.92 (1H, ddd, J = 5.1Hz, 6.4Hz, 15.9Hz) ppm "C- nuclear magnetic resonance scan Bae spectrum:. d (CD ₃ 0D)

 11.7, 18.1, 19.3, 27.3, 29,4, 32.1, 34.9, 36.5, 44.1, 56.7, 60.2, 74.3, 80.2, 82,7, 84.6, 110.3, 116.7, 120.9, 122.6, 131.2, 135.8, 143.6, 146.5, 159.0, 169.5 ppm.

 Soluble angle solubility: Soluble in methanol and dimethyl sulfoxide (DMS0)

 Thin layer chromatography: Rf value 0.12

 Thin layer: Silica gel TLC (Merck)

 Developing solvent: Methanol / chloroform mixture (1: 9) Reference Example 1 Production of compounds 1, 2, and 3

To 29 L of the fermentation culture solution obtained in Example 1, a filter aid (Radiolite 600, manufactured by Showa Chemical Industry Co., Ltd.) was added at a 10% harmful concentration, and then suction filtration was performed to separate the culture filtrate and the bacterial cells. I did. 16 L of methanol was added to the separated cells, and the mixture was sufficiently stirred and extracted. The mixture was again filtered with a suction filter, and the obtained methanol extract was diluted 3-fold with water to obtain a 48 L cell extract.垮 The nutrient filtrate and the bacterial cell extract were separately passed through a column packed with 2 L of Diaion HP-20 to adsorb the active ingredient. After washing each column with a 30% aqueous methanol solution, the active components were eluted with 50% and 75% aqueous methanol solutions. Next, the active fraction was passed through a column packed with 500 mL of Diaion HP-20ss to adsorb the active ingredients.After washing each column with a 50% aqueous methanol solution, 60% and 70% The active ingredient was eluted with a% aqueous methanol solution. The active fraction derived from the culture filtrate was evaporated under reduced pressure to remove the organic solvent, the residue was dissolved in methanol, 35 mL of silica gel was added thereto, and the methanol was distilled off to adsorb the active ingredient onto silica gel. . Next, the silica gel on which the active ingredient was adsorbed was placed on a column filled with 300 mL of silica gel, and developed with a methanol / chloroform-form mixed solution, which was separated into 10% and 20% methanol / chloroform-form fractions. It eluted the active ingredient. The active fraction derived from the bacterial cell extract was similarly adsorbed on 75 mL of silica gel, placed on a 300 mL silica gel that had already been filled, and developed with a methanol / chloroform mixture. The active ingredient was eluted in the formal fraction. The active fraction from the supernatant and the bacterial cell extract are mixed and retained on a reversed-phase silica gel YMC-GEL ODS-AQ 120-S50 400 mL 丽 column (flow rate 2.5 mL / min, mobile phase 55% methanol aqueous solution) Each active fraction (55% methanol) was obtained for a time of 385-480 minutes, 325-385 minutes and 260-325 minutes. After evaporating the solvent of the active fraction (55% methanol) with a retention time of 385 to 480 minutes, the mixture was powdered with dichloromethane to obtain 90.4 mg of Compound 1. The active fraction (55% methanol) with a retention time of 325-385 minutes was further separated by preparative high-performance liquid chromatography (column Inertsil ODS <^ 20 x 250 thigh, column temperature 35 ° C; flow rate 8 mL / min, migration) An active fraction having a retention time of 20 to 21 minutes was collected by using a 30% aqueous phaseacetonitrile solution) to obtain 23.Omg of Compound 2. The active fraction (55% methanol) with a retention time of 260 to 325 minutes was further separated by preparative high performance liquid chromatography (column Inertsil ODS02OX25O mm, column temperature 35 ° C :, flow rate 8 mL / min, mobile phase 25% acetate). An active fraction having a retention time of 28 to 30 minutes was collected by a trilus aqueous solution) to obtain 25.4 mg of compound 3. ,

, Compound 1

 FAB mass spectrum (m / z): 547 (M-1) —

¾-nuclear magnetic resonance spectrum: (5 (500 MHz, DMSO-d ₆ , 323K)

 0.84 (3H, d, J = 6.7 Hz), 0.91 (3H, d, J-6.7 Hz), 1.22 (1H, m), 1.33-1,44 (2H, m), 1.47 (3H, s), 1.54 (1H, m), 1.69 (3H, s), 1.76 (1H, m), 2.13 (1H, m), 2.21 (1H, m), 2.38 (1H, dd, J = 6.5 Hz, 13.8 Hz), 2.41 (1H, m), 2.56 (1H, dd, J = 6.3 Hz, 13.4 Hz), 3.06 (1H, ddd, J = 2.8 Hz, 3.9 Hz, 9.1 Hz), 3.23 (3H, s), 3.28 (1H, m), 3.32 (1H, m), 3.34, (3H, s), 3.65 (3H, s), 4.15 (1H, d, J = 5.5 Hz), 4.89 (1H, d, J = 7,2Hz), 5.33 (1H, d, J = 9.4Hz), 5.90 (1H, br.t, J = 5.7Hz), 6.33 (1H, br.s), 6.34 (2H, br.s), 6.91 (1H, br. s) 9.04 (1H, br s) 6.91 (1H, s) ppm.

¹³ C-nuclear magnetic resonance spectrum: 5 (125 MHz, DMSO-d ₆ , 323 K)

 11.6, 12.8, 15.7, 20.0, 23.6, 29.7, 31.0, 33.5, 34.6, 35.7, 56.3, 58.0, 59.6, 73.9, 79,6, 80.4, 81.1, 107.1, 114.5, 129.6, 132.2, 133.3, 133.5, 134.4, 142.6, 149,6, 156.0, 170.0 ppm.

Compound 2 ESI mass vector (m / z): 517 (Ml) "

 -Nuclear magnetic resonance spectrum: 6 {MHz, CD3OD)

 0.71 (3H, d, J = 6.8Hz), 0.98 (3H, d, J = 6.4Hz), 1.06-1.30 (3H, m), 1.40 (3H, s), 1.58 (1H 'm), 1.77 (3H , s), 1.92 (1H, m), 2.08 (1H, m), 2.23 (1H, m), 2.31-2.37 (2H, m), 2.65 (1H, dd, J = 4.4Hz, 13.2Hz), 3.04 (1H, m), 3.30 (3H, s), 3.39 (3H, s), 3.55 (1H, dd, J = 2.7Hz, 9.2Hz), 5.19 (1H, d, J = 9.9Hz), 5.70 (1H , M), 6.23 (1H, s), 6.32 (1H, m), 6.51 (1H, br. S) ppm.

 ESI mass vector (m / z): 533 (M-1) —

¾-nuclear magnetic resonance spectrum: (5 (300 MHz, DMS0-d ₆ )

 0.75 (3H, d, J = 6.2Hz), .0.90 (3H, d, J = 6.4Hz), 1.04-1.25 (3H, m), 1.38 (3H, s), 1.60 (1H, m), 1.66 ( 3H, s), 1.79 (1H, m), 1.99-2.19 (2H, m), 2.24-2.38 (2H, m), 2.65 (1H, dd, J = 6.1Hz, 13.4Hz), 2.97 (1H, m ), 3.21 (3H, s), 3.31 (3H, s), 4.26 (1H, br. S), 4.83 (1H, d, J = 7.5Hz), 5.21 (1H, d, J = 9.5Hz), 5.70 (1H, br.s), 6.15 (1H, br.s), 6.45 (2H, br.s), 6.67 (1H, br.s), 7.86 (1H, br.s), 9.08 (1H, br.s) s), 9.212 (1H, br. s) ppm. Example 2 Compound 5

Dissolve 1.3 mg (0.0024 bandol) of Compound 1 in 1 mL of methanol / acetonitrile mixture (1: 9), add 100 _/ L (0.2 mmol) 2 mol / L trimethylsilyldiazomethane / hexane solution, and stir at room temperature did. After 2 hours and 15 minutes, the solvent was distilled off. The residue was purified by preparative thin-layer chromatography [methanol / form-form mixture (1: 9)] to obtain 0.3 mg of Compound 5 (yield).

23%). '

 FAB Mass Vector (m / z): 561 (M-1 Square)

 -Nuclear magnetic resonance spectrum: (5 (300 MHz, CDCI3)

0.84 (3H, d, J = 6.8Hz), 1.05 (3H, d, J = 6.6Hz), 1.85 (3H, s), 2.00 (1H, m), 2.25 (1H 'm), 2.35-2.70 (4H , M), 3.12 (1H, m), 3.34 (3H, s), 3.35 (1H, m), 3.400 (3H, s), 3.64 (1H, m), 3.74 (3H, s), 3.86 (3H, s), 4,76 (2H, br.s), 5.18 (1H, m), 5.37 (1H, d, J-9.4Hz), 6.03 (1H, m), 6.40 (1H, d, 'J = 2.4 Hz), 7.90 (1H, br. S) ppm. Example 3 Compound 6, Compound 8

Compound 1 1.2 mg of (0.00 ²² thigh ol) was dissolved in pyridine 500 L, followed by stirring at room temperature was added acetic anhydride 10 ^ L (O.llmmol). Two hours later, water was added, and extraction was performed with ethyl acetate. The organic layer was dried over sodium sulfate, and the solvent was distilled off. The residue was purified by preparative thin-layer chromatography [methanol / chloroform mixture (7:93)] to give 0.5 mg of compound 6 (38% yield). Compound 8 (0.5 mg, yield 36%) was obtained.

 Compound 6

 FAB Mass Spectrum (m / z): 589 (M-1) "

¾ - nuclear magnetic resonance scan Bae _{spectrum: d (300 MHz, CDC1 3} )

 0.85 (3H, d, J = 6.4.4Hz), 1.07 (3H, d, J = 6.8Hz), 1.85 (3H, s), 2.05 (1H, m), 2.317 (3H, s), 2.20-2.60 (5H, m), 3.14 (1H, m), 3.34 (3H, s), 3.40 (3H, s) 3.64 (1H, m), 3.72 (3H, s), 4.80 (2H, br s), 5.20 ( 1H, m), 5.37 (1H, m), 6.02 (1H, in), 6.723 (1H, d, J = 2.9Hz), 7.35 (1H, br.s), 8.05 (1H, br.s) ppm.

 Compound 8

 FAB mass spectrum (m / z): 631 (M-l) "

¾- nuclear magnetic resonance scan Bae _{spectrum: 5 (300 MHz, CDC1 3} )

 1.00 (3H, d, J = 6.8Hz), 1.08 (3H, d, J = 6.8Hz), 1.878 (3H, s), 2.098 (3H, s) 2.312 (3H, s), 2.72-2.80 (2H, m), 3.24 (1H, m), 3.36 (3H, s), 3.42 (3H, s) 3.73 (3H, s), 4.72 (2H, br.s), 4.91 (1H, m), 5.06 (1H, d, J two 6.1Hz) 5.33 (1H, d, J = 9.0Hz), 6.08 (1H, m), 6.69 (1H, d, J = 2.4Hz), 7.75 (1H, br.s), 8.30 (1H , br.s) ppm.Example 4 Compound 7

Compound 8 was dissolved in 500 μL of methanol, and 500 L of a saturated aqueous solution of sodium hydrogen carbonate was added at room temperature, followed by stirring. Twenty minutes later, extraction was carried out using a mixture of form-lmol / L hydrochloric acid and lip. The organic layer is dried over sodium sulfate, and the solvent is distilled off. The mixture was purified with a feed [methanol / chloroform mixture (1: 9)] to give 1.2 mg of compound 7. FAB Mass Spectrum (m / z): 589 (M-1) "

Nuclear magnetic resonance scan Bae _{spectrum: 5 (300 MHz, CDC1 3} )

0.95 (3H, d, J = 6.6Hz), 1.06 (3H, d, J = 6.6Hz), 1.12-1.49 (4H, m), 1.53 (3H, s), 1.82 (1H, m), 1.87 (3H , s), 2.10 (3H, s), 2.23-2.41 (3H, m), 2.65-2.70 (2H, m), 3.25 (1H, m), 3.36 (3H, s), 3.42 (3H, s), 3.76 (3H, s), 4.70 (2H, br s), 4.93 (1H, m), 5.05 (1H, d, J = 6.4Hz) 5.32 (1H, d, J = 9.0Hz), 5.70 (1H, s) ), 6.03 (1H, m), 6.33 (1H, m), 7.49 (1H, br. S), 8.08 (1H, br. S) ppm.

 A tablet having the following composition is prepared by a conventional method. 40 g of Compound 4, 286.8 g of lactose and 60 g of corn starch are mixed, and 120 g of a 10% aqueous solution of hydroxypropylcellulose is added thereto. This mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. 1.2 g of magnesium stearate was added and mixed, and the mixture was tableted with a tableting machine equipped with a 8 mm diameter punch (Clean Press Collect 12 manufactured by Kikusui Seisakusho), and tablets (containing 20 mg of active ingredient per tablet) Get)

Formulation Compound 4 20 mg

 Lactose 143.4 mg

 Corn starch 30 mg

 Hydroxypropyl cellulose 6 mg

 Magnesium stearate 0.6 mg

 200 mg Example 6

A tablet having the following composition is prepared by a conventional method. 40 g of compound 6, 286.8 g of lactose and 60 g of corn starch are mixed, and 120 g of a 10% aqueous solution of hydroxypropyl cellulose is added. This mixture is kneaded by a conventional method, granulated and dried, and then sized to obtain granules for tableting. Add 1.2g of magnesium stearate to this Mix and compress with a tableting machine equipped with a 8 mm diameter punch (Clean Press Collect 12 manufactured by Kikusui Seisakusho) to obtain tablets (each tablet containing 20 mg of active ingredient).

Prescription compound 6 20 mg

 Lactose 143.4 mg

 Corn starch 30 mg

 Hydroxypropyl cellulose 6 mg

 0.6 mg magnesium stearate

 200 fflg Example 7

 An injection having the following composition is prepared by a conventional method. Dissolve lg of compound 7 and 9 g of sodium chloride in 100 mL of distilled water for injection by a conventional method. The resulting solution is aseptically filtered using a disposable membrane filter (0.2), and aseptically filled into glass vials by 2 mL each to contain an injection (containing 2 mg of active ingredient per vial). obtain.

Formulation compound 7 2 mg

 Sodium chloride 18 mg

 Appropriate amount of distilled water for injection

 2.00inL Industrial Applicability

 According to the present invention, there are provided hsp90 family protein inhibitors and the like, which are useful as pharmaceuticals such as antitumor agents and contain a penzenoid ansamycin derivative or a pharmacologically acceptable salt thereof as an active ingredient. You.

Claims

The scope of the claims

 1. Formula (I)

(Wherein, R ² represents hydrogen or methyl, R ¹¹ and R ¹⁸ are the same or different and represent a hydroxy, a substituted or unsubstituted lower alkoxy or a substituted or unsubstituted lower alkanoyloxy, ¹⁷ represents hydrogen, hydroxy, substituted or unsubstituted lower alkoxy or substituted or unsubstituted lower alkanoyloxy), a benzenoid ansamycin derivative or a pharmacologically acceptable salt thereof. Heat shock protein 90 (hsp90) family protein inhibitor contained as an ingredient.

2. Hsp90 Fuamiri one protein inhibitor according to claim 1, wherein R ¹⁷ is hydrogen.

3. The hsp90 family protein inhibitor according to claim 1, wherein R ² is hydrogen. '. Is methyl, hsp9 Q Fuamiri one protein inhibitor in the range 1 or 2, wherein according to R ¹¹ is a lower alkanoyloxy Noi Ruo alkoxy lower alkoxy, or substituted or unsubstituted substituted or unsubstituted.

 ■

5. Hsp90 Fuami Lee protein inhibitor according to any one of claims 1 to 3 wherein R ¹¹ is hydroxy.

6. The hsp90 family protein inhibitor according to claim 1, wherein R ² is methyl, R ¹¹ is hydroxy, R ¹⁷ is hydrogen or hydroxy, and R ¹⁸ is hydroxy.

7. Hsp90 Fuami Lee protein inhibitor according to any one of claims 1 to 5 wherein R ¹⁸ is low grade alk Noi Ruo alkoxy lower alkoxy, or substituted or unsubstituted substituted or unsubstituted.

8. The method of claims 1 to 5, wherein R ¹⁸ is a substituted or unsubstituted lower alkanoyloxy. The hsp90 family protein inhibitor according to any one of the above.

 9. Expression (la)

^Wherein R ^2a , R ^lla , R ^17a and R ^18a are each as defined for R ² , R ^U , R ¹⁷ and R ¹⁸ , but R ^2a is methyl, and R ^lla and R ^18a When both are hydroxy, R is not hydrogen, hydroxy or methoxy) or a pharmaceutically acceptable salt thereof.

10. The benzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to claim 9, wherein R ^2a is hydrogen.

 11. The benzenoid ansamisin derivative or the pharmaceutically acceptable salt thereof according to claim 9 or 10, wherein R is hydrogen.

12. R ^lla and one or both of R ^18a is, benzene Noi de answer according to either the range 9-11 Noi deviation according a lower alkanoyloxy Noi Ruo alkoxy lower alkoxy, or substituted or unsubstituted substituted or unsubstituted A mycin derivative or a pharmacologically acceptable salt thereof.

 13. A medicament comprising the benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to any one of claims 9 to 12 as an active ingredient.

 14. An antitumor agent comprising, as an active ingredient, the benzenoidoansamycin derivative or the pharmaceutically acceptable salt thereof according to any one of claims 9 to 12.

15. An hsp90 family protein or a protein to which an hsp90 family protein binds, comprising as an active ingredient a benzenoid doansamycin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 9 to 12 A therapeutic agent for a disease.

16. An hsp90 family protein inhibitor comprising, as an active ingredient, the benzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to any one of claims 9 to 12.

 17. Inhibiting the hsp90 family protein, which comprises administering an effective amount of a benzenoid ansamycin derivative represented by the formula (I) or a pharmacologically acceptable salt thereof according to claim 1. how to.

 18. A method for treating a malignant tumor, comprising administering an effective amount of the benzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to any one of claims 9 to 12.

 19. An hsp90 family protein or an hsp90 family protein, which comprises administering an effective amount of the benzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to any one of claims 9 to 12. A method for treating a disease associated with a protein to which the protein binds.

 20. A method for inhibiting hsp90 family protein, which comprises administering an effective amount of the benzenoid ansamycin derivative or the pharmaceutically acceptable salt thereof according to any one of claims 9 to 12.

 21. Use of a penzenoid doansamycin derivative represented by the formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof for the production of an hsp90 family one protein inhibitor.

 22. Use of a benzenoidoansamycin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 9 to 12 for the manufacture of an antitumor agent.

 23. The benzenoid doansamycin derivative or pharmacologically thereof according to any one of claims 9 to 12 for the manufacture of a therapeutic agent for a disease associated with an hsp90 family 1 protein or a protein to which the hsp90 family protein binds. Use of acceptable salts.

 24. Use of the benzenoid ansamycin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 9 to 12 for the production of an hsp90 family protein inhibitor.

25. Formula (lb)

A microorganism belonging to the genus Streptomyces, which produces a benzenoid ansamycin derivative represented by the formula:

 26. Streptomyces sp. EH21 (Streptomyces sp. EH21, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary No. FERM BP-08574).

 27. The expression of the formula (lb) according to claim 25, comprising a step of culturing the microorganism according to claim 25 and isolating a compound produced from the obtained culture solution. A method for producing a benzenoidoansamycin derivative.

 28. Streptomyces sp.EH21 (Streptomyces sp.EH21, Independent Administrative Agency, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary Depositary No. FERM BP-08574) The method for producing a benzenoid ansamycin derivative according to any one of claims 9 to 12, comprising a step of isolating.

 29. Streptomyces sp.EH21 (Streptomyces sp.EH21, Independent Administrative Agency National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary Depositary Number FERM BP-08574) A method for producing a benzenoid ansamycin derivative represented by the formula (lb) according to claim 25, comprising a step of isolating.