Tricyclo compounds, a process for their production and a pharmaceutical composition containing the same.

[Detailed Description of the Invention]

This invention relates to the immunosuppressant containing the salt permitted as the new tricyclo compound which has immunosuppressive activity, or its medicine. That is, the object of this invention is to provide the immunosuppressant containing the salt permitted as a tricyclo compound or its medicine as an active ingredient. The immunosuppressant of this invention is useful to the medical treatment and prevention of the rejection by transplant, the graft versus host disease by a bone marrow transplantation, an autoimmune disease, etc. This invention is based on FR-900506 substance which are four sorts of fermentation products, FR-900520 substance, FR-900523 substance, and FR-900525 substance having been found out. More particularly, they are FR-900506 substance, FR-900520 substance, FR-900523 substance, and FR-900525 substance, It dissociates from the cultivation broth obtained by fermenting new Bacterial species belonging to Strept married-woman (Streptomyces) Genus for the first time in a pure form. And in order to clarify chemical structure of FR-900506 substance, FR-900520 substance, FR-900523 substance, and FR-900525 substance, as a result of inquiring wholeheartedly, the artificer of this invention determined those chemical structure, and succeeded in composition of the derivative of those compounds. The following general formula can show the tricyclo compound used by this invention.<img file="JPH0346445B2_D0001.tif" />(Inside of a formula, R)¹A Is hydroxy group or the protected hydroxy group, R²Is hydrogen, a hydroxy group or the protected hydroxy group, R³It was found out that the four following sorts of compounds are produced by fermentation among compound () as which a Is methyl group, an ethyl group, a propyl group or an allyl group, and the sign with which n is expressed by the integer, solid line, and dotted line of 1 or 2 mean a single bond or a double bond. (1) R¹And R²A Respectively hydroxy group, R³Compound () as which a But allyl group and the sign with which n is expressed by integer 2, a solid line, and the dotted line mean a single bond (FR-900506 substance and naming) (2) R¹And R²A Respectively hydroxy group, R³Compound ()[as which a But ethyl group and the sign with which n is expressed by integer 2, a solid line, and the dotted line mean a single bond (FR-900520 substance and naming (alias: WS7238A substance)]) (3) R¹And R²A Respectively hydroxy group, R³Compound () as which a But methyl group and the sign with which n is expressed by integer 2, a solid line, and the dotted line mean a single bond [FR-900523 substance and naming (alias: WS7238B substance)] (4) R¹And R²A Respectively hydroxy group, R³Compound () as which a But allyl group and the sign with which n is expressed by integer 1, a solid line, and the dotted line mean a single bond (FR-900525 substance and naming) In tricyclo compound () contained in the immunosuppressant of this invention, One or more pairs of stereoisomeric form pairs like an optical isomer and geometric isomer resulting from the Cong Houma- or an asymmetric carbon atom, and a double bond may exist, and such Conformer or an opposite-sex object is also included within the limits of this invention. The salt permitted as tricyclo compound () used in this invention or its medicine can be manufactured according to the manufacturing process shown below.

[] The fermenting method Strept To Mrs. Genus Bacterial species which belongs Fermentation -&gt; FR-900506 substance FR-900520 substance FR-900523 FR-900525 substance (Following space) [] Synthetic method<img file="JPH0346445B2_D0002.tif" /><img file="JPH0346445B2_D0003.tif" /><img file="JPH0346445B2_D0004.tif" /><img file="JPH0346445B2_D0005.tif" /><img file="JPH0346445B2_D0006.tif" /><img file="JPH0346445B2_D0007.tif" /><img file="JPH0346445B2_D0008.tif" /><img file="JPH0346445B2_D0009.tif" /><img file="JPH0346445B2_D0010.tif" /><img file="JPH0346445B2_D0011.tif" />(Inside of a formula, R)¹R²R³The sign with which it is expressed by n and a solid line, and the dotted line is the respectively same meaning as a front, and is R.¹_aAnd R²_aThe hydroxy group, R by which Are protection was carried out²_bA Is desorption machine is meant. The example of the various definitions used in tricyclo compound () and its desirable embodiment are explained to details below. Especially the word that is used in this specification and "low-grade" Becoming means 1~6 carbon atoms, if there are no directions. As the suitable hydroxy blocking group in "the protected hydroxy group", For example, methyl thiomethyl, ethyl thiomethyl, propyl thiomethyl, It is C as a 1-(low-grade Alkylthio) (low-grade) Alkyl machine like low-grade Alkyl thiomethyl machines, such as isopropyl thiomethyl, butyl thiomethyl, isobutyl thiomethyl, and hexyl thiomethyl, and a still more desirable thing.₁~C₄It is methyl thiomethyl machine;, for example, trimethylsilyl, as a Alkyl thiomethyl machine and most desirable thing, Triethyl silyl, tributyl silyl, the third class butyl dimethylsilyl, (Tori (low-grade) Alkyl silyl, for example, methyl diphenyl silyl, such as the third class of Tori butylsilyl) He is Tori (C) as the Tori substitution silyl groups, such as low-grade Al kill- diaryl silyl, such as ethyl diphenyl silyl, propyl diphenyl silyl, and the third class butyl diphenyl silyl, and a still more desirable thing.₁~C₄A Alkyl silyl group and C₁~C₄as a An alkyl loosey phenyl silyl group and the most desirable thing -- the third class butyl dimethylsilyl machine and the third -- the fatty series acyl group guided from class butyl diphenyl silyl group; carboxylic acid and sulfonic acid, Acyl group; like the fatty series acyl group replaced with the aromatic series acyl group and the aromatic series machine, etc. are mentioned. As a fatty series acyl group, they are Formyl, Acetyl, and pro Pionyl, for example, Butyryl, iso Butyryl, valeryl, iso valeryl, pivaloyl, Hexa noil, Carboxyacetyl, Carboxypropionyl, Carboxybutyryl, The lower alkanoyl group which may have one or more suitable substitution machines like Carboxy, such as Carboxyhexa noil, For example, cyclopropyloxy Acetyl, cyclobutyloxy pro Pionyl, Cyclo heptyloxy Butyryl, menthyl Oxyacetyl, minced meat Leuoxypropionyl, A cyclo (low-grade) alkoxy (low-grade) alkanoyl group, a camphor sulfonyl machine, etc. which may have one or more suitable substitution machines like low-grade Alkyl, such as menthyl Oxybutyryl, menthyl Oxypenta noil, and menthyl Oxyhexa noil, are mentioned. As an aromatic series acyl group, they are benzoyl, Toluoyl, and Xyloyl, for example, Naphthoyl, nitrobenzoyl, dinitro benzoyl, nitroglycerine naphthoyl, etc., The Aroyl machine which may have one or more suitable substitution machines like nitroglycerine, For example, benzenesulphonyl, toluene sulfo nil, xylene sulfo nil, The Arene sulfonyl group etc. which may have one or more suitable substitution machines like halogen, such as Naphthalene sulfo nil, Fluoro benzenesulphonyl, chloro benzenesulphonyl, Bromo benzenesulphonyl, and iodine benzenesulphonyl, are mentioned. As the fatty series acyl group replaced with the aromatic series machine, For example, phenylacetyl, phenyl prop Onil, phenyl Butyryl, 2-trifluoromethyl 2-methoxy 2-phenylacetyl, 2-ethyl 2-trifluoromethyl 2-phenylacetyl, The Al (low-grade) alkanoyl group etc. which may have one or more suitable substitution machines like low-grade alkoxy And trihalo (low-grade) Alkyl, such as 2-trifluoromethyl 2-propoxy 2-phenylacetyl, are mentioned. C which may have Carboxy as a still more desirable acyl group among the above-mentioned acyl group₁~C₄Into an alkanoyl group and a cycloalkyl portion (C)₁~C₄Cyclo (C) which has two Alkyls₅~C₆Alkyloxy (C)₁~C₄An alkanoyl group, a camphor sulfonyl machine, the benzoyl group that may have one piece or two nitroglycerine, the benzenesulfonyl group which has halogen, C₁~C₄Alkoxy and Trihalo (C)₁~C₄Phenyl (C) which has Alkyl₁~C₄) -- an alkanoyl group is mentioned -- among those -- as the most desirable thing Acetyl, Carboxypropionyl, menthyl Oxyacetyl, camphor sulfonyl, benzoyl, nitrobenzoyl, dinitro benzoyl, iodine benzenesulphonyl, and 2-trifluoromethyl 2-methoxy 2-phenylacetyl are mentioned. As suitable "desorption machine", a hydroxy group, the reed Roxy machine which is what the Acyl portion illustrated above, etc. are mentioned. The manufacturing process of tricyclo compound () used in this invention is explained to details below.

[] The fermenting method FR-900506 substance, FR-900520 substance, FR-900523 substance, and FR-900525 substance of this invention, Streptomyces tsukubaensis (Streptomyces tsukubaensis) No.9993 and Strept Mrs. high gloss Copicus subsp. Yakushima Encis (Streptomyces hygroscopicus) It is producible by fermenting FR-900506 belonging to Strept married-woman Genus like Subsp.yakushimaensisNo.7238, FR-900520, FR-900523, and/or a FR-900525 substance-production strain in a culture medium. Production by the fermentation of FR-900506 substance, FR-900520 substance, FR-900523 substance, and FR-900525 substance is explained below.

[A] FR-900506 substance, FR-900520 substance, and FR-900525 substance of this invention, It is producible by fermenting FR-900506, FR-900520, and/or the FR-900525 substance-production strain belonging to Strept married-woman Genus like Streptomyces tsukubaensis No.9993 in a culture medium. Microorganism A bacillus used for production of FR-900506 substance, FR-900520 substance, and/or FR-900525 substance is FR-900506, FR-900520, and/or a FR-900525 substance-production strain belonging to Strept married-woman Genus. Among those, Streptomyces tsukubaensis No.9993 was newly separated from a soil sample extracted in Tsukuba-gun, Ibaraki Toyosato-cho. The freeze-drying specimen of this newly separated Streptomyces tsukubaensis No.9993, It Deposits to the Fermentation Research Institute, the Agency of Industrial Science and Technology (1-1-3, Yabebe-cho East, Tsukuba-gun, Ibaraki-ken) as deposition number Fermentation Research Institute mycoparasite No. 7886 on October 5, 1984, Subsequently, it was switched to the Budapest Treaty route of the depository institution as new deposition number FERM BP No. 927 on October 19, 1985. In production of FR-900506 new substance, FR-900520 substance, and/or FR-900525 substance, the bacillus written in this specification is hung up only for explanation, and is not limited to it. In this invention, they are a natural mutation bacillus, X ray irradiation, and an ultraviolet exposure again, The artificial mutation bacillus built by processing of common use by N-methyl N'-nitroglycerine N-nitroso guanidine, 2-amino pudding, etc. is also included, It is contained also when using what kind of mutation bacillus which can produce FR-900506 substance, FR-900520 substance, and/or FR-900525 substance. Streptomyces tsukubaensis No.9993 has the following morphological character, the cultivation characteristic, and biological and physiological characterization. [1] Morphological character Mainly, Shirring (Shirling) And a A pot LEAP (To Gottlieb) Method [shirring A- B of a statement And Day- A pot LEAP: Methods folky Yarakutation Seisonobu Strept Mrs. Space (International journal of systematic bacteriology -)(Methods for characterization of Streptomyces ) The 16th volume of species.International Journal of Systematic Bacteriolgy, the 313~340th page, and 1966] were used for taxonomic examination. Using the culture grown for 14 days by 30 degreeC on oatmeal agar, yeast malto extract Tract agar, and mineral salt and starch agar, optics and an electron microscope are used for morphologic observation, and it is I got it. the straight chain shape which has a number of spores with which a mature spore object exceeds 10~50 pieces or 50 pieces to each spore chain -- or -- being wavelike (Rectiflexibiles) -- it formed. According to observation by an electron microscope, the elliptical form of this spore is cylindrical, and a size is 0.5~0.7x0.7~0.8 micrometer. It is smooth and the spore surface is Oh. [2] Cultivation characteristic The cultivation characteristics are the above-mentioned shirring, a A pot LEAP, and Waksman (Waksman)[Waksman S Ey:the Actinomyzetsu, The 2nd volume, classification identification and desk Repetition of, a Generale and Space the Williams and Wilkins company, Walcz More, 1961 (Waksman.S.A.:The actinomycetes,Vol.2:Classification. identification and description of genera and species.The It observed by ten kinds of culture media of the statement to Williams and Wilkins Co., Baltimore, and 1961]. It cultivated for 14 days by 30 degreeC. The color name used by this research was based on fresh color name A book (guide of Tokyo and Japan Color Research Institute). When it was made to grow up on oatmeal agar, yeast malto extract Tract agar, and mineral salt and starch agar, the colony belonged to the gray system. Although yeast malto extract Tract agar generated the soluble pigment, it did not generate in other culture media. A result is shown in Table 1.

[Table]

[Table]

About cell wall composition analysis, they are method [Becker Bi-. and M Py Resieve Arriel, such as Becker, R A- Gordon And h Ey Resieve Arriel: Rapid Diffey Rensiers-Shillon Bitvoy-A Nocardia and Strept Mrs. by paper-chromatography of hole cell hydronalium Lizz: Applied microbiology -(Becker,B.,M.P.Lechevalier.R.E . Gordob and H.A.Lechevalier:Rapid differentiation between Nocardia and Streptomyces by, paper chromatography of whole cell hydrolysates:Appl.Microbiol. -- the 12th volume, the 421~423rd page, 1964], and method [ of Yamaguchi ] [Yamaguchi, Tei-.: party Rhyson's the - cell wall composition Omorphhori curry distinct Actinomyzetsu: -- journal of bacteriology -(Yamaguchi.T.:Comparison of the cell wall composition of morphologically distinct actinomycetes:J.Bacteriol. -- the method of the 89th volume, the 444~453rd page, and 1965] -- I got it. The analysis result of all the cell hydrolysis things of strain No.9993 showed that LL-diaminopimelic acid existed. Therefore, it is thought that the cell wall of this strain belongs to a type.

[3] Biological and a physiological property The physiological property of strain No.9993 was measured by the method indicated to the above-mentioned shirring and a A pot LEAP. The result is shown in Table 2. The growth temperature range and the growth optimum temperature were measured on yeast malto extract Tract agar using the temperature gradient cultivation machine (made by Toyo Chemical industrial incorporated company). The growth temperature range is 18~35degreeC, the growth optimum temperature is 28degreeC, and it is Oh. Peptonization and gelatin liquefaction of milk are positive, and Oh. Production of a melanin is netative and Oh.

[Table]

Availability of the source of carbon Prideaux Ham And method [Prideaux Ham Tei- G [ of a A pot LEAP ] - And Dee Goto LEAP: The Utilization Sequence carbon compounds Bayh, the Sam Actino My Sets Tails As Ann Ade Ho Species determination: Journal of bacteriology -(Pridham.T.G.and D.Gottlieb: The 56th volume of The utilization of carbon compounds by some Actinomycetalesas an aid for species determination:J.Bacteriol, It examined by the 107~114th page and 1948]. The growth situation was observed after cultivation for 14 days by 30 degreeC. The availability of the source of carbon of this strain is summarized in Table 3, and is shown. Glycerin, malt sugar, and monosodium succinate were used by strain No.9993. Some D-glucose, shook sirloins, D-mannose, and Salicins use again.

[Table]

- : don't use. It is Noodle that it is clear from the result of microscope study observation of strain No.9993 and cell wall composition analysis that this strain belongs to Waksman (Waksman), Henrici (Henrici), and Strept married-woman Genus depended in 1943. Namely, the 18th volume of quality [International Journal of Systematic Bacteriology (International Journal of Systematic Bacteriology), the 69~189th page released in this strain, The 279~392nd page, 1968, the 19th volume, the 391~512nd page, The 8th edition of a 1969 and barge AIDS manual Of denaturation native bacteriology (Bergey's Manual of Determinative Bacteriology), It compared with various kinds of Strept married-woman Genus in the light of 1974]. Strain No.9993 is Strept Mrs. Abra viensis Nishimura (Streptomyces aburaviensis Nishimura et.al) etc. as a result of comparison, Strept Mrs. Au "-- it is thought that it is similar to Elaneus Baldatus, grain (Streptomyces avellaneus Baldacci and Grein), and Strept Mrs. Mass Chiemens Nakamura. Therefore, the cultivation characteristic of strain No.9993 was compared with corresponding Strept Mrs. AbraWeenscis IFO12830, Strept Mrs. Avellanéus IFO13451, and Strept Mrs. Mass Chiemens IFO12891. As a result, strain No.9993 was most similar to Strept Mrs. mixer Ensys IFO12891. Therefore, strain No.9993 was compared with Strept Mrs. Mass Chiemens IFO12891 further shown in the above-mentioned tables 1, 2, and 3 in detail. Strain No.9993 is distinguishable from this result in respect of Strept Mrs. Mass Chiemens IFO12891 and the following, Therefore, it was thought that it was a Strept married woman's new species, this microorganism was connected with the soil extracted in Tsukuba-gun, Ibaraki, and strain No.9993 was named the Streptomyces tsukubaensis Sp knob (Streptomyces tsukbaensis, sp nov.). The cultivation characteristic of difference strain No.9993 of Strept Mrs. Mass Chiemens IFO12891, When it cultivates using oatmeal agar, yeast malto extract Tract agar, glucose asparagine agar, Supapec agar, and potato dextrose agar, it differs from Strept Mrs. Mass Chiemens IFO12891. In the case of Strept Mrs. Mass Chiemens IFO12891, it is positive although starch hydrolysis of strain No.9993 is netative. In the case of Strept Mrs. Mass Chiemens IFO12891, it is netative although the gelatin liquefaction of strain No.9993 is positive. In the availability of the source of carbon, although strain No.9993 uses glycerin, malt sugar, and monosodium succinate, Strept Mrs. Mass Chiemens IFO12891 does not use these. Although strain No.9993 does not use D-galactose and inulin, Strept Mrs. Mass Chiemens IFO12891 can use these. FR-900506 new substance of an invention of Production of FR-900506 substance, FR-900520 substance, and FR-900525 substance, FR-900520 substance and FR -- 900525 substances, For example, it can produce by cultivating Streptomyces tsukubaensis No.9993, FR-900506 belonging to Strept married-woman Genus like (deposition number:FERM BP No. 927), FR-900520, and/or a FR-900525 substance production strain in a culture medium. Generally, they are FR-900506 substance, FR-900520 substance, and/or FR-900525 substance, It can produce by cultivating under aerobic conditions, such as shaking cultivation and submerged culture, preferably in the water culture medium containing the source of carbon and the source of nitrogen which can assimilate FR-900506, FR-900520, and/or a FR-900525 substance production strain. As a desirable source of carbon of a culture medium, they are glucose, xylose, and galactose, As other sources of carbon where carbohydrates, such as glycerin, starch, and dextrin, are mentioned, malt sugar, rum north, raffinose, arabinose, mannose, Salicin, monosodium succinate, etc. are mentioned. As a desirable source of nitrogen, they are yeast extract Tract, peptone, and a gluten meal, Inorganic matter, such as ammonium salt, such as ammonium nitrate, such as cottonseed powder, soybean flour, a corn, a Steep liquor, dry yeast, a wheat germ, feathers powder, and peanut powder, ammonium sulfate, and ammonium phosphate, urea, and amino acid, or an organic nitrogen compound is mentioned. Although preferably used in those combination, if the source of carbon and the source of nitrogen contain a little a growth factor and a considerable quantity of mineral nutrients, they can also use a substance with low purity and it is not necessary to necessarily use them in a pure form. Mineral salt, such as sodium carbonate or calcium carbonate, phosphoric acid sodium or phosphoric acid potassium, sodium chloride or potassium chloride, iodination sodium or iodination potassium, magnesium salt, Copper salt, and cobalt salt, may be added to a culture medium by request. When especially a cultivation culture medium foams remarkably, liquefied paraffine, fatty oil, vegetable oil, mineral oil, or an antifoaming agent like silicon may be added as occasion demands. As conditions which mass-produce FR-900506 substance, FR-900520 substance, and FR-900525 substance, depths good mind cultivation is preferred. Shaking cultivation or surface culture is carried out to small-lot production in a flask or a bottle. In growing within a large-sized tank again, in order to avoid the growth delay in the production process of FR-900506 substance, FR-900520 substance, and FR-900525 substance, it is preferred to use pre-cultivation of a microorganism and to inoculate a bacillus into a production tank. That is, it is desirable to move abacterially the pre-cultivation inoculation thing which inoculated the spore or fungal thread of the microorganism into comparatively little cultivation culture media, cultivated the inoculation culture medium, produced the pre-cultivation inoculation thing of the microorganism first, and subsequently cultivated it to a large-sized tank. The different intermediary also with Then the same as substantially as the culture medium used for production of FR-900506 substance, FR-900520 substance, and FR-900525 substance and sufficient of the culture medium which produces this pre-cultivation inoculation thing is also good. Churning and aeration of a cultivation mixture can be performed by various methods. or [ whether churning uses the agitating device according to a propeller or this, or that it rotates a fermentation machine ] -- or it can carry out also by shaking, using various pumping plants, or letting sterilization air pass in a culture medium. A line intermediary is also well ventilated by passing the inside of a fermentation mixture for sterilization air. Although it is the temperature range of about 20 degrees C~40 degreeC, preferably 25~35degreeC and fermentation is usually performed for about 50~150 hours, it may be made to change with fermentation conditions and fermentation scales suitably. Thus, FR-900506 substance, FR-900520 substance, and FR-900525 substance which were produced are recoverable from a cultivation culture medium by the method of the common use usually used for recovery of other known biological active substances. FR-900506 substance, FR-900520 substance, and FR-900525 substance which were produced, It is found out in cultured-bacteria thread and Filtrate, therefore they are FR-900506 substance, FR-900520 substance, and FR-900525 substance, From the fungal thread obtained by filtration of cultivation broth, or centrifugal separation, and Filtrate, Vacuum concentration, freeze-drying, extraction by a solvent in ordinary use, PH adjustment, for example, anion exchange resin, or cationic exchange resin, It can dissociate and refine by the method of common use, such as processing by the adsorbent of daily use of processing by resin of daily use of nonionic adsorption resin etc., for example, activated carbon, silicic acid, silica gel, cellulose, alumina, etc., crystallization, and re-crystallization. FR-900506 substance, FR-900520 substance, and FR-900525 substance which were produced by the physicochemical quality above-mentioned fermenting method of FR-900506 substance, FR-900520 substance, and FR-900525 substance have the following physicochemical quality. FR-900506 substance (1) Shape and color tone: White powder (2) Ultimate analysis value: C:64.72%, H,8.78%, N:1.59% 64.59% 8.74% 1.62% (3) Color reaction: Positivity: A sulfuric acid cerium reaction, a sulfuric acid reaction, the Ehrlich reaction, the Dragendorff reaction, an iodine steamy reaction Negativity: A ferric chloride reaction, a ninhydrin reaction, the Molisch reaction (4) Solubility: Meltable: Methanol, ethanol, acetone, ethyl acetate, chloroform, diethylether, benzene Refractory: Hexane, petroleum ether Insoluble: Water (5) Melting point: 85~90degreeC (6) Specific optical rotation: [alpha]²³_D: -73 degreeC (C= 0.8, CHCl)₃) (7) Ultraviolet absorption spectrum: End absorption (8) Infrared resonance spectrum: nu^CHCl3_nax:3680,3580,3520,2930,2870,2830,1745,1720,1700,1645,1450,1380,1350,1330,1310,1285,1170,1135,1090,1050,1030-1000, 990, 960 (sh),918cm^-1(9)¹³C nuclear magnetic resonance spectrum: delta (ppm, CDCl)₃):212.59(s) 212.45(s)196,18(s) 192.87(s), 169.07(s) 168.90(s)164.90(s) 166.01(s)138.89(s) 139.67(s), 135.73(d) 135.60(d)132.52(s) 131.99(s)130.27(d) 130.21(d) ， 122.87(d) 123.01(d)116.57(t) 116.56(t)97.35(s) 98.76(s), 84.41(d), 77.79(d) 78.22(d)75.54(d) 76.97(d)73.93(d) 73.09(d), 73.72(d) 72.57(d)70.05(d) 69.15(d)56.75(d) ， 53.03(d) 53.13(d)48.85(t) 48.62(t)40.33(d) 40.85(d), 39.40(t),31.58(t),30.79(t), 27.72(t) 26.34(t)26.46(d) 24.65(t) ， 20.45(q) 19.73(q)14.06(q) 14.23(q)9.69(q) 9.98(q) The chart of the above-mentioned spectrum is shown in Drawing 1. (10)¹H nuclear magnetic resonance spectrum: The chart of the above-mentioned spectrum is shown in Drawing 2. (11) Thin layer chromatography: [Table]

(12) Character of a substance: Neutral substance About FR-900506 substance,¹³C Reach.¹In the case of measurement of H nuclear magnetic resonance spectrum, this substance shows one pair of signals in various chemical shifts, respectively. FR-900506 substance which has such a feature has the further following character. (i)¹³When C nuclear magnetic resonance spectrum is measured in 25degreeC and 60 degreeC, the intensity ratio of one pair of signals changes, respectively. (ii) When thin layer chromatography and high speed liquid chromatography are measured, FR-900506 substance appears as a single peak in high speed liquid chromatography as a single spot at thin layer chromatography, respectively. When this white powder of FR-900506 substance is re-crystallized from acetonitrile, it changes to the shape of a crystal and this crystal has the following physicochemical quality. (1) Shape and color tone: A colorless prism-like crystal (2) Ultimate analysis value: C:64.30%,H:8.92%,N:1.77% 64.20%, 8.86%, 1.72% (3) Melting point: 127~129degreeC (4) Specific optical rotation: [alpha]²³_D:-84.4 *(C=1.02,CHCl₃) (5)¹³C nuclear magnetic resonance spectrum: delta (ppm, CDCl)₃):211.98(s) 211.74(s)196.28(s) 193.56(s), 168.97(s) 168.81(s)164.85(s) 165.97 (s) 138.76 (s) 139.51(s), 135.73(d) 135.63(d)132.38(s) 131.90(s)130.39(d) 130.17(d), 122.82(d) 122.96(d)116.43(t) 97.19(s) 98.63(s), 84.29(d) 77.84(d) 78.21(d)77.52(d) 76.97(d), 69.89(d) 69.00(d)56.63(d) 54.87(d)52.97(d) 52.82(d), 48.76(t) 48.31(t)40.21(d) 40.54(d)31.62(t) ， 30.72(t) 24.56(t) 24.12(t) 20.86(t), 20.33(q) 19.74(q)16.17(q) 16.10(q)15.88(q) 45.75(q), 13.89(q) 14.05(q)9.64(q) 9.96(q) The chart of the above-mentioned spectrum is shown in Drawing 3. (6)¹H nuclear magnetic resonance spectrum: The chart of the above-mentioned spectrum is shown in Drawing 4. It is the same as them of white powder, and the character of the color reaction of the colorless prism-like crystal of other physicochemical qualities, i.e., FR-900506 substance, solubility, an ultraviolet absorption spectrum, an infrared resonance spectrum, thin layer chromatography, and physical properties is Oh. As for the above-mentioned physicochemical quality and X ray analysis to FR-900506 substance, it is I and It was to have the following chemical structure.<img file="JPH0346445B2_D0012.tif" />17-ant roux 1, 14-dihydroxy 12-[2-(4-hydroxy 3-methoxy cyclohexyl)-1-methylvinyl]-23.25-dimethoxy 13, 19, and 21, and 27-tetramethyl 11.28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10, 16-tetra--on FR-900520 substance The physicochemical quality of this compound is described later. FR-900525 substance (1) Shape and color tone: White powder (2) Ultimate analysis value: C:65.17%,H:8.53%,N:1.76% (3) Color reaction: Positivity: A sulfuric acid cerium reaction, a sulfuric acid reaction, the Ehrlich reaction, the Dragendorff reaction, an iodine steamy reaction Negativity: A ferric chloride reaction, a ninhydrin reaction, the Molisch reaction (4) Solubility: Meltable: Methanol, ethanol, acetone, ethyl acetate, chloroform, diethylether, benzene Refractory: Hexane, petroleum ether Insoluble: Water (5) Melting point: 85~89degreeC (6) Specific optical rotation: [alpha]²³_D:-88 * (C=1.0₁CHCl₃) (7) Ultraviolet absorption spectrum: End absorption (8) Infrared resonance spectrum: nu^CHCl3_nax:3680,3580,3475,3340,2940,2880,2830,1755,1705,1635,1455,1382,1370,1330,1310,1273,1170,1093,1050,1020,995,970,920,867cm^-1(9)¹³C nuclear magnetic resonance spectrum: delta (ppm, CDCl)₃):212.61(s) 211.87(s)188.57(s) 191.12(s), 168.76(s) 170.18(s)163.11(s) 161.39(s)140,28(s) 139.37(s), 135.62(d) 135.70(d)132.28(s) 131.34(s)130.09(d) 130.00(d), 122.50(d) 123.23(d)116.48(t) 99.16(s) 99.11(s), 84.42 84.48(d)78.60(d) 79.86(d)76.73(d) 77.33(d), 59.97(d) 60.45(d)57.52(q) 56.56(q) 56.48(q), 56.14(q) 55.97(q)53.45(d) 53.26(d)49.15(t) 49.73(t), 48.46(t) 47.62(t)44.47(t) 45.23(t)41.40(d) 40.40(d), 35.19(d) 35.11(d)33.10(d) 34.17(d)32.81(t) 32.29(t), 31.53(t) 31.33(t)30.80(t) 30.66(t)28.60(t) ， 26.03(d) 26.98(d)25.43(t) 24.40(t)18.93(q) 20.57(q), 14.09(q) 13.95(q)9.85(q) 10.00(q) The chart of the above-mentioned spectrum is shown in Drawing 5. (10)¹The chart of the H nuclear-magnetic-resonance-spectrum above-mentioned spectrum is shown in Drawing 6. (11) Thin layer chromatography: [Table]

(12) Character of a substance: Neutral substance About FR-900525 substance,¹³C Reach.¹Although this substance showed one pair of signals in various chemical shifts, respectively at the time of measurement of H nuclear magnetic resonance spectrum, When thin layer chromatography and high speed liquid chromatography were measured, FR-900525 substance showed the single peak by thin layer chromatography at a single spot and high speed liquid chromatography, respectively. By having determined the chemical structure of the above-mentioned physicochemical quality and FR-900506 substance, it became clear that FR-900525 substance had the following chemical structure.<img file="JPH0346445B2_D0013.tif" />16-ant roux 1, 13-dihydroxy 11-[2-(4-hydroxy 3-methoxy cyclohexyl)-1-methylvinyl]-22, 24-dimethoxy 12, 18, and 20, 26-tetramethyl 10, and 27-dioxa 4-azatricyclo [21.3.1.0^4,8] Peptakos- 17-En- 2, 3, and 9 and 15-tetra--on [B] -- FR-900520 substance of this invention, and FR-900523 substance, FR-900520 substance and/or FR-900525 substance . production strain belonging to Strept married-woman Genus like Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238 are producible by making it ferment in a culture medium. Microorganism A bacillus used for production of FR-900520 substance and/or FR-900523 substance is FR-900520 and/or a FR-900523 substance-production strain belonging to Strept married-woman Genus. Among those, Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238 was newly separated from a soil sample extracted in Kagoshima Yaku Islands. The freeze-drying specimen of this newly separated Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238, It Deposited to the Fermentation Research Institute, the Agency of Industrial Science and Technology, on January 15, 1985 as deposition number Fermentation Research Institute mycoparasite No. 8043, and, subsequently was switched to the Budapest Treaty route of the depository institution as new deposition number FERM BP No. 928 on October 19, 1985. In production of FR-900520 new substance and FR-900523 substance, the bacillus written in this specification is what was hung up only for explanation, and is not limited to Then and it. In this invention, they are a natural mutation bacillus, X ray irradiation, and an ultraviolet exposure again, It is contained also when using what kind of mutation bacillus which can produce FR-900520 substance and/or FR-900523 substance also containing the artificial mutation bacillus built by processing of common use of N-methyl N'-nitroglycerine N-nitroso guanidine, 2-amino pudding processing, etc. Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238 has the following morphological character, the cultivation characteristic, and biological and physiological characterization.

[1] Morphological characteristics The method indicated by above-mentioned shirring and A pot LEAP was mainly used for the research on this taxonomy. The culture grown for 14 days by 30 degreeC is used for morphologic observation on oatmeal agar, yeast malto extract Tract agar, and mineral salt and starch agar, and it is I got it by optics and an electron microscope. The mature spore object formed the shape (Retinaculiaperti) of a key and the shape (Spirales) of Spiral which have about 20 spores to short Nuclear and each chain a little. The hygroscopic spore lump was looked at by the mind fungal thread on oatmeal agar, and mineral salt and starch agar. The irregular unevenness on the surface of a spore is the middle shape between a very short and thick prickle and wart, and is Oh.

[2] Cultivation characteristic The cultivation characteristic was observed on the above-mentioned shirring, a A pot LEAP, and ten kinds given in above-mentioned Waksman of culture media. It cultivated for 14 days by 30 degreeC. The color name used by this research was based on the above-mentioned fresh color name A book. When it was made to grow on oatmeal agar, yeast malto extract Tract agar, and mineral salt and starch agar, the colony belonged to the gray system. The soluble pigment was not produced in the examined culture medium. A result is shown in Table 4. Table 4. strain No.7238 and Strept Mrs. anti my Cottex(Streptomyces antimicoticus) IFO12839 -- and -- Strept Mrs. high-gloss Copicus subsp. - prodigal -- Boss (Streptomyces) The cultivation characteristic of hygroscopicus subsp.glebosusIFO13786 [Table]

[Table]

It is I got it by the method of Yamaguchi, such as above-mentioned Becker, about cell wall analysis. The analysis result of all the cell hydrolysis things of strain No.7238 showed existence of LL-diaminopimelic acid. Therefore, it is thought that the cell wall of this strain belongs to Type I.

[3] Biological and a physiological property the physiological property of strain No.7238 -- a method the above-mentioned shirring and given in a A pot LEAP -- therefore, it measured. The result is shown in Table 5. The growth temperature range and the growth optimum temperature were measured on yeast malto extract Tract agar using the temperature gradient cultivation machine (made by Toyo Chemical industrial incorporated company). The growth temperature range is 18~36degreeC, the optimum temperature is 28degreeC, and it is Oh. Starch hydrolysis and gelatin liquefaction are positive, and Oh. The melanin was not produced.

[Table]

[Table]

The availability of the source of carbon was examined by above-mentioned Prideaux Ham and the method of the A pot LEAP. It observed, after cultivating a growth state for 14 days by 30 degreeC. The source availability of carbon of this strain is summarized in Table 6, and is shown. D-glucose, a shook sirloin, lactose, malt sugar, D-trehalose, inositol, inulin, and Salicin were used by strain No.7238.

[Table]

- : don't use. It is Noodle that it is clear from microscope study observation of strain No.7238 and a cell wall composition analysis result that this strain belongs to Waksman, Henrici, and Strept married-woman Genus by 1943. Therefore, the 18th volume of quality [International Journal of Systematic Bacteriology (International Journal of Systematic Bacteriology), 69~189 pages released in this strain, 279~392 pages, 1968, the 19th volume, 391~512 pages, The 8th edition of a 1969 and barge AIDS manual Of denaturation native bacteriology, It compared with the kind which belongs to various Strept married-woman Genus in the light of 1974(Bergey's Manual of Determinative Bacteriology 8th Edition, 1974)]. It is thought as a result of comparison that strain No.7238 reaches Strept Mrs. anti my Cottex Waksman 1957, Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and similar to Boss's Eye Mori etc. therefore, the cultivation characteristic of strain No.7238 -- corresponding Strept Mrs. anti my CottexIFO12839 -- and as it Strept Mrs. high-gloss Copicus subsp. - is prodigal and was indicated in Tables 4, 5, and 6 as Boss IFO13786, it compared. From the result of this Group of small comparison, strain No.7238 was able to reach Strept Mrs. anti my Cottex IFO12839, and could Strept Mrs. high-gloss Copicus subsp. - Be prodigal, and it was able to distinguish it from Boss IFO13786 in respect of the following. (i) The difference with Strept Mrs. anti my Cottex IFO12839 The cultivation characteristic of strain No.7238 is with Strept Mrs. anti my Cottex IFO12839, It differs, when it cultivates using yeast malto extract Tract agar, glucohol asparagine agar, glycerin asparagine agar, potato dextrose agar, and tyrosine agar. In the availability of the source of carbon, although strain No.7238 uses malt sugar, Strept Mrs. anti my Cottex IFO12839 does not use. Although strain No.7238 does not use glycerin, D-fructose, rum north, raffinose, D-galactose, D-mannose, mannitol, and monosodium succinate, Strept Mrs. anti my Cottex IFO12839 uses. (ii) It Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and is the difference with Boss IFO13786. The cultivation characteristics of strain No.7238 differ, when it is prodigal and cultivates with Boss IFO13786 using yeast malto extract Tract agar, potato dextrose agar, and tyrosine agar, a Strept Mrs. high gloss Copicus subsp. and. Although milk peptonization of strain No.7238 is netative, it Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and Boss IFO13786 is positive. Although strain No.7238 can be grown under existence of 7%NaCl, it Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and does not grow Boss IFO13786 under the condition. In use of the source of carbon, stock No.7238 uses lactose, inulin, and Salicin, Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and it does not grow Boss IFO13786 under the condition. In the availability of the source of carbon, although strain No.7238 can use lactose, inulin, and Salicin, it Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and Boss IFO13786 does not use them. Strain No.7238 are glycerin, D-xylose, and D-fructose, Although raffinose, D-galactose, D-mannose, mannitol, and monosodium succinate are not used, it Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and Boss IFO13786 uses them. However, strain No.7238 forms a hygroscopic spore lump in a mind fungal thread with oatmeal agar and mineral salt starch agar, The morphological characteristics and the cultivation characteristic of strain No.7238 Strept Mrs. high-gloss Copicus subsp. - Are prodigal, and similar to Boss IFO13786. Therefore, it is although strain No.7238 belongs to Strept Mrs. high gloss Copicus and this publicly known strain is most similar to strain No.7238 in the subspecies of Strept Mrs. high gloss Copicus, Strain No.7238 Strept Mrs. high-gloss Copicus subsp. - Is prodigal, and differs from Boss IFO13786. It is thought from the above fact that strain No.7238 is a new subspecies of Strept Mrs. high gloss Copicus, and it is connected with a microorganism being separated from the soil of Yaku Islands, It was named Strept Mrs. high gloss Copicus subsp. Yakushima Encis (Streptomyces hygroscopicus subsp.yakushimaensis). FR-900520 new substance and/or FR-900523 substance substance of an invention of Production of FR-900520 substance and FR-900523 substance, For example, the thing for which FR-900520 and/or the FR-900523 substance production strain belonging to Strept married-woman Genus like Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.723 (deposition number: FERM BP No. 928) are cultivated by a culture medium It can produce. Generally, they are FR-900520 substance and/or FR-900523 substance, It can produce by cultivating in a water nutrition culture medium including the source of carbon and the source of nitrogen which can assimilate FR-900520 substance and/or a FR-900523 substance production strain, preferably under aerobic conditions, such as for example, shaking cultivation and submerged culture. As a desirable source of carbon in a culture medium, carbohydrates, such as glucose, a shook sirloin, lactose, glycerin, starch, and dextrin, are mentioned. In addition, as a source of carbon which may be included in the culture medium, malt sugar, D-trehalose, inositol, inulin, Salicin, etc. are mentioned. As a desirable source of nitrogen, they are yeast extract Tract, peptone, and a gluten meal, Inorganic matter, such as ammonium salt, such as ammonium nitrate, such as cottonseed powder, soybean flour, a corn, a Steep liquor, dry yeast, a wheat germ, feathers powder, and peanut powder, ammonium sulfate, and ammonium phosphate, urea, and amino acid, or an organic nitrogen compound is mentioned. Although preferably used in those combination, if the source of carbon and the source of nitrogen contain a little a growth factor and a considerable quantity of mineral nutrients, they can also use a substance with low purity and it is not necessary to necessarily use them in a pure form. Mineral salt, such as sodium carbonate or calcium carbonate, phosphoric acid sodium or phosphoric acid potassium, sodium chloride or potassium chloride, iodination sodium or iodination potassium, magnesium salt, Copper salt, and cobalt salt, may be added to a culture medium by request. When especially a cultivation culture medium foams remarkably, liquefied paraffine, fatty oil, vegetable oil, mineral oil, or an antifoaming agent like silicon may be added as occasion demands. As conditions which mass-produce FR-900520 substance and FR-900523 substance, depths good mind cultivation is preferred. Shaking cultivation or surface culture is carried out to small-lot production in a flask or a bottle. In growing within a large-sized tank again, in order to avoid the growth delay in the production process of FR-900520 substance and FR-900523 substance, it is preferred to use pre-cultivation of a microorganism and to inoculate a bacillus into a production tank. That is, it is desirable to move abacterially the pre-cultivation inoculation thing which inoculated the spore or fungal thread of the microorganism into comparatively little cultivation culture media, cultivated the inoculation culture medium, produced the pre-cultivation inoculation thing of the microorganism first, and subsequently cultivated it to a large-sized tank. The different intermediary also with Then the same as substantially as the culture medium used for production of FR-900520 substance and FR-900523 substance and sufficient of the culture medium which produces this pre-cultivation inoculation thing is also good. Churning and aeration of a cultivation mixture can be performed by various methods. or [ whether churning uses the agitating device according to a propeller or this, or that it rotates a fermentation machine ] -- or it can carry out also by shaking, using various pumping plants, or letting sterilization air pass in a culture medium. A line intermediary is also well ventilated by passing the inside of a fermentation mixture for sterilization air. Although it is the temperature range of about 20 degrees C~40 degreeC, preferably 25~35degreeC and fermentation is usually performed for about 50~150 hours, it may be made to change with fermentation conditions and fermentation scales suitably. Thus, FR-900520 substance and FR-900523 substance which were produced are recoverable from a cultivation culture medium by the method of the common use usually used for recovery of other known biological active substances. FR-900520 substance and FR-900525 substance which were produced, It is found out in cultured-bacteria thread and Filtrate, therefore they are FR-900520 substance and FR-900523 substance, From the fungal thread obtained by filtration of cultivation broth, or centrifugal separation, and Filtrate, Vacuum concentration, freeze-drying, extraction by a solvent in ordinary use, PH adjustment, for example, anion exchange resin, or cationic exchange resin, It can dissociate and refine by the method of common use, such as processing by the adsorbent of daily use of processing by resin of daily use of nonionic adsorption resin etc., for example, activated carbon, silicic acid, silica gel, cellulose, alumina, etc., crystallization, and re-crystallization. It is especially FR-900520 substance and FR-900523 substance, The substance containing both the output produced by fermentation is dissolved in suitable solvents, such as ethyl acetate and n-hexane, Subsequently, it is separable by giving the solution to chromatography, for example, the column chromatography which uses silica gel, and being eluted with ethyl acetate and n-hexane, or a suitable solvent like those mixtures. FR-900520 substance and FR-900523 substance which were separated by doing in this way can further be refined by usual state methods, such as for example, a re-crystal, re-chromatography, and high speed liquid chromatography, respectively. The physicochemical quality of FR-900520 substance and FR-900523 substance FR-900520 substance (1) Shape and color tone: A colorless plate crystal (2) Ultimate analysis value: C:64.81%,H:8.82%,N:1.55% (3) Color reaction: Positivity: A sulfuric acid cerium reaction, a sulfuric acid reaction, the Ehrlich reaction, the Dragendorff reaction, an iodine steamy reaction Negativity: A ferric chloride reaction, a ninhydrin reaction, the Molisch reaction (4) Solubility: Meltable: Methanol, ethanol, acetone, ethyl acetate, chloroform, diethylether, benzene Refractory: N-hexane, petroleum ether Insoluble: Water (5) Melting point: 163~165degreeC (6) Specific optical rotation: [alpha]²³_D:-84.1 *(C=1.0,CHCl₃) (7) Ultraviolet absorption spectrum: End absorption (8) Infrared resonance spectrum: nu^CHCl3_nax: 3680,3575,3520, 2940,2875,2825, 1745,1725,1700, 1647, 1610(sh) '1452,1380,1350, 1330,1285,1170, 1135,1090,1030, 1005,990,980 (sh)960(sh)913,908(sh)cm^-1(9)¹³C nuclear magnetic resonance spectrum: delta (ppm, CDCl)₃):213.04(s) 196.21(s) 193.23(s), 169.07(s) 168.85(s)164.92(s) 165.97(s)138.67(s) 139.53(s), 132.46(s) 131.98(s)130.20(d) 130.08(d)123.42(d) 123.59(d), 97.28(s) 98.75(s)84.37(d) 77.80(d) 78.24(d), 75.53(d) 76.98(d)73.92(d) 73.69(d) ， 73.11(d) 72.72(d)70.11(d) 69.21(d)57.02(q) ， 56.60(q) 57.43(q)56.23(q) 55.98(q)56.72(d) 52.91(d), 55.10(d) 54.90(d)48.90(t) 48.57(t)40.19(d) 40.63(d), 27.67(t) 26.32(t)26.51(d) 26.44(d)24.60(t) ， 21.19(t) 20.86(t)20.47(q) 19.75(q)16.21(q) 15.97(q), 15.83(q) 15.94(q)14.04(q) 14.16(q)11.68(q) ， 9.64(q) 9.93(q) The chart of the above-mentioned spectrum is shown in Drawing 7. (10)¹H nuclear magnetic resonance spectrum: The chart of the above-mentioned spectrum is shown in Drawing 8. (11) Thin layer chromatography: [Table]

(12) Character of a substance: Neutral substance About FR-900520 substance,¹³C Reach.¹Although this substance shows one pair of signals to various chemical shifts, respectively in the case of measurement of H nuclear magnetic resonance spectrum, When thin layer chromatography and high speed liquid chromatography were measured, FR-900520 substance showed the single spot by thin layer chromatography, and showed the single peak by high speed liquid chromatography, respectively. By having determined the chemical structure of the above-mentioned physicochemical quality and FR-900506 substance, it became clear that FR-900520 substance had the following chemical structure.<img file="JPH0346445B2_D0014.tif" />17-ethyl 1 and 14-dihydroxy 12-[2-(4-hydroxy 3-methoxy cyclohexyl)-1-methylvinyl]-23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10, 16-tetra--on FR-900523 substance (1) Shape and color tone: A colorless needle crystal (2) Ultimate analysis value: C:64.57%,H:8.87%,N:1.81% (3) Color reaction: Positivity: A sulfuric acid cerium reaction, a sulfuric acid reaction, the Ehrlich reaction, the Dragendorff reaction, an iodine steamy reaction Negativity: A ferric oxide reaction, a ninhydrin reaction (4) Solubility: Meltable: Methanol, ethanol, acetone, ethyl acetate, chloroform, diethylether, benzene Refractory: N-hexane, petroleum ether Insoluble: Water (5) Melting point: 152~154degreeC (6) Specific optical rotation: [alpha]²³_D-73.0 *(C=0.65,CHCL₃) (7) Ultraviolet absorption spectrum: End absorption (8) Infrared resonance spectrum: nu^CHCl3_nax:3670,3580,3510,2930,2875,2825,1745,1722,1700,1647,1450,1380,1350,1330,1307,1285,1170,1135,1090,1050,1030-1000, 990, 978, 960,930,915,888,870,850cm^-1(9)¹³C nuclear magnetic resonance spectrum: delta (ppm, CDCl)₃):213.82(s) 213.32(s)196,31(s) 193.34(s), 168.96(s) 168.85(s)164.84(s) 165.98(s)137.80(s) 138.41(s), 132.89(s) 131.96(s)129.62(d) 130.03(d)124.51(d) 124.84(d), 97.13(s) 98.67(s)84.38(d) 76.69(d) 78.06(d), 75.45(d) 76.91(d)73.89(d) 73.70(d)73.70(d) ， 73.09(d) 72.84(d)70.40(d) 69.24(d)56.75(d) 52.89(d), 56.93(q) 57.43(q)56.61(q) 56.56(q)56.24(q) 55.94(q), 48.58(t) 48.32(t)47.14(d) 47.38(d)40.23(d) 40.65(d), 27.85(t) 26.32(t)26.48(d) 26.64(d)24.68(t) ， 21.33(t) 20.83(t)20.63(q) 19.77(q)16.24(q) 16.34(q), 15.70(q) 15.96(q)15.51(q) 15.96(q)14.31(q) 14.18(q), 9.64(q) 10.04(q) The chart of the above-mentioned spectrum is shown in Drawing 9. (10)¹H nuclear magnetic resonance spectrum: The chart of the above-mentioned spectrum is shown in Drawing 10. (11) Thin layer chromatography: [Table]

(12) Character of a substance: Neutral substance About FR-900523 substance,¹³C Reach.¹Although this substance shows one pair of signals to various chemical shifts, respectively in the case of measurement of H nuclear magnetic resonance spectrum, In measurement of thin layer chromatography and high speed liquid chromatography, thin layer chromatography showed the single spot and, as for FR-900523 substance, it showed the single peak by high speed liquid chromatography, respectively. By having determined the chemical structure of the above-mentioned physicochemical quality and FR-900506 substance, it became clear that FR-900523 substance had the following chemical structure.<img file="JPH0346445B2_D0015.tif" />1, 14-dihydroxy 12-[2-(4-hydroxy 3-methoxy cyclohexyl)-1-methylvinyl]-23, 25-dimethoxy 13, 17, 19, and 21, 27-pentamethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10, 16-tetra--on [] synthetic method: (1) Manufacturing process 1 : (introduction of a hydroxy blocking group) A compound (b) or its salt can be manufactured by introducing a hydroxy blocking group into a compound (a) or its salt. The thing of the common use as an introductory agent of the hydroxy blocking group used in this reaction, For example, dimethyl sulfoxide, ethyl methyl sulfoxide, propyl methyl sulfoxide, Isopropyl methyl sulfoxide, butyl methyl sulfoxide, isobutyl methyl sulfoxide, (Di(low-grade) Alkyl sulfoxide;, for example, chlorination trimethylsilyl, like low-grade Alkyl methyl sulfoxide, such as hexyl methyl sulfoxide) The Tori (low-grade) Alkyl silyl halogenation things, such as bromination triethyl silyl, chlorination tributyl silyl, and chlorination t-butyldimethylsilyl, For example, chlorination methyl diphenyl silyl, ethyl bromide diphenyl silyl, A 3 substitution silyl compound like low-grade Alkyl- diaryl silyl halogenation things, such as propyl chloride ditolyl silyl and chlorination t-butyl diphenyl silyl; carboxylic acid which can introduce the above-mentioned acyl group, Sulfonic acid, an acylating agent, for example like these reactant derivatives, such as acid halide, an acid anhydride, active amide, and active ester, etc. can be mentioned. As the suitable example of such a reactant derivative, An acid chloride, an acid bromide, for example, Dialkyl phosphoric acid, phenyl phosphoric acid, diphenyl phosphoric acid, Substitution phosphoric acid, such as Dibenzyl phosphoric acid and halogenation phosphoric acid, Dialkyl phosphorous acid, Alkyl carbonic ester, such as sulfurous acid, thiosulfuric acid, sulfuric acid, for example, methyl carbonate, ethyl carbonate, and carbonic acid propyl, For example, pivalate, pentanoic acid, isopentanoic acid, 2-ethyl butyl acid, A mixed acid anhydride with acid like aromatic carboxylic acid, such as aliphatic carboxylic acid, for example, benzoic acid etc., such as trichloroacetic acid and trifluoroacetic acid, A symmetrical type acid anhydride, imidazole, 4-substitution imidazole, dimethylpyrazol, doria -- active acid amide with the heterocyclic compound containing an imino group like Sole and tetra-Sole, For example, p-nitrophenyl ester, 2, 4-dinitrophenyl ester, Trichloro phenyl ester, pentachlorophenyl ester, Mesir phenyl ester, Phenylazo phenyl ester, phenylthio ester, p-nitroglycerine phenylthio ester, p-Cresylchio ester, carboxymethyl Thioester, pyridyl ester, piperidyl ester, 8-Quinolylthio ester or N, and N-Dimethyl hydroxylamine 1-hydroxy 2 -(1H)- Pyridone, N-hydroxysuccinimide, benzo[ N-hydroxy Phthal imide and / 1-hydroxy ] -- doria -- Sole and 1-hydroxy 6-chlorobenzo -- doria -- active ester, such as ester with an N-hydroxy compound like Sole, etc. are mentioned. In this reaction, when Di (low-grade) Alkyl sulfoxide is used as an introductory agent of a hydroxy blocking group, a reaction is performed under existence of anhydrous low-grade alkane acid usually like an acetic anhydride. When a 3 substitution silyl compound is used as an introductory agent of a hydroxy blocking group, it is preferred to perform a reaction under existence of the condensing agent of common use of imidazole etc. It is a reaction when an acylating agent is used as an introductory agent of a hydroxy blocking group, For example, alkali ground metal, such as alkaline metals, for example, calcium etc., such as lithium, sodium, and potassium, For example, alkali ground metal hydrides, such as alkaline metal hydrogenation things, for example, calcium hydride etc., such as sodium hydride, For example, alkali metal hydroxide, such as sodium hydroxide and a potassium hydrate, (For example, alkaline metal carbonate, for example, sodium bicarbonate, such as sodium carbonate and potassium carbonate) Alkaline metal carbonated water matter salt, for example, sodium methoxide, such as potassium bicarbonate Alkaline metal Alkoxide, such as sodium ethoxide and potassium t-butoxide, For example, it is preferred to carry out under existence of organicity like pyridine compounds, such as trialkylamine, for example, pyridine, such as alkaline metal alkane acid, for example, triethyl amine etc., such as acetic acid sodium, Lutidine, picoline, and 4-N,N-dimethylamino pyridine, and quinoline or a mineral salt machine. It is a reaction when an acylating agent uses it in the acid of isolation, or the form of that salt in this reaction, For example, N and N'-dicyclohexyl Carbo diimide, N-cyclohexyl N'-(4-diethylamino cyclohexyl) Carbo diimide, N,N'-Diethylcarb diimide, N,N'-Diiso propyl Carbo diimide, Carbo diimide compounds, such as N-ethyl N'-(3-dimethylaminopropyl) Carbo diimide, For example, an N and N'-carbonyl screw (2-methyl imidazole), pentamethylene ketene N-cyclohexyl imine, Ketene imine compounds, such as diphenyl ketene N-cyclohexyl imine, For example, Ole Huynh or acetylene system ether compounds, such as ethoxy acetylene and beta-cyclo vinyl ethyl ether, benzo[ for example, / 1-(4-chlorobenzene sulfonyloxy)-6-chloro 1H-] -- doria -- benzo[, such as Sole, / N'-hydroxy ] -- doria -- it is preferred to carry out under existence of the condensing agent of common use of the sulfonic ester of a Sole derivative, etc. Reactions are usually water, acetone, and dichloromethane, for example, methanol, It is carried out in the solvent of the common use which does not have a bad influence on this reaction like these mixed solvents, such as alcohol [, such as ethanol, ], tetrahydro franc, pyridine, N, and N-Dimethylform amide, When the introductory agent of a base or a hydroxy blocking group is a fluid, these can also be used as a solvent. Reaction temperature in particular is not limited but is usually performed under cooling thru/or heating. In this reaction, it is R of a compound (a).²Since it converts into the hydroxy group from which it came out and the hydroxy group shown was protected, also in such a case, it is included by this manufacturing process. When bottom Di (low-grade) Alkyl sulfoxide of existence of anhydrous low-grade alkane acid is used as a hydroxy blocking group introduction agent in this reaction, Formula: [Formula]

(Inside of a formula, R)²With a Is hydroxy group being certain -- the compound (a) which has the partial structure shown oxidizes -- formula: [Formula]

(Inside of a formula, R)²The object compound (b) which has the partial structure shown by Is hydroxy Is may be obtained, and, also in such a case, it is included by the range of this manufacturing process. (2) Manufacturing process 2 : (introduction of a hydroxy blocking group) A compound (d) or its salt can be manufactured by introducing a hydroxy blocking group into a compound (c) or its salt. This reaction can be substantially performed in a similar way with manufacturing process 1, therefore reaction conditions, such as a base, a condensing agent, a solvent, and reaction temperature, can use manufacturing process 1, for example. In this reaction, it is R of a compound (c).¹The hydroxy group which can be boiled and set may convert into the protected hydroxy group corresponding with the object compound (d), and, also in such a case, it is included by the range of this manufacturing process. (3) Manufacturing process 3 : (formation of a double bond) A compound (f) or its salt can be manufactured by making a base react to a compound (e) or its salt. As a base used at this reaction, the base illustrated by manufacturing process 1 can be mentioned as it is. This reaction is R again.²It can manufacture also by making a But hydroxy Is compound (e) or its salt react to the bottom acylating agent of existence of a base. This reaction is water, acetone, dichloromethane, for example, methanol, and ethanol, It is carried out in the solvent of the common use which does not have a bad influence on this reaction like those mixed solvents, such as alcohol [, such as propanol, ], tetrahydro franc, pyridine, N, and N-Dimethylform amide, and further, when a base is a fluid, these can also be used as a solvent. Reaction temperature in particular is not limited but is usually performed under cooling thru/or heating. (4) Manufacturing process 4 : (oxidization of a hydroxy ethylene group) A compound (b) or its salt can be manufactured by oxidizing a compound (g) or its salt. As an oxidizer used at this reaction, Di (low-grade) Alkyl sulfoxide as shown according to manufacturing process 1 can be mentioned. Under existence of anhydrous low-grade alkane acid [ usually like an acetic anhydride ] in which this reaction is, Acetone, dichloromethane, ethyl acetate, a tetrahydro franc, pyridine, It is carried out in the solvent of the common use which does not have a bad influence on this reaction like those mixed solvents, such as N and N-Dimethylform amide, and further, when anhydrous low-grade alkane acid is a fluid, these can also be used as a solvent. Reaction temperature in particular is not limited but is usually performed under cooling thru/or heating. This manufacturing process is R of a materials compound (g) in the middle of a reaction.¹It may come out, the hydroxy group shown may convert into 1-(low-grade Alkylthio) (low-grade) alkoxy group in the object compound (h), and, also in such a case, it is included within the limits of this manufacturing process. (5) Manufacturing process 5 : (reduction of an allyl group) A compound (j) or its salt can be manufactured by returning compound (i) or its salt. Reduction in this manufacturing process can be performed by the usual state method which can return allyl groups, such as contact reduction, to a propyl group. As a catalyst used by contact reduction, they are a platinum board, platinum sponge, and platinum black, for example, Platinum catalysts, for example, palladium sponge, such as colloidal platinum, platinum oxide, and a platinum line Palladium black, oxidization palladium, palladium carbon, colloid palladium, The palladium catalyst of palladium and barium sulfate, palladium charcoal acid barium, etc., For example, conventional things, such as a copper catalyst of the iron catalyst of the cobalt catalyst of nickel catalysts, for example, reduction cobalt, such as reduced nickel, nickel oxide, and Raney nickel, Raney cobalt, etc., for example, reduced iron, Raney iron, etc., for example, reduction copper, Raney copper, Ullman copper, etc., are mentioned. This reduction is performed in the solvent of the common use which does not usually have a bad influence on this reaction like water, methanol, ethanol, propanol, pyridine, ethyl acetate, N, and N-Dimethylform amide, dichloro ethane, or its mixed solvent. the reaction temperature in particular of this reduction is not limited -- usually -- warming under cooling -- it is carried out in the lower range. For example by usual state methods, such as extraction, precipitation, fractional-crystallization-izing, re-crystallization, and chromatography, tricyclo compound () obtained by manufacturing process 1~5 explained above can isolate, and it can refine it. Compound () and (a) As the salt in ~ (j), It is salt of the nonpoisonous common use permitted as medicine, for example, is sodium salt, Salt with the inorganic matter or the organic base like amine salt, such as alkali ground metal salt, such as alkali metal salt, for example, calcium salt, such as potassium salt, and magnesium salt, ammonium salt, for example, triethyl amine salt, and N-Benji Lu N-methylamine salt, is mentioned. It is under [ reaction / of the above-mentioned manufacturing process 1~5 /, or post-processing / of the reaction mixture ] setting, The stereoisomeric form by Conformer and asymmetric carbon atom, or double bond of a start compound and the object compound may be converted into other Conformer and stereoisomeric forms, and, also in such a case, it is included by the scope of this invention. Tricyclo compound () used by this invention, Having [ therefore ] an immunodepressive effect, the immunosuppressants of the present invention are the heart, the kidney, and liver, It is useful to the medical treatment and prevention of an autoimmune disease etc. of internal organs, such as marrow and the skin, or the rejection to a transplant of an organization, a graft versus host reaction that occurs by a bone marrow transplantation, rheumatoid arthritis, systemic lupus erythematosus, the Hashimoto thyroiditis, multiple sclerosis, myasthenia gravis, model diabetes, a grape film flame, etc. In order to show the usefulness of this immunosuppressant, pharmacological test data is shown below. Examination 1 the control effect of tricyclo compound () in an in vitro mixed lymphocyte reaction (MLR) -- each -- a well -- C57BL / 6 response cells (H-2)^b5x10⁵The BALB/C stimulus cell which mitomycin C processing (it processes for 30 minutes by 37 degreeC by dynamite Machining C25microg/ml, and washes 3 times by RPMI1640 culture medium) made into Pieces (H-2)^dx5x10⁵A MLR examination is done in the microtiter plate which added Pieces to Fetal bovine serum RPMI1640 culture medium [it is addition about sodium bicarbonate 2mM, penicillin (50 units/(ml)), and streptomycin (50 microg/(ml))] 0.2-ml 10%. A cell is cultured by 37 degreeC for 68 hours within Culture device maintained at 100% of humidity, 5% of carbon dioxide, and 95% of air, and it is 4 hours.³A pulse is carried out by H-thymidine (0.5 microcurie), and cells are collected. The object compound of this invention is dissolved in ethanol, and it dilutes to RPMI1640 culture medium, adds to a culture, and makes the last concentration become [ ml ] less than in 0.1 microg /. A result is shown in Table 7~10. The tricyclo compound of this invention controlled mouse MLR.

[Table]

[Table]

[Table]

[Table]

[Table]

Examination 2 The piece of an abdomen skin transplant of effect donor (Fischer) Rat of tricyclo compound () to the piece survival of an of-the-same-kind skin transplant in Rat is transplanted to the outside chest of recipient (WKA) Rat. A bandage will be removed on the 5th. The piece of a transplant is inspected every day until there is rejection in which 90% or more of the transplant Kataue skin necroses. FR-900506 substance is dissolved in olive oil and continuation intramuscular administration is carried out for 14 days from a transplant day every day. As shown in Table 11, in Rat which carried out continuation intramuscular administration of the olive oil for 14 days, the skin allograft was altogether refused within eight days, but skin allograft survival extended clearly that with which it dealt by FR-900506 substance every day.

[Table]

Examination 3 The effect collagen of tricyclo compound () to the model collagen indecement arthritis in Rat is dissolved in cold 0.01M acetic acid so that it may become the concentration of 2mg/ml. It emulsifies in the imperfect Freund adjuvant of capacity [ solution ]. Intradermal injection of this emulsification thing of 0.5 ml of total amounts is carried out to several places of the back of feminity Lewis system Rat, and 1 or 2 places of a tail. It dissolves in olive oil and administers FR-900506 substance orally. Only olive oil is administered orally to contrast Rat which carried out immunity by an equivalent amount of model collagen. The revelation rate of arthritis is observed. A test result is shown in Table 12. Arthritis was induced by all Rat processed with olive oil for 14 days from the same day as model collagen immunization. When FR-900506 substance was prescribed for the patient every day during 14 days, indecement of arthritis was completely controlled during the observation period for three weeks.

[Table]

Examination 4 The effect of tricyclo compound () to the experimental allergic encephalomyelitis (EAE) in a SJL/J system mouse Spine gay Ginet is adjusted from a SJL/J system mouse. A spine is taken out by the aerating method, and it mixes with the water of capacity, such as abbreviation, and carries out gay Ginet by 4 degreeC. This cold gay Ginet (10mg/(ml)) of Equal capacity is emulsified with the perfect Freund adjuvant (CFA) containing Maiko bacterium Tuberculosis H37RA0.6mg/ml. Eyes will pour 0.2 ml of spine-CFA emulsion into a SJL/J system mouse twice with the 0th day for 13 days, and it will make it induce EAE. The clinical fine season of EAE is evaluated and judged every day about all the mice used for this examination. The grade of EAE is Yoda to the following judging standard. Grade 1: A tail strain fall, grade 2: An awkward walk, exhaustion of the limbs of 3:1 or more grades, grade 4: Pair paralysis or Hemiplegia. FR-900506 substance is dissolved in olive oil and it will administer orally for 19 days from the 0th (day of first time immunity) day. As shown in Table 13, FR-900506 substance prevented the appearance of the fine season of EAE clearly.

[Table]

Examination 5 Raw splenic cells obtained from effect C57BL/6 donor of tricyclo compound () to the local graft versus host reaction (GvHR) kicked to a mouse (1x10)⁷About Pieces, it is BDF.₁a system -- the right rear of a mouse -- a leg -- a hypodermic injection is carried out to a planta. A mouse is slaughtered in seven days and the weight of the popliteal lymph nodes (PLN) of both the right (injected leg) and the left (leg which is not injected) is measured. GvHR is denoted by the weight difference of right and left PLN. FR-900506 substance is dissolved in olive oil and it administers orally for five days from a sensitization day. ED of local graft versus host reaction control of FR-900506 substance₅₀A value is 19mg/kg. Examination 6 Acute toxicity of tricyclo compound () The examination on the acute toxicity of FR-900506 by the intraperitoneal injection in a ddy mouse, FR-900520, FR-900523, and FR-900525 substance was done, and death was observed with the given dose of 100mg/kg at neither of the cases. The immunosuppressant of this invention can contain tricyclo compound () as an active ingredient, and can be mixed with an organic or inorganic carrier or a diluent base suitable for external application, internal use, or parenteral supplementation, for example, can be used in the shape of a solid, the shape of a half-solid, or the form of a liquefied medicine tablet. An active ingredient can be used in a tablet, a pellet agent, a capsule agent, a Sitting agent, liquid medicine, emulsion, suspension, and other suitable forms, for example, usual is nonpoisonous, and it may mix with the carrier permitted as medicine. The carriers which can be used are water, glucose, lactose, acacia gum, and gelatin, It is a carrier suitable for manufacturing mannitol, a starch paste, magnesium trisilicate, talc, cornstarch, keratin, colloid silica, potato starch, urea, the shape of other solid, the shape of a half-solid, or a liquefied tablet. A diluent base, a stabilizer, a consistency-ized agent, colorant, and spice may be used. Sufficient quantity to demonstrate a desired effect contains the object compound in a medicine tablet according to progress or the state of a disease. When using this immunosuppressant for humans, it is preferred to use it by parenteral supplementation or internal use. Although the medical treatment effective amount of tricyclo compound () changes with the age of patient each and the grades of the illness to treat, usually, about 0.01~1000 mg with an active ingredient of one day -- desirable, still more preferably 0.5~100 mg is used for the medical treatment of a disease, and, generally an average of about 0.5 mg, 1 mg, 5 mg, 10 mg, 50 mg, 100 mg, 250 mg, and 500 mg 0.1~500-mg is prescribed once for the patient. Hereinafter, the example of manufacture of tricyclo compound () contained in the immunosuppressant of this invention is shown. Example of manufacture 1 Streptomyces tsukubaensis No.9993 was separated using the monotonous dilution method shown below in separation of Streptomyces tsukubaensis No.9993. About 1 g of soil extracted in Tsukuba-gun, Ibaraki Toyosato-cho is put into a sterility test pipe, sterile water is added, and it is considered as the capacity of 5 ml. At a tube buzzer, it mixes for 10 seconds and neglects a mixture for 10 minutes. Supernatant liquid is diluted with sterile water 100 times one by one. It is thiamin hydrochloride addition Supapec agar (saccharose 30g) about a diluted solution (0.1 ml). It extends on sodium nitrate 3g, 1 g of phosphoric acid 2 potassium, magnesium sulfate 0.5g, 0.5 g of potassium chloride, ferrous sulfate 0.01g, 0.1 g of thiamin hydrochloride, agar 20g, and the petri dish containing tap water 1000 ml;PH7.2. It incubates for 21 days by 30 degreeC, the colony grown on the plate is moved to a slope culture medium [yeast-malt extract agar (ISP culture medium 2)], and it cultivates for ten days by 30 degreeC. Streptomyces tsukubaensis No.9993 was found out in the separated colony. Fermentation Glycerin (1%), soluble starch (1%), glucose (0.5%). The culture medium (160 ml) (it adjusts to PH6.5) containing cottonseed powder (0.5%), dried yeast (0.5%), corn Steie Prika (0.5%), and calcium carbonate (0.2%) is respectively put into 20 500-ml Elren Mayer flasks, and it sterilizes for 30 minutes by 120 degreeC. One platinum loop of the slant culture thing of Streptomyces tsukubaensis No.9993 (deposition number: FERM BP No. 927) is inoculated into each culture medium, and it cultivates for four days by 30 degreeC on a rotation shaker. Soluble starch which put the culture obtained into the 200-liter Jeffrey- mentor which sterilized for 20 minutes by 120 degreeC beforehand (4.5%), Corn Steie Prika (1%), dried yeast (1%), calcium carbonate (0.1%), and the culture medium (150 liters) containing Adekanol (an antifoaming agent, a brand name, the product made from the Asahi electrification) (0.1%) are inoculated, and it is 30degreeC, It cultivates for four days under aeration (150 liters/(minute)) and churning (250 rpm). Isolation and refining Thus, the obtained cultivation broth is filtered using diatomite (5 kg). Methanol (50 liters) extracts the lump of a fungal thread object, and it obtains 50 liters of extraction liquid. The methanol extraction liquid and Filtrate which are obtained from a fungal thread object are united, and it lets it pass in the column of nonionic adsorption resin "Dia ion HP-20" (a brand name, the Mitsubishi Kasei Corp. make) (10 liters). After washing with water (30 liters) and water methanol (30 liters), it is eluted with methanol. It shall be made to evaporate under decompression of elution liquid, and a residue shall be 2 liters. Ethyl acetate (2 liters) extracts this. Ethyl acetate extraction liquid is concentrated in vacuum and an oily residue is obtained. An oily residue is mixed with acid silica gel (special silica gel and grade:12, made in Devi Fuji Son) of weight twice, and this mixture is slurry-ized in ethyl acetate. The dry powder obtained is given to the chromatography using the column which filled up the same acid silica gel (800 ml) as the above with n-hexane after distilling off a solvent. A column is developed with the mixture (9:1 v/v, 3 liter;4:1 v/v, 3 liters) of n-hexane (3 liters), n-hexane, and ethyl acetate, and ethyl acetate (3 liters). A part for the drawing containing the object compound is collected and concentrated in vacuum, and an oily residue is obtained. An oily residue is dissolved in the mixture (1:1 v/v, 30 ml) with n-hexane and ethyl acetate, and the chromatography using the column which filled up silica gel (230~400 meshes, Merck Co. make) (500 ml) with the same solvent system is given. It is eluted with the mixture (1:1 v/v, 2 liter;1:2 v/v, 1.5 liters) of n-hexane and ethyl acetate. A part for the drawing containing the first object compound is collected and concentrated in vacuum, and a yellow oily matter is obtained. This oily residue is mixed with acid silica gel of weight twice, and this mixture is slurry-ized in ethyl acetate. The dry powder obtained is covered over the column which filled up acid silica gel with n-hexane after distilling off a solvent, and it develops by n-hexane. A part for the drawing containing the object compound is collected and concentrated in vacuum, and rough FR-900506 substance (1054 mg) is obtained as white powder. 100 mg of this rough product is given to high speed liquid chromatography. It is eluted using the column (8phix500mm) which makes a carrier Lichrosorb (Lichrosorb)SI60 (a brand name, the Merck Co. make). This chromatography is monitored on the wavelength of 230 nm with UV detector, and methylene chloride dioxane Mixture (85:15 v/v) is used at the 5ml/minute flow velocity, and a mobile phase collects parts for active drawing, and carries out evaporation concentration. This high speed liquid chromatography is repeated and 14 mg of refining FR-900506 substances are obtained as white powder. It carries out by continuing elution with ethyl acetate (1.5 liters), a part for the drawing containing the second object compound is collected and concentrated in vacuum, and rough FR-900525 substance (30 mg) is obtained as a yellow oily matter. Example of manufacture 2 Fermentation Glycerin (1%), cornstarch (1%), glucose (0.5%), The pre-cultivation culture medium (100 ml) of PH6.5 containing cottonseed powder (1%), corn Steie Prika (0.5%), dried yeast (0.5%), and calcium carbonate (0.2%) is put into a 500-ml Elren Mayer flask, and it sterilizes for 30 minutes by 120 degreeC. One platinum loop of the slant culture thing of Streptomyces tsukubaensis No.9993 is inoculated into a culture medium, and it cultivates for four days by 30 degreeC. The culture obtained is moved to the pre-cultivation culture medium (20 liters) in the 30-liter Jihuahua mentor which sterilized for 30 minutes by 120 degreeC beforehand. A culture is incubated for two days by 30 degreeC, and they are 16 liters of front cultures, Soluble starch put into the 2-ton tank which sterilized for 30 minutes by 120 degreeC beforehand (4.5%), Corn Steie Prika (1%), dried yeast (1%), calcium carbonate (0.1%), and the fermentation culture medium (1600 liters) of PH6.8 containing Adekanol (an antifoaming agent, a brand name, the product made from the Asahi electrification) (0.1%) are inoculated, and it cultivates for four days by 30 degreeC. Isolation and refining Thus, the cultivation broth obtained is filtered using diatomite (25 kg). Acetone (500 liters) extracts the lump of a fungal thread object, and it obtains 500 liters of extraction liquid. This acetone extraction liquid and Filtrate (1350 liters) are united, and it lets it pass in the column of nonionic adsorption resin "Dia ion HP-20" (a brand name, the Mitsubishi Kasei Corp. make) (100 liters). After washing with water (300 liters) and 50% water acetone (300 liters), it is eluted with water acetone 75%. It shall be made to evaporate under decompression of elution liquid, and a residue shall be 300 liters. Ethyl acetate (20 liters) extracts this 3 times. Ethyl acetate extraction liquid is concentrated in vacuum and an oily residue is obtained. This oily residue is mixed with acid silica gel (special silica gel, grade 12, made in Devi Fuji Son) of weight twice, and this mixture is slurry-ized in ethyl acetate. A solvent is evaporated and the dry powder obtained is given to the column chromatography which filled up the same acid silica gel (8 liters) as the above with n-hexane. A column is developed with n-hexane (30 liters), n-hexane ethyl acetate Mixture (4:1 v/v, 30 liters), and ethyl acetate (30 liters). A part for the drawing containing the object compound is collected and concentrated in vacuum, and an oily residue is obtained. This oily residue is mixed with acid silica gel of weight twice, and a mixture is slurry-ized in ethyl acetate. The powder obtained is again given to the chromatography using the column of acid silica gel (3.5 liters) filled with n-hexane after distilling off of a solvent. A column is developed with n-hexane (10 liters), n-hexane ethyl acetate Mixture (4:1 v/v, 10 liters), and ethyl acetate (10 liters). A part for the drawing containing the object compound is collected and concentrated in vacuum, and a yellow oily matter is obtained. This oily residue is dissolved in n-hexane ethyl acetate Mixture (1:1 v/v, 300 ml), and the chromatography using the column of silica gel (230~400 meshes, Merck Co. make) (2 liters) filled with the same solvent system is given. It is eluted with n-hexane ethyl acetate Mixture (1:1 v/v, 10 liter;1:2 v/v, 6 liters) and ethyl acetate (6 liters). A part for the drawing containing the first object compound is collected and concentrated in vacuum, and FR-900506 substance is obtained as white powder (34g). It dissolves in acetonitrile and concentrates this white powder in vacuum. A concentrate is neglected by 5 degreeC overnight and a prism-like crystal (22.7g) is obtained. It re-crystallizes with the same solvent and obtains refining FR-900506 substance (13.6g) as a colorless prism-like crystal. A part for the drawing containing the second object compound is collected and concentrated in vacuum, and rough FR-900525 substance (314 mg) is obtained as yellow powder. Example of manufacture 3 Fermentation Glycerin (1%), cornstarch (1%), glucose (0.5%), The culture medium (160 ml) (it adjusts to PH6.5) containing cottonseed powder (1%), dried yeast (0.5%), corn Steie Prika (0.5%), and calcium carbonate (0.2%) is respectively put into ten 500-ml Elren Mayer flasks, and it sterilizes for 30 minutes by 120 degreeC. One platinum loop of the slant culture thing of Streptomyces tsukubaensis No.9993 is inoculated into each culture medium, and it cultivates for four days by 30 degreeC on a rotation shaker. Soluble starch which put the obtained culture into the 200-liter Jihuahua mentor which sterilized for 20 minutes by 120 degreeC beforehand (5%), Peanut powder (0.5%), dried yeast (0.5%), a gluten meal (0.5%), Calcium carbonate (0.1%) and the culture medium (150 liters) containing Adekanol (an antifoaming agent, a brand name, the product made from the Asahi electrification) (0.1%) are inoculated, and it cultivates for four days under aeration (150 liters/(minute)) and churning (250 rpm) by 30 degreeC. Isolation and refining Thus, the cultivation broth obtained is filtered using diatomite (5 kg). Acetone (50 liters) extracts the lump of a fungal thread object, and it obtains 50 liters of extraction liquid. The acetone extraction liquid and Filtrate (135 liters) which are obtained from a fungal thread object are united, and it lets a column pass to nonionic adsorption resin "Dia ion HP-20" (a brand name, the Mitsubishi Kasei Corp. make) (10 liters). It washes with water (30 liters) and 50% water acetone (30 liters), and is eluted with water acetone 75%. Evaporation concentration is carried out under decompression of elution liquid (30 liters), 2 liters of residues are carried out, and ethyl acetate (2 liters) extracts 3 times. Ethyl acetate extraction liquid is concentrated in vacuum and an oily residue is obtained. This oily residue is mixed with acid silica gel (special silica gel, grade 12, made in Devi Fuji Son) of weight twice, and a mixture is slurry-ized in ethyl acetate. The dry powder obtained by distilling off a solvent is given to the chromatography using the column which filled up the same acid silica gel (800 ml) as the above with n-hexane. A column is developed with n-hexane (3 liters), n-hexane ethyl acetate Mixture (4:1 v/v, 3 liters), and ethyl acetate (3 liters). A part for the drawing containing the object compound is collected and concentrated in vacuum, and an oily residue is obtained. This oily residue is dissolved in n-hexane ethyl acetate Mixture (1:1 v/v, 30 ml), and the chromatography using the column of silica gel (230~400 meshes, Merck Co. make) (500 ml) filled with the same solvent system is given. It is eluted with n-hexane ethyl acetate Mixture (1:1 v/v, 2 liter;1:2 v/v, 1.5 liters) and ethyl acetate (1.5 liters). A part for the drawing containing the first object compound is collected and concentrated in vacuum, and rough FR-900506 substance (3g) is obtained as yellow powder. A part for the drawing containing the second object compound is collected and concentrated in vacuum, and an oily residue is obtained. This oily residue is again given to silica gel chromatography, and a yellow oily matter is obtained. This oily residue is mixed with acid silica gel of weight twice, and a mixture is Slurry-ized in ethyl acetate. Dry powder produced by distilling off a solvent is given to the chromatography using the column which filled up acid silica gel (100 ml) with n-hexane, and it develops by n-hexane. A part for the drawing containing the object compound is collected and concentrated in vacuum, and FR-900525 substance is obtained as light yellow powder (380 mg). This powder is dissolved in ethyl Mixture from n-hexane vinegar (1:2 v/v, 5 ml), and the chromatography using the column of the acid silica gel (special silica gel, grade 922, made in Devi Fuji Son) (100 ml) which was filled up with the same solvent system and washed is given. It is eluted with ethyl acetate. Collect parts for active drawing, it is made to evaporate under decompression, and FR-900525 refined substance (230 mg) is obtained as white powder. Example of manufacture 4 With the monotonous dilution method shown below in separation of Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238, Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238 is separated. About 1 g of soil extracted in Kagoshima Yaku Islands shall be put into a sterility test pipe, and capacity shall be 5 ml, adding sterile water. At a tube buzzer, it mixes for 10 seconds and neglects a mixture for 10 minutes. Supernatant liquid is diluted with sterile water 100 times one by one. It is thiamin hydrochloride addition Supapec agar (saccharose 30g) about a diluted solution (0.1 ml). It extends on sodium nitrate 3g, 1 g of phosphoric acid 2 potassium, magnesium sulfate 0.5g, 0.5 g of potassium chloride, ferrous sulfate 0.01g, 0.1 g of thiamin hydrochloride, agar 20g, and the petri dish containing tap water 1000 ml;PH7.2. It incubates for 21 days by 30 degreeC, the colony grown on the plate is moved to a slope culture medium [yeast-malt extract agar (ISP-culture medium 2)], and it cultivates for ten days by 30 degreeC. In the separated colony, Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238 was found out. Fermentation Glycerin (1%), soluble starch (1%), glucose (0.5%), The culture medium (160 ml) (it adjusts to PH6.5) containing cottonseed powder (0.5%), dried yeast (0.5%), corn Steie Prika (0.5%), and calcium carbonate (0.2%) is respectively put into 20 500-ml Elren Mayer flasks, and it sterilizes for 30 minutes by 120 degreeC. One platinum loop of the slant culture thing of Strept Mrs. high gloss Copicus subsp. Yakushima Encis No.7238 (deposition number: FERM BP No. 928) is inoculated into each culture medium, and it cultivates for four days by 30 degreeC on a rotation shaker. About the culture obtained, they are glucose (4.5%) and corn Steie Prika (1%), Dried yeast (1%), a gluten meal (1%), malt (0.5%), calcium carbonate (0.1%), Adekanol (an antifoaming agent, a brand name, the product made from the Asahi electrification) (0.1%) is contained, The culture medium (150 liters) in the 200-liter Jihuahua mentor which sterilized for 20 minutes by 120 degreeC beforehand is inoculated, and it cultivates for four days under 30degreeC, aeration (150 liters/(minute)), and churning (250 rpm). Isolation and refining Thus, the cultivation broth obtained is filtered using diatomite (5 kg). Acetone (50 liters) extracts the lump of a fungal thread object, and it obtains 50 liters of extraction liquid. The acetone extraction liquid and Filtrate (135 liters) which are obtained from a fungal thread object are united, and it lets it pass in the column of nonionic adsorption resin "Dia ion HP-20" (a brand name, the Mitsubishi Kasei Corp. make) (10 liters). After washing with water (30 liters) and water acetone (30 liters), it is eluted with acetone. Evaporation concentration shall be carried out under decompression of elution liquid, a residue shall be 2 liters, and ethyl acetate (4 liters) extracts this. Ethyl acetate extraction liquid is concentrated in vacuum and an oily residue is obtained. This oily residue is mixed with acid silica gel (special silica gel grade 12, made in Devi Fuji Son) of weight twice, and this mixture is slurry-ized in ethyl acetate. After distilling off a solvent, the dry powder obtained is given to the chromatography using the column of the same acid silica gel (800 ml) filled with n-hexane. A column is developed with n-hexane (3 liters), n-hexane ethyl acetate Mixture (4:1 v/v, 3 liters), and ethyl acetate (3 liters). A part for the drawing containing FR-900520 and FR-900523 substance is collected and concentrated in vacuum, and an oily residue is obtained. This oily residue is dissolved in n-hexane ethyl acetate Mixture (1:1 v/v, 50 ml), and the chromatography using the column of silica gel (70~230 meshes, Merck Co. make) (1000 ml) filled with the same solvent system is given. It is eluted with n-hexane ethyl acetate Mixture (1:1 v/v, 3 liter;1:2 v/v, 3 liters) and ethyl acetate (3 liters). A part for the drawing containing the object compound is collected and concentrated in vacuum, and yellow powder (4.5g) is obtained. This powder is dissolved in methanol (20 ml), and it mixes with water (10 ml). The chromatography using the column of the negative phase silica gel "YMC" (60~200 meshes) (500 ml) (a brand name, mountain village Institute for Chemical Research make) which fills up a mixture with methanol Water mixture (4:1 v/v), and it develops now is given. A part for the drawing containing FR-900520 substance is collected and concentrated in vacuum, and the rough product (1.8g) of FR-900520 substance is obtained as light yellow powder. This powder is dissolved in a small amount of diethylether. After neglecting it overnight, the precipitating crystal is separated, and decompression dryness is washed and carried out by diethylether. It re-crystallizes by diethylether and obtains 600 mg of refining FR-900520 substances as a colorless plate crystal. It carries out by continuing the negative phase silica gel chromatography using the same solvent system, the part for the following drawing containing FR-900523 substance is collected and concentrated in vacuum, and the rough product (0.51g) of FR-900523 substance is obtained as light yellow powder. The chromatography using the negative phase silica gel "YMC" (70 ml) which dissolves this rough product in acetonitrile (3 ml), is filled up with acetonitrile tetrahydro franc 50mM phosphate buffer solution Mixture (PH2.0) (3:2:5 v/v), and is developed now is given. Ethyl acetate extracts a part for the drawing containing the object compound. This extraction liquid is concentrated in vacuum and the powder (190 mg) of a yellowish-white color is obtained. The powder of this yellowish-white color is again given to chromatography on negative phase silica gel "YMC", and white powder (80 mg) is obtained. This white powder is dissolved in a small amount of diethylether, and it allows to stand at room temperature overnight, and obtains 56 mg of crystals. It re-crystallizes by diethylether and obtains 34 mg of colorless needle shape crystals of FR-900523 substance. Example of manufacture 5 Pyridine (0.1 ml) and an acetic anhydride (0.05 ml) are added to the solution in dichloromethane (0.2 ml) of FR-900506 substance (10.4 mg) at room temperature, and a mixture is agitated for 5 hours. It removes under decompression of a solvent. A residue is given to silica gel thin layer chromatography (developing solvent: diethylether dichloromethane, 1:2 v/v), 12-[2-(4-acetoxy 3-methoxy cyclohexyl)-1-methylvinyl]-17-ant roux 1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (6.0 mg) are obtained. IRnu (CHCl)₃):3520,1728,1705(sh)1640,1095cm^-1Example of manufacture 6 Pyridine (0.5 ml) and an acetic anhydride (0.3 ml) are added to the solution in dichloromethane (1 ml) of FR-900506 substance (52.5 mg) at room temperature, and a mixture is agitated at room temperature for 9 hours. It removes under decompression of a solvent. A residual substance is given to silica gel thin layer chromatography (developing solvent: the diethylether hexane, 3:1 v/v), 14-acetoxy 12-[2-(4-acetoxy 3-methoxy cyclohexyl)-1-methylvinyl]-17-ant roux 1-hydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10, 16-tetra--on (48.0mg) And 12-[2-(4-acetoxy 3-methoxy cyclohexyl)-1-methylvinyl]-17-ant roux 1-hydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] Octakosa- 14, 18-Diene- 2, 3, and 10, and 16-tetra--on (5.4 mg) are obtained. The above-mentioned compound IRnu (CHCl)₃):1730,1720(sh)1640cm^-1The after-mentioned compound IRnu (CHCl)₃):1730,1690,1640,1627cm^-1Example of manufacture 7 Chlorination benzoyl (50microl) is added to the dichloromethane (0.2 ml) of FR-900506 substance (9.7 mg), and the solution in pyridine (0.1 ml) at room temperature, and a mixture is agitated at room temperature for 2 hours. It removes under decompression of a solvent and obtains a rough oily matter. This oily matter is refined by silica gel thin layer chromatography (developing solvent: the diethylether hexane, 2:1 v/v), 17-ant roux 12-[2-(4-benzoyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (8.0 mg) are obtained. IRnu (CHCl)₃):3500,1735(sh)1710,1640,1600cm^-1Example of manufacture 8 Chlorination p-nitrobenzoyl (about 100 mg) is added to the solution in pyridine (1 ml) of FR-900506 substance (30.5 mg), and a mixture is agitated at room temperature for 2 hours. A reaction mixture is diluted with ethyl acetate, and in saturated sodium bicarbonate solution, water, 1N chloride, water, saturated sodium bicarbonate solution, water, and sodium chloride solution, it washes one by one and dries. The solution obtained is concentrated in vacuum and a residual substance is refined by silica gel column chromatography, 17-ant roux 1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, and 27-tetramethyl 12-[-- 2-[4-(p-nitrobenzoyloxy)-3 - methoxy -- cyclohexyl]-1-methylvinyl]-11, 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (37.7 mg) are obtained. IRnu (CHCl)₃):1720,1640,1610,1530-1520cm^-1Example of manufacture 9 FR-900506 substance (30.6 mg) and chlorination 3,5-Dinitobe b Soil (33 mg) are made to react in pyridine (0.5 ml) according to the method of example 8 of manufacture, 17-ant roux 1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, and 27-tetramethyl 12-[-- 2-[4-(3, 5-dinitro benzoyloxy)-3 - methoxy -- cyclohexyl]-1-methylvinyl]-11, 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (36.0 mg) are obtained. IRnu (CHCl)₃):1730,1640,1610,1530-1520cm^-1Example of manufacture 10 FR-900506 substance (48 mg) is made to react to chlorination 2-l-menthyl Oxyacetyl (0.08 ml) in pyridine (0.5 ml) according to the method of example 8 of manufacture, 17-ant roux 1, 14-dihydroxy 23, and 25-dimethoxy 12-[-- 2-[4-(2-l-menthyl Oxy acetoxy)-3 - methoxy -- cyclohexyl]-1-methylvinyl]-13, 19, 21, and 27-tetramethyl 11, 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (50.9 mg) are obtained. IRnu(neatness): 3520-1760, 1740(sh)1720(sh)1652cm^-1Example of manufacture 11 N and N'-dicyclohexyl Carbo diimide (47 mg) is added to the solution in ethyl acetate (10 ml) of (-)-2-trifluoromethyl 2-methoxy 2-phenylacetic acid (51 mg) at room temperature. After agitating at room temperature for 1.5 hours, FR-900506 substance (25.0 mg) and 4-(N,N-dimethylamino) pyridine (11 mg) are added, and it agitates at room temperature for 3.5 hours. The solution obtained is condensed and a residual substance is obtained, and it takes and ranks second to diethylether and washes this one by one in chloride, sodium bicarbonate solution, and sodium chloride solution. An organic layer is dried and condensed with sodium sulfate, a residual substance is obtained, and this is given to silica gel chromatography (developing solvent: dichloromethane diethylether, 10:1 v/v), 17-ant roux 12-[2-[4-[ (-)-2-trifluoromethyl 2-methoxy 2-phenyl -- acetoxy]-3-methoxy cyclohexyl]-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, and 27-tetramethyl 11, 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10, 16-tetra--on (6.5mg) And 17-ant roux 14-[ (-)-2-trifluoromethyl 2 - methoxy 2-phenyl acetoxy]-12-[2-[4-[(-)-2-trifluoromethyl 2-methoxy 2-phenyl acetoxy]-3-methoxy cyclohexyl]-1-methylvinyl]-1-hydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (20.2 mg) are obtained. The above-mentioned compound IRnu(neatness): 3510-1750, 1730(sh)1710-1652, 1500 cm^-1The after-mentioned compound IRnu(neatness): 1750-1720, 1652 or 1500 cm^-1Example of manufacture 12 The mixture which adds a succinic anhydride (145 mg) and 4-(N,N-dimethylamino) pyridine (7 mg) to this, and is obtained is agitated at room temperature for 18 hours, agitating the solution in pyridine (7 ml) of FR-900506 substance (248 mg). A reaction mixture is concentrated in vacuum and a residual substance is given to the chromatography of silica gel (20g) using ethyl acetate, 17-ant roux 12-[2-[4 - (3-Carboxy propionyloxy)-3-methoxy cyclohexyl]-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (90 mg) are obtained. IRnu (CHCl)₃):3500,3100-2300,1720,1705(sh)1635cm^-1Example of manufacture 13 Chlorination p-iodine benzenesulphonyl (500 mg) is added to the solution in pyridine (3 ml) of FR-900506 substance (100.7 mg), and a mixture is agitated at room temperature for 36 hours. Solution is diluted with ethyl acetate and it washes in saturated sodium bicarbonate solution, water, and sodium chloride solution. An organic layer is dried, filtered and concentrated in vacuum with sodium sulfate. It is silica gel chromatography (developing solvent: Diethylearch roux hexane) about a residual substance. 3:1 v/v is given, 17-ant roux 1 and 14-dihydroxy 12-[2-[4 - (p-iodine benzene sulfonyloxy)-3-methoxy cyclohexyl]-1-methylvinyl]-23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10, 16-tetra--on (61mg) And 17-ant roux 1-hydroxy 12-[2-[4 - (p-iodine benzene sulfonyloxy)-3-methoxy cyclohexyl]-1-methylvinyl]-23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] Octakoser 14, 18-Diene- 2, 3, and 10, and 16-tetra--on (12 mg) are obtained. The above-mentioned compound IRnu (CHCl)₃):3470,1730,1717,1692,1635,1568cm^-1The after-mentioned compound¹H NMR delta ppm (CDCl)₃): 6.15(d,J=15Hz) 6.25(d,J=15Hz)(1H) 6.70(dd,J=15=Hz,10Hz)6.80(dd,J=15Hz,10Hz)(1H) 7.60(2H,m)7.90(2H,m Example of manufacture 14 FR-900506 substance (27 mg) is made to react to chlorination d-camphor sulfonyl (97 mg) in pyridine (0.6 ml) according to the method of example 13 of manufacture, 17-ant roux 12-[2 - (4-d-camphor sulfonyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (34 mg) are obtained. IRnu(neatness): 3500-1747, 1720(sh)1710(sh)1655cm^-1Example of manufacture 15 Imidazole (118 mg) and chlorination t-butyl diphenyl silyl (52.2 mg) are added to this, agitating the solution in dichloromethane (3 ml) of FR-900506 substance (89.7 mg). A mixture is agitated at room temperature for 2 hours, and it dilutes with a saturated ammonium chloride solution, and it is in diethylether and extracts 3 times. Extraction liquid is washed in water and sodium chloride solution, and decompression dryness is dried and carried out with sodium sulfate. It is silica gel column chromatography (developing solvent: ethyl acetate hexane) about a residual substance. It refines by 1:3 v/v, 17-ant roux 12-[2 - (4-t-butyl diphenyl silyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (107 mg) are obtained. IRnu(neatness): 3520-1742, 1705 or 1650 cm^-1Example of manufacture 16 It applies to the method of example 15 of manufacture correspondingly, and is in N and N-Dimethylform amide (1 ml) under existence of imidazole (15 mg), FR-900506 substance (80 mg) is made to react to chlorination t-butyl dimethylsilyl (17 mg), 17-ant roux 12-[2 - (4-t-butyl dimethylsilyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (85 mg) are obtained. IRnu (CHCl)₃):1735,1720(sh)1700,1640cm^-1Example of manufacture 17 An acetic anhydride (1.5 ml) is added to the solution in dimethyl sulfoxide (1.5 ml) of FR-900506 substance (100 mg), and a mixture is agitated at room temperature for 14 hours. A reaction mixture is diluted with ethyl acetate and it washes one by one in saturated sodium bicarbonate solution, water, and sodium chloride solution. With sodium sulfate, an organic layer is dried and filtered and is concentrated in vacuum. A residual substance is given to the thin layer chromatography (developing solvent: diethylether) of silica gel, 17-ant roux 1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 12-[2-(4-Methylthio methoxy 3-methoxy cyclohexyl)-1-methylvinyl]-11, 28-dioxa 4-azatricyclo [22.3.1.0^4,9] Octakoser 14, 18-Diene- 2, 3, and 10, 16-tetra--on (51 mg), 17-ant roux 1-hydroxy 12-[2-(4-hydroxy 3-methoxy cyclohexyl)-1-methylvinyl]-23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] Octakoser 14, 18-Diene- 2,3,10, 16-tetra--on (18 mg) And 17-ant roux 1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 12-[2-(4-Methylthio methoxy 3-methoxy cyclohexyl)-1-methylvinyl]-11, 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (10 mg) are obtained. It is a compound of eyes most. IRnu (CHCl)₃):3470,1730,1635,1630(sh)1580(sh)cm^-1The second compound IRnu (CHCl)₃):1728,1640,1090cm^-1The third compound IRnu (CHCl)₃):3480,1735,1710,1640cm^-1Example of manufacture 18 17-ant roux 12-[2 - (4-t-butyl dimethylsilyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9An acetic anhydride (0.5 ml) is added to the solution in pyridine (1.5 ml) of] octacos 18-En- 2,3,10,16-tetra--on (39.9 mg), and a mixture is agitated at room temperature for 6 hours. It removes under decompression of a solvent, a rough oily matter is obtained, and it is silica gel thin layer chromatography (developing solvent: diethylether hexane) about this. 1:1 v/v is given, 14-acetoxy 17-ant roux 12-[2 - (4-t-butyl dimethylsilyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1-hydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (26.5 mg) are obtained. IRnu (CHCl)₃):1728,1715(sh)1635cm^-1Example of manufacture 19 It applies to the method of example 18 of manufacture correspondingly, Pyridine (0.2 ml) It is 17-ant roux 12-[2 in inside. - (4-t-butyl diphenyl silyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1, 14-dihydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (10.6 mg) are made to react with an acetic anhydride (0.1 mg), 14-acetoxy 17-ant roux 12-[2 - (4-t-butyl diphenyl silyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1-hydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (10 mg) are obtained. IRnu (CHCl)₃):3500,1730,1720(sh)1660(sh)1640,1640(sh)1100cm^-1Example of manufacture 20 14-acetoxy 17-ant roux 12-[2 - (4-t-butyl diphenyl silyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1-hydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9Potassium carbonate (about 100 mg) is added to the solution in a tetrahydro franc (1.5 ml) of] octacos 18-En- 2, 3, and 10 and 16-tetra--on (43.8 mg) at room temperature, and a mixture is agitated by the Warm for 3 hours. The solution obtained by diluting a reaction mixture with diethylether is washed one by one in a saturated ammonium chloride solution, water, and sodium chloride solution, and it dries with sodium sulfate. The solution obtained is concentrated in vacuum and a residual substance is refined by silica gel thin layer chromatography (developing solvent: the diethylether hexane, 3:2 v/v), 17-ant roux 12-[2 - (4-t-butyl diphenyl silyloxy 3-methoxy cyclohexyl)-1-methylvinyl]-1-hydroxy 23, 25-dimethoxy 13, 19, and 21, 27-tetramethyl 11, and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] Octakoser 14, 18-Diene- 2, 3, and 10, and 30 mg of 16-tetra--on are obtained. IRnu (CHCl)₃):1733,1720(sh)1685,1640(sh)1620cm^-1Example of manufacture 21 Contact reduction of the solution in ethyl acetate (2 ml) of FR-900506 substance (50 mg) is carried out for 20 minutes at room temperature under atmospheric pressure using palladium carbon (10 mg) 10%. A reaction mixture is filtered, Filtrate is evaporated to dryness and it refines by thin layer chromatography. It develops by chloroform acetone Mixture (5:1 v/v), 1,14-dihydroxy 12-[2-(4-hydroxy 3-methoxy cyclohexyl)-1-methylvinyl]-23,25-dimethoxy 13,19,21,27-tetramethyl 17-propyl 11 and 28-dioxa 4-azatricyclo [22.3.1.0^4,9] octacos 18-En- 2, 3, and 10 and 16-tetra--on (50.0 mg) are obtained. IRnu (CHCl)₃):3480,1735(sh),1717,1700,1650(sh)1625cm^-1Example of manufacture 22 The rough white powder (1g) of FR-900506 substance obtained by the same fermenting method as example 1 of manufacture is melted in acetonitrile (5 ml), and the high speed liquid chromatography of Shimazu LC4A (a trademark, the Shimadzu Corp. make) is given. It is eluted at the 12ml/minute flow velocity using the steel column (25 mm in inside diameter, and 250 mm in length) filled up with YMC-S343 (ODS) (a trademark, the Shimakyu Yakuhin, Inc. make). UV detects a mobile phase at 210 nm using Mixture of 28% acetonitrile solution, 10%n-butanol, 0.075% phosphoric acid, and 3.75mM dodecyl sodium sulfate (SDS). The sample of 100microper time l is poured in, this high speed liquid chromatography is repeated 50 times, and the sample of all the quantity is given to chromatography. The ethyl acetate (3.6) of capacity, such as collection, extracts a part for the drawing eluted in the retention time for 85 minute~90 minutes. An ethyl acetate layer is isolated preparatively, and after washing with sodium bicarbonate (2) 1%, it condenses to a small quantity by decompression. The rough powder obtained by filtering out depositing SDS is melted in acetonitrile so that it may become the concentration of 100mg/ml, and also high speed liquid chromatography is given. 12.5% acetonitrile solution, 9.75%n-butanol, 0.075% phosphoric acid, and 3.75mMSDS are used as a mobile phase. A column is eluted at the 10ml/minute flow velocity. The ethyl acetate of capacity, such as collection, extracts a part for the drawing eluted in the retention time for 131 minute~143 minutes. Extraction liquid is washed in sodium bicarbonate solution 1%, and it condenses to a small quantity under decompression. SDS which deposits in the case of concentration is filtered out. The rough powder obtained is dissolved in a little ethyl acetate, and the column chromatography using silica gel (Kiesel gel, 230 - 400 meshes) (made by Merck Co.) is given. A column is washed by Mixture (30 ml) (1:1 v/v) of n-hexane and ethyl acetate, and then Mixture (60 ml) (1:2 v/v) of n-hexane and ethyl acetate. A column is eluted with ethyl acetate and it collects parts for 3 ml of each drawing at a time. If a part for the drawing of 18-24 is put together and it condenses under decompression, FR-900520 substance (24 mg) will be obtained.

[Brief Description of the Drawings]

Drawing 1 is the white powder of FR-900506 substance.¹³C nuclear magnetic resonance spectrum is expressed. Drawing 2 is the white powder of FR-900506 substance.¹H nuclear magnetic resonance spectrum is expressed. The prism-like crystal of the colorlessness [ Drawing / 3 ] of FR-900506 substance¹³C nuclear magnetic resonance spectrum is expressed. The prism-like crystal of the colorlessness [ Drawing / 4 ] of FR-900506 substance¹H nuclear magnetic resonance spectrum is expressed. Drawing 5 is FR-900525 substance.¹³C nuclear magnetic resonance spectrum is expressed. Drawing 6 is FR-900525 substance.¹H nuclear magnetic resonance spectrum is expressed. Drawing 7 is FR-900520 substance.¹³C nuclear magnetic resonance spectrum is expressed. Drawing 8 is FR-900520 substance.¹H nuclear magnetic resonance spectrum is expressed. Drawing 9 is FR-900523 substance.¹³C nuclear magnetic resonance spectrum is expressed. Drawing 10 is FR-900523 substance.¹H nuclear magnetic resonance spectrum is expressed.