JP2000169495A - Glycolipid, and production and use of the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound having antibacterial action on fungi, especially on Aspergillus fumigatus and Aspergillus niger, and useful as a therapeutic agent for fungus infection. SOLUTION: This new compound is expressed by the formula [wherein R1-R9 are each (a substituted) hydroxyl group; R10 is an (esterinfied or antidified) carboxyl; and (n) is 0 or 1]. The compound is obtained e.g. by culturing a microorganism (e.g. Cryptococcus sp. AFB-8 strain) belonging to Cryptococcus capable of producing the compound of the formula in culture medium to form and accumulate the compound of the formula, and by collecting it.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to novel glycolipids which have an antibacterial effect on fungi, especially Aspergillus fumigatus and Aspergillus niger, and are useful as therapeutic agents for fungal infections, and It relates to the manufacturing method.

[0002]

[0002] Fungal infections are classified into local infections such as athlete's foot and systemic infections such as cryptococcosis, candidiasis and aspergillosis. It is known that the latter is so deep that the pathogenic fungus infects and proliferates deep into the body, so that even if an antibacterial agent is administered, it is not easily cured and often causes a fatal condition. In addition, patients with a weakened immune system, such as patients using immunosuppressants for organ transplant surgery and AIDS patients, are more susceptible to infection,
The number of patients is increasing worldwide. Conventionally, amphotericin B, an antibiotic derived from actinomycetes, and azole-based chemically synthesized antibacterial agents such as fluconazole and itraconazole have been generally used as therapeutic agents for systemic fungal infections. However, amphotericin B is known to have side effects such as renal and hepatotoxicity, and a low dose has a low healing effect. Azole-based chemically synthesized antibacterial agents have been widely used because they have few side effects and are effective against various fungal infections, but the emergence of resistant bacteria has been reported, and long-term continuous administration treatment will be refrained It is pointed out that it should be.

[0003]

In the future, there is a demand for a substance which has less side effects on the human body and which has a different mechanism of action or a new chemical structure from conventional antifungal agents.
The present invention provides a medicament that can be expected to have great clinical utility against fungal infections.

[0004]

DISCLOSURE OF THE INVENTION In view of the above situation, the present inventors have sought out the source of a therapeutic agent useful for systemic fungal infections in microbial metabolites. As a result of diligent search, it was found that a novel glycolipid having antibacterial activity against fungi, particularly Aspergillus fumigatus and Aspergillus niger, was produced from a culture of AFB-8 belonging to the genus Cryptococcus isolated from soil. . The present inventors have further studied based on these findings and completed the present invention. That is, the present invention provides the following equation (1).

Embedded image [In the formula, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ represent an optionally substituted hydroxyl group, and R ¹⁰ represents an esterified or amidated group. And n represents 0 or 1), or a salt thereof, (2) R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷
There compounds of (1), wherein the hydroxy group which may be substituted with an acyl group, (3) R ⁸ and R ⁹ is a hydroxyl group (1) above the compounds described, (4) R ¹⁰ is a carboxyl group Certain compounds described in the above (1) and (5) are compounds of the formula

Embedded image Wherein R ²¹ represents a hydroxyl group which may be substituted by an acetyl group, n represents 0 or 1, and Ac represents an acetyl group. )
A pharmaceutical composition comprising the compound of the above (1),
(7) an antifungal agent characterized by containing the compound of (1), (8) an anti-aspergillus agent characterized by containing the compound of (1), (9) a genus Cryptocox And culturing a microorganism having the ability to produce the compound according to the above (5) in a medium, producing and accumulating the compound in a culture, and collecting the compound. The production method, (10) the production method according to (9), wherein the microorganism is Cryptocox sp. AFB-8 strain, and (11) Cryptocox sp. A having the ability to produce the compound according to (5).
FB-8 and the like. Further, when the compound (I) or a salt thereof contains an asymmetric carbon in the structure, both the optically active form and the racemic form are included in the scope of the present invention, and the compound (I) or a salt thereof may be a hydrate, an anhydrous form. Any of Japanese products may be used.

Hereinafter, the present invention will be described in detail. Formula (I)
Where R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R
Examples of the optionally substituted hydroxyl group represented by ⁹ include a hydroxyl group, an acyloxy group, and a hydroxyl group substituted with an optionally substituted hydrocarbon group. R ¹ ,
Examples of the “acyl group” of the “acyloxy group” of R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ include, for example, an optionally substituted alkanoyl group and a substituted Aroyl group, an optionally substituted heterocyclic carbonyl group,
An optionally substituted carbamoyl group, an optionally substituted thiocarbamoyl group, an optionally substituted alkylsulfonyl group, an optionally substituted arylsulfonyl group, an optionally substituted sulfamoyl group, And an optionally substituted aryloxycarbonyl group. Among them, for example, an alkanoyl group which may be substituted is preferable. As the “alkanoyl group” of the “optionally substituted alkanoyl group”, for example, a C _1-20 alkanoyl group (eg, formyl, acetyl,
Propionyl, isopropionyl, butyryl, pentanoyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, lauroyl, undecanoyl, myristoyl, palmitoyl, stearoyl, etc.) and the like, particularly C _1-6 alkanoyl group (e.g., formyl, acetyl, propionyl, iso Propionyl, butyryl,
Pentanoyl, hexanoyl and the like). “Aroyl group” of the “optionally substituted aroyl group”
Examples thereof include a C _7-16 aroyl group (for example, naphthoyl such as benzoyl, 1-naphthoyl and 2-naphthoyl) and the like. As the `` heterocyclic carbonyl group '' of the `` optionally substituted heterocyclic carbonyl group '', a heteroatom other than a carbon atom (for example, nitrogen, oxygen,
5- or 6-membered heterocyclic carbonyl group containing 1 to 4 sulfur atoms (eg, pyrrolylcarbonyl such as 3-pyrrolylcarbonyl, imidazolylcarbonyl such as 2-imidazolylcarbonyl, pyrazolylcarbonyl such as 1-pyrazolylcarbonyl, 3 Isothiazolylcarbonyl such as -isothiazolylcarbonyl, isoxazolylcarbonyl such as 3-isoxazolylcarbonyl, pyrazinylcarbonyl, pyrimidinylcarbonyl such as 2-pyrimidinylcarbonyl, and pyrimidinylcarbonyl such as 3-pyrazinylcarbonyl. Inridinylcarbonyl such as radinylcarbonyl, 2-indolizinylcarbonyl, isoindolylcarbonyl such as 2-isoindolylcarbonyl, 1
Indolylcarbonyl, such as indolylcarbonyl,
Furoyl such as 2-furoyl, thenoyl such as 2-thenoyl, nicotinyl, isonicotinyl, morpholinocarbonyl, piperidinocarbonyl, piperazinocarbonyl and the like).

[0005] The "optionally substituted carbamoyl group" includes a carbamoyl group, a mono-substituted carbamoyl group and a di-substituted carbamoyl group. Examples of the substituent include a C _1-6 alkyl group (eg, methyl, ethyl , Propyl, isopropyl, butyl, isobutyl, etc.), C _6-14
An aryl group (eg, phenyl, 1-naphthyl, 2-naphthyl, etc.), a C _7-16 aralkyl group (eg, phenyl-C _1-6 alkyl such as benzyl), a C _1-6 alkanoyl group (eg, acetyl, Propionyl, isopropionyl, butyryl, etc.), C _7-16 aroyl group (for example,
Benzoyl, 1-naphthoyl, 2-naphthoyl, etc.),
5- or 6-membered heterocyclic carbonyl group (for example, pyrrolylcarbonyl such as 3-pyrrolylcarbonyl, imidazolylcarbonyl such as 2-imidazolylcarbonyl, 1-
Pyrazolylcarbonyl such as pyrazolylcarbonyl, 3
-Isothiazolylcarbonyl such as -isothiazolylcarbonyl, isoxazolylcarbonyl such as 3-isoxazolylcarbonyl, pyrazinylcarbonyl, pyrimidinylcarbonyl such as 2-pyrimidinylcarbonyl,
Pyrazinylcarbonyl such as 3-pyrazinylcarbonyl, indolizinylcarbonyl such as 2-indolizinylcarbonyl, isoindolylcarbonyl such as 2-isoindolylcarbonyl, indolylcarbonyl such as 1-indolylcarbonyl, Furoyl such as 2-furoyl, thenoyl such as 2-thenoyl, nicotinyl, isonicotinyl, morpholinocarbonyl, piperidinocarbonyl, piperazinocarbonyl, etc.). Preferable examples of the “optionally substituted carbamoyl group” include, for example, a mono- or di-C _1-6 alkanoyl-carbamoyl group (eg, acetylcarbamoyl and the like). The `` optionally substituted thiocarbamoyl group '' includes a thiocarbamoyl group, a mono-substituted thiocarbamoyl group, a di-substituted thiocarbamoyl group,
Examples of the substituent include a C _1-6 alkyl group (eg, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, etc.) and a C _6-14 aryl group (eg, phenyl, 1-naphthyl, 2-naphthyl) , A C _7-16 aralkyl group (eg, benzyl etc.), a C _1-6 alkanoyl group (eg, acetyl, propionyl, isopropionyl, butyryl etc.), a C _7-16 aroyl group (eg,
Benzoyl, 1-naphthoyl, 2-naphthoyl, etc.),
5- or 6-membered heterocyclic carbonyl group (for example, pyrrolylcarbonyl such as 3-pyrrolylcarbonyl, imidazolylcarbonyl such as 2-imidazolylcarbonyl, 1-
Pyrazolylcarbonyl such as pyrazolylcarbonyl, 3
-Isothiazolylcarbonyl such as -isothiazolylcarbonyl, isoxazolylcarbonyl such as 3-isoxazolylcarbonyl, pyrazinylcarbonyl, pyrimidinylcarbonyl such as 2-pyrimidinylcarbonyl,
Pyrazinylcarbonyl such as 3-pyrazinylcarbonyl, indolizinylcarbonyl such as 2-indolizinylcarbonyl, isoindolylcarbonyl such as 2-isoindolylcarbonyl, indolylcarbonyl such as 1-indolylcarbonyl, Furoyl such as 2-furoyl, thenoyl such as 2-thenoyl, nicotinyl, isonicotinyl, morpholinocarbonyl, piperidinocarbonyl, piperazinocarbonyl, etc.). Preferred examples of the “optionally substituted thiocarbamoyl group” include, for example, a mono- or di-C _1-6 alkanoyl-thiocarbamoyl group (eg, acetylthiocarbamoyl).

The "alkylsulfonyl group" of the "optionally substituted alkylsulfonyl group" includes, for example, a C _1-20 alkylsulfonyl group (eg, methylsulfonyl, ethylsulfonyl, propylsulfonyl, isopropylsulfonyl, etc.). And a C _1-6 alkylsulfonyl group (eg, methylsulfonyl, ethylsulfonyl, propylsulfonyl, etc.) is particularly preferable. The "optionally substituted arylsulfonyl group"
Examples of the “arylsulfonyl group” include a C _6-14 arylsulfonyl group (for example, benzenesulfonyl,
-Naphthylsulfonyl, 2-naphthylsulfonyl, etc.)
And the like. The “optionally substituted sulfamoyl group” includes a sulfamoyl group, a mono-substituted sulfamoyl group, a di-substituted sulfamoyl group and the like, and examples of the substituent include a C _1-6 alkyl group (eg, methyl, ethyl, propyl , Isopropyl, butyl, isobutyl, etc.), C _6-14 aryl group (for example, phenyl, 1
- naphthyl, 2-naphthyl), C _7-16 aralkyl group (e.g., phenyl -C _1-6 alkyl such as benzyl) and the like. Examples of the “alkoxycarbonyl group” of the “optionally substituted alkoxycarbonyl group” include a C _1-20 alkoxy-carbonyl group (for example, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, Butoxycarbonyl and the like, and particularly a C _1-6 alkoxy-carbonyl group (for example,
Methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl and the like). The “aryloxycarbonyl group” of the “optionally substituted aryloxycarbonyl group” is a C _6-14 aryloxy-carbonyl group (for example, phenoxycarbonyl, 1-naphthyloxycarbonyl, 2-naphthyloxycarbonyl and the like) ).

As the "hydrocarbon group" of the "optionally substituted hydrocarbon group", for example, an aliphatic hydrocarbon group or a cyclic hydrocarbon group is used. Examples of the “aliphatic hydrocarbon group” include an aliphatic hydrocarbon group having 1 to 20 carbon atoms (eg, an alkyl group, an alkenyl group, an alkynyl group, etc.). Examples of the “cyclic hydrocarbon group” include a cyclic hydrocarbon group having 3 to 20 carbon atoms (eg, a cycloalkyl group, a cycloalkenyl group, an aryl group, etc.). The “alkyl group” includes, for example, methyl, ethyl, propyl, 2-propyl, 1-ethylpropyl, butyl, 1-methylpropyl, 2-methylpropyl, 1,1-dimethylethyl,
Preferred are C _1-10 alkyl groups such as 1,1-dimethylbutyl, 2,2-dimethylbutyl, pentyl, 3-methylbutyl, 2,2-dimethylpropyl, and hexyl.
The "alkenyl group" is, for example, ethenyl, 2-propenyl, 1-methylethenyl, butenyl, 2-methyl-1-
Propenyl, 2-methyl-2-propenyl, 1-methyl-2-propenyl, 2-pentenyl, 3-pentenyl,
4-pentenyl, 2-hexenyl, 3-hexenyl, 5
_C2-10 alkenyl groups such as -hexenyl and the like are preferable. The "alkynyl group" includes, for example, ethynyl, 2-propynyl, 2-butyn-1-yl, 3-butyn-2-yl, 1-pentyn-3-yl, 3-pentyn-1-yl, 4-pentyn- Preferred are C _2-10 alkynyl groups such as 2-yl and 3-hexyn-1-yl. The “cycloalkyl group” is preferably a C _3-10 cycloalkyl group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like. The “cycloalkenyl group” is preferably a C _3-10 cycloalkenyl group such as cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl and the like. The "aryl group"
Is preferably a C _6-14 aryl group such as phenyl, 1-naphthyl and 2-naphthyl.

R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸
And “acyl group” of “hydrocarbon group” and “acyloxy group” in “hydroxyl group substituted with optionally substituted hydrocarbon group” as an example of “optionally substituted hydroxyl group” for R ⁹ In the above, the alkanoyl group, aroyl group, heterocyclic carbonyl group, alkylsulfonyl group, arylsulfonyl group, alkoxycarbonyl group, aryloxycarbonyl group may have a substituent which achieves the object of the present invention. As long as they are not particularly limited, they may be the same or different, for example, an amino group,
A mono- or di-C _1-6 alkylamino group (eg, methylamino, ethylamino, propylamino, isopropylamino, dimethylamino, diethylamino, etc.),
A mono- or di-C _6-10 arylamino group (eg, phenylamino, diphenylamino, etc.), a mono- or di-C _7-11 aralkylamino group (eg, phenyl-C _1-5 alkylamino such as benzylamino, etc.) , Dibenzylamino and the like, di (phenyl-C _1-5 alkyl) amino and the like), azide group, nitro group, halogen (eg, fluorine, chlorine, bromine, iodine and the like), hydroxyl group, C
_1-6 alkoxy group (eg, methoxy, ethoxy, propoxy, isopropoxy, butoxy, etc.), C _6-10 aryloxy group (eg, phenoxy, 1-naphthyloxy, 2-naphthyloxy, etc.), C _7-11 aralkyl An oxy group (eg, benzyloxy), a formyloxy group, a C _1-6 alkyl-carbonyloxy group (eg, acetoxy, propionyloxy, etc.), a C _6-10 aryl-carbonyloxy group (eg, benzoyloxy, etc.), C _7-11 aralkyl-carbonyloxy group (for example, phenyl-C _1-5 such as benzylcarbonyloxy)
Alkyl-carbonyloxy, etc.), sulfonyloxy group, C _1-6 alkylsulfonyloxy group (eg, methylsulfonyloxy), mercapto group, C _1-6 alkylthio group (eg, methylthio, ethylthio, propylthio, isopropylthio, etc.) , A C _6-10 arylthio group (eg, phenylthio, 1-naphthylthio, 2-naphthylthio, etc.), a C _7-11 aralkylthio group (eg,
A phenyl-C _1-5 alkylthio such as benzylthio), a phosphonooxy group, a cyano group, a carbamoyl group,
Mono- or di-C _1-6 alkylcarbamoyl group (for example, methylcarbamoyl, ethylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, etc.), mono-
Or a di-C _6-10 arylcarbamoyl group (eg, phenylcarbamoyl, diphenylcarbamoyl, etc.),
Mono- or di-C _7-11 aralkylcarbamoyl group (eg, phenyl-C _1-5 alkyl-carbamoyl such as benzylcarbamoyl, di (phenyl-C _1-5 alkyl) -carbamoyl such as dibenzylcarbamoyl), carboxyl Groups, C _1-6 alkoxy-carbonyl groups (eg, methoxycarbonyl, ethoxycarbonyl, etc.), C _6-10 aryloxy-carbonyl groups (eg, phenoxycarbonyl, 1-naphthyloxycarbonyl, 2-naphthyloxycarbonyl, etc.), C _7-11 aralkyloxy-carbonyl group (for example, phenyl-C _1-5 alkyloxy-
Carbonyl, etc.), formyl group, C _1-6 alkyl-carbonyl group (eg, acetyl, propionyl, isopropionyl, butyryl, pentanoyl, hexanoyl, etc.), C _6-10 aryl-carbonyl group (eg, benzoyl, 1-naphthoyl, 2-naphthoyl, etc.), C _7-11
An aralkyl-carbonyl group (eg, phenyl-C _1-5 alkyl-carbonyl such as benzylcarbonyl), a sulfo group, a C _1-6 alkylsulfinyl group (eg, methylsulfinyl, ethylsulfinyl, etc.),
A C _6-10 arylsulfinyl group (eg, benzenesulfinyl, 1-naphthylsulfinyl, 2-naphthylsulfinyl, etc.), a C _1-6 alkylsulfonyl group (eg, methylsulfonyl, ethylsulfonyl, etc.),
_6-10 arylsulfonyl group (eg, benzenesulfonyl, 1-naphthylsulfonyl, 2-naphthylsulfonyl, etc.), C _1-6 alkyl group (eg, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, etc.) A C _2-6 alkenyl group (for example, vinyl, allyl, 2-butenyl and the like), a C _2-6 alkynyl group (for example,
Ethynyl, propargyl, etc.), C _3-6 cycloalkyl group (eg, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.), C _3-6 cycloalkenyl group (eg, cyclobutenyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, etc.) , C _6-10
An aryl group (eg, phenyl, 1-naphthyl, 2-naphthyl, etc.), a monocyclic to tricyclic heterocyclic group (eg, a 5- or 6-membered Monocyclic, bicyclic or tricyclic heterocyclic groups: pyridyl, pyrazyl, pyrimidyl, quinolyl, isoquinolyl, indolyl, isoindolyl, indazolyl, pyridazinyl, imidazolyl, pyrazolyl, pyrrolyl, furyl, benzofuranyl, thienyl, benzothienyl, benzimidazolyl , Quinazolyl, pyrrolidinyl,
Pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, indolizyl, isoindolinidyl, morpholinyl, etc., a monocyclic to tricyclic heterocyclic thio group (for example, a group in which a thio group is bonded to the heterocyclic group, Specifically, pyridylthio such as 4-pyridylthio, pyrimidylthio such as 2-pyrimidylthio, thiadiazolylthio such as 1,3,4-thiadiazol-2-ylthio, tetrazolylthio such as 5-tetrazolylthio, 2-benzothiazolylthio and the like Benzothiazolylthio, quinolylthio such as 8-quinolylthio) and the like. These substituents are the aforementioned “hydrocarbon group” and “alkanoyl group, aroyl group, heterocyclic carbonyl group, alkylsulfonyl group,
The substituent is substituted on the "arylsulfonyl group, alkoxycarbonyl group, aryloxycarbonyl group" in a chemically acceptable range, and the number of the substituents is 1 to 5, preferably 1 to 3. However, when the number of substituents is two or more, they may be the same or different. These substituents, if chemically permissible, may further include an amino group, a mono- or di-C _1-6 alkylamino group, a nitro group, a halogen, a hydroxyl group, a C _1-6 alkoxy group,
C _1-6 alkyl-carbonyloxy group, sulfonyloxy group, C _1-6 alkylsulfonyloxy group, mercapto group, C _1-6 alkylthio group, phosphonooxy group, cyano group, carbamoyl group, mono- or di-C _{1- 6} alkylcarbamoyl group, carboxyl group, C _1-6 alkoxy-
It may be substituted with 1 to 3 substituents selected from a carbonyl group, a formyl group, a C _1-6 alkyl-carbonyl group, a sulfo group, a C _1-6 alkylsulfinyl group and the like.

In the formula (I), in the “carboxyl group which may be esterified or amidated” represented by R ¹⁰ ,
As the “esterified carboxyl group”, for example, those that are pharmacologically acceptable or those that change to pharmacologically acceptable for the first time in vivo are used. As the esterified carboxyl group, for example, a group represented by the formula —COOR ¹¹ [wherein, R ¹¹ represents a hydrocarbon group which may be substituted] is preferable. The hydrocarbon group represented by R ¹¹ is, like is used the same as the hydrocarbon group described above. Such a hydrocarbon group may have, at a substitutable position, one to three substituents similar to the above-described substituents in the hydrocarbon group. In the formula (I), in the “carboxyl group which may be esterified or amidated” represented by R ¹⁰ , the “amidated carboxyl group” includes, for example, those which are pharmacologically acceptable, Alternatively, a substance that changes to a pharmacologically acceptable substance for the first time in a living body is used. As the amidated carboxyl group, for example,
Wherein, R ¹² and R ¹³ are the same or different and each is a hydrogen atom or an optionally substituted hydrocarbon group] formula -CON (R ¹²⁾ R ¹³ and a group represented by are preferred. R
^{As the} optionally substituted hydrocarbon groups represented by ¹² and R ¹³ , those similar to R ¹¹ are used. In the formula (I), n
Represents 0 or 1. In the formula (I), R ¹ , R ² , R ³ ,
R ⁴ , R ⁵ , R ⁶ and R ⁷ are preferably a hydroxyl group optionally substituted with an acyl group, and particularly preferably a hydroxyl group optionally substituted with an acetyl group. Formula (I)
Wherein R ⁸ and R ⁹ are preferably a hydroxyl group. Formula (I)
Wherein R ¹⁰ is preferably a carboxyl group.

Next, a method for producing the compound according to the present invention will be described. In the formula (I), R ¹ , R ⁶ , R ⁷ , R ⁸ and R ⁹ are a hydroxyl group, R ² is a hydroxyl group optionally substituted by an acetyl group, R ³ , R ⁴ and R ⁵ are an acetoxy group, and n is 0
Or the compound AFB-8-A ₁ , A ₂ , B ₁ and B _{2 which is 1} (hereinafter sometimes referred to as “AFB-8s”)
Can be produced by using a microorganism. The microorganism used in this production method belongs to the genus Cryptococcus,
Any microorganism may be used as long as it is capable of producing AFB-8s. An example is the AFB-8 strain newly isolated from the soil in Okinawa Prefecture.

This strain has the following properties. 1. Morphological characteristics AFB-8 strain is a yeast extract-malt extract agar plate (YMaga
r) on top of a circle with a smooth or finely wrinkled surface
Form grayish white colonies. Cells vary in shape and size, exhibiting spheres, lemons, ellipses, oblongs, and are (3-5) × (5-20) μm. Proliferates by germination, and its germination base is relatively wide. It germinates mainly from the long axis poles, but also germinates from the sides. Some cells have a capsule. In the YM liquid medium, a thick pellicle is formed on the liquid surface, and a large amount of precipitate is formed on the bottom. The liquid part is slightly turbid. On corn meal agar, it forms a smooth surface, milky colony. The hypha spreads from the outer edge of the colony like a root and extends into the agar. Cells include oval, oblong, and lemon-shaped cells, pseudohypha in which cells that are slightly elongated are linked, and true but undeveloped, branched mycelium (true hyph).
a) occurs. Clasp connection
Is not seen. When cultured on malt extract agar medium (Malt agar), the shape of colonies and cells is similar to that of YM agar medium culture, but oil droplets (oil dr
op). Yeast extract-malt extract agar medium, malt extract agar medium and cornmeal agar medium 24. C,
After culturing for 6 weeks, no formation of ascospores, ejected spores, and organs indicating the presence of sexual generation is observed in any medium. 2) Physiological properties 28. The physiological characteristics of the AFB-8 strain cultured in C and observed for 2 to 28 days are shown in Table 1. The test method and medium are described in Kurtzmann and Fell, The East, 4th Edition (1998
(CP Kurtzman and JW Fell, The Yeasts, A
Taxonomic Study, 4th edition).

[0012]

[Table 1]

From the above bacteriological properties, it has been revealed that the present strain is a yeast belonging to the genus Cryptococcus, among the anamorphic yeasts of the basidiomycetes. Kurtzman and JW Fell, The Yea, The East, 4th Edition (1998)
sts, A Taxonomic Study, 4th edition), compared with the genus Cryptocox and its related genera.
ccus humicolus). However, Cryptocox fumikols form yellow colonies with deep wrinkles on malt extract agar,
On YM agar, it was different from this strain in that it formed elongated cylindrical cells. In asexual yeast,
The morphological features have been identified as one of the key differentiators for identification, and AFB-8 appears to be a new strain different from Cryptococcus humichorus, but other strains that clearly distinguish the species Since no properties were found, the strain was identified as Cryptococcus sp.

This strain was obtained from the Fermentation Research Institute (IFO) on October 22, 1998 under the accession number IFO10934.
And deposited with the Ministry of International Trade and Industry of Japan on November 24, 1998 under the accession number FERM BP-6581 under the Budapest Treaty at the National Institute of Advanced Industrial Science and Technology (NIBH). Cryptocox spp. Can be mutated naturally or by a mutagen as a general property of microorganisms. For example, irradiation with radiation such as X-rays, gamma rays, and ultraviolet rays, treatment with various drugs or culturing on a medium containing the drugs, or a number of mutant strains obtained by mutation by other means, or naturally obtained strains Any mutant strain having the property of producing the compound AFB-8s can be used in the method of the present invention.

The medium used for culturing in the method of the present invention may be liquid or solid as long as it contains a nutrient that can be used by the strain used, but it is more appropriate to use a liquid medium when treating in large quantities. is there. The medium is appropriately mixed with a carbon source, a nitrogen source, an inorganic substance, and a micronutrient which can be assimilated by the compound-producing bacterium. As a carbon source, for example, glucose,
Lactose, sucrose, maltose, dextrin, starch, glycerin, mannitol, sorbitol, fats and oils (such as cottonseed oil, soybean oil, lard oil, and chicken oil), n-paraffin, and the like. Examples of nitrogen sources include meat extract and yeast. extract,
Malt extract, dried yeast, soy flour, corn steep,
Liquor, peptone, raw soy flour, cottonseed flour, tomato paste, peanut meal, molasses, urea, ammonium salts (ammonium sulfate, ammonium chloride, ammonium nitrate, ammonium acetate, etc.) are used. further,
Salts containing sodium, potassium, calcium, magnesium and the like, metal salts such as iron, manganese, zinc, cobalt and nickel, salts such as phosphoric acid, boric acid and sulfuric acid and salts of organic acids such as acetic acid and propionic acid are appropriately used. Is also good. In addition, amino acids (glycine, glutamic acid, aspartic acid, alanine, lysine, methionine, proline, etc.), peptides (dipeptides, tripeptides, etc.),
Vitamins (B ₁ , B ₂ , nicotinic acid, B ₁₂ , C, etc.), nucleic acids (purines, pyrimidines, derivatives thereof, etc.) may be contained. Of course, an inorganic or organic acid or alkali, a buffer, or the like may be added for the purpose of adjusting the pH in the medium, or an appropriate amount of an oil or fat, a surfactant, or the like may be added for the purpose of defoaming. The culture method is static culture,
Means such as shaking culture or aeration-agitation culture may be used. For a large amount of treatment, it is desirable to use so-called deep aeration stirring culture. Culture conditions vary depending on the state and composition of the medium, the type of strain, and the means of culture.
It is preferable to select a temperature of about 30 to about 30 ° C. and an initial pH of about 4 to 7. In particular, the temperature in the middle stage of the culture is preferably about 25 ° C. to 30 ° C. and the initial pH is about 6 to 7. The cultivation time is not constant depending on the above conditions, but it is desirable to culture until the concentrations of various physiologically active substances are maximized. The time required for this is usually about 1 to 5 days in the case of shaking culture using a liquid medium or aeration and stirring culture.

By culturing as described above, compounds AFB-8 described below are produced and accumulated, and can be extracted and purified from the culture according to their chemical properties. The method for collecting the desired compound AFB-8 from the culture is described below. Since the compound is fat-soluble, general means utilizing this property may be employed. First, an organic solvent that is immiscible with the culture or culture filtrate with water, for example, a halogenated hydrocarbon such as dichloromethane, an ester such as ethyl acetate, a ketone such as methyl isobutyl ketone, or an alcohol such as isobutanol, AF
Extract B-8 class. The obtained organic solvent layer is washed with water and concentrated to obtain a crude substance containing AFB-8s. Various well-known chromatographic methods are advantageously used to further purify the crude material to obtain pure AFB-8s. As the carrier, those utilizing the difference in the adsorptivity of compounds such as activated carbon, silica gel, microcrystalline cellulose, and adsorptive resin, and those utilizing the difference in the molecular weight of the compounds such as molecular sieve resins are advantageously used. In order to elute the target compound from these carriers, combinations vary depending on the type and properties of the carrier, but suitable organic solvents (eg, halogenated hydrocarbons such as dichloromethane and chloroform, aliphatic hydrocarbons such as hexane, etc.) , Aromatic hydrocarbons such as toluene, esters such as ethyl acetate, ketones such as acetone, alcohols such as methanol, ethanol, isopropyl alcohol and isobutanol, and nitriles such as acetonitrile). Organic acid (eg, acetic acid, formic acid, etc.), aqueous solution [eg, water, alkali (eg, sodium hydroxide, potassium hydroxide, sodium hydrogen carbonate, etc.)-Containing aqueous solution, acid (eg, hydrochloric acid, acetic acid, formic acid, phosphoric acid, etc.) ) -Containing aqueous solution, salt-containing aqueous solution (eg, saline, acetate buffer, phosphate buffer, etc.)] or a mixed solvent of an appropriate ratio Used. More specifically, activated carbon for chromatography (Takeda Pharmaceutical Co., Ltd.), Kieselgel 60 (Merck, Germany), microcrystalline cellulose [eg, Avicel (Asahi Kasei Co., Ltd.), Funacell (Funakoshi Co., Ltd.) as carriers.
Adsorbent resin [eg, Diaion HP-20, Sepabeads SP-207 or SP-850 (manufactured by Mitsubishi Chemical Corporation), Amberlite XAD-I or II (manufactured by Rohm and Haas Company, USA), etc.] A molecular sieve resin [eg, Sephadex LH-20 (Pharmacia,
Sweden) etc. are advantageously used. In the present invention, when the target compound is purified, a preparative high performance liquid chromatography (HPLC) method is also advantageously used. When this method is applied, octadecylsilane (ODS) -based, polymer-based and silica gel-based carriers are advantageously used. For example, in the case of ODS,
When YMC gel (YMSC) or TSK gel (Tosoh) is a polymer type, ODP (manufactured by Asahi Kasei Corporation) having an octadecyl group introduced into the polymer or NH2P (manufactured by Asahi Kasei Corporation) having a polyamine introduced into the polymer. As the mobile phase, water, an acid-containing aqueous solution, a salt-containing aqueous solution, a mixed solution of methanol, acetonitrile, or the like alone or in an appropriate ratio is advantageously used. Further, in the present invention, crystallization is advantageously used when purifying the target compound. As a solvent used for crystallization, for example, chloroform, ethyl acetate, ether,
A single solution or a mixed solution of an appropriate ratio such as hexane is used.

The physicochemical properties of AFB-8s obtained in Example 2 described below are shown below. AFB-8-A ₁ 1) Appearance: white powder 2) Molecular weight: FAB-mass spectrum; m / z 823 (M
-H) ^- 3) Elemental analysis: (%) (calculated as water 1 mol) Calculated; C, 54.15; H, 7.89 Found; C, 54.43; H, 7.63 4) Molecular formula: C ₃₈ H ₆₄ O ₁₉ 5) UV spectrum: in methanol; terminal absorption 6) IR spectrum: shows the main absorption in the KBr tablet (wave number, cm ^-1 ). 3430, 2920, 2850, 1750, 1370, 1250, 1230, 1040 7) ¹³ C NMR spectrum: signals at δ ppm or less in 75 Mz, DMSO-d ₆ (FIG. 1) 175.9, 170.2, 169.5, 169.1, 169.0, 102.9, 99.4, 8
0.8, 74.2, 73.5, 73.0, 72.5, 71.1, 69.6, 68.9, 68.
4, 62.9, 59.9, 33.9, 33.0, 29.1, 29.0, 28.9,28.8,
24.9, 24.6, 20.5, 20.3, 20.2, 20.1 8) Color reaction: positive; phosphomolybdic acid, Maurish negative; Dragendorf, ninhydrin 9) High-performance liquid chromatography (HPLC): column; YMC-Pack A-312 , ODS, 6.0 x 150 mm (YYMSHI) Mobile phase; 50% (v / v) acetonitrile / 0.05% aqueous trifluoroacetic acid Flow rate; 1.0 ml / min Detection method; 214 nm retention time; 5 minutes 10) Thin layer chromatography (TLC): carrier; silica gel 60F ₂₅₄ (Merck, Germany) Developing solvent: chloroform / methanol / acetic acid (16:
4: 1) Rf value; 0.31

AFB-8-A ₂ 1) Appearance: white powder 2) Specific rotation: -12 ° (c 0.52, methanol, 2
3) Molecular weight: FAB-mass spectrum; m / z 781 (M
-H) ^- 4) Elemental analysis: (%) (calculated as water 2 mol) Calculated; C, 52.80; H, 8.12 Found; C, 52.86; H, 7.69 5) Molecular formula: C ₃₆ H ₆₂ O ₁₈ 6) UV spectrum: in methanol; terminal absorption 7) IR spectrum: showing the main absorption in KBr tablets (wave number, cm ^-1 ). 3390, 2920, 2850, 1740, 1370, 1250, 1050 8) ¹³ C NMR spectrum: signals of δ ppm or less are observed in 75 Mz, DMSO-d ₆ (FIG. 2) 175.8, 170.2, 169.6, 169.1, 102.9, 99.7, 80.9, 76.
3, 75.2, 74.3, 74.1, 73.0, 71.6, 71.2, 69.5, 68.9,
67.6, 63.0, 60.4, 33.8, 33.0, 29.1, 29.0, 28.9, 2
8.7, 24.9, 24.6, 20.6, 20.5, 20.2 9) Color reaction: positive; phosphomolybdic acid, Maurish negative; Dragendorf, ninhydrin 10) High performance liquid chromatography (HPLC): column; YMC-Pack A-312 , ODS, 6.0 x 150 mm (manufactured by YMC) Mobile phase; 50% (v / v) acetonitrile / 0.05% aqueous trifluoroacetic acid Flow rate; 1.0 ml / min Detection method; 214 nm retention time; 9 minutes 11) thin layer chromatography (TLC): support: silica gel 60F ₂₅₄ (Merck, Germany) Developing solvent: chloroform / methanol / acetic acid (16:
4: 1) Rf value; 0.13

AFB-8-B ₁ 1) Appearance: white powder 2) Molecular weight: FAB-mass spectrum; m / z 779 (M
-H) ^- 3) Elemental analysis: (%) (calculated as water 2 mol) Calculated; C, 54.12; H, 7.82 Found; C, 54.40; H, 7.69 4) Molecular formula: C ₃₆ H ₆₀ O ₁₈ 5) UV spectrum: in methanol; terminal absorption 6) IR spectrum: shows the main absorption in the KBr tablet (wave number, cm ^-1 ). 3450, 2920, 2850, 1750, 1370, 1250, 1230, 1040 7) ¹³ C NMR spectrum: signals at δ ppm or less in 75 Mz, DMSO-d ₆ (FIG. 3) 175.8, 170.2, 169.5, 169.2, 169.0, 102.4, 99.5, 8
0.9, 74.5, 73.5, 72.9, 72.5, 71.1, 69.5, 68.8, 68.
4, 62.9, 59.9, 33.8, 29.2, 29.0, 28.9, 28.8,28.7,
25.4, 24.6, 20.5, 20.3, 20.2, 20.1 8) Color reaction: positive; phosphomolybdic acid, Maurish negative; Dragendorf, ninhydrin 9) High performance liquid chromatography (HPLC): column; YMC-Pack A-312 13. ODS, 6.0 x 150 mm (YYMSHI) Mobile phase; 50% (v / v) acetonitrile / 0.05% aqueous trifluoroacetic acid Flow rate; 1.0 ml / min Detection method; 214 nm retention time; 8 minutes 10) thin layer chromatography (TLC): support: silica gel 60F ₂₅₄ (Merck, Germany) Developing solvent: chloroform / methanol / acetic acid (16:
4: 1) Rf value; 0.41

AFB-8-B ₂ 1) Appearance: white powder 2) Specific rotation: -15 ° (c 0.66, methanol, 2
1) 3) Molecular weight: FAB-mass spectrum; m / z 737 (M
-H) ^- 4) Elemental analysis: (%) (calculated as water 2 mol) Calculated; C, 52.70; H, 8.07 Found; C, 52.65; H, 7.69 5) Molecular formula: C ₃₄ H ₅₈ O ₁₇ 6) UV spectrum: in methanol; terminal absorption 7) IR spectrum: showing the main absorption in KBr tablets (wave number, cm ^-1 ). 3430, 2920, 2850, 1750, 1370, 1250, 1040 8) ¹³ C NMR spectrum: signals at δ ppm or less in 75 Mz, DMSO-d ₆ (FIG. 4) 175.8, 170.2, 169.6, 169.1, 102.4, 99.7, 81.0, 76.
3, 75.2, 74.7, 72.9, 71.6, 71.1, 69.5, 68.8, 67.6,
63.0, 60.4, 33.8, 29.2, 29.0, 28.9, 28.8, 28.7, 2
5.4, 24.6, 20.6, 20.5, 20.2 9) Color reaction: positive; phosphomolybdic acid, Maurish negative; Dragendorf, ninhydrin 10) High performance liquid chromatography (HPLC): column; YMC-Pack A-312, ODS , 6.0 x 150 mm (manufactured by YMC) Mobile phase; 50% (v / v) acetonitrile / 0.05% trifluoroacetic acid aqueous solution; 1.0 ml / min Detection method; 214 nm retention time; 9.0 minutes 11) Thin layer chromatography (TLC): carrier; silica gel 60F ₂₅₄ (Merck, Germany) Developing solvent: chloroform / methanol / acetic acid (16:
4: 1) Rf value; 0.20

The structural formula of AFB-8 is as follows.

Embedded image

In the formula (I), R ¹ , R ² , R ³ , R ⁴ , R
Compounds or salts thereof in which ⁵ , R ⁶ , R ⁷ , R ⁸ or R ⁹ is a hydroxyl group substituted with a hydrocarbon group can be used, for example, in formula (I) for R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ or R ⁹ can be produced by reacting a compound having a hydroxyl group or a salt thereof with a compound having a leaving group in the presence of a base. In the reaction, protection of a functional group that should not be involved in the reaction of the raw material, elimination of the protective group, and the like can be appropriately selected from means known per se. The compound having a leaving group refers to a compound having a functional group which is easily substituted by a chemical reaction, and specifically includes a halide (eg, methyl iodide, ethyl iodide, propyl iodide,
Isopropyl iodide, butyl iodide, pentyl iodide,
Allyl bromide, benzyl bromide, etc.), sulfonates (eg, methyl methanesulfonate, methyl p-toluenesulfonate, ethyl p-toluenesulfonate, methyl benzenesulfonate, methyl trifluoromethanesulfonate, trifluoromethanesulfone) Ethyl sulfate and the like, and sulfates (eg, dimethyl sulfate, diethyl sulfate, and the like). As the base, for example, an alkali metal hydride (eg,
Sodium hydride, potassium hydride, etc.), alkaline earth metal hydrides (eg, calcium hydride, etc.), alkali metal alkoxides (eg, sodium methoxide, sodium ethoxide, etc.), alkali metal hydroxides (eg, , Sodium hydroxide, potassium hydroxide, etc.), alkali metal carbonates (eg, sodium carbonate, potassium carbonate, sodium bicarbonate, etc.), alkaline earth metal hydroxides (eg, calcium hydroxide, etc.), tertiary Amine (eg, trimethylamine, triethylamine, tripropylamine, N-methylpiperidine, N-methylpyrrolidine, cyclohexyldimethylamine, N-methylmorpholine, etc.) Secondary amine (eg, di-n-butylamine, diisobutylamine, dicyclohexylamine ), Aromatic bases (eg, pyridine,
4-dimethylaminopyridine, lutidine, collidine, etc.),
Alkyl lithium (eg, methyl lithium, butyl lithium, etc.) is used. This reaction is usually performed in a solvent. Examples of the solvent include amides (eg, formamide, N, N-dimethylformamide, N, N-dimethylacetamide, N-methylpyrrolidone, etc.), sulfoxides (eg,
Dimethyl sulfoxide, etc.), aromatic bases (eg, pyridine, etc.), halogenated hydrocarbons (eg, chloroform, dichloromethane, etc.), ethers (eg, diethyl ether, tetrahydrofuran, dioxane, etc.), nitriles (eg, acetonitrile) And the like (eg, ethyl acetate, ethyl formate, etc.), or a mixture of these at an appropriate ratio. The reaction temperature is not particularly limited as long as the reaction proceeds, but is usually from -70 ° C to 150 ° C.
C., preferably at -30.degree. C. to 80.degree. The reaction time varies depending on the starting materials, the base, the reaction temperature and the type of the solvent to be used, but the reaction is usually carried out for several tens to several tens of hours.

In the formula (I), R ¹ , R ² , R ³ , R ⁴ , R
^The compound or a salt thereof in which ⁵ , R ⁶ , R ⁷ , R ⁸ or R ⁹ is an acyloxy group is, for example, a compound represented by the formula (I) wherein R ¹ ,
It can be produced by subjecting a compound wherein R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ or R ⁹ is a hydroxyl group or a salt thereof to an acylation reaction. In the reaction, protection of a functional group that should not be involved in the reaction of the raw material, elimination of the protective group, and the like can be appropriately selected from means known per se.
The hydroxyl group can be acylated by reacting the starting compound with an acylating agent, for example, an organic acid (eg, organic carboxylic acid or the like) or a reactive derivative thereof in a solvent. As the reactive derivative of the organic acid, for example, an acid halide, an acid anhydride, an active ester and the like are used, and such a reactive derivative is specifically described as follows. 1) Acid halide As the acid halide, for example, acid chloride, acid bromide and the like are used. 2) Acid anhydride, mixed acid anhydride Here, examples of the acid anhydride include a mixture of an aliphatic carboxylic acid (eg, acetic acid, valeric acid, hexanoic acid, etc.) or an aromatic carboxylic acid (eg, benzoic acid, etc.) An acid anhydride or a symmetric acid anhydride is used. 3) Active ester Here, examples of the active ester include methyl ester,
Ester such as ethyl ester, methoxymethyl ester, propargyl ester, 4-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, and 1-hydroxy-1H-2-pyridone Esters with N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole (HOBT) and the like are used. When directly reacting with a carboxylic acid, for example, N, N'-dicyclohexylcarbodiimide (D
CC), condensing agents such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (WSC) and N, N'-diisopropylcarbodiimide (DIC). These condensing agents include the active esters described above, for example, 1-hydroxy-1H-2-
Pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole (HOB
It may be used for ester synthesis with T). The reaction may be carried out in the presence of a base. Examples of the base used include tertiary amines (eg, trimethylamine,
Triethylamine, tripropylamine, N-methylpiperidine, N-methylpyrrolidine, cyclohexyldimethylamine, N-methylmorpholine, etc., and secondary amines (eg,
-n-butylamine, diisobutylamine, dicyclohexylamine, etc.), aromatic bases (eg, pyridine, 4-dimethylaminopyridine, lutidine, collidine, etc.), hydroxides of alkali metals (eg, sodium hydroxide, potassium hydroxide, etc.) ), Alkali metal carbonates (eg, sodium carbonate,
Potassium carbonate, sodium hydrogen carbonate, etc.), hydroxides of alkaline earth metals (eg, calcium hydroxide, etc.) and the like are used. This reaction is usually performed in a solvent. Examples of the solvent include amides (eg, formamide, N, N-dimethylformamide, N, N-dimethylacetamide, N-methylpyrrolidone, etc.), sulfoxides (eg, dimethyl sulfoxide, etc.), aromatic bases (eg, , Pyridine, etc.), halogenated hydrocarbons (eg, chloroform, dichloromethane, etc.), ethers (eg, diethyl ether, tetrahydrofuran, dioxane, etc.), nitriles (eg, acetonitrile, etc.), esters (eg, ethyl acetate, Ethyl formate, etc.), or a mixture thereof at an appropriate ratio. The reaction temperature is not particularly limited as long as the reaction proceeds, but the reaction is usually carried out at about -50 ° C to 150 ° C, preferably at -30 ° C to 80 ° C. The reaction time varies depending on the starting materials, the base, the reaction temperature and the type of the solvent to be used, but the reaction is usually carried out for several tens to several tens of hours. General formula (I)
Wherein R ¹⁰ is a carboxyl group esterified or a salt thereof, for example, can be produced by subjecting a compound or a salt thereof wherein R ¹⁰ is a carboxyl group in general formula (I) to an esterification reaction. it can. The esterification reaction is performed by a method known per se, for example, the following method. (1) A starting compound is reacted with a diazoalkane (eg, diazomethane, phenyldiazomethane, diphenyldiazomethane, etc.). (2) reacting the starting compound with an activated alkyl halide (eg, methyl iodide, benzyl bromide, etc.); (3) The starting compound is reacted with an alcohol (eg, methanol, ethanol, benzyl alcohol, etc.) in the presence of an acid catalyst or a condensing agent. Examples of the acid catalyst include hydrochloric acid,
Examples of the condensing agent include sulfuric acid and camphorsulfonic acid, and N, N′-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (WSC), N, N′- Diisopropylcarbodiimide (D
IC) is used. (4) The starting compound is introduced into an active ester and reacted with an alcohol (eg, methanol, ethanol, benzyl alcohol, etc.). As the active ester, for example, esters with 1-hydroxy-1H-2-pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole and the like are used. (5) The starting compound is reacted with an acid halide (eg, ethyl chloroformate, benzyl chloroformate, etc.), led to an acid anhydride, and reacted with an alcohol (eg, methanol, ethanol, benzyl alcohol, etc.). The reaction may be carried out in the presence of a base. Examples of the base used include tertiary amines (eg, trimethylamine, triethylamine, tripropylamine, N-methylpiperidine, N-methylpyrrolidine, cyclohexyldimethylamine, -Methylmorpholine, etc.), secondary amines (eg, di-n-butylamine, diisobutylamine, dicyclohexylamine, etc.), aromatic bases (eg, pyridine,
Lutidine, collidine, etc.), alkali metal hydroxides (eg, sodium hydroxide, potassium hydroxide, etc.), alkali metal carbonates (eg, sodium carbonate, potassium carbonate,
Examples thereof include sodium hydrogen carbonate and the like, and hydroxides of alkaline earth metals (eg, calcium hydroxide and the like). This reaction is usually performed in a solvent. Examples of the solvent include amides (eg, formamide, N, N-dimethylformamide, dimethylacetamide, N-methylpyrrolidone, etc.),
Sulfoxides (eg, dimethyl sulfoxide, etc.), aromatic bases (eg, pyridine, etc.), halogenated hydrocarbons (eg, chloroform, dichloromethane, etc.), ethers (eg, diethyl ether, tetrahydrofuran, dioxane, etc.), nitrile (Eg, acetonitrile, etc.), esters (eg, ethyl acetate, ethyl formate, etc.), alcohols (eg, methanol, ethanol, etc.), or a mixture of these at an appropriate ratio. The reaction temperature is not particularly limited as long as the reaction proceeds, but is usually about-
50 ° C to 150 ° C, preferably about -30 ° C to 8 ° C
Performed at 0 ° C. The reaction time depends on the starting materials used, the base,
The reaction is usually performed for several tens of minutes to several tens of hours, depending on the reaction temperature and the type of the solvent. The compound of the formula (I) in which R ¹⁰ is an amidated carboxyl group or a salt thereof is obtained, for example, by subjecting a compound of the formula (I) in which R ¹⁰ is a carboxyl group or a salt thereof to an amidation reaction. Can be manufactured. The amidation reaction, for example,
A carboxylic acid or a salt thereof or a reactive derivative thereof, wherein R ¹⁰ is a carboxyl group in the general formula (I);
The reaction can be carried out by reacting with a compound represented by the general formula HNR ¹² R ¹³ (III) (wherein each symbol has the same meaning as described above) or a salt thereof. Examples of the reactive derivative of a carboxylic acid include, for example,
Acid halides, active esters and the like are used, and such reactive derivatives are specifically described as follows. (1) Acid halide As the acid halide, for example, acid chloride, acid bromide and the like are used. (2) Active ester Here, examples of the active ester include methyl ester, ethyl ester, methoxymethyl ester, propargyl ester, 4-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester,
Pentachlorophenyl ester, other esters such as mesylphenyl ester, 1-hydroxy-1H-2-pyridone,
Esters with N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole (HOBT) and the like are used. When directly reacting with a carboxylic acid, for example, N, N′-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride (WSC), N, N′-diisopropylcarbodiimide A condensing agent such as (DIC) is used. These condensing agents are useful for the synthesis of esters with the above-mentioned active esters such as 1-hydroxy-1H-2-pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole (HOBT), and the like. May be used. The reaction may be carried out in the presence of a base. Examples of the base used include tertiary amines (eg, trimethylamine, triethylamine, tripropylamine, N-
Methylpiperidine, N-methylpyrrolidine, cyclohexyldimethylamine, N-methylmorpholine, etc., secondary amines (eg, di-n-butylamine, diisobutylamine, dicyclohexylamine, etc.), aromatic bases (eg, pyridine, lutidine, collidine) ), Alkali metal hydroxides (eg, sodium hydroxide, potassium hydroxide, etc.), alkali metal carbonates (eg, sodium carbonate, potassium carbonate,
Examples thereof include sodium hydrogen carbonate and the like, and hydroxides of alkaline earth metals (eg, calcium hydroxide and the like). This reaction is carried out usually in a solvent using 0.5 to 10 molar equivalents of the compound represented by the general formula (III) with respect to the compound in which R ¹⁰ is a carboxyl group in the general formula (I). Examples of the solvent include amides (eg, formamide, N, N-dimethylformamide, dimethylacetamide, N-methylpyrrolidone, etc.), sulfoxides (eg, dimethylsulfoxide, etc.), aromatic bases (eg, pyridine, etc.) ), Halogenated hydrocarbons (eg, chloroform, dichloromethane, etc.), ethers (eg, diethyl ether,
Tetrahydrofuran, dioxane, etc.), nitriles (eg, acetonitrile, etc.), esters (eg, ethyl acetate, ethyl formate, etc.), and mixtures of these at an appropriate ratio are used. The reaction temperature is not particularly limited as long as the reaction proceeds, but is usually about -50 ° C to 15 ° C.
It is carried out at 0 ° C, preferably at about -30 ° C to 80 ° C.
The reaction time varies depending on the starting materials, the base, the reaction temperature, and the type of the solvent to be used. A compound or a salt thereof in which R ¹ is an acyloxy group and R ² is a hydroxyl group in the general formula (I) is, for example, a compound or a salt thereof in which R ¹ is a hydroxyl group and R ² is an acyloxy group in the general formula (I). It can be produced by subjecting to a transfer reaction under acidic conditions. Preferable examples of the acid used include, for example, organic acids (eg, formic acid, acetic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.) and inorganic acids (eg, hydrochloric acid, sulfuric acid, hydrobromic acid) Etc.). This reaction is usually performed in a solvent. Examples of the solvent include amides (eg, formamide, N, N-dimethylformamide, dimethylacetamide, N-methylpyrrolidone, etc.), sulfoxides (eg, dimethylsulfoxide, etc.), halogenated hydrocarbons (eg, chloroform,
Dichloromethane, etc.), ethers (eg, diethyl ether, tetrahydrofuran, dioxane, etc.), nitriles (eg, acetonitrile, etc.), esters (eg, ethyl acetate, ethyl formate, etc.), ketones (eg, acetone, methyl isobutyl ketone) And the like, or a mixture thereof at an appropriate ratio. The reaction temperature is not particularly limited as long as the reaction proceeds, but is usually about -50 ° C to 1 ° C.
It is carried out at 50 ° C, preferably at about 0 ° C to 120 ° C.
The reaction time varies depending on the starting material, acid, reaction temperature, and type of solvent used, but the reaction is usually carried out for tens of minutes to tens of hours.

In the general formula (I), R ¹ , R ² , R ³ ,
A compound wherein R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ or R ⁹ is a hydroxyl group or a salt thereof is, for example, a compound represented by the formula (I) wherein R ¹ , R ² ,
It can be produced by subjecting a compound or a salt thereof in which R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ or R ⁹ is an acyloxy group to a hydrolysis reaction under basic conditions. Preferred examples of the base used include, for example, alkali metal hydroxides (eg, sodium hydroxide, potassium hydroxide, etc.),
Examples thereof include alkali metal carbonates (eg, sodium carbonate, potassium carbonate, sodium hydrogen carbonate, etc.) and alkaline earth metal hydroxides (eg, calcium hydroxide, etc.).
This reaction is generally performed in an aqueous solution or a solvent containing water. Examples of the solvent include alcohols (eg, methanol, ethanol, etc.), amides (eg, formamide, N, N-dimethylformamide, dimethylacetamide, N-methylpyrrolidone, etc.), sulfoxides (eg, dimethyl sulfoxide, etc.) , Ethers (eg, tetrahydrofuran, dioxane, etc.), nitriles (eg, acetonitrile, etc.), and mixtures of these at an appropriate ratio. The reaction temperature is not particularly limited as long as the reaction proceeds, but the reaction is usually carried out at about -50 ° C to 150 ° C, preferably at about -30 ° C to 80 ° C. The reaction time varies depending on the starting materials, the base, the reaction temperature, and the type of the solvent to be used.

The structural formulas of the compounds obtained by the following Examples are shown below.

Embedded image

[0026] Compound No. ^{n R 1 R 2 R 3 R} 4 R 5 1 (AFB-8-A 1) 1 OH OAc OAc OAc OAc 2 (AFB-8-A 2) 1 OH OH OAc OAc OAc 3 (AFB- 8-B ₁ ) 0 OH OAc OAc OAc OAc 4 (AFB-8-B ₂ ) 0 OH OH OAc OAc OAc 5 1 OAc OH OAc OAc OAc 6 0 OAc OH OAc OAc OAc 71 1 OH OH OH OH OH (However, OAc represents an acetoxy group.) The compound represented by the formula (I) or a salt thereof may be hereinafter referred to as a compound (I).

The salt of the compound (I) of the present invention includes pharmacologically acceptable base addition salts and acid addition salts. Examples of the base addition salt include a salt with an alkali metal (eg, sodium, potassium, etc.) and a salt with an alkaline earth metal (eg, calcium, magnesium, etc.). Examples of the acid addition salts include salts with inorganic acids (eg, hydrochloric acid, hydrogen bromide, hydrogen iodide, sulfuric acid, phosphoric acid, etc.) and organic acids (eg, acetic acid, propionic acid, lactic acid, succinic acid, maleic acid, And tartaric acid, citric acid, gluconic acid, ascorbic acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid, cinnamic acid, fumaric acid, malic acid, oxalic acid and the like. Compound (I) of the present invention
Can be isolated and purified by a means known per se, for example, solvent extraction, liquid conversion, phase transfer, crystallization, recrystallization, chromatography and the like. The starting compound of the compound (I) of the present invention or a salt thereof can be isolated and purified by the same known means as described above, but can be used as a reaction mixture without isolation as a starting material for the next step. May be served as When the compound (I) of the present invention contains an optical isomer, a stereoisomer, a positional isomer or a rotamer, these are also included in the compound of the present invention, and can be synthesized by a method known per se. Each can be obtained as a single item by the method. For example, when the compound of the present invention has an optical isomer, the optical isomer resolved from the compound is also included in the present invention. The optical isomer can be produced by a method known per se. Specifically, an optical isomer is obtained by using an optically active synthetic intermediate or by subjecting a final racemic mixture to optical resolution according to a conventional method.

As the optical resolution method, a method known per se, for example, the following fractional recrystallization method, chiral column method, diastereomer method and the like are used. (1) Fractional recrystallization method A method in which a salt is formed between a racemate and an optically active compound, which is separated by a fractional recrystallization method, and a free optical isomer is obtained through a neutralization step, if desired. (2) Chiral column method A method in which a racemate or a salt thereof is applied to an optical isomer separation column (chiral column) to separate the racemate or a salt thereof. For example, in the case of liquid chromatography, a mixture of optical isomers is added to a chiral column such as ENANTIO-OVM (manufactured by Tosoh Corporation), and water,
Various buffers (eg, phosphate buffer, etc.), organic solvents (eg, ethanol, methanol, acetonitrile, etc.)
Is developed as a single or mixed solution to separate optical isomers. For example, in the case of gas chromatography, separation is performed using a chiral column such as CP-Chirasil-DeXCB (manufactured by GL Sciences). (3) Diastereomer method A racemic mixture is converted into a diastereomer mixture by a chemical reaction with an optically active reagent, and the mixture is subjected to a conventional separation means (for example, fractional recrystallization, chromatography, etc.) to form a single mixture. A method of obtaining an optical isomer by separating the optically active reagent site by a chemical treatment such as a hydrolysis reaction after forming the substance. For example, when the compound of the present invention has a hydroxyl group or a primary or secondary amino group in the molecule,
The compound and an optically active organic acid (for example, MTPA [α-
Methoxy-α- (trifluoromethyl) phenylacetic acid], (−)-menthoxyacetic acid, etc.) to give a diastereomer of an ester form or an amide form, respectively. On the other hand, when the compound of the present invention has a carboxylic acid, by subjecting the compound and an optically active amine or alcohol reagent to a condensation reaction,
An amide or ester diastereomer is obtained, respectively. The separated diastereomer is converted into an optical isomer of the original compound by subjecting it to acid hydrolysis or basic hydrolysis reaction.

The compound (I) of the present invention has low toxicity and has an excellent bioactive activity for pharmaceutical use, and is effective for preventing or treating fungal infections, for example. Therefore, the preparation of the present invention containing the compound (I) of the present invention can be used as a safe antifungal agent or the like for mammals such as humans (for example, humans, dogs, cats,
(Monkey, rat, mouse, horse, cow, etc.) orally or parenterally, and usually oral administration is preferred. Examples of dosage forms for oral administration include, for example, tablets (including sugar-coated tablets and film-coated tablets), pills, granules, powders, capsules (including soft capsules),
Syrups, emulsions, suspensions and the like can be mentioned. Examples of the dosage form for parenteral administration include injections, infusions, drops, suppositories and the like. The content of compound (I) in the preparation of the present invention is usually 2 to 85% by weight, preferably 5 to 70% by weight. As a method for producing the compound (I) into the above-mentioned dosage form, a known production method generally used in the art can be applied. In the case of producing the above-mentioned dosage form, if necessary, an excipient, a binder, a disintegrant, a lubricant, a sweetener, an interface, which are usually used in the field of formulation when producing the dosage form It can be produced by appropriately containing an appropriate amount of an activator, a suspending agent, an emulsifier and the like. For example, when compound (I) is manufactured into a tablet,
It can be manufactured by containing excipients, binders, disintegrants, lubricants and the like, and in the case of making pills and granules, containing excipients, binders, disintegrants and the like Can be manufactured. In the case of preparing powders and capsules, excipients and the like, in the case of preparing syrups, sweeteners and the like, in the case of preparing emulsions and suspensions, suspending agents, It can be produced by incorporating a surfactant, an emulsifier and the like. Examples of excipients include lactose, sucrose, glucose, starch, sucrose, microcrystalline cellulose, licorice powder, mannitol, sodium hydrogen carbonate, calcium phosphate, calcium sulfate and the like. Examples of binders include 5-10% by weight
Examples include starch paste, 10-20% by weight gum arabic solution or gelatin solution, 1-5% by weight tragacanth solution, carboxymethyl cellulose solution, sodium alginate solution, glycerin and the like. Examples of disintegrants include starch, calcium carbonate and the like. Examples of lubricants include magnesium stearate, stearic acid, calcium stearate, purified talc, and the like. Examples of sweetening agents include glucose, fructose, invert sugar, sorbitol, xylitol, glycerin, simple syrup and the like. Examples of the surfactant include sodium lauryl sulfate, polysorbate 80, sorbitan monofatty acid ester, polyoxyl 40 stearate and the like. Examples of suspending agents include gum arabic, sodium alginate, sodium carboxymethylcellulose, methylcellulose, bentonite and the like. Examples of the emulsifier include gum arabic, tragacanth, gelatin, polysorbate 80 and the like. When the compound (I) of the present invention is produced in the above-mentioned dosage form, if necessary, a coloring agent, a preservative, a fragrance, a flavoring agent, a stabilizer, a thickener and the like which are usually used in the field of pharmaceutical preparations may be appropriately added. , Can be added in an appropriate amount. The preparation of the present invention containing compound (I) or a salt thereof,
Stable, low toxicity and safe to use. Part 1
The daily dose varies depending on the condition and weight of the patient, the type of the compound, the administration route and the like. However, when administered orally, the compound (I) or a salt thereof is usually used as an active ingredient in an adult patient with a daily fungal infection ( Weight 50Kg) About 2mg to 1 per person
g, especially about 10 mg to 500 mg, are preferably used for the treatment of the disease. These formulations are 1-3 times daily
It can be administered in divided doses. When the compound of the present invention or a salt thereof is parenterally administered subcutaneously, intravenously or intramuscularly as an injection, the dose is about 1 mg to 200 mg, preferably 2 mg to 200 mg / day for each adult patient with a fungal infection. 100 mg.

[0030]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described more specifically with reference to Examples, Experimental Examples and Preparation Examples, but the present invention is not limited thereto. The percentage (%) in the medium represents weight / volume percent unless otherwise specified. The numerical value indicating the mixing ratio in the mixed solvent is the volume mixing ratio of each solvent. NMR spectra were measured using a Bruker AM300 spectrometer. Using tetramethylsilane as internal standard,
All δ values are given in ppm.

[0031]

Example 1 Culture of AFB-8 Cryptococcus sp. AFB-8 strain cultured on peptone-yeast extract-glucose slant agar medium was used in a 2 L Sakaguchi flask with 2% glucose, 2% malt extract, 0.1% polypeptone. Inoculate 500 ml of seed medium (pH 6.4) containing 28%.
C. The cells were cultured on a reciprocating shaker for 2 days. 1 L of this culture
2% glucose in a 200 L tank, malt extract 2
% L, 0.1% polypeptone, and 0.02% Actol as antifoaming agent, 0.02% silicone
H 6.2) at 28 rpm at 100 revolutions per minute while injecting 50 L of sterile air per minute. C. The cells were cultured for 3 days to obtain a main culture solution.

Example 2 AFB-8-A ₁ (compound 1),
Isolation of A ₂ (Compound 2), B ₁ (Compound 3) and B ₂ (Compound 4) After adjusting the pH of the culture solution (120 liters) obtained in Example 1 to 3.0, ethyl acetate (100 Liter) and filtered using a filter aid (Radiolite # 600, manufactured by Showa Chemical Industry Co., Ltd.). The obtained organic layer was washed with water (30 liters) and then concentrated to obtain a dried product (79 g). The obtained dried product is subjected to silica gel (Kieselgel 60, 500).
g), chloroform / methanol / acetic acid [98: 2: 1 (6 liters), 9
5: 5: 1 (8 liters), 90: 10: 1 (6 liters), 80: 20: 1 (6 liters) and 50: 5
0: 1 (4 liters)]. Fr. Concentrate 8 and 9 to give AFB-8-B
A fraction containing ₁ (19.3 g) was obtained. Similarly, F
r. A ₁ containing fractions from the 16 11 (14.2g), F
r. The B ₂ -containing fraction (9.2 g) from 17 to 22 and
Fr. It was obtained from 24 of 28 A ₂ containing fraction (6.9 g). The A ₁ -containing fraction was dissolved by heating in ethyl acetate (600 ml) and left at room temperature. The resulting precipitate is collected by filtration and AF
B-8-A ₁ (compound ₁ , 9.2 g) was obtained. A _2-containing fractions were dissolved in methanol (50ml), LH-20
(2.4 liter) column chromatography, and eluted with methanol (100 ml × 30).
Concentrated active fraction (Fr.9 from 11) to afford A ₂ containing dry solid of (4.2 g). The dried product containing A ₂ was divided into three portions by preparative HPLC [column: LH50HJ 50 mm I.Dx 210 mm (manufactured by Kurita Kogyo), filler: YMC ODS-AM120-S10 (manufactured by YMC), mobile phase: 45 % (V / v) acetonitrile /
0.1% trifluoroacetic acid, flow rate: 40 ml / min], fractions having an elution volume of 1500 to 2300 ml were collected, acetonitrile was distilled off under reduced pressure, and the mixture was extracted with ethyl acetate.
The extract was washed with water, dried over anhydrous sodium sulfate and concentrated.
A concentrate (1.9 g) was obtained. The concentrate was divided into three portions and again subjected to preparative HPLC [column: YMC-Pack SH-363-15, ODS (manufactured by YMC), mobile phase: 45% (v / v) acetonitrile / 0.1% trifluoroacetic acid , Flow rate: 12 ml / min]
The fractions having an elution volume of 700 to 840 ml were collected, acetonitrile was distilled off under reduced pressure, and the mixture was extracted with ethyl acetate.
The extract was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain AFB-8-A2 (Compound ₂ , 790 mg). The B ₁ -containing fraction was dissolved in methanol (80 ml), subjected to two portions of column chromatography on LH-20 (2.4 liter), and eluted with methanol (100 ml × 30). Effective fraction (1st time: Fr.
14 to 19, 2nd time: Fr. Was concentrated to 14 17), to give B ₁ containing powder (12.1 g). The B ₁ -containing powder was subjected to column chromatography on silica gel (Kieselgel 60, 160 g) to give chloroform / methanol /
Acetic acid [98: 2: 1 (400 ml), 95: 5: 1 (2
000 ml), 90: 10: 1 (400 ml)]. Fr. 9 to 14 were collected to obtain a powder (4.7 g) containing concentrated B _1. The obtained powder was dissolved by heating in ethyl acetate (200 ml), and the resulting precipitate (2.4 g) was collected by filtration after cooling. The resulting precipitate was dissolved by heating in ethyl acetate again (100 ml), collected by filtration and allowed to cool after the resultant precipitate was obtained AFB-8-B ₁ (compound 3,1.3g). The filtrate was concentrated, similarly dissolved in ethyl acetate (300 ml, 100 ml) by heating, and then precipitated. AFB-8-B ₁ (compound 3, compound 3)
1.1 g). 4.2 g of the B ₂ -containing fraction was divided into six portions and subjected to preparative HPLC [column: YMC-Pack SH-363-15,
ODS (manufactured by YMC), mobile phase; 50% (v / v) acetonitrile / 0.1% trifluoroacetic acid, flow rate: 12 ml
/ Min]. Fractions having an elution volume of 480-520 ml were collected, acetonitrile was distilled off under reduced pressure, and the mixture was extracted with ethyl acetate. The extract was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain AFB-8-B ₂ (compounds 4, 65).
0 mg) as a white powder.

Example 3 Preparation of Compound 5 AFB-8-A ₁ (Compound ₁ , 824 mg) was prepared by adding 4-methyl-2-pentanone (225 ml) and acetic acid (25 ml).
And stirred at 100 ° C. for 4 hours. After cooling, the reaction solution was concentrated under reduced pressure, and the resulting dried product (1.2 g) was subjected to preparative HPLC [column; YMC-Pack SH-363-15, ODS (manufactured by YMC), mobile phase: 50% ( v / v) acetonitrile /
0.1% trifluoroacetic acid, flow rate: 12 ml / min]. Fractions having an elution volume of 560-620 ml were collected, acetonitrile was distilled off under reduced pressure, and the mixture was extracted with ethyl acetate. In the fraction having an elution volume of 620 to 700 ml, acetonitrile was distilled off under reduced pressure, extracted with ethyl acetate, and again subjected to preparative HPLC under the same conditions. Fractions having an elution volume of 600 to 660 ml were collected, acetonitrile was distilled off under reduced pressure, and extracted with ethyl acetate. The ethyl acetate extracts obtained by two preparative HPLCs were combined, washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain Compound 5 (210 mg) as a powder. FAB- mass spectrum; m / z823 (M-H ) - 13 C NMR spectrum: 75 MHz, in DMSO-d _6, δppm, the signal is observed or less. 175.8, 170.2, 170.1, 169.6, 169.1, 102.9, 99.1, 8
0.3, 74.7, 74.1, 73.7, 73.1, 71.3, 71.2, 69.5, 68.
9, 67.6, 62.9, 62.8, 33.8, 33.0, 29.1, 29.0, 28.9,
28.7, 24.9, 24.6, 20.6, 20.5, 20.2

Example 4 Preparation of Compound 6 AFB-8-B ₁ (Compound 3, 800 mg) was prepared by adding 4-methyl-2-pentanone (225 ml) and acetic acid (25 ml).
And stirred at 115 ° C. for 10 hours. After cooling,
The reaction solution was concentrated under reduced pressure, and the resulting dried product (1.08 g) was
Preparative HPLC [column; YMC-Pack SH-363-15, ODS (manufactured by YMC), mobile phase: 55% (v / v) acetonitrile / 0.1% trifluoroacetic acid, flow rate: 12 ml / min]
Attached to Collect fractions with an elution volume of 420-500 ml,
Acetonitrile was distilled off under reduced pressure, and extracted with ethyl acetate. The extract was washed with water, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain Compound 6 (206 mg) as a powder. FAB- mass spectrum; m / z779 (M-H ) - 13 C NMR spectrum: 75 MHz, in DMSO-d _6, δppm, the signal is observed or less. 175.9, 170.2, 170.2, 169.6, 169.1, 102.4, 99.2, 8
0.5, 74.8, 74.1, 73.2, 73.1, 71.3, 71.2, 69.5, 68.
8, 67.7, 63.0, 62.8, 33.9, 29.2, 29.0, 29.0,28.8,
28.8, 25.4, 24.7, 20.6, 20.5, 20.2

Example 5 Preparation of Compound 7 AFB-8-A ₁ (Compound ₁ , 412 mg) was dissolved in methanol (40 ml), and a 0.1N aqueous sodium hydroxide solution (10 ml) was added. Stirred for hours. After neutralizing the reaction solution with 0.1N hydrochloric acid, it was concentrated under reduced pressure to distill off methanol. After adding water (100 ml) and adjusting the pH to 3 with 0.1N hydrochloric acid, the resulting precipitate was collected by filtration and the compound 7 was obtained.
(280 mg) of a white powder was obtained. FAB- mass spectrum; m / z655 (M-H ) - 13 C NMR spectrum: 75 MHz, in DMSO-d _6, δppm, the signal is observed or less. 175.8, 103.1, 80.4, 76.7, 76.3, 74.7, 74.6, 74.2,
73.2, 69.9, 69.5, 68.9, 60.9, 60.3, 33.8, 33.0, 2
9.1, 29.0, 28.9, 28.7, 25.0, 24.6

Experimental Example 1 In Vitro Antibacterial Test: In Vitr against Fungi and Bacteria
o Antibacterial activity test The antibacterial activity of AFB-8-related compounds was tested using Aspergillus fumigatus TIMM1728 as a test fungus.
migatus TIMM1728) and Aspergillus niger IFO4
414 bacteria (Aspergillus niger IFO4414), Escherichia coli IFO3301 (Escherichia coli)
IFO3301) and Bacillus subtilis NIHJPCI219 (Bac
illus subtilis NIHJ PCI219), and measured by the following method (agar dilution method).
Test samples were prepared by dissolving the compound in methanol and serially diluting with methanol two-fold.
As a medium, Aspergillus fumigatus and Aspergillus niger are malt extract medium (malt extract 2).
%, Glucose 2%, polypeptone 0.1%, agar 0.8
%, PH 6.4), Escherichia coli and Bacillus.
As a subtilis, a bouillon medium (meat extract 1%, polypeptone 1%, salt 0.1%, agar 0.8%, pH 7.0) was used. Prepare a spore or cell suspension prepared in advance in a measurement medium, and add 10 ⁵ cfu / ml for Aspergillus fumigatus and Aspergillus niger to Escherichia.
E. coli and Bacillus subtilis were inoculated at 10 ⁷ cfu / ml and 28 after adding serially diluted test samples. C and cultured for 4 days. After the culture, the growth of the strain was visually observed, and the lowest concentration at which no growth was observed was taken as the MIC value (minimum inhibitory concentration) of the test sample. The MIC values of the compounds were as shown in Table 2.

[0037]

[Table 2]

EXPERIMENTAL EXAMPLE 2 In Vivo Test Example A mouse (Cr) whose immunosuppressive agent cyclophosphamide was intraperitoneally administered (200 mg / kg) 4 days prior to infection to reduce the infection protective ability
j: CDF1, female, 6 weeks old) was inoculated intravenously with 1.5 × 10 ⁵ cfu (150,000 cfu) of A. fumigatus TIMM1728 cells per mouse. Specimens were suspended in a 0.5% methylcellulose solution and administered first time 2 hours after infection, and orally 3 times a day for 2 days from the day after infection, for a total of 7 times. The progress of death of the mice was observed, and the surviving number and the average surviving days of each group on day 10 of infection are shown in Table 3.

[0039]

[Table 3]

As shown in Table 3, the non-administration group died in all cases (7/7) by day 8 of the infection, but the compound 1, 2 or 5
In the treated group, 4/7, 5/7, and 4/7 cases survived on day 10 of infection, respectively, and a significant survival advantage was observed in the survival time analysis (Log-Rank test) compared to the non-treated group. Was. The above results indicate that the preparation of the present invention is effective in preventing and treating the onset of mycosis caused by Aspergillus.

Formulation Examples When used as a therapeutic agent for fungal infection containing the compound of the present invention or a salt thereof as an active ingredient, it can be produced by the following formulation. 1. Capsules (1) AFB-8-A 1 100mg (2) Lactose 90 mg (3) Microcrystalline cellulose 70 mg (4) Magnesium stearate 10 mg 1 capsule 270 mg (1), the total amount and (4 (2) and (3) )), And granulate. The remaining (4) is added to this, and the whole is encapsulated in a gelatin capsule. 2. Tablets (1) AFB-8-A 1 100mg (2) Lactose 35 mg (3) Corn starch 150 mg (4) Microcrystalline cellulose 30 mg (5) Magnesium stearate 5 mg 1 tablet 320 mg (1), and (2) (3) Total amount and 2/3 of (4) and 1/2 of (5)
And then granulating. The remaining (4) and (5) are added to the granules and pressed into tablets.

[0042]

The compound (I) or a salt thereof has excellent antifungal activity and is therefore useful as a safe preventive and therapeutic agent for fungal infections and the like.

[0043]

[Brief description of the drawings]

Figure 1 is a AFB-8-A ₁ obtained in Example 2
³ shows a ¹³ C-NMR spectrum.

FIG. 2 shows AFB-8-A ₂ obtained in Example 2.
³ shows a ¹³ C-NMR spectrum.

FIG. 3 is a diagram of AFB-8-B ₁ obtained in Example 2.
³ shows a ¹³ C-NMR spectrum.

FIG. 4 is a diagram of AFB-8-B ₂ obtained in Example 2.
³ shows a ¹³ C-NMR spectrum.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme court ゛ (Reference) C12P 19/44 C12P 19/44 // (C12P 19/44 C12R 1:01) (72) Inventor Ikedo 4F term of 6F Kobayashi, Bessho-cho, Himeji-shi, Hyogo Pref. AA04 EA04 MA01 MA04 NA14 ZB35

Claims (11)

[Claims] (1) Formula (1) [Wherein, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ⁹ represent an optionally substituted hydroxyl group, and R ¹⁰ represents an esterified or amidated group. And n represents 0 or 1], or a salt thereof. 2. R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ and R ⁷
Is a hydroxyl group which may be substituted with an acyl group. 3. The compound according to claim 1, wherein R ⁸ and R ⁹ are hydroxyl groups. 4. The compound according to claim 1, wherein R ¹⁰ is a carboxyl group. (5) a compound represented by the formula: The compound according to claim 1, wherein R ²¹ represents a hydroxyl group which may be substituted with an acetyl group, n represents 0 or 1, and Ac represents an acetyl group. 6. A pharmaceutical composition comprising the compound according to claim 1. 7. An antifungal agent comprising the compound according to claim 1. [8] An anti-aspergillus agent comprising the compound according to [1]. 9. A microorganism which belongs to the genus Cryptococcus and has the ability to produce the compound according to claim 5, which is cultured in a medium, the compound is produced and accumulated in the culture, and collected. A method for producing the compound according to claim 5. 10. The microorganism is Cryptocox SP A.
The method according to claim 9, which is a strain FB-8. (11) Cryptocox sp. AFB-8 capable of producing the compound according to (5).