JPH078872B2 - New tetracyclo compounds and their production method - Google Patents

Description

[Detailed Description of the Invention] The present invention relates to novel tetracyclo compounds.

More particularly, the present invention relates to novel tetracyclo compounds or salts thereof having pharmacological activity and methods for preparing them.

Therefore, one object of the present invention is to provide novel tetracyclo compounds having pharmacological activities such as antitumor activity and antibacterial activity, and pharmaceutically acceptable salts thereof.

Another object of the present invention is to provide a process for producing said tetracyclo compound or a salt thereof, which comprises a fermentation process and a synthesis process.

The present invention is based on the first discovery of a novel compound, substance FR-900482.

More specifically, the above-mentioned FR-900482 substance was isolated for the first time in pure form from a culture broth obtained by fermenting a new species belonging to the genus Streptomyces.

As a result of extensive research to clarify the chemical structures of FR-900482 substance and its triacetyl derivatives, the inventors of the present invention determined the unique chemical structures of these compounds and further succeeded in synthesizing the tetracyclo compound of the present invention, thereby completing the present invention.

The novel tetracyclo compound of the present invention is represented by the following general formula:

[wherein R ¹ is a formyl group, a protected formyl group, a hydroxymesyl group, a protected hydroxymethyl group, an arylaminomethyl group, a carboxy group, a protected carboxy group or a substituted iminomethyl group; R ² is a hydroxy group, an alkoxy group or a protected hydroxy group; R ³
is hydrogen, and ^R4 is a methyl group, a hydroxymethyl group or a protected hydroxymethyl group, or ^R3 and ^R4 are bonded to each other to form a methylene group or an oxo group, ^R5 is a hydroxy group, an alkoxy group or a protected hydroxy group, and ^R6 is hydrogen, an imino-protecting group or ^an alkyl ^{group .}
are each a hydroxy group, ^R3 and ^R6 are each a hydrogen atom,
The compound in which ^R4 is a carbamoyloxymethyl group was produced by fermentation and named substance FR-900482.

In the tetracyclo compound (I) of the present invention, one or more stereoisomers such as optical and geometric isomers due to asymmetric carbon atoms and double bonds may exist, and such isomers are also included within the scope of the present invention.

Suitable salts of the object compound (I) are conventional pharmaceutically acceptable salts, including inorganic base salts [for example, alkali metal salts (e.g., sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g., calcium salts, magnesium salts, etc.),
ammonium salts], salts with bases such as organic base salts [for example, organic amine salts (e.g., triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc.)]; salts with acids such as inorganic acid addition salts (e.g., hydrochloride, hydrobromide, sulfate, phosphate, etc.), organic acid addition salts (e.g., formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.); basic or acidic amino acids (e.g., arginine, aspartic acid, glutamic acid, etc.).
and salts thereof.

In the present invention, the target tetracyclo compound (I) can be produced by the following method.

[I] Fermentation method: Streptomyces species [II] Synthesis Method: (1) Production Method 1 (Introduction of an imino-protecting group) (2) Production Method 2 (Introduction of a hydroxy-protecting group into the 6-hydroxy group) (3) Production Method 3 (Introduction of a hydroxy-protecting group into the 9-hydroxy group) (4) Production Method 4 (Introduction of a hydroxy-protecting group into an 8-hydroxymethyl group) (5) Production Method 5 (Introduction of a hydroxy-protecting group into a 4-hydroxymethyl group) (6) Production Method 6 (Reaction with Base) (7) Production Method 7 (Reduction of Methylene Group) (8) Production Method 8 (Reduction to Hydroxymethyl Group) (9) Production Method 9 (Introduction of a Formine Protecting Group) (10) Production Method 10 (Alkylation of 6-hydroxy group) (11) Production Method 11 (Alkylation of 9-hydroxy group) (12) Production Method 12 (Reaction with Substituted Amine) (13) Production Method 13 (reduction of alinomycin imino group) (14) Production Method 14 (Hydrolysis of Protected Hydroxymethyl Group) (15) Production Method 15 (Removal of imino protecting group) (16) Production Method 16 (Oxidation of Formyl Group) (17) Production Method 17 (Oxidation of Methylene Group) (18) Production Method 18 (Introduction of a carboxy-protecting group) (19) Production Method 19 (Removal of Formyl Protecting Group) (20) Production Method 20 (Alkylation of imino group) (21) Production Method 21 (Hydrolysis of Protected 6-Hydroxy Group) (22) Production Method 22 (Introduction of a hydroxy-protecting group into a hydroxyiminomethyl group) wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ and R ⁶ are the same as above, ▲R ¹ _a ▼ and ▲R ⁴ _a ▼ are each a protected hydroxymethyl group, ▲R ¹ _b ▼ is a formyl group or a protected formyl group, ▲R ¹ _c ▼ is a protected formyl group, ▲
R ¹ _d ▼ is a substituted iminomethyl group, ▲R ¹ _e ▼ is an aryliminomethyl group, ▲R ¹ _f ▼ is an arylaminomethyl group, ▲R
¹ _g ▼ is a protected carboxy group, ▲R ¹ _h ▼ is a protected hydroxyiminomethyl group, ▲R ² _a ▼ and ▲R ⁵ _a ▼ are protected hydroxy groups, ▲R ² _b ▼ and ▲R
⁵ _b ▼ represents an alkoxy group, R ⁶ _a ▼ represents an imino-protecting group, and R ⁶ _b ▼ represents an alkyl group. Specific examples of the above definitions and their preferred embodiments are described in detail below.

Unless otherwise indicated, "lower" as used in this specification
means 1 to 6 carbon atoms;
The term "higher" is intended to mean 7 to 20 carbon atoms.

Suitable "imino protecting groups" and suitable hydroxy protecting groups in "protected hydroxy groups" and "protected hydroxymethyl groups" and "protected hydroxyiminomethyl groups" include carbamoyl groups resulting from carboxylic acids, carbonic acids, sulfonic acids and carbamic acids;
Acyl groups include aliphatic acyl groups, aromatic acyl groups, heterocyclic acyl groups, and aliphatic acyl groups substituted with aromatic or heterocyclic groups.

Examples of the aliphatic acyl group include saturated or unsaturated acyclic or cyclic aliphatic acyl groups, for example, alkanoyl groups such as lower or higher alkanoyl groups (e.g., formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, heptanoyl, octanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, etc.), alkyl groups such as lower alkylsulfonyl groups (e.g., mesyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, pentylsulfonyl, hexylsulfonyl, etc.), and alkyl groups such as lower alkylsulfonyl groups (e.g., mesyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, butylsulfonyl, isobutylsulfonyl, pentylsulfonyl, hexylsulfonyl, etc.). Examples include sulfonyl groups; N-alkylcarbamoyl groups (e.g., methylcarbamoyl, ethylcarbamoyl, etc.); alkoxycarbonyl groups such as lower alkoxycarbonyl groups (e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, t-butoxycarbonyl, etc.); alkenoyl groups such as lower alkenoyl groups (e.g., acryloyl, methacryloyl, crotonoyl, etc.); cycloalkanecarbonyl groups such as cyclo(lower)alkanecarbonyl groups (e.g., cyclopropanecarbonyl, cyclopentanecarbonyl, cyclohexanecarbonyl, etc.); and the like.

Examples of the aromatic acyl group include an aroyl group (e.g., benzoyl, toluoyl, xylyl, etc.), an N-arylcarbamoyl group (e.g., N-phenylcarbamoyl, N-
tolylcarbamoyl, N-naphthylcarbamoyl, etc.),
Examples include arenesulfonyl groups (for example, benzenesulfonyl, tosyl, etc.).

Examples of the heterocyclic acyl group include heterocyclic carbonyl groups (for example, furoyl, thenoyl, nicotinoyl, isonicotinoyl, thiazolylcarbonyl, thiadiazolylcarbonyl, tetrazolylcarbonyl, etc.).

Examples of the aliphatic acyl group substituted with an aromatic group include phenyl (lower) alkanoyl groups (e.g., phenylacetyl, phenylpropionyl, phenylhexanoyl, etc.).
aralkanoyl groups such as phenyl(lower)alkoxycarbonyl groups (e.g., benzyloxycarbonyl,
aralkoxycarbonyl groups such as phenethyloxycarbonyl, etc.; and aryloxyalkanoyl groups such as phenoxy(lower)alkanoyl groups (for example, phenoxyacetyl, phenoxypropionyl, etc.).

Examples of the aliphatic acyl group substituted with a heterocyclic group include heterocyclic (lower) alkanoyl groups such as thienylacetyl, imidazolylacetyl, furylacetyl, tetrazolylacetyl, thiazolylacetyl, thiadiazolylacetyl, thienylpropionyl, and thiadiazolylpropionyl.

These acyl groups may be further substituted with one or more suitable substituents such as lower alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, etc.), halogen (e.g., chlorine, bromine, iodine, fluorine), lower alkoxy (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, pentyloxy, hexyloxy, etc.), lower alkylthio (e.g., methylthio, ethylthio, propylthio, isopropylthio, butylthio, benzylthio, hexylthio, etc.), nitro, etc., and preferred acyl groups having such substituents are methylthio, ethylthio, propylthio, isopropylthio, butylthio, benzylthio, hexylthio, etc. Examples of the acyl group include mono- (or di- or tri-)haloalkanoyl groups (e.g., chloroacetyl, bromoacetyl, dichloroacetyl, trifluoroacetyl, etc.), mono- (or di- or tri-)haloalkoxycarbonyl groups (e.g., chloromethoxycarbonyl, dichloromethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, etc.), and nitro (or halo or lower alkoxy)aralkoxycarbonyl groups (e.g., nitrobenzyloxycarbonyl, chlorobenzyloxycarbonyl, methoxybenzyloxycarbonyl, etc.).

More preferred examples of the imino-protecting group and hydroxy-protecting group defined above include a carbamoyl group, a lower alkanoyl group, a higher alkanoyl group, an aroyl group which may have a halogen atom, a lower alkylsulfonyl group, a phenyl(lower)alkoxycarbonyl group, and a N-
and most preferred examples include a carbamoyl group, an acetyl group, a propionyl group, an octanoyl group, a p-bromobenzoyl group, a mesyl group, a benzyloxycarbonyl group and an N-phenylcarbamoyl group.

Suitable "alkoxy groups" include straight-chain or branched-chain alkoxy groups such as lower alkoxy groups (e.g., methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentyloxy, isopentyloxy, hexyloxy, etc.), and among these, more preferred examples are _C1 - _C4 alkoxy groups, and most preferred examples are methoxy groups.

Suitable "protected formyl groups" include acyclic or cyclic acetals of formyl groups, dialkoxymethyl groups such as di(lower)alkoxymethyl groups (e.g., dimethoxymethyl, diethoxymethyl, dipropoxymethyl, diisopropoxymethyl, dibutoxymethyl, dibenzyloxymethyl, dihexyloxymethyl, etc.), lower alkylenedioxymethyl groups (e.g., 1,3-
Among these, more preferred examples are di(lower)alkoxymethyl groups, and most preferred examples are dimethoxymethyl and diethoxymethyl groups.

Suitable "protected carboxy groups" include lower alkoxycarbonyl groups (e.g., methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, t-butoxycarbonyl, etc.), mono (or di- or tri)phenyl (lower) alkoxycarbonyl groups optionally having nitro (e.g., benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, phenethyloxycarbonyl,
and esterified carboxy groups such as benzhydryloxycarbonyl, trityloxycarbonyl, etc., among which more preferred examples include C ₁ -
A _C4 alkoxycarboxy group is the most preferred example, and a methoxycarbonyl group is the most preferred example.

Suitable aryl moieties of the "arylaminomethyl group" include phenyl, tolyl, xylyl, cumenyl, mesityl, naphthyl, etc., and among these, the preferred example is phenyl.

Suitable "alkyl groups" include straight-chain or branched-chain alkyl groups such as lower alkyl groups (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, neopentyl, hexyl, etc.), and among these, more preferred examples include _C1 - _C4 alkyl groups, and the most preferred example is a methyl group.

Suitable substituted imino moieties in the "substituted iminomethyl group" include aryl imino (e.g., phenyl imino,
tolylimino, xylylimino, cumenylimino, mesitylimino, naphthylimino, etc.), hydroxyimino,
Alkoxyimino such as lower alkoxyimino (e.g., methoxyimino, ethoxyimino, propoxyimino, isopropoxyimino, butoxyimino, hexyloxyimino, etc.); lower alkanoyloxyimino (e.g., formyloxyimino, acetoxyimino,
and acyloxyimino (e.g., propionyloxyimino, butyryloxyimino, pentanoyloxyimino, hexanoyloxyimino, etc.), semicarbazono, arylsemicarbazono (e.g., 4-phenylsemicarbazono, 4-tolylsemicarbazono, 4-naphthylsemicarbazono, etc.), and more preferred examples among these include phenylimino, hydroxyimino, C ₁
Examples of the alkoxyimino group include C 1 -C ₄ alkoxyimino, C ₁ -C ₄ alkanoyloxyimino, semicarbazono, and phenylsemicarbazono, and the most preferred examples include phenylimino, hydroxyimino, methoxyimino, acetoxyimino,
semicarbazono and 4-phenylsemicarbazono.

The method for producing the tetracyclo compound (I) of the present invention will be explained in detail below.

[I] Fermentation method: The FR-900482 substance in the present invention is produced by fermentation of Streptomyces
Sandaensis No. 6897 (Streptomyces Sandaensis No.
FR-900482 belonging to the genus Streptomyces, such as 6897
The substance can be produced by fermenting a substance-producing bacterium in a medium.

The details of the microorganisms used to produce the FR-900482 substance are described below.

The microorganism that can be used for producing the FR-900482 substance is a strain that belongs to the genus Streptomyces and produces the FR-900482 substance. Streptomyces sandaensis No. 6897, newly isolated from a soil sample collected in Sanda City, Hyogo Prefecture, Japan, is used.
Examples include:

A freeze-dried specimen of this newly isolated Streptomyces sandaensis No. 6897 was deposited on June 1, 1984, with the Fermentation Research Institute, Agency of Industrial Science and Technology (305, 1-1-3 Higashi, Yatabe-cho, Tsukuba-gun, Ibaraki Prefecture) under deposit number 7654, and this deposit was transferred to the Budapest Treaty route on May 18, 1985, under the new deposit number 792.

The production of the novel FR-900482 substances is not limited to the use of the specific microorganisms described herein, which are shown for illustrative purposes only.

The present invention also encompasses the use of any mutant strains capable of producing the FR-900482 substance, including naturally occurring mutant strains as well as artificial mutant strains that can be produced from the above microorganisms by conventional methods such as X-ray irradiation, ultraviolet irradiation, or treatment with N-methyl-N'-nitro-N-nitrosoguanidine, 2-aminopurine, etc.

Streptomyces sandaensis No. 6897 has the following morphological, cultural, biological and physiological characteristics:

[1] Morphological characteristics: In principle, the method described by Scherling and Gottlieb, "Scherling, E. B. and D. Gottlieb: Identification of Streptomyces species," International Journal of Systematic Bacteriology, Vol. 16, No. 31
3-340, 1966 (Shirling, EBand D. Gottlieb:Me
thods for characterization of Streptomyces species.
International Journal of Systematic Bacteriology, 1
6 , 313-340, 1966) was used for this taxonomic study.

Morphological observations were performed by light and electron microscopy on cultures grown on yeast-malt extract agar, oatmeal agar, or mineral salts-starch agar at 30°C for 14 days. Mature sporophytes formed rectiflexibiles and retinaculia perti with 10-20 spores in each spore chain. The spores were cylindrical and approximately 0.5 mm in size under electron microscopy.
The size of the spores was about 0.7 x 0.6-0.8 μm. The spore surface was smooth.

[2] Cultivation characteristics The cultivating characteristics were as described by Scherling and Gottlieb, supra, and Waxman, "Waxman, SA: Actinomycetes, 2nd Edition."
Volume: Classification, Identification and Characterization of Genera and Species, Williams and Wilkins, Baltimore, 1961 (Wa
ksman,SA:The actinomycetes,vol.2:Classification,
identification and description of genera and speci
The observations were made on 10 different media described in (The Williams and Wilkins Co., Baltimore, 1961).

Incubation was continued at 30°C for 14 days. The color names used in this study were based on the Shinshiki Meicho (Nippon Shikisai Co., Ltd.). When grown on oatmeal agar, yeast-malt extract agar, and inorganic salts-starch agar, the colonies belonged to the gray color range. Soluble pigments were produced on yeast-malt extract agar but not on other media. The results are shown in Table 1.

Analysis of cell wall composition was carried out by Becker et al., "Becker, B., MP Rechevalier, RE Gordon and HA Rechevalier: Rapid Differentiation of Nocardia and Streptomyces by Paper Chromatography of Whole-Cell Hydrolysates: Applied Microbiology, 12 , 421-423, 1964" (Becke
r,B.,MPLechevalier,REGordon and HALechevalie
r:Rapid differentiation bet−ween Nocardia and Str
eptomyces by paper chromatography of whole−cell h
Hydrolysates: Appl. Microbiol., 12 , 421-423, 1964) and Yamaguchi, T.: Comparison of cell wall compositions of morphologically distinct actinomycetes: Journal of Bacteriology, 8
9,444−453,1965 ” (Yamaguchi, T.: Comparison of the
cell−wall composition of morphologically distinct
The cell wall of this strain was analyzed by the method of the microbial cell wall cloning method (J. Bacteriol., 89 , 444-453, 1965). Analysis of the whole-cell hydrolysate of strain No. 6897 revealed the presence of LL-diaminopimelic acid. Therefore, the cell wall of this strain is considered to be Type I.

[3] Biological and physiological properties The physiological properties of strain No. 6897 were examined by the method of Shirling and Gottlieb. The results are shown in Table 2.

The growth temperature range and optimum temperature were examined using a temperature gradient incubator (manufactured by Toyo Kagaku Sangyo Co., Ltd.) on yeast-malt extract agar.

The growth temperature range was 10-35° C., with the optimum temperature being 30-32° C. Starch hydrolysis was positive, and black pigment production was negative.

The utilization of carbon sources was determined by the method of Pridham and Gottlieb. "Pridham, T.G. and D. Gottlieb: Utilization of carbon compounds by Actinomycetes as an aid to species identification: Journal of Bacteriology, 56 , 107-114, 1948"
(Pridham, T. G. and D. Gottlieb: The utilization of ca.
rbon compounds by some Actinomycetales as an aid f
or species determination:J.Bacteriol., 56 ,107−114,
Growth was observed after 14 days of incubation at 30°C.

The carbon source utilization of this strain is summarized in Table 3. Sucrose and D-fructose were not utilized by strain No. 6897.

Microscopic studies and cell wall composition analysis of strain No. 6897 confirmed that this strain was Streptomyces Waksman and Henrici 1943 (Streptomyces Waksman and Henrici 1943).
It was discovered that it belongs to the genus (1943).

Therefore, this strain and various Streptomyces species were compared based on their published properties [International Journal of Systematic Bacteriology 18 , 69-189, 279
-392 (1968) and 19 , 391-512 (1969), and Burgess's Manual of Determinative Bacteriology, 8th ed. (1974)] [International Journal
of Systematic Bacteriology, 18 ,69 to 189,279 to 392
(1968) and 19,391 to 512 (1969), and Bergy's Manu
al of Determinative Bacteriology 8th Edition (197
4)] and compared.

As a result of the comparison, strain No. 6897 was found to be a strain of Streptomyces aburaviensis (Nishimura et al.)
ensis Nishimura et al.) and Streptomyces nitrosporeus (by Okami) (Streptomyces nitro
sporeus Okami).
As shown in 2 and 3, strain No. 6897 was further compared with Streptomyces aburaviensis IFO12830 and Streptomyces nitrosporeus IFO12803. As a result, strain No. 6897 was distinguishable from these two strains in the following respects. Therefore, strain No. 6897 was considered to be a new species of Streptomyces and was named Streptomyces sandaensis sp. nov. after the soil collected in Sanda City from which this microorganism was isolated.

Differences from Streptomyces aburaviensis IFO12830 The cultural characteristics of strain No. 6897 differ from those of Streptomyces aburaviensis on glucose-asparagine agar, sucrose-nitrate agar, and potato dextrose agar.

The gelatin liquefaction test of strain No. 6897 was positive, but the gelatin liquefaction test of Streptomyces aburaviensis was negative.

In terms of NaCl resistance, strain No. 6897 can grow in the presence of 5% NaCl, but Streptomyces aburaviensis cannot grow under the same conditions.

Regarding carbon source utilization, strain No. 6897 was able to utilize D-xylose, rhamnose, raffinose, D-galactose,
It utilizes L-arabinose, D-mannose, and salicin, whereas Streptomyces aburaviensis does not. Furthermore, strain No. 6897 cannot utilize sodium acetate, whereas Streptomyces aburaviensis can.

Differences from Streptomyces nitrosporeus IFO12803 The cultural characteristics of strain No. 6897 differ from those of Streptomyces nitrosporeus on glucose-asparagine agar, glycerol-asparagine agar, and tyrosine agar.

The nitrate reduction and urease activities of strain No. 6897 were negative, but those of Streptomyces nitrosporeus were positive.

Regarding NaCl tolerance, strain No. 6897 cannot grow in the presence of 7% NaCl, but Streptomyces nitrosporeus can grow under the same conditions.
Strain No. 6897 cannot utilize lactose and sodium citrate, but Streptomyces nitrosporeus can. Strain No. 6897 can utilize raffinose, but
Streptomyces nitrosporeus is not available.

Production of FR-900482 substance The novel FR-900482 substance of this invention can be produced by culturing an FR-900482 substance-producing strain belonging to the genus Streptomyces (for example, Streptomyces sandaensis No. 6897; Microbial Research Institute No. 792) in a medium.

In general, the FR-900482 substance can be produced by culturing (e.g., shaking culture, submerged culture, etc.) an FR-900482 substance-producing strain in an aqueous medium containing assimilable carbon and nitrogen sources, preferably under aerobic conditions.

Preferred carbon sources in the medium are glucose, xylose,
Carbohydrates such as galactose, glycerin, starch, dextrin, etc. Other carbon sources include maltose, rhamnose, raffinose, arabinose, mannose, salicin, sodium succinate, etc.

Preferred nitrogen sources are yeast extract, peptone, gluten meal, cottonseed flour, soybean flour, corn steep liquor, dry yeast, malt, feather meal, peanut flour, etc., as well as ammonium salts (e.g., ammonium nitrate, ammonium sulfate,
These are inorganic and organic nitrogen compounds such as ammonium phosphate, urea, amino acids, etc.

Carbon and nitrogen sources are conveniently used in combination, but it is not necessary to use them in pure form, as less pure materials containing trace amounts of growth factors and significant amounts of inorganic nutrients are also suitable for use. Optionally, sodium or calcium carbonate, sodium or potassium phosphate, sodium or potassium chloride, sodium or potassium iodide, magnesium salts, copper salts,
Inorganic salts such as cobalt salts may be added to the medium.
If necessary, an antifoaming agent such as liquid paraffin, fatty oil, vegetable oil, mineral oil or silicone may be added, especially when the culture medium foams significantly.

Submerged aerobic culture is preferred for the mass production of FR-900482. For small-scale production, shaking culture in flasks or bottles or surface culture is used. Furthermore, when growth is carried out in large tanks, FR-900482 is used.
To avoid growth delays during the production process, it is preferable to inoculate a production tank with a pre-culture of microorganisms. Therefore, it is desirable to first inoculate a relatively small amount of culture medium with spores or mycelia of the microorganisms and cultivate the inoculation medium to produce a pre-culture inoculum of the microorganisms, and then aseptically transfer the cultivated pre-culture inoculum to a larger tank. The medium for producing this pre-culture inoculum is preferably FR
- The medium may be substantially the same as or different from the medium used for the production of the 900482 substance.

Agitation and aeration of the culture mixture may be accomplished in a variety of ways. Agitation may be accomplished with a propeller or similar mechanical agitation device, by rotating or shaking the fermenter, by various pumping devices, or by passing sterile air through the medium. Aeration may be accomplished by passing sterile air through the fermentation mixture.

Fermentation is usually carried out at a temperature of 20°C to 40°C, preferably 25°C to 35°C, for about 50 to 150 hours, but these can be varied depending on the fermentation conditions and scale.

The FR-900482 substance produced can be recovered from the culture medium by conventional means commonly used to recover other known biologically active substances.
The majority of the 0482 material is present in the culture filtrate.
The FR-900482 substance can be isolated and purified from the filtrate obtained by filtering or centrifuging the culture broth by conventional methods such as vacuum concentration, freeze-drying, extraction with conventional solvents, pH adjustment, treatment with conventional resins (e.g., anion or cation exchange resins, nonionic adsorption resins, etc.), treatment with conventional adsorbents (e.g., activated carbon, silicic acid, silica gel, cellulose, alumina, etc.), crystallization, recrystallization, etc.
The 82 substances have the following physical and chemical properties and show two Rf values by silica gel thin layer chromatography.

(1) Form and color: Colorless powder (2) Elemental analysis: C: 49.73%, H: 4.83%, N: 12.52%, S: 0.00% (3) Color reaction: Positive: sulfuric acid reaction, potassium permanganate reaction, ninhydrin reaction, 2,4-dinitrophenylhydrazine reaction, iodine vapor reaction, ferric chloride-potassium ferricyanide reaction. Negative: ferric chloride reaction, Sakaguchi reaction (4) Solubility: Soluble: Water, methanol. Insoluble: Acetone, ethyl acetate, chloroform (5) Melting point: Around 175°C: Begins to show pale yellow color. Around 210°C: Turns brown and caramelizes. Around 300°C: Does not melt (6) Specific optical rotation: ▲[α] ²³ _D ▼: +8° (c=1.0, _H2O ) ▲[α] ²³ _D ▼: -26.5° (c=1.0, 0.1N HCl) (7) Ultraviolet absorption spectrum: (8) Infrared absorption spectrum: (9) Molecular weight: SI mass spectrometry: m/z (measured value) 322 (10) ¹³ C nuclear magnetic resonance spectrum δ (ppm, DCl + D ₂ O, pD = ca.1): 195.4 (d), 159.1
(s),156.3(s),147.4(s),136.6(s),118.3
(s), 114.3 (d), 111.2 (d), 90.8 (s), 60.7
(t),49.4(t),44.1(d),36.3(d),36.3
(d), (11) ^1H nuclear magnetic resonance spectrum δ (ppm, DCl + _D2O , pD = ca.1): 9.67 (1H, s), 7.00 (2H,
s),5.21(1H,dd,J=11,6Hz),4.6(1H,d,J=11Hz),4.
16 (1H, d, J = 17Hz), 4.04 (1H, dd, J = 17,5Hz), 3.74 (2
H,m), 3.65 (1H,d,J=6Hz) (12) Thin-layer chromatography: The FR-900482 substance further has the following chemical characteristics:

(a) A single compound (hereinafter referred to as FR-900482A substance) having the following physical and chemical properties can be obtained by treating FR-900482 substance with hydrochloric acid.

(1) Shape and color: Colorless powder (2) Solubility: Soluble in water and methanol; Insoluble in acetone, ethyl acetate, and chloroform (3) Melting point: Around 160°: Begins to show a pale yellow color Around 200°: Turns brown 200-300°: Turns dark (does not melt) (4) Specific optical rotation: ▲[α] ²³ _D ▼: -27° (c=1.0, 0.1N HCl) (5) Ultraviolet absorption spectrum: (6) Infrared absorption spectrum: The single FR-900482A substance thus obtained can be converted into the original FR-900482 substance, which shows two Rf values in silica gel thin layer chromatography, by neutralization with sodium hydroxide solution.

(b) A simple triacetyl derivative of FR-900482 substance having the following physical and chemical properties can be obtained by treating FR-900482 substance with an excess of acetic anhydride:

(1) Shape and color: Colorless plate _- like crystals (2) Elemental analysis: Measured values: C: 53.85%, H: 4.91%, N: 9.19% Measured values: (as _C20H21O9N3 ) C: _53.69 %, H: _4.73 %, N: 9.39% (3) Color reaction: Positive: Cerium sulfate reaction, iodine vapor reaction, phosphomolybdic acid reaction Negative: Sakaguchi reaction (4) Solubility: Soluble: Chloroform, acetone, methanol Insoluble: Water (5) Melting point: 170°-173° (6) Specific optical rotation: ▲[ _α]23D ^▼ : +66.6° (c=1.0, methanol) (7) Ultraviolet absorption spectrum: (8) Infrared absorption spectrum: (9) Thin Layer Chromatography: (10) Molecular weight: EI mass spectrometry: m/z 447 (M ⁺ ) High-resolution mass spectrometry: M ⁺ observed: 447.1286 M ⁺ calculated: (as C ₂₀ H ₂₁ N ₃ O ₉ ) 447.1277 (11) ¹³ C nuclear magnetic resonance spectrum: δ (ppm, CDCl ₃ ): 21.0 (q), 21.7 (q), 22.9 (q), 3
1.6(d),39.8(d),40.2(d),52.9(t),63.2
(t), 96.4 (s), 115.5 (d), 119.0 (d), 124.2
(s),136.2(s),149.4(s),149.7(s),155.9
(s),168.5(s),169.1(s),180.8(s),190.3
(d) (12) ^1H nuclear magnetic resonance spectrum: δ (ppm, CDCl ₃ ): 9.9 (1H, s), 7.31 (1H, d, J = 1.3Hz),
7.14 (1H, d, J = 1.3Hz), 4.72 (2H, broad), 4.4 (1H, dd,
J=12,7Hz),4.35(1H,dd,J=12,3.8Hz),4.06(1H,dd,
J=15,2Hz),3.89(1H,dd,J=7,3.8Hz),3.74(1H,d,J
= 15Hz), 3.41 (1H, d, J = 6.3 Hz), 2.84 (1H, dd, J = 6.3,
2Hz), 2.40 (3H,s), 2.24 (3H,s), 1.98 (3H,s) From the above physical and chemical properties and X-ray diffraction, the FR-
The triacetyl derivative of substance 900482 was found to have the following chemical structure:

11-acetyl-8-carbamoyloxymethyl-4-formyl-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-6,9-
The single triacetyl derivative of the FR-900482 substance thus obtained was treated with an aqueous sodium bicarbonate solution to give two Rf values on silica gel thin layer chromatography, similar to the original FR-
It can be converted into 900,482 substances.

(c) The nuclear magnetic resonance spectrum shown in Figures 1 and 2, or silica gel thin-layer chromatography using a chromatoscanner, demonstrated that the FR-900482 substance exists as a mixture of two components near neutral pH and as a single component near pH 1.

(d) In the silica gel thin layer chromatography, Rf
When the separated component showing an Rf value of 0.55 was further subjected to silica gel thin layer chromatography, it showed two Rf values corresponding to the Rf values of the FR-900482 substance. Similarly, when another separated component showing an Rf value of 0.65 was further subjected to silica gel thin layer chromatography, it showed two Rf values corresponding to the Rf values of the FR-900482 substance.
Two Rf values corresponding to the Rf value of the -900482 substance are shown.

From the above chemical characteristics of FR-900482 substance, it was identified as having the following planar structural formula: Furthermore, the two components revealed by silica gel thin layer chromatography are considered to be a mutually equilibrated mixture having the same planar structural formula.

4-formyl-6,9-dihydroxy-14-oxa-1,11
-Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate [II] Synthesis Method: (1) Production Method 1 (Introduction of Imino Protecting Group) Compound (I-b) or a salt thereof can be produced by introducing an imino protecting group into compound (I-a) or a salt thereof.

Suitable salts of compounds (Ia) and (Ib) include the same as salts of compound (I).

Suitable introducing agents used in this reaction include conventional acylating agents capable of introducing the acyl group, such as carboxylic acids, carbonic acids, sulfonic acids, and reactive derivatives thereof (e.g., acid halides, acid anhydrides, activated amides, activated esters, etc.). Preferred examples of such reactive derivatives include acid chlorides; acid bromides;
Mixed acid anhydrides with acids such as substituted phosphoric acids (e.g., dialkyl phosphoric acid, phenyl phosphoric acid, diphenyl phosphoric acid, dibenzyl phosphoric acid, halogenated phosphoric acids, etc.), dialkyl phosphorous acids, sulfurous acid, thiosulfuric acid, sulfuric acid, alkyl carbonates (e.g., methyl carbonate, ethyl carbonate, propyl carbonate, etc.), aliphatic carboxylic acids (e.g., pivalic acid, pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.), aromatic carboxylic acids (e.g., benzoic acid, etc.); symmetrical acid anhydrides; imidazole, 4-substituted imidazole, dimethylpyrazole, triazole, and tetrazole. activated amides with heterocyclic compounds containing an imino function such as aryl; activated esters (e.g., p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyridyl ester, piperidinyl ester, 8-quinolyl thioester, or N,N-dimethylhydroxylamine, 1-
esters with N-hydroxy compounds such as hydroxy-2-(1H)-pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole, and 1-hydroxy-6-chlorobenzotriazole, etc.

This reaction can be carried out using alkali metals (e.g., lithium, sodium, potassium, etc.), alkaline earth metals (e.g., calcium, etc.), alkali metal hydrides (e.g., sodium hydride, etc.), alkaline earth metal hydrides (e.g., calcium hydride, etc.), alkali metal hydroxides (e.g.,
sodium hydroxide, potassium hydroxide, etc.), alkali metal carbonates (e.g., sodium carbonate, potassium carbonate, etc.),
The reaction can be carried out in the presence of an organic or inorganic base such as an alkali metal bicarbonate (e.g., sodium bicarbonate, potassium bicarbonate, etc.), an alkali metal alkoxide (e.g., sodium methoxide, sodium ethoxide, potassium t-butoxide, etc.), an alkali metal alkanoic acid (e.g., sodium acetate, etc.), a trialkylamine (e.g., trimethylamine, etc.), a pyridine compound (e.g., pyridine, lutidine, picoline, 4-dimethylaminopyridine, etc.), or quinoline.

In this reaction, when the agent for introducing an imino-protecting group is used in the free form or in the form of a salt, the reaction can be carried out by using a carbodiimide compound [e.g., N,N'-dicyclohexylcarbodiide, N-
cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide, N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide, N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide, etc.], ketenimine compounds [for example, N,N'-carbonylbis(2-methylimidazole), pentamethyleneketene-N-cyclohexylimine, diphenylketene-N-cyclohexylimine, etc.], olefinic or acetylenic ether compounds (for example, ethoxyacetylene, β-chlorovinylethylene ether),
Sulfonic acid esters of N-hydroxybenzotriazole derivatives [e.g., 1-(4-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole, etc.], combinations of trialkyl phosphites or triphenylphosphine with carbon tetrachloride, disulfides or diazenedicarboxylates (e.g., diethyl diazenedicarboxylate, etc.), phosphorus compounds (e.g., ethyl polyphosphate, isopropyl polyphosphate, phosphoryl chloride, phosphorus trichloride, etc.), thionyl chloride, oxalyl chloride, N-ethylbenzisoxazolium salts, N-ethyl-5-phenylisoxazolium-3
It is preferable to carry out the reaction in the presence of a condensing agent such as a reagent (the so-called "Vilsmeier reagent") prepared by reacting an amide compound such as N,N-di(lower)alkylformamide (e.g., dimethylformamide) with a halogen compound such as thionyl chloride, phosphoryl chloride, or phosgene.

The reaction is usually carried out in a conventional solvent that does not adversely affect the reaction, such as water, acetone, methylene chloride, alcohol (e.g., methanol, ethanol, etc.), tetrahydrofuran, pyridine, N,N-dimethylformamide, etc., or a mixture thereof. When the imino-introducing agent is liquid, this agent can also be used as the solvent.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

In this production method, the hydroxyiminomethyl group or hydroxymethyl group represented by ^R1 , the hydroxymethyl group represented by ^R4 , and
The case where at least one of the hydroxy groups represented by ^R2 and ^R5 is converted into the corresponding protected hydroxyiminomethyl group, protected hydroxymethyl group, and protected hydroxy group, respectively, in the target compound (I-b) is also included within the scope of the present invention.

(2) Production method 2 (Introduction of a hydroxy-protecting group into the 6-hydroxy group) (3) Production method 3 (Introduction of a hydroxy-protecting group into the 9-hydroxy group) (4) Production method 4 (Introduction of a hydroxy-protecting group into the 8-hydroxymethyl group) (5) Production method 5 (Introduction of a hydroxy-protecting group into the 4-hydroxymethyl group) Compounds (I-d), (I-f), (I-h) and (I-i) were prepared.
The compounds (I-c), (I--j) or salts thereof can be used in the preparation of
They can be prepared by introducing a hydroxy protecting group into (Ie), (Ig) and (Ii) or a salt thereof, respectively.

Suitable salts of compounds (Ic) to (Ij) include the same as those shown for compound (I).

Suitable agents for introducing a hydroxy-protecting group used in Production Methods 2 to 5 include the above-mentioned conventional acylating agents capable of introducing an acyl group, as shown in Production Method 1.

These reactions can be carried out in substantially the same manner as in Production Method 1, and therefore the explanations for Production Method 1 can be used for the reaction methods and reaction conditions (for example, solvents, reaction temperatures, etc.).

During the reaction in these Production Methods 2 to 5, (i) in Production Methods 2 to 4, the starting compound (I-
(c), (I-e) and (I-g) are optionally converted into the corresponding protected hydroxyiminomethyl or hydroxymethyl groups, ^respectively , of the starting compounds (I-
(iii) in Production Methods 2, 3 and 5, a hydroxymethyl group represented by ^R4 in starting compounds (I-c), (I-e) and (I-i) is optionally converted to a corresponding protected hydroxymethyl group, (iv) in Production Methods 2, 4 and 5, a hydroxy group represented by ^R5 in starting compounds (I-c), (I-g) and (I-i) is optionally converted to a corresponding protected hydroxy group, (v) in Production Methods 2 to ⁵ , starting compounds (I-c), (I- ^e ),
When the free imino groups of (I-g) and (I-i) are optionally converted to protected imino groups, the following may be obtained, and these cases are also included in the scope of Production Methods 2 to 5.

(6) Production Method 6 (Reaction with Base) The compound (Ik) or a salt thereof can be produced by reacting the compound (Ih) or a salt thereof with a base.

Suitable salts of compounds (Ih) and (Ik) include the same as those for compound (I).

The base used in this reaction includes conventional bases such as those shown in Production Method 1, which are capable of converting a protected hydroxymethyl group into a methylene group.

This reaction is usually carried out in a conventional solvent which does not adversely influence the reaction, such as water, an alcohol (e.g., methanol, ethanol, etc.), pyridine, N,N-dimethylformamide, etc., or a mixture thereof. Furthermore, when the base used is a liquid, the base can also be used as the solvent.

The reaction temperature is not particularly limited, and the reaction is usually carried out at room temperature or under warming.

This production method also includes within its scope the case where the formyl group represented by R ¹ is optionally converted to a hydroxymethyl group during the reaction.

(7) Production Method 7 (Reduction of Methylene Group) The compound (I-l) or a salt thereof can be prepared by reducing the compound (I-k) or a salt thereof.

Suitable salts of compound (I-1) include the same as those shown for compound (I).

The reduction in this production method can be carried out by a conventional method capable of reducing a methyl group to another methyl group, such as catalytic reduction.

Suitable catalysts for use in catalytic reduction include platinum catalysts (e.g., platinum plate, platinum sponge, platinum black, colloidal platinum, platinum oxide, platinum wire, etc.), palladium catalysts (e.g.,
Examples of such catalysts include conventional catalysts such as palladium sponge, palladium black, palladium oxide, palladium-carbon, colloidal palladium, palladium-barium sulfide, palladium-barium carbonate, etc.), nickel catalysts (e.g., reduced nickel, nickel oxide, Raney nickel, etc.), cobalt catalysts (e.g., reduced cobalt, Raney cobalt, etc.), iron catalysts (e.g., reduced iron, Raney iron, etc.), copper catalysts (e.g., reduced copper, Raney copper, Ullmann copper, etc.), etc.

The reduction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, methanol, ethanol, propanol, pyridine, N,N-dimethylformamide, methylene chloride, or a mixture thereof.

The reaction temperature for this reduction is not particularly limited, and the reaction is usually carried out under cooling or heating.

(8) Production Method 8 (Reduction to Hydroxymethyl Group) Compound (I-i) or a salt thereof can be prepared by reducing compound (I-m) or a salt thereof.

Suitable salts of compound (Im) include the same as those shown for compound (I).

The reduction in this production method can be carried out by a conventional method capable of reducing a formyl group or a protected formyl group to a hydroxymethyl group, such as catalytic reduction or reduction using a reducing agent.

The catalytic reduction can be carried out in substantially the same manner as in Production Method 7, and therefore the explanations in Production Method 7 can be used for suitable contact and reaction conditions for this reduction (for example, solvent, reaction temperature, etc.).

Suitable reducing agents include alkali borohydrides (e.g.,
Examples of suitable amalgam include sodium borohydride, sodium cyanoborohydride, and the like, and alkali amalgam (for example, sodium amalgam, and the like).

The reduction using a reducing agent is usually carried out in a conventional solvent which does not adversely influence the reaction, such as those shown in the catalytic reduction of Production Method 7.

The reaction temperature for this reduction is not particularly limited, and the reaction is usually carried out under cooling or heating.

This reduction also includes within its scope the case where at least one of the hydroxy protecting groups R ² , R ⁴ and R ⁵ and the imino protecting group R ⁶ are optionally eliminated simultaneously during the reaction.

Furthermore, this reduction can also be carried out using an alkali metal hydroxide as shown in Production Method 1. In this case, the protected hydroxymethyl group represented by ^R4 may be converted to a methylene group in some cases.

(9) Production Method 9 (Introduction of a Formyl Protecting Group) Compound (I-o) or a salt thereof can be produced by introducing a formyl protecting group into compound (In) or a salt thereof.

Suitable salts of the compounds (In) and (Io) include the same as those for the compound (I).

Suitable agents for introducing a formyl protecting group used in this reaction include alcohols such as alkanols (e.g., methanol, ethanol, propanol, isopropanol, butanol, pentyl alcohol, hexyl alcohol, etc.), alkanediols (e.g., ethylene glycol, propylene glycol, 1,3-propanediol, etc.), etc.

The reaction is preferably carried out under anhydrous conditions in the presence of an acid such as a hydrogen halide (eg, hydrogen chloride, hydrogen bromide, etc.).

Furthermore, this reaction is usually carried out in a conventional solvent that does not adversely influence the reaction, and in many cases, the alcohols listed as agents for introducing a formyl protecting group can be used as they are as the solvent.

The reaction temperature for this reduction is not particularly limited, and the reaction is usually carried out under cooling or heating.

(10) Production Method 10 (Alkylation of 6-Hydroxy Group) (11) Production Method 11 (Alkylation of 9-Hydroxy Group) Compounds (I-p) and (I-q) or salts thereof can be produced by reacting compounds (I-c) and (I-e) or salts thereof with an alkylating agent, respectively.

Suitable salts of the compounds (Ip) and (Iq) include the same as those shown for the compound (I).

Suitable alkylating agents for use in this reaction include those capable of alkylating a hydroxy group to an alkoxy group, such as dialkyl sulfates (e.g., dimethyl sulfate, diethyl sulfate, etc.), alkyl sulfonates (e.g., methyl sulfonate, etc.), alkyl halides (e.g., methyl iodide, ethyl ethoxide, propyl bromide, etc.), and diazoalkanes (e.g., diazomethane, diazoethane, etc.).

This reaction is preferably carried out in the presence of an inorganic or organic base as described in the explanation of Production Method 1.

Furthermore, this reaction is usually carried out in a conventional solvent which does not adversely influence the reaction, such as water, acetone, methylene chloride, methanol, ethanol, propanol, pyridine, N,N-dimethylformamide, or a mixture thereof.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

Production Method 10 also encompasses within its scope the case where at least one of the hydroxy group represented by _R5 in the starting compound (I-c) and the imino group formed when _R6 is hydrogen is optionally converted during the reaction to the corresponding alkoxy group and alkylimino group, respectively.

Furthermore, within its scope, process 11 includes the starting compound (I-
In e), the case where at least one of the hydroxy group represented by _R2 and the imino group formed when _R6 is hydrogen is optionally converted into the corresponding alkoxy group and alkylimino group, respectively, during the reaction is also included.

(12) Production Method 12 (Reaction with Substituted Amine) The compound (Ir) or a salt thereof can be prepared by reacting the compound (Im) or a salt thereof with a substituted amine or a salt thereof.

Suitable salts of compound (Ir) include the same as those shown for compound (I).

Suitable salts of the substituted amine include salts with acids such as those shown in compound (I).

Suitable substituted amines for use in this reaction include arylamines (e.g., aniline, toluidine, xylidine, cumenylamine, mesitylamine, naphthylamine, etc.), hydroxylamine, O-alkylhydroxylamine (e.g., O-methylhydroxylamine, etc.), semicarbazide, arylsemicarbazide (e.g., 4-phenylsemicarbazide, etc.), and the like.

The reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, methylene chloride, methanol, ethanol, propanol, pyridine, N,N-dimethylformamide, or a mixture thereof.

When the substituted amine is used in the form of a salt, the reaction is preferably carried out in the presence of a base as described in the explanation of Production Method 1.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

(13) Production Method 13 (Reduction of Arylimino Group) The compound (It) or a salt thereof can be prepared by reducing the compound (Is) or a salt thereof.

Suitable salts of compound (It) include the same as those shown for compound (I).

The reduction in this production method can be carried out by a conventional method capable of reducing an arylimino group to an arylamino group, such as catalytic reduction.

The catalytic reduction can be carried out in substantially the same manner as in Production Method 7, and therefore the explanation of Production Method 7 can be used for the catalyst and reaction conditions (for example, solvent, reaction temperature, etc.) suitable for this reduction.

(14) Production Method 14 (Hydrolysis of Protected Hydroxymethyl Group) Compound (Ig) or a salt thereof can be prepared by hydrolyzing compound (Ih) or a salt thereof.

The hydrolysis can be carried out in the presence of a base or an acid,
Suitable bases include inorganic bases such as those shown in Production Method 1. Suitable acids include organic acids (e.g., formic acid,
acetic acid, propionic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.) and inorganic acids (e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.).

This reaction is usually carried out in a conventional solvent which does not adversely influence the reaction, such as water, acetone, methylene chloride, methanol, ethanol, propanol, pyridine, N,N-dimethylformamide, or a mixture thereof.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

(15) Production Method 15 (Removal of Imino Protecting Group) Compound (Ia) or a salt thereof can be prepared by removing the imino protecting group from compound (Ib) or a salt thereof.

This removal reaction can be carried out by a conventional method such as reduction or hydrolysis.

The reduction reaction applicable to this removal reaction includes catalytic reduction as explained in Production Method 7, and therefore the explanation of Production Method 7 can be used for the reaction method and reaction conditions (for example, reaction temperature, solvent, etc.).

The hydrolysis is preferably carried out in the presence of a base or an acid. Suitable bases include alkali metal hydroxides (e.g., sodium hydroxide, potassium hydroxide, etc.), alkaline earth metal hydroxides (e.g., magnesium hydroxide, calcium hydroxide, etc.), alkali metal carbonates (e.g., sodium carbonate, potassium carbonate, etc.), alkaline earth metal carbonates (e.g., magnesium carbonate, calcium carbonate, etc.), alkali metal bicarbonates (e.g., sodium bicarbonate, potassium bicarbonate, etc.), etc.

Suitable acids include organic acids (e.g., formic acid, acetic acid, propionic acid, trifluoroacetic acid, benzenesulfonic acid, p-
toluenesulfonic acid, etc.) and inorganic acids (e.g., hydrochloric acid,
Acid hydrolysis using trifluoroacetic acid is usually promoted by the addition of a cation scavenger (e.g., phenol, anisole, etc.).

The solvent and reaction temperature for this hydrolysis may be the same as those for the reduction.

The removal method can be selected depending on the type of imino-protecting group to be removed.

This process includes within its scope the case where the protected formyl group represented by R ¹ in the starting compound (I-b) is optionally converted to a formyl group in the target compound (I-a).

(16) Production Method 16 (Oxidation of Formyl Group) The compound (Iu) or a salt thereof can be prepared by oxidizing the formyl group of the compound (In) or a salt thereof.

Suitable salts of compound (Iu) include the same as those shown for compound (I).

Suitable oxidizing agents for the formyl group used in this reaction include:
Potassium permanganate, chromium compounds (e.g., chromium trioxide, chromic acid, sodium chromate, dichromic acid,
sodium dichromate, pyridine dichromate, etc.), etc.

This oxidation can be carried out in the presence of an acid, such as those described in the explanation of Process 14, preferably sulfuric acid.

This reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, acetone, dioxane, dimethylformamide, pyridine, etc., or a mixture thereof.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

This production method includes within its scope the case where, when the oxidation is carried out in the presence of an acid, the imino protecting group represented by ^R6 in the starting compound (In) is optionally eliminated during the reaction.

(17) Production Method 17 (Oxidation of Methylene Group) The compound (I-v) or a salt thereof can be prepared by oxidizing the compound (I-k) or a salt thereof.

Suitable salts of compound (I-v) include the same as those for compound (I).

This oxidation can be carried out by reacting compound (I-k) with ozone and then, if necessary, decomposing the resulting ozonide by a conventional method.

The ozonide produced by reacting the starting compound (Ik) with ozone is usually decomposed by reduction.

The reduction can be carried out in substantially the same manner as in Production Method 7,
Therefore, the reaction conditions for this reduction (for example, solvent, reaction temperature, etc.) can be determined from the explanation of Production Method 7.

Further, examples of suitable reducing agents include trialkyl phosphites (for example, trimethyl phosphite, etc.), triphenylphosphine, dimethyl sulfide, sodium bisulfite, sodium sulfite, sodium iodide, stannous chloride, and the like.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

(18) Production Method 18 (Introduction of a Carboxy Protecting Group) The compound (I-w) or a salt thereof can be produced by introducing a carboxy protecting group into the compound (I-u) or a salt thereof.

Suitable salts of compound (I-w) include the same as those for compound (I).

Suitable agents for introducing a carboxy protecting group used in this reaction include conventional esterifying agents capable of converting a carboxy group into an esterified carboxy group, such as alcohols or halides (e.g., chlorides,
Examples of suitable reactive compounds include reactive equivalents of alcohols such as diazoalkane compounds (e.g., diazomethane, diazoethane, etc.), sulfonates, sulfates, and the like.

This reaction can be carried out in the presence of a base or an acid, and suitable bases include inorganic bases such as those mentioned in Production Method 1. Suitable acids include inorganic or organic acids such as those mentioned in Production Method 14.

This reaction is usually carried out in a conventional solvent which does not adversely influence the reaction, such as water, methanol, ethanol, propanol, diethyl ether, pyridine, N,N-dimethylformamide, etc., or a mixture thereof.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

(19) Production Method 19 (Removal of Formyl Protecting Group) The compound (In) or a salt thereof can be prepared by removing the formyl protecting group from the compound (Io) or a salt thereof.

This removal reaction can be carried out by a conventional method such as hydrolysis.

The hydrolysis is usually carried out by the conventional methods usually used for deacetalization of so-called acetal functional groups to the corresponding carbonyl functional groups, for example by hydrolysis with an inorganic acid (e.g.
It is preferably carried out by acid hydrolysis in the presence of an acid such as hydrochloric acid, sulfuric acid, or an organic acid (for example, formic acid, acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, or the like), or an acidic ion exchange resin.

This reaction is usually carried out using water, acetone, methyl ethyl ketone,
The reaction can be carried out in a conventional solvent which does not adversely influence the reaction, such as dioxane, methanol, ethanol, N,N-dimethylformamide, dimethyl sulfoxide, etc., or a mixture thereof.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

This production method includes within its scope the case where the imino-protecting group represented by R ⁶ in the starting compound (Io) is optionally eliminated during the reaction.

(20) Production Method 20 (Alkylation of Imino Group) The compound (I-x) or a salt thereof can be produced by reacting the compound (I-a) or a salt thereof with an alkylating agent.

Suitable salts of compound (I-x) include the same as those for compound (I).

Suitable alkylating agents for use in this reaction include conventional alkylating agents capable of alkylating an imino group to an alkylimino group, such as those mentioned in Preparations 10 and 11.

This reaction is preferably carried out in the presence of an inorganic or organic base such as those mentioned in the explanation of Production Method 1.

This reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, methanol, ethanol, propanol, pyridine, N,N-dimethylformamide, etc., or a mixture thereof.

The reaction temperature is not particularly limited, and the reaction is usually carried out under cooling or heating.

This production method includes within its scope the case where at least one of the hydroxy groups represented by ^R2 and ^R5 in the starting compound (Ia) is optionally converted to an alkoxy group during the reaction.

(21) Production Method 21 (Hydrolysis of Protected 6-Hydroxy Group) Compound (I-c) or a salt thereof can be prepared by hydrolyzing compound (I-d) or a salt thereof.

This hydrolysis can be carried out in substantially the same manner as in Production Method 14, and therefore the explanation for Production Method 14 can be used for the suitable base or acid used in the hydrolysis and the reaction conditions (e.g., solvent, reaction temperature, etc.).

(22) Production Method 22 (Introduction of a hydroxy-protecting group into a hydroxyiminomethyl group) Compound (I-z) or a salt thereof can be produced by introducing a hydroxy-protecting group into compound (I-y) or a salt thereof.

Suitable salts of compounds (Iz) and (Iy) include the same as those for compound (I).

Suitable hydroxy-protecting group-introducing agents for use in this reaction include conventional acylating agents capable of introducing an acyl group, such as those mentioned in Production Method 1.

This reaction can be carried out in substantially the same manner as in Production Method 1, and therefore the reaction method and reaction conditions (e.g., solvent,
The explanation for Production Method 1 can be applied to the reaction conditions (reaction temperature, etc.).

This production method includes, within its scope, ^the steps ^{of :}
and an imino group formed when ^R6 is hydrogen, are optionally converted during the reaction into the corresponding protected hydroxymethyl group, protected hydroxy group, and protected imino group, respectively, in the target compound (I-z).

The target tetracyclo compound (I) obtained by the fermentation method and synthesis methods 1 to 22 described above can be separated and purified by conventional methods such as extraction, precipitation, fractional crystallization, recrystallization, chromatography, etc.

During the above reactions or post-treatment of the reaction mixtures in Synthetic Methods 1 to 22, the configuration of the starting or target compound may be converted to another configuration, and such cases are also within the scope of the present invention.

The tetracyclo compound (I) of the present invention and its pharmaceutically acceptable salts are novel and exhibit pharmacological activities such as antitumor activity and antibacterial activity, and are therefore useful in the treatment of tumors, infectious diseases, etc. in mammals including humans.

To demonstrate the usefulness of the tetracyclo compound (I), the pharmacological test data of representative compounds of compound (I) are shown below.

Test 1 Antitumor activity of FR-900482 substance The antitumor activity of FR-900482 substance was examined in an experimental tumor system in mice.

Lymphocytic leukemia p388 was transplanted intraperitoneally into _BDF1 mice at an inoculation dose of 1 x ¹⁰⁶ cells/mouse. 24 hours after tumor cell transplantation, FR-900482 substance at each dose level was intraperitoneally administered. Administration was performed once a day on days 1, 2, 3, and 4 after tumor inoculation. Control animals were intraperitoneally administered saline. The injection volume was 0.2 ml in each experiment. Six mice were included in each experimental group.
We used 1000 pieces of fish.

Antitumor activity was evaluated based on the mean survival time of each mouse group.
The results were also expressed as T/C % values (mean survival time of drug-administered group/mean survival time of control group x 100).

The results are shown in Table 4.

FR-900482 was highly effective against leukemia p388, and significantly increased the lifespan of mice at doses of 3.2-18 mg/kg.

Test 2: Antibacterial activity of FR-900482 substance The antibacterial activity of FR-900482 substance against several types of bacteria was examined by serial broth dilution method in bacterial broth medium. The minimum inhibitory concentration (MIC) was expressed in μg/ml after overnight incubation at 37°C.

The FR-900482 substance showed antibacterial activity against various pathogenic bacteria: for example, Bacillus subtilis
s) ATCC6633 (MIC: 50 μg/ml), Escherichia coli (E
scherichia coli NIHJJC-2 (MIC: 50 μg/ml) and Pseudomonas aeruginosa
sa) NCTC-10490 (MIC: 25 μg/ml) Test 3 Acute toxicity of FR-900482 substance Acute toxicity tests of FR-900482 substance were carried out in ddY mice by intraperitoneal and intravenous injection. The _LD50 values were both 3.
The dose was 2mg/kg.

Test 4: In vitro cytotoxicity (i) Test method Seven days after tumor implantation, the ascites was diluted with Hank's solution to prepare a suspension of mouse lymphocytic leukemia p388 tumor cells, which was then centrifuged at 1000 rpm for 15 minutes at low temperature. The cell suspension was incubated in MFM Dulbecco's medium supplemented with 10% fetal bovine serum, penicillin G (60 μg/ml), and streptomycin (20 μg/ml).
o) The cells were adjusted to a concentration of 2.5 x ¹⁰⁶ cells/ml in the medium. The cell suspension was then placed in a plastic tissue culture dish, treated with appropriately diluted test compounds at 37°C, and cultured for 72 hours in a humidified atmosphere containing 5% carbon dioxide.
Concentration of substance required for 50% inhibition (IC ₅₀ ) (μg/ml)
was determined by plotting the logarithm of the pharmacological concentration against the viability (percentage of control) of drug-treated cells.

(ii) Test compound 11-acetyl-4-formyl-8-carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.
4.1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-6,9
-diyl diacetate (hereinafter referred to as Compound R), 11-acetyl-4-formyl-9-hydroxy-8-
Methylene-14-oxa-1,11-diazatetracyclo[7.
4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6-
11-acetyl-4-acetoxymethyl-8-carbamoyloxymethyl-9-hydroxy-14-oxa-1,11-yl acetate (hereinafter referred to as Compound S),
-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-6-yl acetate (hereinafter,
Compound T), 4-acetoxymethyl-11-acetyl-8-carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (hereinafter referred to as Compound U), 11-acetyl-8-carbamoyloxymethyl-4-
Formyl-6-methoxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-
Trien-9-yl acetate (hereinafter referred to as Compound V), 11-acetyl-4-formyl-9-hydroxy-6-
Methoxy-14-oxa-1,11-diazatetracyclo[7.
4.1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-8-
Iylmethyl carbamate (hereinafter referred to as Compound W).

(iii) Test results The pharmaceutical composition containing the compound of the present invention or a pharmaceutically acceptable salt thereof can be used in the form of a pharmaceutical agent such as a solid, semi-solid or liquid by mixing with an organic or inorganic carrier or excipient suitable for external, oral or parenteral administration. The active ingredient can be mixed with a generally non-toxic pharmaceutically acceptable carrier and used in the form of a tablet, pellet, capsule, suppository, solution, emulsion,
It can be made into a suspension or any other suitable form for use. Carriers that can be used include water, glucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea and other solid, semi-solid or liquid carriers suitable for pharmaceutical manufacture, and can also contain auxiliary substances, stabilizers,
Thickening agents, coloring agents, and flavoring agents may also be used.The desired active compound is included in the pharmaceutical composition in an amount sufficient to produce the desired effect upon the process or condition of diseases.

When this composition is administered to humans, it is preferably administered parenterally or orally. The therapeutically effective amount of the tetracyclo compound (I) varies depending on the age and condition of the individual patient, but generally, for the treatment of a disease, about 0.1-1000 mg, preferably 0.5-500 mg, more preferably 1-100 mg of the active ingredient is administered per day, and the average single dose is usually about 1 mg, 5 mg, 10 mg, 50 mg, 100 mg.
g, 250 mg and 500 mg are administered.

The present invention will be described below with reference to examples.

Example 1 Streptomyces sandaensis No. 6897
Isolation of Streptomyces sandaensis No. 6897 Streptomyces sandaensis No. 6897 was isolated by the dilution plating method described below.

Approximately 1 g of soil collected from Sanda City, Hyogo Prefecture, was placed in a sterile test tube and the volume was adjusted to 5 ml with sterile water. The mixture was mixed for 10 seconds using a tube buzzer and allowed to stand for 10 minutes. The supernatant was serially diluted 100-fold with sterile water, and the diluted solution (0.1 ml) was used.
l) in a Petri dish containing Ksapek agar containing thiamine hydrochloride (sucrose 30 g, sodium sulfate 3 g, dipotassium hydrogen phosphate 1 g, magnesium sulfate 0.5 g, potassium chloride 0.5 g, ferrous sulfate 0.01 g, thiamine hydrochloride 0.1 g, agar 20 g, and tap water 1000 ml
After culturing at 30°C for 21 days, grown colonies that appeared on the plate were transferred to a slant [yeast-malt extract agar (ISP-medium 2)] and cultured at 30°C for 10 days.
Streptomyces sandaensis No. 6897 was identified in the isolated colony.

A seed medium (160 ml) containing fermented soluble starch (2%), glucose (0.5%), cottonseed flour (1%), dry yeast (1%), corn steep liquor (0.5%), and calcium carbonate (0.2%) (adjusted to pH 7.0 with aqueous sodium hydroxide solution) was added to each of eight 500 ml Erlenmeyer flasks and sterilized at 120°C for 30 minutes. A loopful of a slant culture of Streptomyces sandaensis No. 6897 (Microbiological Research Institute No. 792) was inoculated into each medium and incubated at 30°C for 72 hours on a rotary shaker at 200 rpm with a 3-inch throw. This culture was inoculated into the seed medium in a 30-volume jar fermenter previously sterilized at 120°C for 30 minutes and aerated (20°C).
The seed culture was cultured at 30°C for 48 hours under constant speed (rpm) and stirring (200 rpm).
In a stainless steel fermenter, a production medium (1760 ml) containing soluble starch (8%), dry yeast (1%), peanut powder (3%), and soybean meal (0.5%) (adjusted to pH 6.2 with sulfuric acid) was inoculated and cultured at 31°C for 96 hours under aeration (880/min) and agitation (130 rpm).

Separation and Purification The culture broth thus obtained is filtered using diatomaceous earth (125 kg) as a filter aid. The resulting filtrate (160
0) was adsorbed onto the non-ionic adsorption resin "Diaion HP-20"
The column was then washed with water (400) and then 50%
Elute with aqueous methanol (1200).
The active fraction is concentrated under reduced pressure to 0. This active fraction is applied to a column packed with ion exchange resin "Amberlite IRC-50" (trademark, manufactured by Rohm and Haas Chemical Co.) (H ⁺ ) (300). The column is washed with deionized water (600) and eluted with 0.1N hydrochloric acid (1200). The eluate is neutralized with 12N aqueous sodium hydroxide solution and eluted with non-ionic adsorption resin "Diaion
The column is loaded with HP-20 (200). The column is washed with deionized water (200) and eluted with 50% aqueous methanol (600). The active eluate is concentrated under reduced pressure to 10, to which n-butanol (15) is added and stirred for 10 minutes. This extraction procedure is repeated four times, and the resulting extracts are combined (60). Next, the butanol extract is diluted with n-
Hexane (240) was added and stirred for 10 minutes. After separating the aqueous layer, deionized water (15) was added to the butanol/n-hexane layer and stirred for 10 minutes. The aqueous layers were combined and concentrated under reduced pressure to a volume of 6, after which the mixture was applied to a column packed with "Alumina Oxide AC-11" (trademark, manufactured by Sumitomo Chemical Co., Ltd.) (100). Elution was performed with 80% aqueous isopropyl alcohol, and the resulting active eluate was concentrated under reduced pressure to 300 ml. This active fraction was applied to a column chromatograph packed with "Toyo Pearl HW-40 Fine" (trademark, manufactured by Toyo Soda Kogyo Co., Ltd.) (7) and eluted with deionized water.
The fraction (30) containing the active substance was concentrated under reduced pressure to 500 ml.
A crude powder is obtained by freeze-drying. This crude powder is dissolved in deionized water to a final concentration of 50 mg/ml and purified by high performance liquid chromatography (hereinafter referred to as HPLC). HPLC was performed using a "Waters Model 6000A" pump (trademark, manufacturer: Waters Model U6K) equipped with an injector.
Chromatography was performed using a UV detector "Waters Model 440" (Waters Associates).
(trademark, manufactured by Waters Associates, Inc.) was used to measure the absorption at 254 nm. A steel column (inner diameter 7.9 mm, length 30 cm) was fitted with "Micro Bondapak C18"
The column was filled with a water column (trademark, manufactured by Waters Associates) and used at a flow rate of 6 ml/min.
A mixture of distilled water (1:9, v/v) was used. HPLC was performed under the above conditions, and the active fractions were collected (retention time: 8 minutes). The solvent was removed under reduced pressure to obtain 1 g of colorless powder of FR-900482 substance.

Example 2: FR-900482 substance (57 mg) was dissolved in water (2 ml) and 1N hydrochloric acid (0.3 ml) was added. The solution was left at room temperature for 30 minutes.
The resulting powder is then dried under vacuum in the presence of phosphorus pentoxide, and then under vacuum in the presence of potassium hydroxide pellets to obtain a colorless powder of FR-900482A substance (60 mg).

This FR-900482A substance (5 mg) was dissolved in water (0.4 ml),
Add 100 μl of 0.1N aqueous sodium hydroxide solution. After leaving the mixture at room temperature for 1 hour, it is spotted on a silica gel plate and developed with several solvent systems. The resulting material is chromatographed on silica gel thin layer chromatography (FR-90).
The Rf value was the same as that of the 0482 substance.

Example 3: To FR-900482 substance (25 mg) were added pyridine (2 ml) and acetic anhydride (1 ml). After stirring for 20 minutes, the resulting solution was left at room temperature overnight. Excess solvent was removed under reduced pressure using a high-vacuum pump. The residue was purified by silica gel preparative thin-layer chromatography and developed with a mixture of methanol and chloroform (5:95, v/v) to obtain a colorless powder (20 mg) of the triacetyl derivative of FR-900482 substance. The resulting powder was crystallized from a mixture of ethyl acetate and diethyl ether to obtain colorless platelets of the above substance.

The triacetyl derivative of FR-900482 (20 mg) was dissolved in methanol (1 ml) and diluted with 10% sodium bicarbonate solution (1
After stirring overnight at room temperature, the resulting mixture was subjected to preparative thin layer chromatography, developed with a mixture of chloroform and methanol (4:1, v/v), and FR-900
Obtain 482 substances (10 mg).

This substance was identified by silica gel thin layer chromatography (developing solvent: a mixture of chloroform and methanol and a mixture of isopropyl alcohol and water) and by comparison of infrared absorption spectra.

Example 4 FR-900482 substance (100 mg) was dissolved in water (3 ml) and acetic anhydride (30 μg) was added. After stirring at room temperature for 3 hours, the resulting mixture was subjected to preparative thin layer chromatography and developed with a mixture of chloroform and acetone (1:1, v/v).
- The monoacetyl derivative of substance 900482 (58.5 mg) is obtained.

(1) Comparative luminosity: ▲[α] ²³ _D ▼: +41° (c=1.02, methanol) (2) Infrared absorption spectrum: (3) Molecular weight: SI mass spectrometry: m/z 364 (M ⁺ +1), 386 (M ⁺ +23) (4) ¹ H nuclear magnetic resonance spectrum: δ (ppm, CD ₃ OD): 9.9 (1H, s), 7.0 (1H, d, J = 2Hz), 6.9
(1H,d,J=2Hz),4.7−4.5(2H,m),3.85−3.75(2H,
m), 2.9 (1H, d, J = 6 Hz), 1.85 (3H, s) Example 5 Measurement of the Mixing Ratio of the Two Components of FR-900482 Substance at Various pH Values FR-900482 substance was dissolved in the following various buffer solutions to a concentration of 10 mg/ml. After leaving to stand for 2 hours, 4 μl of each of these solutions was subjected to silica gel thin-layer chromatography and developed with a mixture of chloroform and methanol (4:1, v/v). After development, the mixing ratio of the two components was calculated using a chromatoscanner at 254 nm. The results are shown below.

Example 6 4-formyl-6,9-dihydroxy-14-oxa-1,11
11-acetyl-4-formyl-6,9-dihydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (100 mg) was dissolved in 3 mL of water, 30 μL of acetic anhydride was added, and the mixture was stirred at room temperature for 3 hours. The reaction mixture was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and acetone (1:1, v/v) to give 11-acetyl-4-formyl-6,9-dihydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,
4,6-trien-8-ylmethyl carbamate (80.5m
g) was obtained. ¹ H NMR δ (ppm, CD ₃ OD): 9.9 (1 H, s), 7.00 (1 H, d, J=2
Hz), 6.9 (1H, d, J = 2Hz), 4.7-4.5 (2H, m), 3.85-3.7
5 (2H, m), 3.00−2.85 (1H, m), 1.85 (3H, s) SI Mass Spectrometry: m/z 364 (M ⁺ +1) Example 7 4-formyl-6,9-dihydroxy-14-oxa-1,11
-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (5
4-Bromobenzoyl chloride (170 mg) was added to a solution of 11-(4-bromobenzoyl)-4-formyl-2-methylbenzoyl chloride (100 mg) in pyridine (2 ml) and stirred at room temperature for 1 hour. The resulting solution was left standing at room temperature overnight, and the solvent was removed by evaporation under reduced pressure. Aqueous sodium bicarbonate solution was added to the residue, and the mixture was extracted with ethyl acetate. The ethyl acetate solution was evaporated to dryness under reduced pressure, and the resulting residue (52 mg) was subjected to preparative thin-layer chromatography. The mixture was developed with a mixture of methanol and chloroform (1:9, v/v) to obtain 11-(4-bromobenzoyl)-4-formyl-2-methylbenzoyl chloride (170 mg).
6,9-Dihydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbate (26 mg) was obtained. ¹ H NMR δ (ppm, CD ₃ OD + CDCl ₃ ): 9.88 (1H, s), 7.50 (2
H,d,J=8.5Hz),7.33(2H,d,J=8.5Hz),7.02(1H,d,J
= 1.3Hz), 6.99 (1H, d, J = 1.3Hz), 4.65 (2H, m), 3.97
(1H, dd, J = 2,15Hz), 3.09 (1H, d, J = 15Hz), 3.40 (2H,
m),2.90 (1H,dd,J=2,7Hz) SI Mass Spectrometry: m/z 506 (M ⁺ +3), 504 (M ⁺ +1) Example 8 4-formyl-6,9-dihydroxy-14-oxa-1,11
Acetic anhydride (1 ml) was added to a solution of 20 mg of tetradeca-2,4,6 ^- trien-8 ^- ylmethyl carbamate in 2 ml of methanol, and the mixture was stirred at room temperature for 1 hour. The solvent and excess acetic anhydride were removed under reduced pressure, and the resulting residue was subjected to preparative thin-layer chromatography. A 1:1 mixture of methanol and chloroform was used.
9, v/v) mixture to give 11-acetyl-8-carbamoyloxymethyl-4-formyl-9-hydroxy-
14-Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-trien-6-yl acetate (9 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.9 (1 H, s), 7.31 (1 H, d, J=
1.5Hz), 7.17 (1H, d, J = 1.5Hz), 6.70 (1H, s), 5.06 (1
H,dd,J=2,12.7Hz),4.78(2H,broad),4.35(1H,dd,J
=5,12.7Hz),3.96(1H,dd,J=2,14.7Hz),3.71(1H,d,
J=14.7Hz),3.19(1H,d,J=6.5Hz),3.15(1H,dd,J=
2,5Hz), 2.81 (1H, dd, J = 2,6.5Hz), 2.40 (3H, s), 1.90
(3H,s) SI Mass Spectrometry: m/z 406 (M ⁺ +1) Example 9 6,9-dihydroxy-8-methylene-14-oxa-1,11
Acetic anhydride (16 μ) was added to a solution of 20 mg of tetradeca-2,4,6 ^- triene-4 ^- carbaldehyde in 500 μl of pyridine, and the mixture was left standing overnight at room temperature. The reaction mixture was evaporated to dryness under reduced pressure, and the resulting oil was subjected to silica gel column chromatography using a mixture of methanol and chloroform (5:95, v/v).
The desired fractions were combined and evaporated to dryness under reduced pressure to give 11-acetyl-4-formyl-9-hydroxy-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-
Trien-6-yl acetate (19.5 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.85 (1H, s), 7.27 (1H, d, J=
1.5Hz), 7.20 (1H, d, J = 1.5Hz), 6.21 (1H, s), 6.14 (1
H,s),4.50(1H,broad s),3.87(1H,dd,J=15,2Hz),
3.61 (1H, dd, J = 15, 6.4 Hz), 2.93 (1H, dd, J = 6.4, 6.4,
2Hz), 2.77 (1H, d, J = 6.4Hz), 2.31 (3H, s), 2.18 (3H,
s) EI mass spectrometry: m/z 344 (M ⁺ ) Example 10 6,9-dihydroxy-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
A solution of tetradeca-2,4,6-trien-8-ylmethyl carbamate (35 mg) in pyridine (1 ml) was diluted with acetic anhydride (1
The reaction mixture was evaporated to dryness under reduced pressure, and the residual oil was subjected to preparative thin layer chromatography using a mixture of methanol and chloroform (1:9, v/v).
v), and the mixture was developed with a mixture of 11-acetyl-8-carbamoyloxymethyl-9-hydroxy-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.
0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-6-yl acetate (22 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ + CD ₃ OD): 6.88 (1 H, d, J = 2 Hz).
6.77 (1H, d, J = 2Hz), 5.37 (1H, s), 4.53 (1H, dd, J = 1
2,3Hz),3.87(1H,dd,J=15,2Hz),3.73(1H,d,J=15H
z),3.30(6H,s),3.20(1H,d,J=6Hz),2.83(1H,dd,J
=6,2Hz),2.33(3H,s),1.89(3H,s) EI mass spectrometry: m/z 451 (M ⁺ ). Additionally, the same preparative thin layer chromatography yielded 11-acetyl-8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-6,9-
Diyl diacetate (11 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.92 (1 H, d, J = 2 Hz), 6.77 (1
H,d,J=2Hz),5.37(1H,s),4.37(2H,d,J=6Hz),4.03
(1H,dd,J=15,2Hz),3.83(1H,t,J=6Hz),3.73(1H,
d, J = 15Hz), 3.41 (1H, d, J = 6Hz), 3.30 (6H, s), 2.80
(1H,dd,J=6,2Hz),2.33(3H,s),2.23(3H,s),1.97
(3H,s) EI mass spectrometry: m/z 493 (M ⁺ ) Example 11 4-diethoxymethyl-6,9-dihydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
A solution of tetradeca-2,4,6-trien-8-ylmethyl carbamate (17 mg) in pyridine (1 ml) was diluted with acetic anhydride (2
The reaction mixture was evaporated to dryness under reduced pressure, and the residual oil was subjected to preparative thin layer chromatography using a mixture of methanol and chloroform (5:
95, v/v), and the resulting solution was eluted with 11-acetyl-8-carbamoyloxymethyl-4-diethoxymethyl-9-hydroxy-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-6-yl acetate (7 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.87 (1 H, d, J = 2 Hz), 6.77 (1
H,d,J=2Hz),5.43(1H,s),4.90(1H,dd,J=12,2Hz),
4.33 (1H,dd,J=12,6Hz),3.87(1H,dd,J=15,2Hz),3.
63 (1H, d, J = 15Hz), 3.53 (4H, q, J = 7Hz), 3.10 (1H, d,
J=6Hz),3.07(1H,dd,J=6,2Hz),2.73(1H,dd,J=6,2
Hz),2.33(3H,s),1.83(3H,s),1.17(6H,t,J=7Hz) EI mass spectrometry: m/z 479 (M ⁺ ) Example 12 6,9-dihydroxy-8-hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
Tetradeca-2,4,6-triene-4-carbaldehyde (4
mg) was dissolved in pyridine (1 ml) and acetic anhydride (50 μg)
The reaction mixture was evaporated to dryness under reduced pressure, and the residual oil was subjected to preparative thin layer chromatography using a mixture of methanol and chloroform (5:95, v/v) to obtain 8-acetoxymethyl-11-acetyl-
4-formyl-9-hydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-
2,4,6-Trien-6-yl acetate (1.5 mg) is obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.90 (1 H, s), 7.31 (1 H, d, J=
2Hz), 7.15 (1H, d, J = 2Hz), 5.65 (1H, s), 4.97 (1H, d
d,J=12,2Hz),4.32(1H,dd,J=12,6.7Hz),3.94(1H,d
d,J=15,2Hz),3.72(1H,d,J=15Hz),3.25(1H,dd,J=
6.7,2Hz), 3.18 (1H, d, J = 6,5Hz), 2.80 (1H, dd, J = 6.
5,2Hz),2.39(3H,s),2.07(3H,s),1.88(3H,s) EI mass spectrometry: m/z 404 (M ⁺ ) Example 13 6,9-dihydroxy-4-hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
A solution of tetradeca-2,4,6-trien-8-ylmethyl carbamate (21 mg) in pyridine (1 ml) was diluted with acetic anhydride (3
The reaction mixture was evaporated to dryness under reduced pressure, and the residual oil was subjected to preparative thin layer chromatography using a mixture of methanol and chloroform (5:95, v
/v), and the resulting solution was eluted with a mixture of 4-acetoxymethyl-11-acetyl-8-carbamoyloxymethyl-9-hydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0
^{2,7.0 10,12} ]tetradeca-2,4,6-trien-6-yl acetate (24 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.77 (1 H, d, J = 2 Hz), 6.63 (1
H,d,J=2Hz),5.03(2H,s),4.87(1H,dd,J=12,2Hz),
4.33 (1H, dd, J = 12,5Hz), 3.90 (1H, dd, J = 15,2Hz), 3.
63 (1H, d, J = 15Hz), 3.20−3.05 (2H, m), 2.77 (1H, dd,
J=6,2Hz),2.33(3H,s),2.10(3H,s),1.87(3H,s) EI mass spectrometry: m/z 449 (M ⁺ ) Example 14 4-hydroxymethyl-8-methylene-14-oxa-1,
11-Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diol (10 mg) was dissolved in pyridine (1 mL), acetic anhydride (100 μL) was added, and the mixture was left at room temperature overnight. The reaction mixture was evaporated to dryness under reduced pressure.
The residual oil was subjected to preparative thin-layer chromatography and developed with a mixture of methanol and chloroform (5:95, v/v) to give 4-acetoxymethyl-11-acetyl-9-hydroxy-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-6-yl acetate (6 mg). ^1H NMR δ (ppm, _CDCl3 ): 6.80 (1H, d, J = 2 Hz), 6.73 (1H, d, J = 2 Hz).
H,d,J=2Hz),6.07(1H,s),6.03(1H,s),5.00(2H,
s),3.83(1H,dd,J=15,2Hz),3.63(1H,dd,J=15,6H
z),3.00(1H,ddd,J=6,6,2Hz),2.80(1H,d,J=6Hz),
2.30(3H,s),2.20(3H,s),2.07(3H,s) EI mass spectrometry: m/z 388 (M ⁺ ) Example 15 6,9-dihydroxy-8-methylene-14-oxa-1,11
-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde (5 mg) was dissolved in methanol (2 ml) and heated to 200°C in hydrogen gas at room temperature and atmospheric pressure.
The reaction mixture was filtered and the filtrate was evaporated to dryness under reduced pressure to give 4-hydroxymethyl-8-methyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,
An oily residue of 4,6-triene-6,9-diol is obtained. This oil is dissolved in pyridine (1 ml) and acetic anhydride (20 μg) is added.
) and leave it at room temperature overnight. The resulting mixture is evaporated to dryness under reduced pressure to give an oil, which is subjected to preparative thin layer chromatography.
/v) mixture, 4-acetoxymethyl-11-acetyl-9-hydroxy-8-methyl-14-oxa-1,
11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-6-yl acetate (3 mg)
^1H NMR δ (ppm, _CDCl3 ): 6.74 (1H, d, J = 2 Hz), 6.61 (1
H,d,J=2Hz),5.02(2H,s),3.90(1H,dd,J=14.5,2.3H
z),3.68(1H,d,J=14.5Hz),3.13(1H,d,J=6Hz),3.0
4 (1H,q,J=7Hz),2.82(1H,dd,J=6,2Hz),2.33(3H,
s), 2.10 (3H, s), 1.94 (3H, s), 1.31 (3H, d, J = 7Hz) EI mass spectrometry: m/z 390 (M ⁺ ) Example 16 4-formyl-6,9-dihydroxy-14-oxa-1,11
A solution of 140 mg of tetradeca- ^2,4,6 -trien-8 ^- ylmethyl carbamate in 10 ml of methanol was subjected to catalytic hydrogenation with 10% palladium on carbon under hydrogen gas at room temperature and atmospheric pressure for 2 hours. The reaction mixture was filtered, and the filtrate was evaporated to dryness under reduced pressure to give 6,9-dihydroxy-4-hydroxymethyl-14-
Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (105 mg). ^1H NMR δ (ppm, DCl + _D2O , pD ≈ 1): 6.68 (1H, s), 6.55
(1H,s),5.22(1H,dd,J=11.5,6Hz),4.67(1H,dd,J=
11.5, 1.5Hz), 4.54 (2H, s), 4.16 (1H, d, J = 16Hz), 4.0
6 (1H, dd, J = 16, 5.7Hz), 3.80−3.64 (3H, m) ¹³ C NMR δ (ppm, DCl + D ₂ O, pD≒1): 159.3 (s), 155.
7 (s), 146.6 (s), 142.3 (s), 110.8 (d), 110.4
(d), 110.3 (s), 90.9 (s), 63.6 (t), 61.0
(t),49.6(t),43.6(d),36.3(d),36.2(d) SI Mass Spectrometry: m/z 324 (M ⁺ +1) Example 17 11-acetyl-4-formyl-8-carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-6,9-
Diyl diacetate (20 mg) was dissolved in ethyl acetate (5 mL) and subjected to catalytic hydrogenation using 10% palladium-carbon at room temperature and atmospheric pressure for 2 hours. The reaction mixture was filtered, and the filtrate was evaporated to dryness under reduced pressure. The residual oil was subjected to preparative thin-layer chromatography using methanol and chloroform (5:95, v/v).
v) and develop the solution with a mixture of 11-acetyl-8-carbamoyloxymethyl-4-hydroxymethyl-14-oxa-
1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (17 mg) is obtained. ¹ H NMR δ (ppm, CDCl ₃ + CD ₃ OD): 6.80 (1 H, s), 6.70 (1 H, s).
H,s),4.57(2H,s),4.10−3.70(3H,m),3.50(1H,d,J
=6Hz),2.83(1H,dd,J=6,2Hz),2.33(3H,s),2.23
(3H,s), 1.97(3H,s) EI mass spectrometry: m/z 449 (M ⁺ ) Example 18 4-formyl-6,9-dihydroxy-14-oxa-1,11
Sodium borohydride (40 mg) was added to a solution of 30 mg of tetradeca- ^2,4,6 -trien-8 ^- ylmethyl carbamate in 3 ml of methanol, and the mixture was stirred at room temperature for 1 hour. After the solvent was removed under reduced pressure, the residue was applied to a column chromatograph packed with the nonionic adsorption resin "Diaion HP-20" (trade name, manufactured by Mitsubishi Chemical Industries, Ltd.). The mixture was eluted with 50% methanol, and the fractions containing the target compound were collected. The 6,9-dihydroxy-4-hydroxymethyl-1-methyl-1-propanol obtained by removing the solvent was purified.
4-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^[10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate was dissolved in pyridine (1 mL). Acetic anhydride (0.5 mL) was added to the solution, and the resulting solution was allowed to stand at room temperature overnight. The solvent was removed using a high vacuum pump to give an oil, which was subjected to preparative thin-layer chromatography.
The gel was developed with a mixture of methanol and chloroform (1:9, v/v) to give 4-acetoxymethyl-11-acetyl-8-carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-
Triene-6,9-diyl diacetate (12 mg) is obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.78 (1 H, dd, J=1.3 Hz), 6.62
(1H,d,J=1.3Hz),5.02(2H,s),4.65(2H,broad s),
4.33 (2H, d, J = 6.2Hz), 4.01 (1H, dd, J = 2.1, 15Hz), 3.
83 (1H, t, J = 6.2Hz), 3.68 (1H, d, J = 15Hz), 3.40 (1H,
d, J = 6.5Hz), 2.81 (1H, dd, J = 2.1, 6.5Hz), 2.38 (3H,
s),2.23(3H,s),2.21(3H,s),2.00(3H,s) EI mass spectrometry: m/z 491 (M ⁺ ) Example 19 4-formyl-6,9-dihydroxy-14-oxa-1,11
A solution of diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (50 mg) in anhydrous methanol (20 ml) was added to anhydrous methanol containing hydrogen chloride (hydrogen chloride content: 10% w/w) (200 μg).
) and allowed to stand overnight at room temperature. Triethylamine (75 μl) was added to this solution, and the resulting solution was evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of methanol and chloroform (1:4, v/v) to obtain 6,9-dihydroxy-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0
^{2,7.0 10,12} ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (51 mg) was obtained. ¹ H NMR δ (ppm, pyridine-d ₅ ): 7.13 (1H,s), 6.93 (1H,s).
H,s),6.00(1H,m),5.50(1H,m),5.43(1H,s),4.20
−3.70 (3H, m), 3.33 (6H, s), 3.10 (1H, m), 2.20 (1H,
m) SI Mass Spectrometry: m/z 368 (M ⁺ +1) Example 20 4-formyl-6,9-dihydroxy-14-oxa-1,11
A solution of diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (75 mg) in absolute ethanol (20 ml) was diluted with anhydrous hydrogen chloride-containing ethanol (hydrogen chloride content: 10% w/v) (100 μg).
) was added and the mixture was left standing at room temperature for 2 hours. The reaction mixture was evaporated to dryness under reduced pressure, and the residue was subjected to silica gel column chromatography.
The desired fractions were collected and evaporated to dryness under reduced pressure to give 4-diethoxymethyl-6,9-dihydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0
^{2,7.0 10,12} ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (90 mg) is obtained.

SI Mass Spectrometry: m/z 396 (M ⁺ +1) Example 21 4-formyl-6,9-dihydroxy-14-oxa-1,11
Diazatetetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (40 mg) was dissolved in 1N aqueous sodium hydroxide (4 mL) and heated at 50°C for 100 minutes under an argon atmosphere. The reaction mixture was cooled and neutralized with 1N hydrochloric acid (4 mL), and the resulting aqueous solution was lyophilized. The residue was subjected to silica gel column chromatography and purified by elution with a mixture of methanol and chloroform (1:4,
The desired fractions were collected and evaporated to dryness under reduced pressure to give 6,9-dihydroxy-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.
0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde (23 mg) was obtained. ¹ H NMR δ (ppm, CD ₃ OD): 9.81 (1 H, s), 7.05 (1 H, d, J=
1.3Hz), 6.90 (1H, d, J = 1.3Hz), 6.80 (1H, d, J = 1.6H
z),6.07 (1H,d,J=1.6Hz),3.76−3.68(2H,m),2.40
(1H,m), 2.30 (1H,m) SI Mass Spectrometry: m/z 261 (M ⁺ +1) Example 22 4-formyl-6,9-dihydroxy-14-oxa-1,11
100 mg of 4-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate was dissolved in 10 mL of 1N aqueous sodium hydroxide, and the resulting pale yellow solution was allowed to stand at room temperature for 72 hours. The reaction mixture was neutralized with 1N hydrochloric acid and then lyophilized. The residue was subjected to silica gel column chromatography and eluted with a mixture of methanol and chloroform (1:9, v/v). The desired fractions were collected and evaporated to dryness under reduced pressure to give 4-hydroxymethyl-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-
Triene-6,9-diol (21 mg) is obtained. ¹ H NMR δ (ppm, CD ₃ OD): 6.60 (1 H, s), 6.58 (1 H, d, J=
2Hz), 6.36 (1H, s), 5.87 (1H, d, J = 2Hz), 4.47 (2H,
s),3.77−3.57(2H,m),2.43−2.20(2H,m) SI Mass Spectrometry: m/z 263 (M ⁺ +1). Further elution with the same solvent system gave 6,9-dihydroxy-8
-hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde (4.5 mg) was obtained. ¹ H NMR δ (ppm, CD ₃ OD): 9.87 (1 H, s), 7.00 (1 H, d, J=
2Hz), 6.83 (1H, d, J = 2Hz), 4.50 (1H, dd, J = 12,3Hz),
4.10 (1H, dd, J = 12,6Hz), 3.80−3.60 (2H, m), 2.60−
2.33 (2H,m) SI Mass Spectrometry: m/z 279 (M ⁺ +1) Example 23 11-acetyl-4-formyl-6,9-dihydroxy-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (55 mg) was dissolved in a mixture of acetone (2 mL) and methyl iodide (2 mL). Potassium carbonate (150 mg) was added to the solution, and the mixture was stirred overnight at 45°C under a nitrogen atmosphere. The mixture was cooled to room temperature and filtered, and the filter cake was washed with a mixture of chloroform and acetone (1:1, v/v, 10 mL). The filtrate and washings were combined and evaporated to dryness under reduced pressure. The residue was subjected to silica gel column chromatography.
Elute with a mixture of chloroform and acetone (4:1, v/v). Collect the desired fractions, evaporate to dryness under reduced pressure, and subject the residue to preparative thin-layer chromatography. Develop with a mixture of chloroform and methanol (1:1, v/v) to obtain 11-
Acetyl-4-formyl-9-hydroxy-6-methoxy-14-oxa-1,11-diazatetracyclo[7.4.1.0
^{2,7.0 10,12} ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (11 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.95 (1H, s), 7.23 (1H, s),
6.93(1H,s),5.08(1H,dd,J=1.4,13Hz),4.79(2H,s
like), 4.58 (1H, dd, J = 4.1, 13Hz), 3.99 (1H, dd, J = 2.
3,14.9Hz),3.96(3H,s),3.67(1H,d,J=14.9Hz),3.2
8 (1H, d, J = 4.1Hz), 3.22 (1H, d, J = 6.3Hz), 2.80 (1H,
dd,J=2.3,6.3Hz),1.88(3H,s) EI mass spectrometry: m/z 377 (M ⁺ +1). Additionally, the same preparative thin layer chromatography yielded 11-acetyl-4-formyl-6,9-dimethoxy-14-oxa-
1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (20 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.93 (1H, s), 7.13 (1H, s),
6.91 (1H, s), 4.66 (1H, dd, J = 6.3, 11.3Hz), 4.64 (2H,
s), 4.31 (1H, dd, J = 2.7, 11.3Hz), 3.93 (3H, s), 3.66
(3H,s),3.44(1H,dd,J=2.7,6.3Hz),3.25(1H,d,J=
6.3Hz), 2.86 (1H, dd, J = 2.3, 6.3Hz), 1.87 (3H, s) EI Mass Spectrometry: 391 (M ⁺ ) Example 24 4-formyl-6,9-dihydroxy-14-oxa-1,11
50 mg of tetradeca-2,4,6 ^- trien-8-ylmethyl carbamate was dissolved in 5 ml of methanol. 0.05 ml of aniline was added to the solution, and the mixture was left at room temperature for 3 hours to give 6,9-dihydroxy- ⁴ -phenyliminomethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 2,7 .0 10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate.
^{2,7.0 10,12} ] tetradeca-2,4,6-trien-8-ylmethyl carbamate.
% palladium-carbon (100 mg) was added, and the mixture was heated under a hydrogen atmosphere (4.5
The mixture is stirred at room temperature under atmospheric pressure for 2 hours. The mixture is filtered using cellulose powder as a filter aid, and the filter cake is washed with methanol (20 ml). The filtrate and washings are combined and evaporated to dryness under reduced pressure, and the residue is subjected to preparative thin-layer chromatography. A 65:25 mixture of chloroform, methanol, and water (4 v/v/
v), and the solution was developed with a mixture of 4-anilinomethyl-6,9-dihydroxy-14-oxa-1,11-diazatetracyclo[7.
4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-8-
25 mg of methyl methyl carbamate was obtained. ¹ H NMR δ (ppm, CD ₃ OD): 7.0-7.2 (2H), 6.3-6.5 (5
H),5.2−4.4(3H,m),4.18(2H,s),3.9−3.2(3H,
m), 2.6 (1H, m) SI Mass Spectrometry: m/z 399 (M ⁺ +1), 421 (M ⁺ +23) Example 25 4-formyl-6,9-dihydroxy-14-oxa-1,11
Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (50 mg) was dissolved in mesyl chloride (12 μg) in pyridine (1.19
The resulting solution was dissolved in 100 ml of methyl 4-formyl-6,9-dihydroxy-11-mesyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 2,7 .0 10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (47 mg). The solution was stirred in a dry ice-carbon tetrachloride bath under a nitrogen atmosphere for 1 hour. The solvent was removed under reduced pressure, and the residue was subjected to preparative thin-layer chromatography. Developed with a mixture of chloroform and methanol, the resulting product was 4-formyl-6,9-dihydroxy-11-mesyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (47 mg).

SI Mass Spectrometry: m/z 400 (M ⁺ +1) Example 26 4-formyl-6,9-dihydroxy-14-oxa-1,11
1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (32 mg) was dissolved in a mixture of N,N-dimethylformamide (1.0 mL) and pyridine (1.0 mL). Octanoyl chloride (17 μg) was added to the solution while stirring in an ice-water bath under a nitrogen atmosphere. The mixture was stirred in an ice-water bath under a nitrogen atmosphere for 2 hours and then evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and methanol to give 4-formyl-6,9-dihydroxy-11-octanoyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-
Trien-8-ylmethyl carbamate (33 mg) is obtained.

SI Mass Spectrometry: m/z 448 (M ⁺ +1) Example 27 6,9-dihydroxy-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
Tetradeca-2,4,6-trien-8-ylmethyl carbamate (22 mg) and sodium bicarbonate (12 mg) were dissolved in water (0.5 ml). Benzyl chloroformate (0.02 ml) was added to the solution while stirring in an ice-water bath. The mixture was stirred in an ice-water bath for 30 minutes and then subjected to preparative thin-layer chromatography. The mixture was developed with a mixture of chloroform and methanol.
11-benzyloxycarbonyl-6,9-dihydroxy-
4-Dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (15 mg) is obtained.

EI mass spectrometry: m/z 501 (M ⁺ ) Example 28 11-benzyloxycarbonyl-6,9-dihydroxy-
4-Dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (15 mg) was dissolved in a mixture of pyridine (1.0 mL) and acetic anhydride (0.5 mL) and stirred at room temperature for 1 day. 4-Dimethylaminopyridine (catalytic amount) was added to the solution and stirred at room temperature for 12 hours. The reaction mixture was evaporated to dryness under reduced pressure, and the residue was subjected to preparative thin-layer chromatography. The product was developed with a mixture of chloroform and methanol to give 11-benzyloxycarbonyl-8-
Carbamoyloxymethyl-4-dimethoxymethyl-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
The diacetate (17 mg) is obtained.

SI Mass Spectrometry: m/z 586 (M ⁺ +1) Example 29 11-benzyloxycarbonyl-8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-
Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (270m
g) in methanol (10 ml).
Palladium-carbon (300 mg) was added, and the mixture was stirred under a hydrogen atmosphere (20 p
The mixture was stirred at room temperature for 1 hour. The reaction mixture was filtered using cellulose powder as a filter aid, and the filtrate was evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of methanol and chloroform to give 8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .
0 ^10,12 ］tetradeca-2,4,6-triene-6,9-diyl＝
The diacetate (80 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.95 (1H, s), 6.77 (1H, s),
5.37 (1H, s), 4.63 (2H, broad s), 4.34 (2H, d, J = 6H
z),3.99(1H,dd,J=3,15Hz),3.83(1H,t,J=6Hz),3.
61 (1H, d, J = 15Hz), 3.30 (6H, s), 2.35 (3H, s), 2.22
(3H,s) SI Mass Spectrometry: m/z 452 (M ⁺ +1) Example 30 11-acetyl-4-formyl-6,9-dihydroxy-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (25 mg) in tetrahydrofuran (2.5 m
The solution was added to a suspension of sodium hydroxide (40% oil content, 3 mg) in tetrahydrofuran (0.5 ml).
A solution of 108 μm of tetrahydrofuran (108 μm) was added,
Stir in an ice-water bath for 1 hour. After stirring at room temperature for 36 hours,
Octanoyl chloride (50 μl) is added and stirred at room temperature for 3 hours. The reaction mixture is then subjected to preparative thin layer chromatography.
v) and develop the solution with a mixture of 11-acetyl-8-carbamoyloxymethyl-4-formyl-9-hydroxy-14-
Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-trien-6-yl octanoate (7 mg).

SI Mass Spectrometry: m/z 490 (M ⁺ +1) Example 31 11-acetyl-8-carbamoyloxymethyl-4-formyl-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-6,9-
A mixture of chromium trioxide (26 mg), water (76 μ), and sulfuric acid (22 μ) was added to a solution of diyl diacetate (29 mg) in acetone (2 mL) in an ice-water bath. After stirring for 30 minutes in the ice-water bath, isopropyl alcohol (2 mL) was added. The resulting mixture was added to water (10 mL) and extracted with a mixture of chloroform and methanol (2:1, v/v) (10 mL x 5). The extracts were combined, dried over magnesium sulfate, and
The residue was then evaporated to dryness under reduced pressure, and the residue was subjected to preparative thin layer chromatography, developing with a mixture of chloroform, methanol, and water (65:25:4, v/v/v) to obtain 6,9-diacetoxy-8-carbamoyloxymethyl-14-oxa-1,11
-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carboxylic acid (5 mg) was obtained. ^1H NMR δ (ppm, _CD3OD ): 7.34 (1H, s), 7.19 (1H, s),
4.25 (1H, dd, J = 8.1, 11.7Hz), 4.1−3.5 (4H), 2.95 (1
H,d,J=7.2Hz),2.30(3H,s),2.13(3H,s) SI Mass Spectrometry: m/z 444 (M ⁺ +23), 422 (M ⁺ +1) Example 32 6,9-diacetoxy-8-carbamoyloxymethyl-1
4-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
To a solution of ¹² mg of tetradeca-2,4,6-triene-4-carboxylic acid in 2 ml of methanol was added a solution of excess diazomethane in diethyl ether at room temperature. The mixture was left at room temperature for 20 minutes and then evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and methanol (15:1, v/v) to give 6,9-diacetoxy-8-carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0].
^{2,7.0 10,12} ] methyl tetradeca-2,4,6-triene-4-carboxylate (5 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 7.47 (1H, s), 7.30 (1H, s),
4.62 (2H, broad s), 4.32 (2H, d, J = 5.4Hz), 3.86 (4H,
s), 2.37 (3H,s), 2.20 (3H,s) Example 33 6,9-dihydroxy-8-methylene-14-oxa-1,11
-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde (30 mg)
To the N,N-dimethylformamide (1.0 mL) solution, a suspension of sodium hydride (40% oil content, 4.6 mg) in N,N-dimethylformamide (0.5 mL) was added in an ice-water bath. To this mixture, a solution of methyl iodide (7.2 μg) in N,N-dimethylformamide (0.1 mL) was added in an ice-water bath. After stirring in the ice-water bath for 1 hour, the reaction mixture was evaporated to dryness under reduced pressure.
The residue was subjected to preparative thin layer chromatography and developed with a mixture of chloroform and methanol (8:1, v/v).
-hydroxy-6-methoxy-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
Tetradeca-2,4,6-triene-4-carbaldehyde (1
9 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.90 (1H, s), 7.13 (1H, d, J=
2Hz), 6.97 (1H, d, J = 2 Hz), 6.63 (1H, d, J = 2 Hz), 6.17
(1H,d,J=2Hz),3.97(3H,s),3.89(1H,dd,J=2,14.9
Hz), 3.58 (1H, dd, J=4.5, 14.9Hz), 2.3−3.0 (4H) SI Mass Spectrometry: m/z 275 (M ⁺ +1). Additionally, the same preparative thin layer chromatography yielded 6-methoxy-11-methyl-8-methylene-14-oxa-1,11-
Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde (2 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.89 (1 H, s), 7.12 (1 H, d, J=
1.4Hz), 6.96 (1H, d, J = 1.4Hz), 6.61 (1H, d, J = 1.4H
z),6.16(1H,d,J=1.4Hz),3.99(3H,s),3.83(1H,d
d,J=1.9,15Hz),3.64(1H,dd,J=6.2,15Hz),2.43(3
H,s),1.89(1H,ddd,J=1.9,6.2,6.8Hz),1.79(1H,d,J
= 6.8 Hz) Example 34 6,9-diacetoxy-8-carbamoyloxymethyl-1
4-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] A solution of methyl tetradeca-2,4,6-triene-4-carboxylate (5 mg) in pyridine (1.0 ml) was diluted with acetic anhydride (0.
The reaction mixture was evaporated to dryness under reduced pressure, and the residue was subjected to preparative thin-layer chromatography using a mixture of chloroform and methanol (30:1, v/v) to give 6,9-diacetoxy-11-acetyl-
8-Carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-
Methyl 2,4,6-triene-4-carboxylate (5 mg) is obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 7.44 (1 H, d, J = 1.1 Hz), 7.31
(1H,d,J=1.1Hz),4.63(2H,broad s),4.39(1H,dd,J
=7.0,11.9Hz),4.33(1H,dd,J=4.3,11.9Hz),4.04(1
H,dd,J=2.4,14.9Hz),3.89(3H,s),3.86(1H,dd,J=
4.3,7.0Hz),3.74(1H,d,J=14.9Hz),3.40(1H,d,J=
6.8Hz), 2.82 (1H, dd, J = 2.4, 6.8Hz), 2.38 (3H, s), 2.
24 (3H,s), 1.97 (3H,s) SI Mass Spectrometry: m/z 478 (M ⁺ +1) Example 35 6,9-dihydroxy-8-methylene-14-oxa-1,11
To a solution of 151 mg of tetradeca-2,4,6 ^- triene-4 ^- carbaldehyde in 15 ml of methanol was added 0.5 ml of 6% hydrogen chloride in methanol. The mixture was stirred at room temperature for 1 hour and evaporated to dryness under reduced pressure to give 4-dimethoxymethyl-8-(2-methyl-2-propanol).
-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-
The crude product (286 mg) of 6,9-diol was obtained. This crude product (260 mg) was dissolved in pyridine (5.0 mL) and acetic anhydride (2.5 mL).
The solution was dissolved in a mixture of 11-acetyl-4-dimethoxymethyl-8-methyl-2-propanol (11-acetyl-4-dimethoxymethyl-8-methyl-2-propanol). 4-Dimethylaminopyridine (25 mg) was added to the solution, which was stirred at room temperature for 2 days and evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and acetone (15:1, v/v) to obtain 11-acetyl-4-dimethoxymethyl-8-methyl-2-propanol (11-acetyl-4-dimethoxymethyl-8-methyl-2-propanol).
-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-
Isomer A of 6,9-diyl diacetate (100 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.82 (1H, s), 6.77 (1H, s),
6.17(1H,s),5.78(1H,s),5.35(1H,s),4.08(1H,d
d,J=2.7,14.9Hz),3.72(1H,d,J=14.9Hz),3.35(1H,
d,J=6.3Hz),3.29(6H,s),2.99(1H,dd,J=2.7,6.3H
z), 2.34 (3H,s), 2.24 (3H,s), 1.87 (3H,s). SI Mass Spectrometry: m/z 433 (M ⁺ +1). Furthermore, the same preparative thin layer chromatography yielded 11-acetyl-4-dimethoxymethyl-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ].
100 mg of the B isomer of tetradeca-2,4,6-triene-6,9-diyl diacetate was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.96 (1 H, d, J=2 Hz), 6.90 (1
H, d, J = 2Hz), 6.18 (1H, s), 5.53 (1H, s), 5.34 (1H,
s),3.97(1H,dd,J=2.7,14.4Hz),3.60(1H,dd,J=5.
4,14.4Hz),3.30(6H,s),2.90−3.15(2H),2.32(3H,
s),2.25(3H,s),2.22(3H,s) SI mass spectrometry:m/z 433(M ⁺⁺ 1) Example 36 11-acetyl-4-dimethoxymethyl-8-methylene-
14-Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
The diacetate B isomer (12 mg) was dissolved in methylene chloride and ozone was bubbled through it in a dry ice-carbon tetrachloride bath for 30 minutes, followed by nitrogen bubbling through it for 3 minutes.
Dimethyl sulfide (0.5 ml) was added in a dry ice-carbon tetrachloride bath under a nitrogen atmosphere, and the resulting mixture was stirred for 10 minutes. After standing at room temperature for 15 minutes, the mixture was evaporated to dryness under reduced pressure.
The residue was subjected to preparative thin layer chromatography. It was developed with a mixture of n-hexane and ethyl acetate (1:2, v/v) to obtain 11
-acetyl-4-dimethoxymethyl-8-oxo-14-
Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
2 mg of the diacetate B isomer are obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.97 (1 H, d, J=1.6 Hz), 6.969
(1H,d,J=1.6Hz),5.35(1H,s),4.05(1H,dd,J=1.4,
14.6Hz), 3.65 (1H, dd, J = 6.2, 14.6Hz), 3.33 (3H, s),
3.32(3H,s),3.35−3.29(1H),3.16(1H,dt,J=1.4,
6.2Hz),2.38(3H,s),2.26(3H,s),2.22(3H,s) SI Mass Spectrometry: m/z 435 (M ⁺ +1) Example 37 11-acetyl-4-dimethoxymethyl-8-methylene-
14-Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
Diacetate A isomer (45 mg) was dissolved in methylene chloride (15 m
The mixture was dissolved in 1 ml of dimethyl sulfide and ozone was bubbled through it in a dry ice-carbon tetrachloride bath for 10 minutes, followed by a nitrogen sparge for 3 minutes. Dimethyl sulfide (0.5 ml) was added to the mixture in a dry ice-carbon tetrachloride bath under a nitrogen atmosphere, the mixture was allowed to stand at room temperature for 20 minutes, and the mixture was evaporated to dryness under reduced pressure. The residue was subjected to preparative thin layer chromatography using a mixture of n-hexane and ethyl acetate (1:2,
v/v), the A isomer of 11-acetyl-4-dimethoxymethyl-8-oxo-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (34m
g) obtain. ¹ H NMR δ (ppm, CDCl ₃ ): 6.86 (1H, d, J = 0.8 Hz), 6.84
(1H,d,J=0.8Hz),5.35(1H,s),4.10(1H,dd,J=2.7,
14.6Hz), 3.75 (1H, d, J = 14.6Hz), 3.31 (3H, s), 3.29
(3H,s),3.23(1H,d,J=6.2Hz),3.04(1H,dd,J=1.9,
6.2Hz),2.36(3H,s),2.23(3H,s),1.94(3H,s) SI Mass Spectrometry: m/z 435 (M ⁺ +1) Example 38 11-acetyl-4-dimethoxymethyl-8-oxo-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
To a solution of the diacetate A isomer (6 mg) in acetone (0.5 ml) was added p-toluenesulfonic acid (1 mg). The mixture was stirred at room temperature for 1 hour and then subjected to preparative thin-layer chromatography using chloroform and acetone (5:1, v/v).
v) and develop the mixture of 4-formyl-8-oxo-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
The diacetate (1 mg) is obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 7.20 (1 H, d, J=1.4 Hz), 7.05
(1H, d, J = 1.4Hz), 3.77 (1H, dd, J = 2.7, 13.5Hz), 2.94
(1H,d,J=13.5Hz),2.38(3H,s),2.26(1H,d,J=6.8H
z),2.15−2.20(1H),2.17(3H,s) SI Mass Spectrometry: m/z 369 (M ⁺ 23) Example 39 4-formyl-6,9-dihydroxy-14-oxa-1,11
To a solution of 100 ^mg of tetradeca-2,4,6 ^- trien-8-ylmethyl carbamate in 5 mL of methanol was added 90 mg of hydroxylamine hydrochloride and 106 mg of sodium bicarbonate. The mixture was stirred at room temperature for 1 hour and evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography using a 6:4:1 mixture of chloroform, methanol, and water (v/v/).
The mixture was developed with a mixture of 6,9-dihydroxy-4-hydroxyiminomethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (105 mg). ¹ H NMR δ (ppm, CD ₃ OD): 7.94 (1H, s), SI Mass Spectrometry: m/z 337 (M ⁺ +1), 359 (M ⁺ +23) Example 40 4-formyl-6,9-dihydroxy-14-oxa-1,11
- Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (50 mg) in methanol (5 ml) was added sodium bicarbonate (27 mg) and semicarbazide hydrochloride (35 mg),
After stirring at room temperature for 1 hour, the mixture was evaporated to dryness under reduced pressure. The residue was subjected to preparative thin layer chromatography and developed with a mixture of chloroform, methanol, and water (6:4:1, v/v/v) to give 6,9
-dihydroxy-4-semicarbazonomethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
59 mg of tetradeca-2,4,6-trien-8-ylmethyl carbamate are obtained.

SI Mass Spectrometry: m/z 379 (M ⁺ +1) Example 41 4-formyl-6,9-dihydroxy-14-oxa-1,11
A solution of 50 mg of diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate in 5 ml of methanol was added with 27 mg of sodium bicarbonate and 59 mg of 4-phenylsemicarbazide hydrochloride.
mg) was added. The mixture was stirred at room temperature for 1 hour and evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and methanol (3:1, v/v) to give 6,9-dihydroxy-4-(4-phenylsemicarbazonomethyl)-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,
4,6-trien-8-ylmethyl carbamate (64m
g) is obtained. ¹ H NMR δ (ppm, CD ₃ OD): 5.0−5.3(1H),4.4−4.8(2H),3.5−3.8(2H),2.4−
2.8 (2H) SI Mass Spectrometry: m/z 455 (M ⁺ +1) Example 42 6,9-dihydroxy-8-hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]
Tetradeca-2,4,6-triene-4-carbaldehyde (9
Acetic anhydride (40 μl) was added to a solution of 11-acetyl-6,9-dihydroxy-8-(2-hydroxy-1,3-dihydroxybenzoyl)propan-1-one (8 mg) in methanol (3 mL). The mixture was stirred at room temperature for 1 hour and evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and methanol (10:1, v/v) to give 11-acetyl-6,9-dihydroxy-8-(2-hydroxy-1,3-dihydroxybenzoyl)propan-1-one ( ...
Hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde (71 mg) is obtained.

SI Mass Spectrometry: m/z 321 (M ⁺ +1) Example 43 11-acetyl-6,9-dihydroxy-8-hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.
0 ^2,7 .0 ^10,12 ]Tetradeca-2,4,6-triene-4-carbaldehyde (18 mg) in N,N-dimethylformamide (1.0
ml) solution in an ice-water bath.
A solution of 11-(2-methyl-2-propanol) in N,N-dimethylformamide (1.25 ml) was added. The mixture was stirred in an ice-water bath for 30 minutes and then at room temperature for an additional hour. The mixture was evaporated to dryness under reduced pressure and subjected to preparative thin-layer chromatography, developing with a mixture of chloroform and methanol (15:1, v/v) to obtain 11-(2-methyl-2-propanol).
Acetyl-4-formyl-9-hydroxy-6-(N-
phenylcarbamoyloxy)-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-
2,4,6-Trien-8-ylmethyl N-phenylcarbamate (12 mg) is obtained.

SI Mass Spectrometry: m/z 559 (M ⁺ +1) Example 44 11-benzyloxycarbonyl-6,9-dihydroxy-
To a solution of 4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (400 mg) in pyridine (10 mL) and acetic anhydride (5 mL) was added 4-dimethylaminopyridine (10 mg). The mixture was stirred overnight at room temperature and evaporated to dryness under reduced pressure. The residue was subjected to silica gel column chromatography, eluting with a mixture of chloroform and methanol (50:1, v/v). The desired fractions were collected and evaporated to dryness under reduced pressure to give 11-benzyloxycarbonyl-8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (458 mg).

SI Mass Spectrometry: m/z 586 (M ⁺ +1) Example 45 11-benzyloxycarbonyl-8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-
Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (100m
To a solution of 8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 2,7.0 10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (31 mg) was added 10% palladium-carbon (50 mg) and stirred under a hydrogen atmosphere (20 psi) at room temperature for 1 hour. The reaction mixture was filtered using cellulose powder as a filter aid, and the filtrate was evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and methanol (10:1, v/v) to give isomer A of 8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^{2,7.0 10,12} ]tetradeca-2,4,6-triene-6,9-diyl diacetate (31 mg). ^1H NMR δ (ppm, _CDCl3 ): 6.95 (1H, s), 6.77 (1H, s).
5.37 (1H, s), 4.63 (2H, broad s), 4.34 (2H, d, J = 6H
z),3.99(1H,dd,J=3,15Hz),3.83(1H,t,J=6Hz),3.
61 (1H, d, J = 15Hz), 3.30 (6H, s), 3.0−3.2 (1H), 2.3
5 (3H,s), 2.22 (3H,s) SI Mass Spectrometry: m/z 452 (M ⁺ +1) Furthermore, the same preparative thin layer chromatography yielded 8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .
0 ^10,12 ］tetradeca-2,4,6-triene-6,9-diyl＝
The diacetate B isomer (41 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.94 (1H, s), 6.86 (1H, s),
5.30 (1H, s), 4.5−5.0 (4H), 4.25 (1H, dd, J = 3.2, 7.2
Hz), 3.91 (1H, dd, J = 2.7, 14.4 Hz), 3.61 (1H, dd, J = 6.
3,14.4Hz),3.31(6H,s),2.77(1H,d,J=6.3Hz),2.3
−2.5(1H),2.31(3H,s),2.12(3H,s) SI Mass Spectrometry: m/z 452 (M ⁺ +1) Example 46 A suspension of sodium hydride (40% oil content, 4 mg) in tetrahydrofuran (0.5 ml) was heated in an ice-water bath with 2.0 mg of 8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate B isomer (20 mg).
To this solution, methyl iodide was added in an ice-water bath, and the mixture was stirred at room temperature for 3 hours. Then, acetic acid (0.1 ml) was added to the reaction mixture in an ice-water bath. The resulting mixture was evaporated to dryness under reduced pressure, and the residual oil was subjected to preparative thin-layer chromatography using chloroform and methanol (8:1, v/v).
The mixture was developed with 8-carbamoyloxymethyl-4
-dimethoxymethyl-11-methyl-14-oxa-1,11-
Diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (11m
g) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 6.92 (1 H, d, J=1 Hz), 6.84 (1
H,d,J=1Hz),5.32(1H,s),4.82(1H,dd,J=3.2,11.9H
z),4.76(1H,dd,J=6.8,11.9Hz),4.59(2H,broad
s),4.13(1H,dd,J=3.2,6.8Hz),3.93(1H,dd,J=2.7,
14.9Hz), 3.66 (1H, dd, J = 6.8, 14.9Hz), 3.323 (3H,
s),3.315(3H,s),2.41(3H,s),2.32(3H,s),2.25−
2.35 (2H), 2.19 (3H, s) SI Mass Spectrometry: m/z 466 (M ⁺ +1), 488 (M ⁺ +23) Example 47 11-acetyl-8-carbamoyloxymethyl-4-formyl-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-6,9-
To a solution of diyl diacetate (52 mg) in methanol (5 ml) was added sodium bicarbonate (10 mg) in an ice-water bath.
The mixture was stirred in an ice-water bath for 30 minutes and at room temperature for a further 2.5 hours. The pH of the reaction mixture was adjusted to approximately 6 by adding acetic acid in an ice-water bath, and the mixture was evaporated to dryness under reduced pressure. The residue was extracted with a mixture of chloroform and methanol (5:1, v/v) (5 ml x 3). The extracts were combined and evaporated under reduced pressure. The resulting residue was subjected to silica gel column chromatography, eluting with a mixture of chloroform and methanol (50:1, v/v) to obtain 11-acetyl-8-carbamoyloxymethyl-4-(2-methyl-4-methyl-2-propanol).
-formyl-6-hydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,
4,6-trien-9-yl acetate (26 mg) is obtained. ¹ H NMR δ (ppm, CD ₃ OD): 9.80 (1 H, s), 7.00 (1 H, d, J=
2Hz), 6.83 (1H, d, J = 2Hz), 4.46 (1H, dd, J = 7.2, 11.7H
z),4.14(1H,dd,J=2,11.7Hz),3.7−4.0(3H,m),3.6
0 (1H, d, J = 6.3Hz), 2.90 (1H, m), 2.18 (3H, s), 1.88
(3H,s) Example 48 In the same manner as in Example 47, 9-acetoxy-8-carbamoyloxymethyl-6-hydroxy-14-oxa-1,11
-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carboxylate methyl ester. ¹ H NMR δ (ppm, CDCl ₃ ): 7.29 (1 H, d, J = 1.4 Hz), 6.97
(1H,d,J=1.4Hz),4.86(2H,broad s),4.26(1H,dd,J
=7.3,13Hz),4.20(1H,dd,J=2.4,13Hz),4.08(1H,d
d,J=2.4,7.3Hz),3.97(1H,dd,J=2.2,14Hz),3.86(3
H,s),3.68(1H,d,J=14Hz),3.10(1H,m),2.26(1
H), 2.24 (3H, s) Example 49 4-formyl-6,9-dihydroxy-14-oxa-1,11
To a solution of 100 mg of tetradeca-2,4,6 ^- trien-8 ^- ylmethyl carbamate in 5 mL of methanol was added 105 mg of O-methylhydroxylamine hydrochloride and 106 mg of sodium bicarbonate, the mixture was stirred at room temperature for 1.5 hours, and evaporated to dryness under reduced pressure. The residue was subjected to preparative thin-layer chromatography.
The mixture was developed with a mixture of chloroform and methanol (4:1, v/v) to obtain 6,9-dihydroxy-4-methoxyiminomethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0
^{2,7.0 10,12} ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (80 mg) is obtained.

Example 50 6,9-dihydroxy-4-hydroxyiminomethyl-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (88 mg) was dissolved in a mixture of pyridine (3 ml) and acetic anhydride (1.5 ml). The solution was stirred at room temperature for 20 minutes.
The mixture was stirred for 1 hour, evaporated to dryness under reduced pressure, and then subjected to preparative thin-layer chromatography, developed with a mixture of chloroform and methanol (12:1, v/v), to obtain 4-acetoxyiminomethyl-11-acetyl-8-carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.
0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate (90 mg) is obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 8.23 (1 H, s), 7.15 (1 H, d, J=
2Hz), 7.02 (1H, d, J = 2Hz), 4.68 (2H, broad s), 4.2−
4.6 (2H, m), 4.02 (1H, dd, J=3,14Hz), 3.7−3.9 (1H,
m), 3.70 (1H, d, J = 14Hz), 3.39 (1H, d, J = 6Hz), 2.80
(1H,dd,J=3,6Hz),2.34(3H,s),2.20(6H,s),1.95
(3H,s) Example 51 11-acetyl-8-carbamoyloxymethyl-4-formyl-6-hydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-
To a mixture of trien-9-yl acetate (25 mg) in acetone (1 ml) and methyl iodide (1 ml) was added potassium bicarbonate (50 mg), and the mixture was stirred at 45°C for 3 hours.
The reaction mixture was subjected to preparative thin-layer chromatography and developed with a mixture of chloroform and methanol (15:1, v/v) to obtain 11-acetyl-8-carbamoyloxymethyl-4-(2-methyl-4-methyl-2-propanol).
-formyl-6-methoxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6
25 mg of 1H-trien-9-yl acetate was obtained. ^1H NMR δ (ppm, _CDCl3 ): 9.89 (1H, s), 7.06 (1H, d, J=
1Hz), 6.87 (1H, d, J = 1Hz), 4.73 (2H, broad s), 4.52
(1H,dd,J=7,11Hz),4.23(1H,dd,J=2,11Hz),4.03
(1H,dd,J=2,14Hz),3.92(3H,s),3.87(1H,dd,J=2,
7Hz), 3.68 (1H, d, J = 14Hz), 3.44 (1H, d, J = 6Hz), 2.8
2 (1H, dd, J = 2.6Hz), 2.20 (3H, s), 1.90 (3H, s) Example 52 6,9-dihydroxy-4-methoxyiminomethyl-14-
Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (55 mg) was dissolved in a mixture of pyridine (3 mL) and acetic anhydride (1.5 mL) and stirred at room temperature for 12 hours. The mixture was evaporated to dryness under reduced pressure and then subjected to preparative thin-layer chromatography using a mixture of chloroform and methanol (20:1, v/v) to give 11-acetyl-8
-carbamoyloxymethyl-4-methoxyiminomethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0
^{2,7.0 10,12} ]tetradeca-2,4,6-triene-6,9-diyl diacetate (62 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 7.90 (1 H, s), 7.00 (1 H, d, J=
1Hz), 6.82 (1H, d, J = 1Hz), 4.80 (2H, broad s), 4.32
(2H, d, J = 6Hz), 4.00 (1H, dd, J = 2,14Hz), 3.92 (3H,
s), 3.80 (1H, t, J = 6Hz), 3.67 (1H, d, J = 14Hz), 3.37
(1H, d, J = 7Hz), 2.77 (1H, dd, J = 2,7Hz), 2.31 (3H,
s), 2.17 (3H, s), 1.94 (3H, s) Example 53 11-acetyl-8-carbamoyloxymethyl-4-formyl-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ］tetradeca-2,4,6-triene-6,9-
To a solution of diyl diacetate (40 mg) in methanol (2 ml) was added hydroxylamine hydrochloride (10 mg) and sodium bicarbonate (8 mg). The mixture was stirred at room temperature for 2 hours, evaporated to dryness under reduced pressure, and the residue was subjected to preparative thin-layer chromatography.
0:1, v/v) mixture to obtain 11-acetyl-8-carbamoyloxymethyl-4-hydroxyiminomethyl-
14-Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
The diacetate (13 mg) was obtained. ¹ H NMR δ (ppm, CD ₃ OD): 8.00 (1 H, s), 7.03 (1 H, d, J=
2Hz), 6.92 (1H, d, J = 2Hz), 4.0−4.6 (3H, m), 3.5−3.
9 (3H, m), 2.87 (1H, d, J = 6Hz), 2.35 (3H, s), 2.20 (3
H,s), 1.93 (3H,s) Example 54 In the same manner as in Example 38, 8-carbamoyloxymethyl-4-dimethoxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6
2,4,6-Triene-6,9-diyl diacetate is reacted with p-toluenesulfonic acid to give 8-carbamoyloxymethyl-4-formyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate. This product is then subjected to catalytic reduction in the same manner as in Example 17 to give 8-carbamoyloxymethyl-4-hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .
0 ^10,12 ］tetradeca-2,4,6-triene-6,9-diyl＝
The diacetate is obtained. ^1H NMR δ (ppm, _CDDl3 ): 6.77 (1H, s), 6.61 (1H, s),
4.51 (2H, s), 2.97 (1H, d, J=6.3Hz), 2.30 (3H, s), 2.
14 (3H,s) SI Mass Spectrometry: m/z 408 (M ⁺ +1) Example 55 4-formyl-6,9-dihydroxy-14-oxa-1,11
-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate (50 mg) and 4-dimethylaminopyridine (150 mg)
In an ice-water bath, propionyl chloride (0.1 ml) was added to a solution of the above in dimethylformamide (2 ml). The mixture was stirred at room temperature for 20 hours, evaporated to dryness under reduced pressure, and the residue was subjected to preparative thin-layer chromatography using a mixture of chloroform and methanol (10:1, v/v) to give 8-carbamoyloxymethyl-4-formyl-9-hydroxy-11-propionyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^{2,7.0 10,12} ]tetradeca-2,4,6-triene-
6-yl propionate (30 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.96 (1H, s), 7.37 (2H, s),
5.00 (2H, broad s), 4.58 (1H, dd, J=5,11Hz), 4.2−4.
5 (1H), 4.17 (1H, dd, J = 6, 11Hz), 3.5-3.8 (3H), 3.2
7 (1H,t,J=6Hz),2.82(2H,q,J=7.2Hz),2.34(2H,q,
J=7.2Hz),1.26(3H,t,J=7.2Hz),1.19(3H,t,J=7.2
Hz) Example 56 In a similar manner to Example 52, 6,9-dihydroxy-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.
1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde (6 mg) was reacted with acetic anhydride (0.2 ml) at 80 °C to give 11-acetyl-4-formyl-8-methylene-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ] tetradeca-2,4,6-triene-6,9-diyl =
The diacetate (4 mg) was obtained. ¹ H NMR δ (ppm, CDCl ₃ ): 9.97 (1 H, s), 7.40 (1 H, d, J=
1Hz), 7.31 (1H, d, J = 1Hz), 6.37 (1H, s), 5.70 (1H,
s),3.5−4.2(2H),2.9−3.2(2H),2.37(3H,s),2.2
5 (3H,s), 2.22 (3H,s) EI mass spectrometry: m/z 386 (M ⁺ )

[Brief explanation of the drawings]

1 and 2 show ^{the 13} C nuclear magnetic resonance spectra of the FR-900482 substance obtained in the present invention measured near neutral (measurement solvent: D ₂ O) and near pH=1 (measurement solvent: D ₂ O+DCl), respectively.

───────────────────────────────────────────────────────── Continued from the front page (51) Int.Cl. ⁶ Identification symbol Internal reference number FI Technical indication location C12R 1:465) (72) Inventor Takase Shigehiro 61-9 Namase, Shiose-cho, Nishinomiya-shi, Hyogo Prefecture Examiner Taniguchi Hiroshi

Claims (20)

[Claims] [Claim 1] Formula: [wherein R ¹ is a formyl group, a protected formyl group, a hydroxymethyl group, a protected hydroxymethyl group, an arylaminomethyl group, a carboxy group, a protected carboxy group or a substituted iminomethyl group; R ² is a hydroxy group, an alkoxy group or a protected hydroxy group; R ³
is hydrogen, and ^R4 is a methyl group, a hydroxymethyl group or a protected hydroxymethyl group, or ^R3 and ^R4 are bonded to each other to form a methylene group or an oxo group, ^R5 is a hydroxy group, an alkoxy group or a protected hydroxy group, and ^R6 is hydrogen, an imino-protecting group or an alkyl group, respectively, or a pharmaceutically acceptable salt thereof. Claim 2: R ¹ is a formyl group, a dialkoxymethyl group,
a hydroxymethyl group, an acyloxymethyl group, an arylaminomethyl group, a carboxy group, an esterified carboxy group, an aryliminomethyl group, a hydroxyiminomethyl group, an alkoxyiminomethyl group, an acyloxyiminomethyl group, a semicarbazonomethyl group, or an arylsemicarbazonomethyl group; ^R2 is a hydroxy group, an alkoxy group, or an acyloxy group; ^R3 is hydrogen; and
The compound according to claim 1, wherein ^R4 is a methyl group, a hydroxymethyl group, or an acyloxymethyl group, or ^R3 and ^R4 are bonded to each other to form a methylene group or an oxo group, ^R5 is a hydroxy group, an alkoxy group, or an acyloxy group, and ^R6 is a hydrogen atom, an acyl group, or an alkyl group. Claim 3: R ¹ is a formyl group, a di(lower)alkoxymethyl group, a hydroxymethyl group, a lower alkanoyloxymethyl group, an arylaminomethyl group, a carboxy group, a lower alkoxycarbonyl group, an aryliminomethyl group,
The compound according to claim 2, wherein ^R2 is a lower alkoxyiminomethyl group, a lower alkanoyloxyiminomethyl group, a semicarbazonomethyl group, or an arylsemicarbazonomethyl group; R2 is a hydroxy group, a lower alkoxy group, a lower alkanoyloxy group, a higher alkanoyloxy group, or an arylcarbamoyloxy group; ^R3 is hydrogen and ^R4 is a methyl group, a hydroxymethyl group, a carbamoyloxymethyl group, a lower alkanoyloxymethyl group, or an arylcarbamoyloxymethyl group; or ^R3 and ^R4 are bonded to each other to form a methylene group or an oxo group; ^R5 is a hydroxy group, a lower alkoxy group, or a lower alkanoyloxy group; and ^R6 is hydrogen, a lower alkanoyl group, a higher alkanoyl group, an aroyl group optionally having a halogen atom, a lower alkylsulfonyl group, an aryl(lower)alkoxycarbonyl group, or a lower alkyl group. 4. 4-Formyl-6,9-dihydroxy-14-
Oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate or its hydrochloride salt. 5. The compound according to claim 3, which is 11-acetyl-8-carbamoyloxymethyl-4-formyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate. 6. The compound according to claim 3, which is 11-acetyl-4-formyl-9-hydroxy-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-6-yl acetate. 7. 4-Acetoxymethyl-11-acetyl-8
-carbamoyloxymethyl-9-hydroxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .
0 ^10,12 ]tetradeca-2,4,6-trien-6-yl acetate. 8. 4-Acetoxymethyl-11-acetyl-8
-carbamoyloxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,
The compound according to claim 3, which is 4,6-triene-6,9-diyl diacetate. 9. 11-acetyl-8-carbamoyloxymethyl-4-formyl-6-methoxy-14-oxa-1,11
3) The compound according to claim 3, which is diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-9-yl acetate. 10. The compound according to claim 3, which is 11-acetyl-4-formyl-9-hydroxy-6-methoxy-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-8-ylmethyl carbamate. 11. 6,9-dihydroxy-8-methylene-14
-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0
^10,12 ]tetradeca-2,4,6-triene-4-carbaldehyde. 12. 4-Acetoxymethyl-11-acetyl-
9-hydroxy-8-methylene-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-
The compound according to claim 3, which is 2,4,6-trien-6-yl acetate. 13. The compound according to claim 3, which is 8-carbamoyloxymethyl-4-hydroxymethyl-14-oxa-1,11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-triene-6,9-diyl diacetate. 14. 11-acetyl-8-carbamoyloxymethyl-4-formyl-6-methoxy-14-oxa-1,
3) The compound according to claim 3, which is 11-diazatetracyclo[7.4.1.0 ^2,7 .0 ^10,12 ]tetradeca-2,4,6-trien-9-yl acetate. 15. FR-900482 belonging to the genus Streptomyces
The substance-producing strain is cultured in a medium, and the resulting culture is extracted with the compound of the formula: By isolating and collecting the FR-900482 substance shown in
-900482 Methods for obtaining substances. [Claim 16] Formula: wherein R ¹ is a formyl group, a protected formyl group, a hydroxymethyl group, a protected hydroxymethyl group, an arylaminomethyl group, a carboxy group, a protected carboxy group or a substituted iminomethyl group; R ² is a hydroxy group;
^R3 is hydrogen and ^R4 is a methyl group, a hydroxymethyl group or a protected hydroxymethyl group; or ^R3
and ^R4 bond together to form a methylene group or an oxo group, and ^R5 represents a hydroxy group, an alkoxy group, or a protected hydroxy group], to obtain a compound represented by the formula: [wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are as defined above, and R ⁶ _a represents an imino-protecting group] or a salt thereof. [Claim 17] Formula: [wherein R ¹ , R ³ , R ⁴ and R ⁵ are the same as above, R ⁶
represents hydrogen, an imino-protecting group, or an alkyl group], to obtain a compound represented by the formula: [wherein R ¹ , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined above, and ▲R ² _a ▼ represents a protected hydroxy group] or a salt thereof. [Claim 18] Formula: [wherein R ¹ , R ² , R ³ , R ⁴ and R ⁶ are as defined above] or a salt thereof, to obtain a compound represented by the formula: [wherein R ¹ , R ² , R ³ , R ⁴ and R ⁶ are as defined above, and ▲R ⁵ _a ▼ represents a protected hydroxy group] or a salt thereof. [Claim 19] Formula: wherein R ¹ , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined above, or a salt thereof, is reacted with an alkylating agent to obtain a compound represented by the formula: [wherein R ¹ , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined above, and ▲R ² _b ▼ represents an alkoxy group] or a salt thereof. [Claim 20] Formula: wherein R ¹ , R ² _a , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined above, or a salt thereof, is hydrolyzed to obtain a compound represented by the formula: [wherein R ¹ , R ³ , R ⁴ , R ⁵ and R ⁶ are as defined above] or a salt thereof.